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'ibrarD of tb IJJwseum

OF

COMPARATIVE ZOOLOGY,

AT HARVARD COLLEGE, CAMBRIDGE, MASS.

The gift of tfisiJiijimfnzjCjLcjuut Jkjxcisirnuj

No.

^3^0

<^lJr.l3., I'^CflJ JjjL. ICj. iSp^

TRANSACTIONS

OF THE

OONNEOTICUT ACADEMY

OF

ARTS AND SCIENCES.

VOLUME VIII.

NEW HAVEN:
PUBLISHED BY THE ACADEMY.

•'1888 to 1892.

TUTTLE, MOREHOUSE & TAYLOR, PRINTERS.

6FFICER? 0F THE /IC^DEPY, )S9^-95.

President.
WILLIAM H. BREWER.

Vice-President.

CHARLES S. HASTINGS.

Vorrespondinff Secretary.

ADDISON VAN NAME.

Mecordint/ Secretury.

SAMUEL L. PENFIELD.

JAbrnriitn.

ADDISON VAN NAME.

TreiLfitrer.

WILLIAM W. FARNAM.

J'liltlishiny Coniniittee.

HUBERT A. NEWTON, CHARLES S. HASTINGS,

GEORGE J. BRUSH, ADDISON E. VERRILL,

RUSSELL H. CHITTENDEN, EDWARD S. DANA,

ADDISON VAN NAME.

Aiiditi H g Committee.

ADDISON E. VERRILL, ADDISON VAN NAME.

CON TENTS.

I'AGB

List of Additions to the Library, v

Art. I. — Some Experiments on the Physiological Action
OP Uranium Salts. By R. H. Chittenden and
A. Lambert, 1

II. — Elastine and Elastose Bodies. By R. H. Chit-
tenden and H. S. Hart, . . 19

in. — The Influence of Urethan, Paraldehyde, Anti-

PY^RIN, AND AnTIFEBRIN ON PrOTEID MeTABOLISM.

By R. H. Chittenden, 39

IV. — The Influence of skveral new Therapeutic
Agents on Amyloly^tic and Proteoly^tic Action.
By R. H. Chittenden and C. W. Stewart, 60

V. — Caseoses, Casein Dyspkptone, and Casein Pep-
tone. By R. H. Chittenden, 66

VI. — Some Experiments on the Influence of Arsenic
AND Antimony on Glycogenic Function and
Fatty Degeneration of the Liver. By R. H.
Chittenden and J. A. Blake, 106

VII. — The Nature and Chemical Composition of the
Myosin of Muscle Tissue. By R. H. Chittenden
and G. W. Cummins, 115

VIII. — Myosinoses. By W. Kuhne and R. H. Chittenden. 139

IX. — The Relative Absorption of Nickel and Cobalt.

By R. H. Chittenden and C. Norris, Jr., 148

X. — Results obtained by Etching a Sphere and Crys-
tals OF Quartz with Hydrofluoric Acid. By
O. Meyer and S. L. Penfield. Plates 1, 2, 158

^I. — New England Spiders of the Families Drassid^,
Agalenid^ and Dy^sderid^. By J. H. Emekton.
Plates 3-8, 166

XII. — The Development of a Paleozoic Poriferous

Coral. By C. E. Beecher. Plates 9-13, 207

IV CONTENTS.

PAGE

XIII. — Symmetrical Cell Development ijst the Favosi

TiD.E. By C. E. Beecher. Plates 14, 15, 215

XIV. — New England Spiders of the Family Attid^.

By J. H. Emerton. Plates 16-21, .-- 220

XV. — A Provisional List of the Hepatic.e of the
Hawaiian Islands. By A. W. Evans. Plates

22, 23, - - - - 253

— XVI. — An Arrangement of the Genera of Hepatic^.

By A. W. Evans, 262

XVII. — On the Ferments contained in the Juice of the
Pineapple (Ananassa sativa). By R. H. Chit-
tenden, E. P. JosLiN and F. S. Meara, . 281

^VIII. — The Nephrostomes of Rana. By O. C. Farring-

TON. Plate 24, 309

XIX. — Notes on the Fauna of the Island of Dominica,
British West Indies. By G. E. Verrill. Plates
25-27, .^ 315

XX. — On a Collection of Land Moli.usca from the
Island of Dominica, West Indies. By H. A.

PiLSBRY, 356

XXI. — New England Spiders of the Family Thromsid^.

By J. H. Emerton. Plates 28-32, 359

0- XXIL — The Marine Nemerteans of New England and
Adjacent Waters. By A. E. Verrill. Plates
33-39, 382

XXIII. — D1N0PHILID.E OF New England. By A. E. Ver-
rill. Plate 36, Figures 6, Qa, 457

XXIV. — Marine Planarians of New England. By A. E.

Verrill. Plates 40-44, 459

ADDITIONS TO THE LIBRARY

Connecticut Academy of Arts and Sciences,

By Gift and Exchange, from Aug. 1, 1888, to Dec. 31, 1890.

American Association for the Advancenie^it of Science.

Proceedings. Meeting XXXVII, XXXVIII, 1888-89. Salem, 1889-90. 8°.
Albany.— TVeu) York State Library.

Annual report. LXX-LXXII, 1887-89. 8°.
New York State Museum of Natural History.

Annual report. XL-XLIII, 1886-89. 8°.

Bulletin. No. 1-10, 1887-90. 8°.

Memoirs. Vol. I. 1, 1889. 4°.
Annapolis. — United States Naval InstittUe.

Proceedings. Vol. XIV. 3, 4, XV, XVI. 2-4, 1888-90. 8°.

Index to vol. I-XV.
Baltimore. — Johns Hopkins University.

American chemical journal. Vol. X. 5, 6, XI. XII, 1888-90. 8°.

Studies from the biological laboratory. Vol. IV 5-7, 1889-90. 8°.

University circulars. No. 80, 1890. 4°.
Maryland Academy of Sciences.

Transactions. Vol. I, pp. 1-68, 1888-89. 8°.
Boston. — American Academy of Arts and Sciences.

Proceedings. Vol. XXIII, XXIV, 1887-89. 8°.
Society of Natural History.

Memoirs. Vol. IV. 7-9, 1890 4°.

Proceedings. Vol. XXIII. 3, 4, XXIV, 1888-90. 8°.
Brooklyn. — Entomological Society.

Entomologica Americana. Vol. IV. .5-12, V, VI, 1888-90. 8°.
Cambridge. — Harvard College.

Annual reports of the president and treasurer. 1887-88, 1888-89. 8°.
Astronomical Observatory of Harvard College.

Annals. Vol. XVIII. 6-10, XIX. 1, XX, XXI, XXII, XXIV, XXXI. 1,
1888-90. 4°.

Annual report. XLIII, XLIV, 1888-89. 8°.

Henry Draper Memorial Annual report. Ill, IV, 1889-90. 4°.

History, 1840-90. 8°.
Museum of Comparative Zoology at Harvard College.

Memoirs. Vol. XIV. 1, pt. 2\ XVI. 3, XVII. 1, 1889-90. 4°.

Bulletin. Vol. XVI. 2-9, XVII, XX. 1-5, 1889-90. 8°.
Annual report. 1887-88, 1888-89. 8°.

-Entomological Club.

Psyche. No. 138-40, 147-176, 188.5-90. 8°.
-New England Meteorological Society.

Investigations. 1889. 4°.

Bulletin. 1889 appendix. 4°.

vi Additions to the Library.

Chapel Hill. — Elisha Mitchell Scientific Society.

Journal. Vol. V, VI, VII. 1, 1888-90. 8°.
Cincinnati. — Observatory.

Publications. No. X, 1882-86. 8°.
Society of Natural History.

Journal. Vol. XI. 2-4, XII, XIII. 1-3, 1880-90. 8°.
Colorado Springs. — Colorado College Scientific Society.

Colorado College studies. I, 1890. 8°.
Davenport. — Academy of Natural Sciences.

Proceedings. Vol. V. 1, 1884-89. 8°.
Frankfort. — Kentucky Geological Survey.

Chemical report. Vol. A, pt. III. 1888. 8°.

Report on the geological and economic features of the Jackson Purchase
region. By R. H. Loughridge. 1888. 8°.

Reports on the geology of Bath, Fleming, Henry, Shelby, Oldham and
Mason counties. By W. McLinney. [1886-88]. 8°.

Kentucky fossil shells. By Henry Nettelroth. 1889. 4°.
Granville. — Dennison Utiiversity.

Bulletin of the scientific laboratories. Vol. IV, V. 1888-90. 8°.
Harrisborg. — Second Geological Survey of Pennsylvania.

Annual report for 1886. Pt. IV and atlas. 8°.

Atlas of northern anthracite field. Pt. II-V.' 1888-89. 8°.

Atlas of eastern anthracite field. Pt. II, III. 1888-89. 8°.

Atlas of southern anthracite field. Pt. II. 1889. 8''.

South Mountain atlas. 8°.

Catalogue of the geological museum. Pt. III. 1889.

Dictionary of fossils. Vol. I. 1889. 8o.
Little Rock. — Arkansas Geological Survey.

Annual report, 1888. Vol. I-III. 8°.
Madison. — Was?>bur7ie Observatory.

Publications. Vol. VI, VII. 1, 1890. 8°.
Wisconsin Academy of Sciences^ Arts and Letters.

Transactions. Vol. Vli, 1883-87. 8°.
University of Wisconsiri.

Agricultural station. Annual report. V, 1887-88. 8°.
Mebiden. — Scientific Association.

Transactions. Vol. Ill, 1887-88. 8°.
Middletown. — Wesleyaji University.

Annual report of the curators of the museum. XVIII, XIX, 1888-90. 8°.
Minneapolis. — Geological and Natural History Survey of Minnesota.

Geology of Minnesota. Final rei)ort. Vol. II, 1888. 4°.
Minnesota Academy of Natural Sciences.

Bulletin. Vol. III. 1, 1885-86. 8°.
New Orleans. — Academy of Sciences.

Papers. Vol. I. 2, 1887-88. o°.
Neav York. — Academy of Sciences.

Annals. Vol. IV, 5-12, V. 1-8, 1888-90. 8°.

Ti-ansactions. Vol. VII, VIII, IX. 1-8, 1887-90. 8°.
American Geographical Society.

Bulletin. Vol. XX. 3, 4, .and supp't, XXI, XXII. 1-3, 1888-90. 8°.
American Museum of Natural History.

Bulletin. Vol. II. 2-4, III, pp. 1-194, 1889-90. 8°.

Annual report. 1888-89, 1889-90. 8°.
Astor Library.

Annual report. XXXIX-XLI, 1887-89. 8°.

Additions to the Lihrary. vii

New York. — Microsropkal Society.

Journal. Vol. IV. 3, 4, V, VI, 1888-90. 8°.

Torrey Botanical Club.

Bulletin. Vol. XV. 8-12, XVI, XVII, 1888-90. 8°.
Philadelphia. — American Entomological Society.

Transactions. Vol. V-XVI, XVII. 1, 2, 1874-90. 8°.

Franklin Institute.

Journal. Vol. CXXVI. 3-P., CXXVII-CXXX, 1888-90. 8°.

Wag7ier Free Institute of Science.

Transactions. Vol. II, III, 1889-90. 8°.
Rochester. — Academy of Science.

Proceedings. Vol. I. 1, 1889-90. 8°.
Sacramento. — California State Mining Bureau.

Annual report of the state mineralogist. IX, 1889. 8°.
Bulletin. No. 1, 1888. 8°
Salem. — Fssex Institute.

Bulletin. Vol. XX, XXI, XXII. 1-3, 1888-90. 8°.
San Francisco. — California Academy of Sciences.
Memoirs. Vol. II. 2, 1888. 4°.
Proceedings. Ser. II. Vol. I, II, 1889. 8°.
Occasional papers. 1, II, 1890. 8°.

Technical Society of the Pacific Coast.

Transactions and proceedings. Vol. V. 2-4, VI, VII. 1-3, 1888-90. 8°.
ToPEKA. — Kansas Academy of Science.

Transactions. Vol. X, XII. 1, 1885-89. 8°.

Washburn College Laboratory of Natural History.

Bulletin. Vol. I. 9-11, 1889-90. 8°.
Trenton, N. J. — Natural History Society.

Joumal. Vol. II. 1, 1889. 8°.
Washington. — Bureau of Education.

Report of the commissioner of education. 1886-87, 1887-88. 8°.
Circulars of information. 1888, 1889, 1890, i. 8°.

Chief Signal Officer.

Annual report. 1887 pt. 2, 1888, 1889. 8°.

Departmeiit of Agriculture.

Division of Entomology. Bulletin. No. 1, 10-10. 19-22, 1888-90. 8°.

Periodica] bulletin. Vol. I, II, III. 1-3, 1888-90. 8°.

United States Geological Survey.

Annual report. VII, VIII, IX, 1885-86, 1886-87, 1887-88. 8°.
Bulletin. No. 40-61, 63^, 66, 1887-90. 8°.

Monographs. Vol. I. XIII-XVI and atlas of vol. XII, 1888-90. 4°.
Mineral resources of the United States. 1887, 1888. 8°.
Mineral products of the United States. 1882-87.

United States Naval Observatory .

Astronomical and meteorological observations. 1884. 4°.
Report of the superintendent. 1888, 1889. 8°.

Smithsonian Institution.

Annual report of the Bureau of Ethnology. V, VI, 1883-84, 1884-85. 8°.
Bibliography of the Iroquoian languages. By James C. Pilling. 1888. 8"..
Bibliography of the Muskhogean languages. By James C. Pilling.

1889. 8°.
Textile fabrics of ancient Peru. By William H. Holmes. 1889. 8°.
The circular, square and octagonal earthworks of Ohio. By Cyrus

Thomas. 1889. 8°.
The problem of the Ohio mounds. By Cyrus Thomas. 1889. 8°.

viii Additions to the Library.

Washington. —SmitliKonian InstUution.

Work in mound exploration of the Bureau of Ethnology. By Cyrus

Thomas. 1887. 8°.
Perforated stones from California. By Henry W. Henshaw. 1887. 8°.

Surgeon GeneroVs Office.

Medical and surgical history of the war of the rebellion. Pt. III. Vol. I.
1888. 4°.
Worcester. — American Antiquarian Society.

Proceedings. New series. Vol. V. 2, 3, VI, 1888-90. 8°.

Amiens. — Societe Linneenne du Nbrd de la France.

Bulletin. No. 175-310, 1887-89. 8°.

M^moires. Tome VII, 1886-88. 8°.
Amsterdam. — Kon. Akademie van Wetenschappen.

Jaarboek. 1886-89. 8°.

Verslagen en mededeelingeu. Afdeel. natuurkunde. 3de reeks. Deel III-
VII, 1887-90. 8°.
Kott. Zoologisch Genootschap ^'- Natura Artis Magistra."

Bijdragen tot de dierkunde. Aflev. 14-16, and festuummer. 1886-88. 4°.
Auxerre. — Societe des Sciences Historiques et Nafurelles de f Yonne.

Bulletin. Tome XLII-XLIII, 1888-89. 8°.
Barcelona. — Real Academia de Ciencias Naturales y Aries.

Meraoria inaugural. Por D. Lauro Clariana y Rycart. 1889. 8°.
Basel. — Naturforschende Gesellschaft.

Verhandlungen. Theil VIII. 3, IX. 1, 1890. 8°.
Batavia. — Kon. Natnurkundige Vereeniging in Nederlandsch-lndie.

Natuurkundige tijdschrift. Deel XLVIII, XLIX, 1889-90. 8°.
Magnetical and Meteorological Observatory.

Observations. Vol. VIII, X, XI, 1883-88. 4".
Bergen. — Museum.

Aarsberetning. 1887-89. 8°.
Berlin . — Konigliche Sterntuarte.

Berliner astronomisches Jahrbuch. 1891, 1892. 8°.
Bologna. — R. Accademia delle Scienze deW Istituto di Bologna.

Rendiconto. Anno 1887-88, 1888-89. 8°.
Bombay. — Bombay Branch of the Royal Asiatic Society.

Journal. Vol. XVII. 2, 1889. 8°.
Government Observatory.

Magnetical and meterological observations. 1886, 1887. 4°.
Bonn. — Naturhistorischer Verein der preussischen Rheinlande, Westfalens nnd des
Reg.-Bezirks Osnabriick.

Verhandlungen. Jahrg. XLV, XLVI, XLVII. 1, 1888-90. 8°.
Bordeaux. — Acade'mie Nationale des Sciences, Belles- Lettres et Arts.

Actes. Annde XLVIII, 1886. 8°.
Societe Linneenne.

Actes. Tome XXIX, XXX, 1873-7.5. 8°.

Proces-verbaux. 1887, 1889. 8°.
-Societe des Sciences Physiques et NatureUes.

M^moires. 3« s^r. Tome III. 2 and appendice, IV, V. 1 and appendice
1887-89. 8°.
Braunschweig. — Verein fiir Naturwissenschaft.

Jahresbericht. V, 1886-87. 8°.
Bremen. — NdtJirwissenschaftlicher Verein.

Abhandlungen. Bd. X. 3, XI. 1, 2, 1889-90. S^.

Additions to the Library. ix

Brkslau.-— >Sc7jZeaiscAe Oesellschaft fiir vaterldndische Cultur.

Jahres Bericht. LXV-LXVII, 1887-89. 8°.
Brunn. — NatUrforscher Verein.

Verhandlungen. Bd. XXVI, XXVII, 1887-88. 8°.

Bericht der meteorologischen Commission. VI, VII, 1886-87. 8°.
Bruxelles. — Academie Royale des Sciences, des Lettrea et des Bemix-ArU de Belgique.

Memoires couronnes et raemoires des savants Strangers. Tome XLIX,
1888. 4°.

Memoires couronnes et autres memoires. Tome XL-XLII, 1887-89. 8°.

Bulletins. 3« ser. Tome XIII -XVI, 1887-88. 8°.

Annuaire. Annee LIV, LV, 1888-89. 8°.
Mufiee Boyal de VHistoire Naturelle.

Bulletin. Tome V. 1, 1887-88. 8°.
Observatoire Royal.

Annales. 2« s^r. Annales astronomiques. Tome V. 3, VI, 1885-87. l*.

Annales meteorologiques. Tome II, 1885. 4°.

Societe Entomolcgique de Belgique.

Annales. Tome XXXI-XXXIII, 1887-89. 8°.
Societe Royale Beige de Geographic.

Bulletin. Annee XII, XIII, XIV. 1, 2, 18SS-90. 8°.
Societe Royale de Botanique.

Bulletin. Tome XXVI. 2, XXVII, XXVIII, 1887-89. 8°.

Tables generales, tome I-XXV. 1890. 8°.
Societe Royale Malacologiqiie de Belgique.

Annales. Tome XXII, XXIII,' 1887-88. 8°.

Proces-verbaux. Tome XVI, pp. 81-141, XVII, XVIII, pp. 1-133, 1887-89.
8°.
BuCAREST. — Instifut Ifeteorologique de Roimianie.

Annales. Tome III, 1887. 4°.
Budapest. — Kdn. ung. Ceiitral-AnMalt far Meteorologie utid Erdmagnetisnmn.

Jahrbiicher. Jahrg. XVI, XVII, 18S6-87. 4°.
Buenos Aires. — Museo Publico.

Anales. Tome III. 3, 1888. 4°,

Los caballos fosiles de la pampa Argentina descriptos par Dr. G. Burmei-
ster. 1887. f°.
Sociedad Cientiflca Argentina.

Anales. Tome XXV. 2-fi, XXVI-XXIX, XXX. 1-5, 1888-90. 8°.
Province de Buenos-Ayres.

Annuaire statistique. Annee VIII, 1888. 8°.
Caen. — Societe Linneenne de Nbrmandie.

Bulletin. 4" s^r. Vol. II, 1887-88. 8°.
Calcutta. — Asiatic Society of Bengal.

Journal. Vol. LVII, pt. i, ii, no. 2-5 ; LVIII, pt. i, ii ; LIX, pt. i, no. 1-2,
pt. ii, no. 1 and supp't, 1888-90. 8°.

Proceedings. 1888, no. 4^10, 1889, 1890, no. 1-3. 8°.
Geological Survey of India.

Palseontologia Indica. Ser. XIII, vol. iv, pt. 1, 1889. 4°.

Memoirs. Vol. XXIV. 2, 1890. 8°.

Records. Vol. XXI. 3, 4, XXII, XXIII. 1-3, 1888-90. 8°.

Bibliographj- of Indian geology, compiled by R D. Oldham. Preliminary
issue. 1888. 8°.
Meteorological Department of the Oovernmejit of India.

Indian meteorological memoirs. Vol. III. 3-4, IV. 5, 6, 1888-89. f°.

Report on the meteorology of India. 1887, 1888. f°.

Report on the administration of the meteorological department. 1887-88
f°.

X Additions to the Library.

Calcutta. — Meteorological Department of the Government of India.

Meteorolotfical observations recorded at seven stations in India, 1888. f°.

Cyclone memoirs. Pt. II, 1890. 8°.

Meteorological observations at Simla, 1841-45. Vol. II, Lond., 1877. 4°.

Handbook of the cyclone storms in the Bay of Bengal. By John Eliot.
1890. 8°.

Charts of the Arabian Sea and the adjacent portion of the North Indian
Ocean, showina; the pressure, winds and currents in each month of the
year [1888]. f°.
Cambridge. — Philosophical Society.

Transactions. Vol. XIV. 3, 4, 1889. 4°.

Proceedings. Vol. VI. 4^6, VII. 1-3, 1888-90. 4°.
Catania. — Accadenda Gioenia di Scienze Naturali.

Atti Ser. III. Tomo XX. Ser. IV. Tomo I. 1888-90. 4°.

Bullettino mensile. Nuova serie. Fasc. 1, 4-18, 1888-90. 8°.
Cherbourg. — Societe Rationale des Sciences Naturelles.

Meraoires. Tome XXVI, 1889. 8°.
Christiania. — KiJn. Universitiits-Stervwarte.

Zonenbeobachtungen der Sterne zwischeu 64° 50' und 70° 10' ncirdlicher
Declination. 1888. 4°.
Norwegisches meteorologisches Institnt.

Jahrbuch. 1886, 1887. 4°.
Nm-wegian North- Atlantic Expedition, 1876-78.

Publication XIX. 4°.
Vidensknbs SelsJcabet.

Forhandliugar. 1888, 1889. 8°.
Qh0r_ — Naturforschende Gesellschaft Granbiindens.

Jahres-Bericht. Neue Folge. Jahrg. XXXI-XXXIII, 1886-89. 8°.
Cordoba. — Academia Nacional de Ciencias.

Actas. Tomo VI and atlas, 1889. 4°.

Boletin. Tomo X. 2, 3, XI. 1-3, 1887-89 8°.
Da>zig. — Naturforschende Gesellschaft.

Schriften. Neue Folge. Bd. VII. 1, 2, 1888-89. 8°.
Dijon. — Academic des Sciences, Arts et Belles-Lettres.

Memoires. 3« s(5r. Tome X. 4 ser. Tome I. 1887-89. 8°.
DoRPAT.— Ge/e/irfe Estnische Gesellschaft.

Sitzungsberichte. 1888-89. 8°.
Naturforscher-Geselhchaft bei der Universitiit Dorpat.

Archiv fiir die Naturkunde Liv-Elist-und Kurlands. Ser. I. Bd. IX. 5,
1889. 8°.

Sitzungsberichte. Bd. VIII. 3, IX. 1, 1888-89. 8°.

Schriften. V, 1890. 8°.
Dresden. — Naturwissenschaftliche Gesellschaft Isis.

Sitzungsberichte und Abhandhmgen. 1888, 1889, 1890. 8°.
Verein filr Erdkunde.

Mittheilungen. 1888. 8°.

Festschrift zur Jubelfeier des 25jahrigen Bestehens. 1888. 8°.

Jubileumschrift. Litteratur des Landes-und Volkskunde des Konigreich
Sachsen. Bearb. von Paul Emil Richter. 1889. 8°.
Dublin.— i?o?/ai Irish Academy.

Transactions. Vol. XXIX. 1-13. 1887-90. 8°.

Proceedings. Ser. II. Polite lit. and antiq. Vol. II. 8. Science. Vol.
IV. 6. Ser. III. Vol. I. 1-3. 1888-90. 8°.

Cunningham memoirs. No, V, 1890. 4°.

Todd lecture series. Vol. I. 1, II, 1887-89. 8°.

Additions to the Library.

Dublin. — Royal Irish Academy.

Irish manuscript series. Vol. II. 1, 1890. 8°.

List of papers published in the transactions, Cunningham memoirs, and
Irish manuscript series, 1786-1886. 4°.

Royal Geological Society of Ireland.

Journal. Vol. XVII. 2, 1S8.5-87. 8°.
Edinburgh. — Botanical Society.

Transactions and proceedings. Vol. I. 1-3, III. 2, IV, V, X, XI, XII. 3,

XIII. 2, .3, XIV-XVII, 1841-1889. 8°.
Annual report and proceedings. I- VIII, 1836-44, 1855. 8°.

Geological Society.

Transactions. Vol. V. 4, VI. 1, 1888-90.

Royal Physical Society.

Proceedings. Vol. IX. 2, 3, X. 1, 1886-89. 8°.

Royal Society.

Proceedings. Vol. XV, XVI, 1887-89. 4°.
Emden. — Naturforschende Gesellschaft.

Jahresbericht. LXXII-LXXIV, 1886-89. 8°.
Erfurt. — Kdn. Akadeinie gemeinniiiziger Wissenschaften.

Jahrbiicher. Neue Folge. Heft XVI, 1890. 8°.
FiRENZE. — Biblioteca Nazionale Centrale.

BoUettino delle pubblicazioni Italiane ricevute per diritto di stampa.
No. 63-119, 1888-90. 8°.

R. Istituto di Studi Superiori Practici e di Perfezionamento.

Pubblicazioni. Sezione di filosofla e filologia.
Le seconde nozze del conjuge superstite. Studio storico di Alberto Del

Vecchio. 1885. 8°.
I pin antichi frammenti del costituto Fiorentino raccolti e pubblicati
da Guiseppe Rondoni. 1882. 8°.

Sezione di Scienze fisiche e naturali.

Osservazioni continue della elettricitd atmosferica istiluite a Firenze

dal Prof. Antonio Roiti. 1884. 8°.
Linee generali della tisiologia del cervelletlo. Prima memoria del Prof.
Luigi Luciani. 1884. 8°.
Sezione di medicina e chirurgia.

Archivio della scuola d'anatomia patologica. Vol. II, 1882. 8°.
Esegesi medico legale sul methodus testificandi di Giovan Battista Co
dronchi. Pel Prof. Angiolo Filippi. 1883. 8°.
Frankfurt a. M. — Deutsche malakozoologische Gesellschaft.

Nachrichtsblatt. Jahrg. XX. 7-12, XXI, XXII, 1888-90. 8°.
Senckenbergische naturforschende Gesellschaft.

Abhandluugen. Bd. XV. 3, XVI. 1, 1888-90. 4°.

Bericht. 1888-90. 8°.
Freiburg in B. — Naturforschende Gesellschaft.

Berichte. Bd. il-IV, 1887-89. 8°.
Geneve. — Institut National Genevois.

Bulletin. Tome XXIX, 1890. 8°.

Mcimoires. Tome XVII, 1886-89. 4°.
Societe de Physique et d''IIlstoire Naturelle.

Memoires. Tome XXX, 1888-90. 4°.
Museo Civico di Storia Naturale.

Annali. Ser. II. Vol. Ill -VI, 1886-89. 8°.
GiESSEN. — Oberhessische Gesellschaft ficr Natur-und Heilkuude.

Bericht. XXVI, XXVII, 1889-90. 8°.
Glasgow. — Geological Society.

Transactiojis. Vol. VIII. 2, 1886-88. 8°.

xii Additions to the Library.

Glasgow. — Natural History Society.

Proceedings and transactions. New series. Vol. II, III. 1, 1886-89. 8°.
Philosophical Society.

Proceedings. Vol. XIX, XX, 1888-89. 8°.
GoTTiNGEN. — Konigl. Oesellschaft der Wissetischaftcn.

Nachrichten. 18S8, 1889. 8°.
GusTROW. — Verein der Freunde der Naturgeschichte in Mecklenburg.

Archiv. Jahrg. XLII, XLIII, 1888-89. 8°.
Habana. — Real Colegio de Belen.

Observaciones magnetieas y meteorologicas. 1886. iv, 1887, 1888. i 4°.
Halifax. — Xova Scotian Institute of Natural Science.

Proceedings and transactions. Vol. I. 4, V. 3, 4, VI. 2-4, VII. 1-3,
1865-89. 8°.
Department of Mines.

Report. 1888, 1889. 8°.
Halle. — Kais. Leopoldinisch-VaroUniscJie deutsche Akademie der Naturforscher.

Leopoldina. Heft XXIV, XXV, 1888-89. 4°.
Naturforschende Oesellschaft.

Abhaudlungen. Bd. XVII. 1, 3, 1888. 4°.

Bericht. 1887. 8°.
'■Naturwissenschaftlicher Verein fiir Sachsen und Thiiringen.

Zeitschrift fiir Naturwissenschaften. Bd. LXI, LXII, LXIII. 1-3, 1889-90.
8°.
Hamburg. — Deutsche Seewarte.

Archiv. Jahrg. X-XII, 1887-89. 4°.

Monatliche Uebersicht der AVitterung. 1888 Miirz-Dec, 1889, 1890 Jan.-
Mai. 8°.

Deutsches meteorologisches Jahrbuch. 1887, 1888. 4°.

Ergebnisse der meteorologischen Beobachtungen, 1876-1885. 4°.
Naturwissenschaftlicher Verein.

Abhandlungen. Bd. XI. 1, 1889. 4°.
Wissenschaftliche Anstalten.

Jahrbuch. Jahrg. VI, 1888.
Hannover. — Naturhistorische Oesellschaft.

Jahresbericht. XXXVIII, XXXIX, 1887-89. 8o.
Harlem. — Musee Teyler.

Archives. S(^rie II. Vol. II. 1, III. 2-4, 1884-90. 8°.

Catalogue de la bibliotheque. Vol. I. 7, 8, II. 1, 3. 1887-89. 8°.
Societd Ilollandaise des Scietices.

Archives nt^erlandaises des sciences exactes et uaturelles. Tome XXIII,
XXIV, 1888-90. 8°.
Helsingfors. — Societas Sclentiarum Fennica.

Acta. Tom. XV, XVI, 1888. 4°.

Ofversigt af forhandlingar. XXVIII-XXXI, 1885-89. 8©.

Bidrag till kannedom af Finlauds natur och folk. Hiift. XLV-XLVIII,
1SS7-89. 8o.

Finska Vetensliaps-Societeten, 1838-1888, dess organisation och verksam-
het. Af. A. E. Arppe. 1888. 8°.
Societas pro Fauna et Flora Fennica.

Acta. Vol. Ill, IV, V. 1, 1886-88. 8°.

Meddelauden. Hiift. XIV, XV, 1888-89.

Herbarium musei fennici. Ed. 2. I. Plantae vasculares, 1889. 8°.

Notae conspectus florae fennicae. Auctore H. Hjelt. 1888. 8°.
Hermannstadt.— ;S'ie6«nA;Mr(7i';cAer Verein fiir Naturwissenschaften.

Verhandlungen und Mittheilungen. Jahrg. XXVIII. XXIX, 1888-89. 8°.

Additions to the Library. xiii

HoBART. — Royal Society of Tasmania.

Papers and proceedings. 1887, 1888. 8°.
Jena. — Medicinisch-naturwiisenscliaflliche Oesellschafl.

Jenaische Zeitschrift fiir Naturwisseuschaft. Bd. XXII. 3, 4. XXIII,
XXIV, 1888-90. 8°.
Kharkow. — Societe des Sciences Experimeittales annexee d V UniversiU de Kharkow.

Travaux de la section medicale. 1886-87, 1888. i, 1889. i-iii. 8°.
Ki EL. — Konigl. Christian AlbrecJits- Universitdt.

Sehrifteu aus dem Jahre 1SS7-88, 1888-89, 1889-90. 8° and 4°.
Naturwissenschaftlicher Verein fiir Schleswig-Holstein.

Schriften. Bd. VII. 2, VIII. 1, 1889. 8°.
KiKV. — Kievskie Obshcliestvo lestestvoispytatelei.

Zapiski. Tom. IX. 1, 3, X. 1-3, XI. 1, 1888-88. 8°.
Kjobenhavn. — Kon. Danske Videnskabernes Selskdb.

Oversigt over forhandlinger. 1887. iil, 1888, 1889, 1890. i. 8°.
KoNiGSBERG. — KowigL pJiysikalisch-okonomische Oesellschaft.

Scliriften. Jahrg. XXVIII-XXX, 1887-89. 4°.
Krakow.^JST. k. Sternwarie.

Materyaly do klimatograUi Galicyi. Rok 1887, 1888. 8°.
La Plata.— 3/Mseo.

La musee de La Plata. Rapide coup d'reil sur sa fondatioii et son d6-
veloppement. Par F. P. Moreno. 1890. 8°.
Lausanne. — Societe Vaudoise des Sciences Natnrelles.

Bulletin. 3« ser. No. 97-101, 1888-90. 8°.
Leeds. — Yorkshire Geological and PolytecJinic Society.

Proceedings. New series. Vol. X, XI. 1-:.', 1888-89. 8°.
Leiden. — Nederlandsche Dierkuiidige Vereeniging.

Tijdscrifl. Ser. II. Deel II. 3, 4, supplement deel II, 1888-89. 8°.
Leipzig. — Astronomische Oesellschaft.

Vierteljaiirsschrift. Jahrg. XXIII, XXIV, XXV. 1, 2, 1888-90. 8».

Publication. XIX, 1889 4°.

Catalog. Abth. I. Stuck IV, XIV, 1890. 4°.
Kon. sdchsische Oesellschaft der Wissenschaften.

Berichte. Math.-physische Classe. Bd. XL, XLI, XLII. 1, 1888-90.

Register zu den Jahrg. 1866-85 der Berichte und Bd. I-XII der Abhand-
lungen. 1889. 8°.
Nat urforschende Oesellschaft.

Sitzungsberichte. Jahrg. XIII, XIV, 1886-87. 8°.
Verein fiir Erdkunde.

Mittbeilungen. 1887-89. 8°.

Zoologischer Anzeiger. No. 283-350, 1888-90. 8°.

Liege. — Societe Oeologique de Belgique.

Aunales. Tome XIII. 2, XIV. 2, XV. 2. 3, XVI. 1, 1888-89. 8°.
Societe Royale des Sciences.

Memoires. 2'^ s»5r. Tome XV, XVI, 1888-90. 8°.
LiSBOA, — Sociedade de Geographia.

Boletin. Serie VII. 9-12, VIII, IX. 1-6, 1887-90. 8°.

Catalogos e indices. As publicayoes. 1889. 8°.

Indices e catalogos. A biblioteca. I. 1890. 8°.

Historia do infante D. Duarte irmao de el-rei D. Joao IV. Por Jos^
Ramos-Coelho. Tomo I. 1889. 8°.
Liverpool. — Literary and Philosophical Society.

Proceedings. No. XLI-XLIll, 1887-89. 8°.
London. — Geological Society.

Quarterly journal. Vol. XLIV. 3, 4, XLV, XLVI, 1888-90. 8°,

xiv Additions to the Library.

London. — Linnean Society.

Journal. Zoology. No. 118-123, 130-144, 1888-90. 8°.

—Botany. No. 150, 152-157, 159-174, 181-182, 1887-90. 8°.

Proceedings. 1887-88. 8°.

List. 1888-90. 8°.
MatJiematical Society.

Proceedings. No. 321-390, 1888-90. 8°.
Royal Meteorological Society.

Quarterly journal. New series. No. 67-76, 1888-90. 8°.

List of fellows. 1889, 1890. 8°.
Royal 3Iicroscopical Society.

Journal. 1888. iv-vi, 1S89, 1890. i-v. 8°.
Royal Society.

Philosophical transactions. Vol. CLXXVIII, CLXXIX. A, B, CLXXX.
A, B. 1887-89. 4°.

Proceedings. No. 270-294, 1888-90. 8°.

List of council and members. 1887-89. 4°.
LouvAiN.— La Cellule. Tome I-V, VI. 1, 1881-90. 8°.
Lund. — Universitet.

Acta. Tom. VIII, XVIII, XXIV, XXV, 1871-89. 4°.
Lton. — Acadimie des Sciences, Belles-Lettres et Arts.

Mi^moires. Classe des sciences. Tome XXVIII, XXIX, 1886-88. 8°.

Histoire des herbiers. Par le Dr. Saint-Lager. Paris, 1885. 8°.

Recherches sur les anciens herbaria. Par le Dr. Saint-Lager. Paris, 1886.
8°.

Viciscitudes onomastiques de laglobulaire vulgaire. Paris, 1889. 8°.
Mus4e Guimet.

Aunales. Tome XIII, XV-XVIL 1888-89. 4°.

Revue de I'histoire des religions. Tome XVI, 2, 3, XVII-XXI, 1887-90.
8°.
Madras. — Government Observatory.

Observations made with the meridian circle, 1865-67. 4°.
Madrid. — Comision del Ilapa Geologico de Espana.

Bolctin. Tomo XIV, XV, 1887-88. 8°.

Memorias. Descripcion flsica, geologica y minera de la provincia de
Huelva. Por D. Joaquin Gonzalo y Tarin. Tome I, II, 1886-88. 8°.

Mapa geologico de Espafia. Hoja 6, 8, 12, 16, 19, 20, 23, 24, 27, 28, 31, 32.
1889.
Ohservatorio.

Observaciones meteorologicas. 1886-87. 8°.

Resumen da las observaciones cfectuadas en la peninsula. 1884, 1885. 8°.
Sociedad Espanola de Historia Natural.

Anales. Tomo XVII. 2, 3, XVIII, 1888-89. 8°.
Real icademia de Ciencias Exactas, Fisicas y Naturales.

Memorias. Tomo XIII. 2, 3, 1889. 4°.

Revista de los progresos de las ciencias exactas. Tomo XXII. 5-7,
1888-89. 8°.

Anuario, 1889. 8°.
Manchester. — Literary and Philosophical Society.

Memoirs and proceedings. Series IV. Vol. I-III, 1888-90. 8°.
Marburg. — Gesellschaft zur Beforderung der gesammten Natnrwissenschaften.

Sitzungsberichte. Jahrg. 1888, 1889. 8°.
Melbourne. — National Museum.

Prodromus of the zoology of Victoria. Decade XVI-XX. 1888-90. 8"
Metz. — Academic.

M^moires. 3« s^r. Ann^e XV, 1885-86. 8°.

Additions to the Lihrary. xv

Mexico. — Observatorio Meteorologico-Magnetico Central.

Boletin mensuel. Tomo I. 6-13, II, 1888-89. 4°.

Sociedad CienUfica '■'■Antonio Alzate."

Memorias. Tomo II, III, IV. 1-2, 1888-90. 8°.

Sociedad de Oeographia y Estadistica.

Boletin. Epoca IV. Tomo I. 1-4, 6, 1888-89. 8°.

Sociedad Mexicana de Historia Natural.

La iiaturaleza. Ser. II. Tomo 1. 4-9. 1888-90. 4°.
MiDDELBtiRG. — ZeeiiwHch Genootschap der Weteniichaxipen.
Zelandia illustrata. Vervolj;,-. 1885. 8°.
Levensberichten van Zeeiiwen. Afl. I, II, 1888-89. 8°.
De stadsrekeniu^en van Middelbuij^-. III. 1500-1549. Door H. M. Keste-

loo. 8°.
Vlnchtbergen in Walcheren. Door Dr. J. C. Man. 1888. 8°.
MiLANO. — Real Istitiito Lombardo di Scieme e Lettere.

Rendiconto. Serie II. Vol. XX, XXI, 1887-88. 8°.

Real Osservatorio di Brera.

Pubblicazioni. No. XXXIII-XXXVII, 1888-91. 4°.

Societd Italiana di Scienze JSfaturali.

Atti. Vol. XXIV-XXXII, 1881-90. 8°.
MoDENA. — Regia Accademia delle Scienne, Lettere ed Arti.
Memorie. Serie II. Tomo V, VI, 1887-88. 4°.

Societd del Ndturalisti.

Memorie. Ser. III. Vol. VII. 1, VIII, IX. 1, 1888-90. 8°.
MoNTPELLiER. — Academic des Sciences et Lettres.

M(3moires. Section des lettres. Tome VIII. 2, 3, 1888-89. 4».
Moscou. — Societe Imperiale des Nat^iralides.

Bulletin. Annexe 1888, 1889, 1890. i. 8°.
Nouveaux memoires. Tome XV. 6, 1889. 4°.

Meteorologische Beobachtungen am Observatorium der landwirth. Aka-
demie bei Moskau. Jahr. 1887. ii, 1888, 1889. i. 4°.
MiJNCHEN. — Kon. bayerische Akademie der Wissenschajten.

Sitzungsberichte. Philosph.-philolog. und histor. Classe. 1888, 1889, 1890.
Bd. I. 1, 2. 8*".

Mathemat.physikal. Classe. 1888, 1889, 1890 Heft 1 , 2. 8°.

Ueber die Molekularbeschaffenheit der Krystalle. Festrede von Dr. Paul

Groth. 1888. 4°.
Ueber die historische Methode auf dem Gebiet des deutsehen Civilprocess-

rechts. Festrede von J. W. v Planck. 1889. 4°.
Georg Simon Ohm's wisseuscliaftliche Lelstungen. Festrede von Eugen

Lommel. 1889. 4°.
Die Anfiinge einer politischeii Literatur bei den Griechen. Festrede von

Rudolph Scholl. 1890. 4°.
Gediichtnisrede auf Karl von Prautl. Von W. v. Christ. 1889. 4°.
Gedjichtnisrede auf J. von Dollinger. Von C. A. Cornelius. 1890. 4°.

Kbii. Sternwarte.

Neue Annalen. Bd. I, 1890. 4°.
MiJNSTER. — Westfdli&cher Provincial- Vereinfiir Wissenschaft und Kimst.

Jahresbericht. XVI, XVII, 1887-88. 8°.
Nancy. — Academic de Stanislas.

Memoires. 5« ser. Tome V-VII, 1888-89. 8°.
Napoli. — R. Accademia delle Scienze Flsiche e Matmiatiche.
Atti. Ser. II. Vol. I-lII, 1887-89. 4°.
Rediconto. Ser. II. Vol. I-III, 1887-89. 4°.
Neuchatel. — Societe des Sciences Nattirelles.
Bulletin. Tome XVI, 1888. 8°.

\

;ivi Additions to the Library.

Newcastle-upon-Tyne. — No7-th of England Institute of Mining and Mechanical
Engineers.

Transactions. Vol. XXXVII. 5, 6, XXXVIII. 1-5, 1888-90. 8°.
Report of the French commission on the use of explosives in the pres-
ence of flre-dnmp in mines. Pt. 1, 2. 1890. 8°.
NOhnberg. — Naturhistoriache Qesellschaft.

Jahresbericht. 1888, nebst Abhandlungen, Bd. VIII. 8°.
Odessa. — Societe des Naturalistes de la Nouvelle Russie.

Zapiski. Tom. XII. 2, XIII, XIV, 1888-89. 4°.
Matematicheskoe otdielenie. Tom. VIII-X, 1888-89. 8°.
Ottawa. — Geological and Natural History Survey of Canada.

Annual report. New series. Vol. Ill, with maps, 18S7-88. 8°.
Contributions to palaeontology. Vol. I. 2. 1889. 8°.

Meteorological Service of the Dominion of Canada.

Report. 1885. 8°.
Oxford. — Badcliffe Library.

Catalogue of books added during 1888, 1889. 8°.

Haddiffe Observatory.

Results of astronomical and meteorological observations. Vol. XLIII,
XLIV, 1885, 1886. 8°.
Palermo. — R. Accademia di Scienze, Lettere e Belle Arti.

Atti. Nuova serie. Vol. X, 1888-89. 4°.
Paris. — Ecole Normale Superieure.

Annales scientifiques. 3' s6r. Tome V. 8-12, VI, VII. 1-10, 1888-90. 4°.

J^cole Polytechnique.

Journal. Cahier LVIII, LIX, 1889. 4°.

Observatoire.

Rapport annuel. 1888, 1889. 4°.

Societe Nationale d' Acclimatation.

Revue des sciences naturelles appliquees. 4e serie. Tome V. 16-24, VI,
VII. 1888-90. 8°.

Societe Geologique de France.

Bulletin, 'se s«5r. Tome XIV. 9, XV. 9, XVI. 4-11, XVII. 1-9, XVIII. 1-4,
1886-90. 8°.

Societe 3fathematiqne de France.

Bulletin. Tome XVI. 4, XVII, XVIII, 1888-90. 8°.
Penzance. — Royal Geological Society of Cornwall.
Transactions. Vol. XI. 3, 4, 1888-90. 8°.
Perugia. — Accademia Medico- Chirvrgica.

Atti e rendiconti. Vol. I l-A, 1889. 8°.
Pisa. — Societd Toscana di Scienze Naturali.
Memorie. Vol. IX, X, 1888-89, 8°.

Processi verbali. Vol. VI, pp. 105-302, VII, pp. 1-170, 1888-90. 8°.
Commemorazionc di Giuseppe Menenghini fatta nell' aula magna dell'
Universita Pisana ai 24 Marzo, 1889. 8°.
Potsdam. — Astrophysikalisches Observatoriu7n.

Publicationen. Bd.'lV. 2, VI, 1889. 4°.
Pkag. — Kon. hiihm.ische Ge.ielhchaft der Wissenschaften.

Abhandlungen der math.-naturwiss. Classe. Folge VII Bd. II, III,

1888-90. 4°.
Sitzungsberichte. 1888, 1889, 1890. 8°.
Jahresbericht. 1887-89. 8°.

K. k. Sternwarte.

Magnetische und meteorologische Beobachtungen. Jahrg. XLIX, L,

1888-89. 4°.
Astronomische Beobachtungen. Appendix zu Jahrg. XLVI-XLVIII. 1890.
4°.

Additions to the Library. xvii

PuLKOVA. — Nicolai-Hauptsternwarte.

Jahresbericht. 1887. 8°.

Tabulae quantitatum Besselianarum, 1890-94. 8°.
Quebec — Literary and Historical Society.

Transactions. No. 19, 18S7-89. 8°.
Regensbukg. — Naturwissenschaftlichcr Verein.

Berichte. Heft I, 1886-87. 8°.
Historincher Vtrein von Oberpfalz unci Regensburg.

Verhaudlungen. Bd. XLI, XLII, 1887-88. 8°.
Richmond, Surrey. — Keiv Observatory.

Report of the committee. 1888, 1889. 8°.
Riga. — Naturforscher Verein.

Correspondenzblatt. Jahrg. XXXI-XXXIII, 1888-90. 8°.
La Rochelle. — Academic des Belles- Lettres, Sciences et Arts.

Annales de la soci6t^ des sciences naturelles de la Charente-Inferieure.
Vol. XXIV, XXV, 1887-88. 8°.
Roma. — Biblioteca Nazionale Centrale Vittorio Emajiuele.

Bollettino delle opere moderne straniere acquistate dalle biblioteche
pubbliche governative del regno d'ltalia. Vol. Ill, IV, V. 1-3, 1888-90.
8°.
Meale Accademia dei Lincei.

Atti. Ser. IV. Rendiconti. Vol. IV, i. 11-13, ii, V, VI. i, ii. 1-8,
1888-90. 4°.

Memorie della classe dl scienze morali, storiehe e filologiche. Vol.

II, III, V, 1886-88. 4°.

Memorie della classe di scienze flsiche, matematiche e natural!.

Vol. III-V, 1886-88. 4°.
Accademia Pontifica de'' Nuovi Lincei.

Atti. Tomo XLII, XLIII. 1-3, 1888-90. 4°.
Reale Comitaio Geologicq d'ltalia.

Bollettino. Vol. XVIII-XX, 1887-89. 8°.
Socieid Italiana delle Scietize.

Memorie di matematica e di fisica. Ser. III. Tomo IV, V and appendice,
VII, 1882-90. 4°.
St. Gallen. — Naturwissenschaftliche Gesellschaft.

Bericht. Jahrg. 1886-87, 1887-88. 8°.
San Jose. — Institnto Jleteorologico Nacional.

Boletin trimestral. No. 4, 1888. 4°.
S. Paolo. — Commissao Geographica e Geologica da Provincia de S. Paolo.

Boletin. No I -III, 1889. 8°.
St. Peteksburg. — Vomite Ge'ologique.

Memolres. Vol. II. 2-5, III. 1-4, IV. 1, V. 3-4, VI, VII. 1, 2, VIII. 1, IX. 1,
XI. 1, 1885-89. 4°.

Bulletins. Vol. IV. 7-10, V, VII. 1-10, VIII. 1-8, 1885-89. 8°.

Bibliotheque geologique de la Russie. 1885-88. 8°.
Hortus Petropolitanus.

Acta. Tom. X. 2, XI. 1, 1889-90. 8°.
Lnp. Russ. Geograf. Obshtchestvo.

Otchet. God 1887-89. 8°.
Kais. Akademie der Wissenschqften.

Bulletin. Tome XXXII. 2^, XXXIII. 1, 1888-89.

Repertorium der Meteorologie. Bd. XI, XII, 1888-90. 4°.

Neue Reduction der Bradley'schen Beobachtungen, 1750-62. Von A.
Auwers. Bd. Ill, 1888. f°.
St. Petersburg. — Physikalisches Centralobservatorium.

Annalen. Jahrg. 1887, 1888, 1889. 4°.

xviii Additions to the Lihrary.

Santiago. — Deutscher wisseriHchafUicher Verein.

Verhandlungen. Hefl VI, 1888. 8°.
Schxueizerische naturforschende Gesellschaft.

Verhandlungen. Jahresversammlung LXXI, LXXII, 1888-89. 8°.
Stockholm. — Entomologisk Forening.

Entomologisk tidskrift. Arg. IX, X, 18S8-89. 8°.
Kong. Svenxka Vetenskaps Akademie.

Haudlingar. Ny foljd. Bd. XX, XXI, 1882-85. 4°.

Bihang till haudlingar. Bd. IX-XIII, 1884-88. 8°.

Ofversiiit af forliandlingar. Arg. XLI-XLV, 1884-88. 8°.

Lefnad.steckningar. Bd. II. 3, 1885. 8°.

Meteorologi^ka iakttagelser. Bd. XXII-XXVI, 1S80-84. 4°.

FiJrteckning ofver innehalet i skrifter, 1820-1883. 8°.

Ledamoter, 1885-89. 8°.
Stuttgart.— F«/em far vateiianduche Naturkunde hi Wilrttemberg.

Jahreshef.e. Jahrg. XLV, XLVI, 1889-90. 8o.
Sydney. — Oovemnieid Observatory.

Results of meteorological observations made in New South Wales during
1887. 8°.

Results of rain, river and evaporation observations during 1888, 1889. 8°.
Linnean Society of New South Wales.

Proceedings. Series II. Vol. II. 1, 4, 1887-88. 8°.
Royal Society of New South Wales.

Journal and proceedings. Vol. XXII-XXIII, 1888-89. 8°.
Tacubaya. — Observatorio Astronomico Nacional.

Anuario. Afio IX-XI, 1889-91. 8°.
Thkondhjem. — Kon. Norske Videnskabers Selskab.

Skrifter. 1886, 1887. 8°.
TiFLis. — Physicalisches Observo.torium.

Magnetische Beobachtungcn. 1886-87, 1887-88. 8°.
Tokyo. — Imperial Utiiversity of Japan.

Journal of the college of science. Vol. II. 4, 5, III, 1888-90. 4°.

Calendar. 1889-90. 8°.
Seismological Society of Japan.

Transactions. Vol. XIII, 1889. 8°.
Torino. — Musei di Zoologia ed Anatomia Comparata.

Bollettino. No. 49-86, 1888-90. 8°.
Toronto. — Canadian Institute.

Proceedings. Ser. III. Vol. VI, VII, 1889-90. 8°.

Annual report. 1887-88, 1888-89. 8°.
Toulouse. — Academic des Sciences., Inscriptions et Belles- Lettres.

Memoires. 8« ser. Tome IX, X. g" S6r. Tome I. 1887-89. 8°.'
Tromso. — Mnsenm.

Aarshefter. I-XII, 1878-89. 8°.

Aarsberetning. 1873-1888 8°.
Upsala. — Hegia Societas Scientianmn.

Nova acta. Ser. III.- Vol. XIV. 1, 1890. 4°.

Catalogue methodique des acta et nova acta, 1744-1889. 4°.
Utrecht. — Kon. Nederlandsch Meteorologisch Instituut.

Nederlandsch meteorologisch jaarboek. Jaarg. XXXI. 2, XL, XLI,

1879-90. 4°.
Provinciaal Utrechtsch Genootschap van Kunste^i en Wetenschappen.

Verslag van het verhaudelde in de algemeene vergadering. 1887-89. 8°.

Aanteekeningen van het verhandelde in de sectie-vergaderingen. 1887-89.
8°.
Vknezia. — Istituto Veneto di Scienze, Lettere ed Arti.

Atti. Ser. VI. Vol. V. 10, VI, VII, 1887-89. 8°.

Additions to the Lihrary. xix

ViCENZA. — Accademia Olvmpica.

Atti. Vol. XXT, 1886. 8°.
Wellington. — New Zealand Institute.

Transactions and proceedings. Vol. XXI, 1888. 8°.
WiEN. — Kais. Akademie der Wissenschaften.

Sitzungsberichte. Matheraat.-naturwiss. Classe. Abth. I. Bd. XCV-
XCVII, XCVIII. 1-3, 1887-89. 8°.
K. k. Central- Anstalt filr Meteorologie und Erdmagnetistnus.

Jahrbiicher. Neue Folge. XXIV, XXV, 1887-88. 4°.
K. k. geologische JEteichxaiistalt.

Abliandlungen. Bd. XIII. 1, XV. 1, 2, 1889-90. 4°.

Jahrbucli. ' Bd. XXXVII. 3, 4, XXXVIII, XXXIX, XL. 1, 2, 1887-90. 8°.

Verhandlungen. Jahrg. 1887, no. 10-18, 1888, 1889, 1890, no. 10-13. S*.
K. k. naturhistorisches HofmuMimi.

Annalen. Bd. III. 3, 4, IV. 1, 4, V. 1-3, 1888-90. 8°.
K. k. Universitats-Sternwarte.

Annalen. Bd. V, VI, 1885-86. 4°.
K. k. zoologisch-botanische Oenellschaft.

Verhandlungen. Bd. XXXVIII, XXXIX, XL. 1, 2. 1887. 8«"
Wiesbaden. — Nassauuchei' Verein fur Naturkunde.

Jahrbiicher. Jahrg. XLI, XLII, 1888-89. 8°.
WuRZBURG. — Phy&ikalisch-medicinisclie Gesellschaft.

Sitzungsberichte. Jahrg. 1888, 1889. 8°.
Z URICH. — Naturforschende Gesellschaft.

Vierteljahrschrift. Jahrg. XXXI. 3, 4, XXXII, XXXIII, XXXIV. 1, 2,
1886-89. 8«.

Recent discussions on the abolition of patents for inventions. Lond. 1869. 8*.

From R. A. Macfie, Esq.
De la mesure de la simplicitc dans les sciences mathematiques. Par E. Lemoine.

Paris, 1888. 8°.
Notes sur diverses questions de la gi^ometrie du triangle. Par E. Lemoine. Paris,

1888. S°. From the Author.

Atmospheric economy of solar radiation. By Arthur Searle. 8°. (From Proe.

Amer. Acad, of Arts and Sci., 1888.) Frotii the Author.

Eskimo of Hudson's Strait. By F. F. Payne. 8°. (From Proc. Canad. Inst.

1889.) From the Autfior.

Echinoderms from the northern coast of Yucatan and the harbor of Vera Cruz.

By J. E. Ives. 8°. (From Proc. Acad. Nat. Sci. of Philad., 1890.)

From the Author.
Ueber Feuerbestattung. Vortrag von Prof. Dr. Friedrich Goppelsroeder. Miil-

hausen, 1890. 8°. From the Author.

Die geologischen Horizonte der fossilen Kohlen. Von C. F. Zincken. Leipzig.

1884. 8°. From the Author

I, — Some Experiments on the Physiological Action op Uranium
Salts. By R. H. Chittenden and Alexander Lambert, M.D.

In 1885, experiments were commenced in the laboratory of phys-
iological chemistry at Yale University, to ascertain something
regarding the physiological and toxical action of uranium salts. At
that time there was little accurate knowledge concerning uranium.
Gmelin* had, in 1824, performed a few experiments with the nitrate
from which he concluded that this salt is a feeble poison ; thus he
states that 15 grains had no eifect on a dog, that 1 drachm merely
caused vomiting after more than an hour's interval, and that 34
grains killed a rabbit in 52 hours by stopping the iiTitability of the
heart, while 3 grains injected into the jugular vein of a rabbit
caused instant death. Later, in 1851, there appeared a statement
in the British and Foreign Medico-chirurgical Review that Lecoute
always found sugar in the urine of dogs slowly poisoned by small
doses of uranium nitrate. This statement was commented upon by
Hughes in his manual of pharmico dynamics (p. 866), and has been
made the basis of a claim by the so-called homa?opathic school that
uranium nitrate is a remedy for diabetes. Hughes also refersf to a
monograph by Edward Blake on urapium, where three persons and
nineteen animals were experimented on. In none of Blake's subjects,
however, human or brute, was sugar eliminated in the urine.
L^lceration of the pyloric end of the stomach and of the duodenum
was found well marked in several of the animals, although in no
case was the drug introduced directly into the stomach. Hughes,
likewise, refers to several cases of diabetes which he considers were
cured by the exhibition of small doses of uranium nitrate,J one-sixth
to one-third of a grain three times a day. This constitutes all the
matter bearing on uranium that we have been able to find.

Our work was commenced by a series of experiments on the in-
fluence of a variety of soluble uranium salts on the action of the
araylolytic and proteolytic ferments occurring in the animal organ-

* Edinburgh Medical and Surgical Journal, vol. xxvi, p. 136.

t Ibid., p. 867.

1^ Lancet, June 13, 1874.

Trans. Conn. Acad., Vol. VI [I. 1 Nov., 1888.

2 Chittenden and Lambert — Experiments on the

ism. The results, already published,* show plainly that all of the
uranyl salts, with one or two exceptions, have a more or less marked
inhibitory influence on amylolytic and proteolytic action. With
the salivary ferment, even 0-010 per cent, of uranyl nitrate was suffi-
cient to completely stop all action, while with pepsin-hydrochloric
acid and with alkaline trypsin solutions, 0*5 per cent, of the same
salt was required to produce an equal effect. In this case the
inhibitory action of the uranium salt was, in part at least, due to the
formation of a more or less definite and indigestible compound of
uranium with the proteid matter to be digested.f

On the excretion of carbonic acid, uranyl nitrate, by a later series
of experiments,! was also found to have a marked influence. With
rabbits, the hypodermic injection of this salt was followed by a slight
rise in body temperature and a decided increase in the elimination
of carbonic acid. The action of the salt was somewhat slow, but
repetition of the experiment always led to an increase of body
temperature and a decided increase in the amount of carbonic acid
excreted. 0*7 gram of the salt in divided doses was required to
produce the result stated, the rabbit not suffering any apparent ill
effects from this quantity.

The object of the present series of experiments has been : 1st, to
ascertain the influence of uranium salts on proteid metabolism ; 2d,
to ascertain something regarding the toxic action of uranium salts ;
and 3d, whether uranium has any influence on the production of
glycosuria.

Influence on proteid metabolism.

In this experiment a mongrel bitch weighing 18*8 kilos was em-
ployed. The animal was confined in a convenient cage suitably
arranged for the collection of the excreta, and was fed during the
experiment upon a constant diet of known composition. A large
quantity of fresh, lean beef finely chopped, was dessicated at a low
temperature until it had lost about 75 per cent, of water. It then
contained 11 "88 per cent, of nitrogen, as determined by Kjeldahl's
method. A large quantity of ordinary soda crackers were obtained
and when sampled were found to contain 0*69 per cent, of nitrogen.
40 grams of this prepared beef and 25 grams of the crackers, with

* Chittendea and Hutchinson. Studies from the Laboratory of Physiological
Chemistry, Yale University, vol. ii.

f See Chittenden and Whitehouse. Studies, vol. ii, p. 111.
X Chittenden and Cummins. Studies, vol. ii.

Physiological Action of XJranium Salts.

400 c. c. of water were fed to the animal twice daily, making a total
daily income of 10-054 grams of nitrogen. As soon as nitrogenous
equilibrium was established, the 24 hours' urine was analyzed for
nine consecutive days, thus giving the average composition of the
normal excretion. Uranyl nitrate was then administered for ten
consecutive days in gradually increasing quantities.

The accompanying tables give the analytical results. Nitrogen
was determined by the Kjeldalil method, sulphur and phosphorus
by fusion of a given volume of the urine with potassium hydroxide
and potassium nitrate in a silver crucible, and precipitation of the
sulphur as barium sulphate and of the phosphorus first as phospho-
ammonium molybdate and then as ammonio-raagnesium phosphate.*

Normal Urine— Without uranium.

Date

VoUirae.

ao'lLlsP-^^-

Nitrogen

Sulphur.

Phos-
phorus.

Dose of
Uranyl nitrate.

May
31

June

1

2
3
4
5
6
7
8

c. c.
565

610
630
600
630
570
460
590
640

acid

1016

1017
1016
1018
1018
1019
1020
1018
1016

grams.

8-414

10-380
10-040

9-618
10-770
10-325

8-738
10-091
10-763

gram.
0-441

0-516
0-558
0-589
0-661
0-608
0-628
0-576
0-604

gram.
0-572

0-637
0-665
0-595
0-715
0-632
0-746
0-712
0-645

j 0-025 gram.
( 0-025 ■

Total

5395

----

89-139

5-181

5-919

Daily
average

588

----

1017-5 9-904

0-575

0-657

Examination of the first table shows that the animal was in nitro-
genous equilibrium, the average daily excretion of nitrogen for the
normal period being 9-904 grams while the daily amount of nitrogen
taken was 10-054 grams, thus showing a fairly close agi-eement, espe

* See studies from Laboratory of Physiological Chemistry, Tale University, vol. ii,

p. 88.

Chittenden and Lambert — Experiments on the

cially as there would be a slight loss of nitrogen through the hair shed
daily. On commencing the administration of uranium nitrate, small
doses were at first given two or three times daily in gelatin capsules,
at times not to interfere with digestion. On the second day, uranium
was detected in the urine. On June 12th, the 5th day the uranium
salt was given, a trace of albumin appeared in the urine and on the
day following, the 24 hours' urine contained 1*74 grams of albumin.

With Uranium.

Date.

Volume.

c. c.
600

600

602

680

640

710

720

830

640

Re-
action.

acid

<t

((
((

<<

Sp. Gr.

Total
nitrogen.

Nitro-
gen-
nitrogen

of
albumin.

Sulphur.

Phos-
phorus.

Dose of
Uranyl
nitrate.

June
9

10

11

12

13

14

15

16

17

1017
1015
1018
1017
1022

1020

1023

1022

1025

grams.
10-285

8-895

8-925

8-380

11-391

8-847

10-608

11-767

11-178

grams.
10-285

8-895

8-925

8-380

11-093

8-439

10-141

11-446

10-955

gram.

0-587

0-404
0-574
0-525
0-724

0-634

0-571

0-738

0-606

gram.
1-226

0-556

0-612

0-603

0-621

0-628

0-628

0-704

0-626

gram.
j 035
\ 0-035
j 0-050
\ 0-025
i 0-050
'{ 0'025

0-050
( 0-025
\ 0-025
( 0-025
J 0-025
\ 0-050
( 0-050
\ 0-050
( 0050
( 0-075
\ 0-075
( 0-075
( 0-150
\ 0-150
(0-150

Total.

6022

90-276

88-559

5-363

6-20Jf.

1-295
grams.

Daily
average

669
650

1019-9

10-031

9-839

0-596

0-689

18

acid

1026

10-179

9-772

0-611

0-542

On the 16th of June, the urine contained 7'7 grams of sugar. The
appearance and disappearance of both albumin and sugar were fol-
lowed quantitatively and the results will be discussed later on.
Naturally, the appearance of albumin in the urine compels a correc-
tion in the table of nitrogen results on those days when albumin
was excreted. This has been done by subtracting from the total
nitrogen found, the nitrogen equivalent to the albumin, on the
assumption that the latter contains 15-5 per cent, of nitrogen.

Physiological Action of Uranium Salts. 5

Examination of the analytical results shows that the uranium salt
has a marked influence on the excretion of water, the increase
amounting on an average for the nine days period to over 80 c. c.
per day. There is also a very marked increase in the specific
gravity of the daily excretion, but this is without doubt in great
part due to the presence of albumin and sugar, as the increase is
most noticeable on those days when the largest amounts of sugar and
albumin were excreted.

Regarding nitrogen, a comparison of the total amounts excreted
for the two periods, after correction for the nitrogen of the albumin,
indicates that small doses of the uranium salt have little, if any,
influence on proteid metabolism. On the last three days of the
uranium period, however, when the amount administered had not
only been increased, but there was doubtless also an accumulative
effect, the excretion of nitrogen appeared to be considerably above
the normal, certainly enough to warrant the assumption that com-
paratively large doses of uranium salts may increase somewhat pro-
teid metabolism, and this view is sustained by the increase in both
the total and daily average amounts of sulphur and phosphorus
excreted. During the uranium period of ten days, 1*295 grams of
the nitrate, or 19'98 grains, were given without any apparent ill
effects being immediately produced.

Excretion of sugar atid albumin.

After about 0'4 of a gram of uranium nitrate had been given, the
urine began to show traces of albumin and five days after this, sugar
made its appearance. The amounts excreted are shown in the accom-
panying table. Sugar was determined by AUihn's gravimetric
method, and albumin by boiling with acetic acid and collecting the
coagulum on a weighed filter. On the 18th, the weighed diet was
discontinued, but the urine was kept under close scrutiny with the
results shown in the table. The sugar first disappeared and four
days after this the albumin, likewise. On again administering
uranium, and in much larger doses than in the first series, both
sugar and albumin failed to appear until after a single dose of over
4 grams of the salt had been administered, when considerable albu-
min showed itself in the urine. We were then compelled to stop
the experiment, and the dog was chloroformed and a ])Ost-mortem
immediately made.

The liver was excessively congested and appeared abnormally
large. Microscopic examination of hardened sections showed an

Chittenden and Lambert— Experiments on the

Table showing the amounts of Albumin and Sugar in the Dog's

Ueine.

1

Dose of

Date.

Volume.

Reaction.

Sp. Gr.

Albumin.

Sugar.

Uranyl
nitrate.

June

c. c.

grams.

grams.

grams.

8

640

acid

1016

0-050

9

600

"

1017

0-070

10

600

"

1015

0-075

11

602

"

1018

0-075

12

680

"

1017

trace

0-050

13

640

"

1022

1-741

0-075

14

710

"

1020

2-634

0-050

15

720

"

1023

3-009

150

16

830

"

1022

2-066

7-735

0-225

17

640

"

1025

1-441

7-449

0-450

18

650

weighed

1026
diet dis

2-633
continued.

7-476

19

1280

acid

1016

3-604

14-387

20

((

considerable

considerable

21*

750

"

1027

3-048

0-525

22

410

alkaline

1035

1-756

23t

. . -

acid

some

24

460

"

1018

0-578

25

430

alkaline

1030

trace

26

27

0-4

28

0-5

29

30

Julyl

1-0

2

3

. . .

1-5

4

4-4

5

----

considerable

infiltration of cells around the blood vessels, but aside from this and
an apparent tendency of the liver cells to separate into stringy
masses there was nothing abnormal. The kidneys both had adher-
ent capsules and the medullary portion was large in ratio to the
cortex, which, besides being smaller than normal, was striated. The
lesion as made out microscopically was acute parenchymatous neph-

* On this day granular casts were found, but they were not present on the follow-
ing day.

f Dog- vomited freely.

Physiological Action of Uranium Salts, 7

ritis. The endothelial cells of the malphigian tufts were swollen
and proliferated, so that frequently the tuft of vessels was com-
pressed by these proliferating cells and by the deti'itus and infiltrat-
ing pus cells. In the convoluted tubules the cells were swollen,
granular and very much broken down, while the lumen of the tubes
themselves was often filled with detritus and cast matter. In the
straight tubules, the cells were also occasionally broken down and
frequently swollen and granular. The stroma was found in places
infiltrated with pus cells. Stomach and intestines were normal.

Further data regarding the excretion of albumin and sugar,
through the action of uranium, are given in the description of the
toxic action of this substance.

The toxic action of uranium.
Our experiments on the toxic action of uranium have been confined
wholly to the action of pure uranyl nitrate on rabbits, nine in number.
The animals were confined in cages with proper outlets for collecting
the excreta, and were fed on grass, spinach, and other green food.

Experiment I.

Large, vigorous buck. In this experiment small doses of uranium,
gradually increased, were given in gelatin capsules, and the urine
examined each day for albumin and sugar. The accompanying table
gives the details of dose and the amounts of sugar and albumin
found.

Outside of changes in the urine there were no indications of toxic
action until on the ninth day, when a slight weakness was noticeable,
especially in the hind legs. After this, the animal gradually grew
weaker and emaciated, with total loss of appetite and with eyes dull
and Avatery. On the 8th of June, a large quantity of thick tenacious
mucus mixed with pin-head faeces was passed. Motion of all kinds
was difficult. On the 9lh of June, the animal could not sit up and
during poi'tions of the day there appeared to be paralysis of both
hintl legs. Later in the day the power of motion returned, and when
placed on his back the rabbit could kick out feebly with both legs.
On the next day his weakness and dullness were still more pro-
nounced, and when stirred from a squatting posture he trembled vio-
lently. His eyes were watery and glazed, with the pupils widely
dilated. On touching the cornea he did not offer to close the eye-
lids, but endeavored to withdraw his head. All jjower of vision was
apparently lost. Later, he lost the power of coordination in his legs,
was unable to make any coordinate movements, though still retaining

Chittenden and Lambert — Experiments on the

the power to move each limb separately. All faecal discharges had
stopped and the animal refused all food.

Date.

Volume.

May

c, c.

25

73

36

—

37

70

38

85

39

75

30

—

31

June

1

75
83

3

75

3

80

4

18

5

13

6

—

7

45

8

30

9

20

10

Reaction.

Sp. Gr.

Albumin.

Sugar.

gram.

gram.

alkaline.

1020

((

—

(1

1020

a

1016

0-035

'•

1022

0-189

trace.

"

1023

0-140

0-492

"

1023

0-125

"

1021

0-101

"

1020

0-173

neutral.

—

0-317

alkaline.

—

0-155

-

1025

0-518

"

—

0-301

consid-
erable.

Dose of Uranyl nitrate.

gram.

i 0-035
j 0-050
l 0-025

j 0-050
j 0-050
j 0-050

I 0-050

j 0-050
\ 0-025
j 0-050
] 0-050
I 0-050
\ 0-050
j 0-050
\ 0-050
J 0-050
/ 0-050

-i 0-050

j 0-100

j 0-100
\ 0-100
S 0-150
} 0-150
j 0-200
} 0-200
j 0-200
\ 0-150

2.175 grams.

On the morning of the 11th found dead. Post-mortem showed the
heart distended on the right side and the lungs normal. The liver
was much smaller than normal and the gall-bladder full of very black
bik'. On a microscopic examination of the liver, the only noticeable
feature was the extreme congestion. In the stomach, the mucous
membrane seemed somewhat disintegrated. In the intestines, brown
colored spots were seen where Peyer's patches are situated, and mi-
croscopic examination showed that the latter were deeply inflamed.
Further, from the large amount of mucus in the mucous glands, it
was evident that there was acute catarrhal inflammation of the intes-
tines. The cfficum was full of black matter. There were no normal
faeces in the colon or rectum, only the pin-head variety.

Physiological Action of Uranium Salts.

9

The kidneys were very mncli diminished in size and extremely con-
gested. Sections showed that the epithelial cells of the malpighian
tul'ts were not swollen or prolil'erated, though in some places the
tufts of vessels were slightly compressed by detritus. The cells of
the convoluted tubules were swollen, granular and broken down or
gone entirely, as were also the cells of the straight tubes, the latter
being also filled in places with casts and detritus. The vessels be-
tween the tubes were filled with blood. The stroma was swollen
and infiltrated with pus cells and in j^laces there were extravasations
of blood.

Brain and cord were slightly congested, but otherwise normal.

Examination of the table of urine tests shows that sugar appeared
first in the urine, but continued only lor three days, while albumin
showed itself immediately after the first appearance of the sugar and
continued throughout the experiment. The amount of ui'ine voided
grew gradually smaller as the animal began to feel the full effects of
the uranium,

Ex23erinie7it II.
In this experiment, uranium was given as indicated in the follow-
ing table, and the urine examined each day.

Date.

Volume.

Reaction.

Sp Gr.

Albumin.

Sugar.

Dose of uranyl nitrate.

June

c. c.

gram.

gram.

gram.

1

. —

alkaline.

—

0-050

2

100

"

1014

trace.

\ 0-050
■/ 0-050

3

55

-

1025

0-192

\ 0-050
I 0-050

4

41

"

—

0-227

0-406

\ 0-050
} 0-050

5?
6f

125

acid.

1024

1-095

1-078

j 0-050
/ 0-050

7

120

alkaline.

1024

0-523

0-917

3 0-050
/ 0-050

8

100

acid.

1022

0-237

trace.

( 0-100
] 0-100

9

55

alkaline.

—

0-351

\ 0-100
} 0-100

10

125

"

1022

0-687

0-430

j 0-100

I 0-100

0-100

11

75

a

—

cuiisiderabie.

12

75

a

1022

0-332

0-100

1.350 grams.

On June 4th, the animal began to show weakness, particularly
noticeable in his hind legs, but this disappeared and the animal
Trans. Conn. A.cad., Vol. VIII. 2 Nov., 1888,

10 Chittenden and Lambert — Exfperhnents on the

appeared better. On the 11th, it had lost its appetite, become emaci-
ated and showed general weakness and loss of muscular power. On
the following day at noon it was found dead, without having shown
any marked symptoms other than general depression of the muscular
system.

The heart was found greatly distended on the right side, Inngs
normal and the liver small and congested, especially near the lower
edges of the left lobe. Kidneys were also small and congested. The
bladder contained quite a little urine, with considerable albumin and
sug^ar in it.

Ex2yeriment III.

This experiment was practically a duplicate of the preceding. An
albino buck was used and the uranium salt was forced down the
throat in gelatin capsules, as in the preceding experiments. The
table shows tlie amount of nitrate given and the character of the
urine each day.

Date.

Volume.

Reaction.

Sp. Gr.

Albumin.

Sugar.

Dose of iiranyl nitrate.

May
25

26

c. c.
171

alkaline,
acid.

1014

gram.

trace.

gram.

gram.

0-025

\ 0-050

/ 0-025

27 1

28 f

29 1

30 f

31

no urin

e passed.

] 0-050

75

alkaline.

1026

0-494

0.275

i 0-050
\ 0-050
j 0-050
I 0-025

no urin

e passed.

0-300 gram.

The first symptom noticeable was the suppression of urine for over
forty-eight hours. On the 2Vth, the animal was quite dull and showed
signs of general weakness. As he walked, he moved as if it caused
him pain in the lumbar region. On the next day he showed great
thirst, and he finally died on June 1st, a loose diarrhcea setting in a
few hours before death.

Post-mortem examination showed that the heart had stopped in
diastole, and that it was much distended with blood. Lungs were
normal. The liver was congested, as were also the kidneys. The
latter had non-adherent capsules. The stomach contained consider-
able undigested food, and its mucous membrane was partly disinte-
grated. Portions of the duodenum were somewhat congested. The

Physiological Action of Tlranium Salts. ' 11

bladder contained a few cubic centimeters of urine, which had in it
considerable sugar and some albumin.

The most noticeable feature of this experiment was the suppression
of urine.

Experiment IV.

In this experiment, a large white buck was used and the uranium
was administered in capsules as follows :

May 17

0-050 gram uranyl nitrate.

18

0-075 "

19

0-075 "

20

0-025 "

0-225

There were no symptoms of toxic action until the morning of the
20th. The rabbit was then dull and weak. He trembled violently
as he hopped about; the hind legs appeared j^artly paralyzed. The
pupils were dilated and the appetite gone. On the 21st, weakness
was still more pronounced. Any movement was accompanied by
tremblings and great difficulty was experienced while walking, in
making the dilferent movements correctly. There was severe diar-
rluea and the animal showed extreme emaciation. On the 22d, the
diarrhoea had passed into an involuntary defecation more or less
continuous. The animal was too weak to move and lay all the fore-
noon breathing heavily, though the number of respirations per minute
did not go much beyond normal, 45-54 per minute. On the afternoon
of this day, two days after the last dose of uranium had been admin-
istered, the animal appeared stronger, the involuntary defecations
had ceased, though a loose diarrhoea still continued, and the animal
appeared to have recovered the use of its locomotive muscles. On the
23d, however, the animal was again unable to move and finally died
at noon, the diarrhoea having continued more or less up to death.

During the last three days there was complete suppression of
urine, but on the 20th, 59 c. c. of urine were passed, of specific gravity
1022, and which contained 0-656 gram of sugar.

On post-mortem examination, the heart was found to have stopped
in diastole and engorged with blood. The lungs were congested.
The kidneys had non-adherent capsules and the cortex and medulla
showed severe congestion. The stomach contained no food, but con-
siderable tenacious mucus. The small intestines from duodenum to
caecum were very much congested.

12

Chittende7i and Lambert — Experiments on the

Experiment V.

Gray and white buck. This experiment was a repetition of No.
IV, the same amounts of uranium salt being given and at the same
intervals of time. The animal died three days after the last dose of
uranium had been administered. The symptoms were not very
marked, simply loss of muscular power, and a gradual wasting away.
On post mortem, the heart was found in diastole and engorged with
blood. Lungs congested. The liver was black with the blood in it,
especially on the edges, and when pressed between the fingers was
extremely pliable and of a soft, pulpy consistency, showing marked
parenchymatous degeneration. The gall bladder was distended with
very dark bile. Both kidneys had non-adherent capsules. The left
kidney was slightly congested. The intestines had an inflamed patch
in the jejunum and were congested for a foot above it. The bladder
contained a little urine, which gave reactions for both sugar and
albumin. Sugar and albumin were also found in the urine several
days before death.

Experiment VI.

In this experiment still smaller amounts of uranium were adminis-
tered, a total of only 0-175 gram being given in three days. The
foUowino; table shows the dosase and tlie chanoes in the urine :

Date.

Volume.

Reaction.

Sp. Gr.

Albumin. Sugar.

Dose of
iiraiiyl nitrate.

June

14

15

16

17

18

19/

20 f

21

22

c. c.

110

75
30

60
50

alkaline

1043

1027
1022

1015

gram.

0-104
0-337
0-126

0-154

some
trace

gram.

I

gram.

0-050
j 0-050
\ 0-025

0-050

0-175

There were no noticeable symptoms of toxic action aside from the
changes in the urine, and even here there was no sugar at any time
present. Just as the albumin had almost disappeared from the urine
and we looked for speedy recovery, the animal Avas found dead (on
the 23d). Post-mortem showed the heart in diastole, engorged with
blood. Lungs normal. Liver normal, aside from a slight congestion.

Physiological Action of Tlraniimi Salts. 1 3

Sections under the microscope showed no pathological changes. The
kidneys were congested. Under the microscope, the epithelial cells
were proliferated and occasionally the tufts wei-e seen slightly com-
pressed by these proliferating cells and detritus. The convoluted
tubules showed swollen and granular cells, even broken down, in
places. In the straight tubules, the cells were also swollen and gran-
iilar and sometimes were detached from the tubes. In places, there
was cast matter and detritus in the tubes. The stroma was normal.
Hence, here, as in many of the preceding experiments, it is a case of
acute parenchymatous nephritis. The stomach was full of undi-
gested food and apparently normal ; the duodenum was congested
and the small intestines throughout were nearly empty, except for a
little mucus. The bladder had in it about 10 c. c. of urine, which
contained a trace of albumin but no sugar.

Experiment VII.

An albino rabbit. A single dose of 0*3 gram of uranyl nitrate
was given in a gelatin capsule by mouth on June 13th. Outside of
changes in the urine there were no symptoms whatever until the 17th,
when in the early morning the animal was found weak and power-
less, all motor power coinpletely gone. It rapidly grew weaker and
died in the afternoon of the same day.

The right side of the heart was found much distended. The lungs
were normal, the liver small and congested. Kidneys were also
small and congested and of a cloudy appearance. Microscopic exam-
ination showed acute parenchymatous nephritis. Stomach was full
of undigested food, but the intestines were empty. Both appeared
normal.

On June loth, 160 c. c. of alkaline urine were passed, of specific
gravity 1023. It contained 0-748 gram of albumin and 1-069 grams
of sugar. On the 16th, 30 c. c. were passed of 1022 specific gravity
and containing 0-412 gram of albumin and 0-354 gram of sugar.
On the 17th, no urine was passed and on making the post mortem the
bladder was found empty.

With this rabbit, an attempt was made to ascertain how much
carbohydrate matter there was in the liver at the time of death.
40 grams of the sampled and finely ground liver were thoroughly
extracted Avith boiling water (continuous extraction for three days),
frequently replaced. The several decoctions were ultimately united
and finally brought to a volume of 500 c. c. Two portions of 200 c. c.
each were placed in suitable flasks and suflicient hydrochloric acid

14 Chittenden and Lambert — Experiments on the

added to each, to make the fluid contain 2 per cent. HCl. The acid
fluids were then heated on water-baths for 15 hours, in order to con-
vert all carbohydrate matter into dextrose. The fluids were then
neutralized, evaporated and finally tested for dextrose with Feh-
ling's solution, by Allihn's gravimetric method. Both solutions failed
to give any reducing action whatever, thus showing a total lack of
carbohydrate matter in the liver. This is in strong contrast to the
normal condition of a rabbit's liver, which contains an abundance of
carbohydrate matter, both sugar and glycogen; on an average 10"35
per cent, of total carbohydrates, as determined by methods similar
to the one just described.* Glycogenic function is then destroyed by
uranium, the same as in phosphorus poisoning, but unlike the action
of phosphorus there is apparently no fatty degeneration of the organs.

The liver here experimented with was small for the size of the rab-
bit, a large white buck. It was also noticeable, as in many of the
other rabbits experimented on, that while the stomach was full of
food, sometimes even distended by it, there was nothing at all in the
intestines below the pylorus until the ca3cum was reached, which was
again lull, and the intestines below were either almost free fi'om
faeces or else contained only the pin-head variety. The food in the
stomach was wholly undigested. As has been already pointed out, a
small ])ercentage of a soluble uranium salt is sufficient to completely
stop gastric and pancreatic digestion. This being the case it is quite
probable that the emaciation, etc., so noticeable in uranium poison-
ing, is the direct result of the action of the salt on the digestive func-
tions. Nothing being digested there would be no matter for absorp-
tion, and hence no sugar-forming material for the liver. All the
carbohydrate matter stored up would in a little time be completely
consumed and as the portal blood could bring no new nutritive mat-
ter, the liver would naturally diminish in size and the animal become
emaciated and eveniually die from that cause alone, even if the uran-
ium gave no other direct cause of death. In this connection we also
need to recall the general increased metabolism of both nitrogenous
and non-nitrogenous matter, under the influence of uranium.

The uranium salt may also act specifically on the liver cells, afiect-
ing their metabolic power, preventing any storage of carbohydrate
matter or moi'e probably causing a degeneration of the cells, by
which they may be led to give up to the blood in abnormal abund-
ance all the carbohydrate matter previously stored up. This possible

* Post-mortem formation of sugar in the liver. Studies from Laboratory of Physio-
logical Chemistry, Yale Uuiversity, vol. I, p. 171.

Physiological Action of ZTranium Salts. 15

sjiecific action of the poison on the liver cells would in a measure ex-
plain the temporary glycosuria, which appears and then disappears so
frequently in uranium poisoning. Further, by a combination of this
specific action with the non-absorption of nutritive matter through
retarded digestion, it would be easy to explain the alternate appear-
ance, disappearance and re-appearance, of sugar in the urine in these
cases. Where the symptoms run their course wnth a fair degree of
rapidity, as in continued dosing with uranium, sugar appears in the
urine for a few days and then disappears, although the amount of
uranium administered may be steadily increased. In fact, the in-
creased dose of uranium is doubtless the cause of the further non-
appearance of the sugar, since the stored up carbohydrate material
having been wholly used up, and at the same time digestion and ab-
sorption being prevented, there is no more carbohydrate-producing
material available, either directly or indirectly. On the other hand,
if, after the sugar at first present has disappeared from the urine,
there comes an interval of a day or so when uranium is not adminis-
tered and digestion again starts up, then the specific action may
again come into play and a temporary glycosuria again result. This
condition was noticed in experiment No. II.

In some of the experiments, there seems to have been a visible
change in the hepatic cells, as in the experiment next to be described,
where under the microscope the cell bodies appeared as if collected
into small granules or even broken down. Again, in some experi-
ments, as in No. V, there was a noticeable pulpy degeneration of the
liver cells.

Experiment VIII.

In this experiment, the uranium salt was introduced by hypodermic
injection. 0'23 gram of uranyl nitrate in a little water was injected
beneath the skin of the leg of a good-sized rabbit, on June 20th at 4.0
p. M. Two days after he showed marked weakness, although his
appetite remained good. On the following day he appeared quite
dormant and could be roused only with difficulty ' power of motion
seemed to be nearly gone. He died the next day at noon. On June
21st, the day following the administration of the uranium, 60 c.c. of
alkaline urine were passed containing 0"234 gram of albumin and
0*660 gram of sugar. After this, there was complete suppression of
urine till death.

Post-mortem. — Heart had stopped in diastole ; lungs nornial. The
liver was mottled and congested slightly in areas. Under the micro-

1 6 Chittenden Mid Lambert — Experhnents on the

scope, sections showed the cell bodies strongly granulated, with their
outlines mostly clear, but in places broken down.

There was also an infiltration of pus cells around the blood vessels.
The kidneys were normal in size and appearance, with non-adherent
capsules. Under the microscope, however, all the conditions charac-
teristic of acute parenchymatous nephritis were to be seen. The
epithelial cells of the malpighian tufts were proliferated, so much so
as to compress the vessels. Both convoluted and straiglit tubules
had their cells swollen, granular and broken down. The stroma was
normal. Some adipose tissue in the pelvis of the kidney was infil-
trated with pus cells. There was also considerable detritus between
the papillae. The bladder was empty and contracted. In the abdom-
inal cavity there was about 30 c.c. of a coaguiable, clear fluid. The
stomach was full of food, the intestines empty. No foeces in colon
and rectum. No signs of congestion in the alimentary tract, except
in the rectum, where there were red blotches which proved to be the
beginning of inflammation and infiltration of pus cells. The animal
appeared to have died from suppression of urine.

Experiment IX.

In this experiment, one large dose of uranium (TO gram of the
nitrate) was given by mouth on June 27th, at 4.30 p. m. On July 5th.
the animal was still alive, but weak and emaciated. There were no
symptoms other than those already described. Owing to lack of
time we were not able to continue the experiment, but wishing to see
any changes which might have occurred in the organs the animal was
chloroformed and a post mortem made. On June 28th, the urine
passed contained some albumin, but no sugar; on July 1st, considera-
ble albumin, but still no sugar.

The liver was found to be small but not congested. The cell sub-
stance, as seen iinder the microscope, was collected into small gran-
ules. Nuclei and nucleoli were quite distinct. Kidneys had non-
adherent capsules, but they were congested and the cortex looked
striated. Under the microscoi^e, the epithelial cells of the glomeruli
were seen to be swollen and proliferated. In the convoluted tubes,
the cells were slightly swollen and granular and occasionally broken
down. The straight tubes were also broken down in places and con-
tained cast matter. The stroma was normal. Stomach was filled
with food, while the intestines were entirely empty. Peyer's patches
were swollen and infiltrated with pus cells. The cgecum was par-
tially filled witli matter and the end of it looked honey-combed. It

Physiological Action of Uranium Salts. 17

proved to be slightly inflamed, and with cells infiltrated into it more
or less. Colon and rectum were empty.

From these results, collectively, it is to be seen that uranium is an
irritant poison and, like other metallic irritants, produces gastro-intes-
tinal irritation of more or less intensity, as shown by the acute diar-
rhoea and other symptoms met with in this lorm of poisoning. In
the majority of cases, the action of the uranium salt on the intestines
results in a simple enteritis, but this is liable to pass eventually into
acute catarrhal inflammation.

As ordinarily administered, it is not in any sense a rapid poison;
the ingestion of a fatal dose of a uranium salt is not followed by any
noticeable effects for some time. The action of a small amount (150
milligrams) is apparently as rapid and pronounced as that of large
quantities (I gram). The first noticeable symptom in rabbits is gen-
eral weakness, lack of motor power, loss of coordination and occa-
sional temporary paralysis of the locomotor muscles. Introduced
into the stomach in suflicient amounts, it checks digestion and even
stops it altogether. On the other hand, it appears to increase some-
what proteid metabolism and also to increase the elimination of car-
bonic acid and raise the body temperature. Hence, it is to be consid-
ered as having a direct action on nutrition, the disturbance of which
is also plainly indicated by the rapid emaciation which follows the
administration of uranium.

Its most marked lesions are its destructive action on the kidneys,
and its destruction of the kidney tissue itself. It causes here an acute
parenchymatous nephritis of the same kind as found in arsenic, mer.
cury, and phosphorus poisoning. Further, the quantity of albumin
found in the urine shows plainly how greatly the blood vessels are
involved in the inflammation. The albuminuria produced is severe and
constant, and when the uranium is given in a single large dose, as in ex-
periment No. IX, or in a small dose by hypodermic injection, as in ex-
periment No. VIII, then albumin may appear in the urine within 24
hours. The uranium must have some specifically destructive action on
the kidney epithelium cells, causing them to swell and break down. At
first, with small doses of uranium, the urine is decidedly increased in
volume, but later on, when toxic action is more pronounced, there may
be a partial or even complete suppression of the urine. This latter
condition is naturally more quickly produced by large doses of ura-
nium. In several cases, suppression of urine would seem to have been
the cause of death. The urine, too, in a short time after the admin-
Traxs. Conn. Acad., Vol. VIII. 3 Nov., 1888.

18 Chittenden and Lamhert — Experhnents on TJraniuiin.

istration of uranium contains more or less sugar; as a rule the sugar
does not make its appearance until after the albumin. Salkowski has
shown that mercury will also cause diabetes in rabbits, but it is
questionable whether it is as constant a symptom as in uranium pois-
oning. It is also stated that in phosphorus poisoning the urine some-
times contains sugar.* In the hypodermic injection of uranium, ex-
periment No. VIII, sugar appeared in the rabbit's urine within 24
hours. The production of glycosuria is a very characteristic symptom
in uranium poisoning. The urine also contains invariably a large
amount of crystallized calcium oxalate, which would also point to de-
cided malnutrition and help explain the marked emaciation so com-
monly seen.

In those cases where the poisoning becomes in a measure chronic,
the nervous symptoms sometimes predominate, as shown in loss of
sight and power of coordination.

* H. C. Wood. Therapeutics, p. 110.

II. — Elastin and the Elastose Bodies. By R. H. Chittenden
AND Horace S. Hart, B.A., Ph.B.

Elastin, the basis of the so-called elastic tissue, has generally been
considered one of the most indigestible of the albuminoid substances.
In fact, the older writers looked on it as almost insoluble in the
digestive juices. Recently, however, the experiments of Etzinger,*
Horbaczewskijf and Morochowetz,J have shown that both the ligamen-
turn nucliOi of the ox when finely divided, and purified elastin when
powdered, are fairly soluble both in pepsin-hydrochloric acid and in
natural gastric juice, as Horbaczewski's observations on a man with
stomach fistula have fully indicated.

It has been our object, therefore, to study somewhat in detail the
primary cleavage products of elastin, so far as they may be formed?
making use of the methods§ which have yielded such fruitful results
with many of the proteid bodies.

Preparation of Elastin.

As usually described, elastin is a body free from sulphur, insoluble
in water even after several days boiling, likewise in cold dilute alkali,
acetic acid, dilute hydrochloric acid, alcohol, and ether. Naturally,
therefore, the method of preparing such an insoluble body has con-
sisted in removing from a tissue rich in elastin, all extraneous sub-
stances by successive treatment with the above-mentioned reagents.
In fact, so vigorous is the method of treatment, as usually described,
that it appears almost questionable whether a body belonging to a
group noted for ease of decomposition might not sufi'er some change
in such a long process of preparation.

We have employed two methods, the first one of which is practi-
cally identical with that followed by Horbaczewski. The neck bands
of a number of recently killed oxen were thoroughly freed from all
adhering fat and muscle and then chopped quite fine ; they weighed

* Zeitschrift fiir Biologie, Band x, p. 84.

f Ueber das Verhalteu des elastins bei der Pepsinverdauung, Zeitschrift fiir physiol-
ogische chemie, Band vi, p. 330.

\ Verdauiingsgesetze, Abstract in Jahresbericht fiir Thierchemie, 1886, p. 271.

§ Kiihiie and Chittenden, Zeitschrift fiir Biologie, Band xx, xxii; also Studies from
Laboratory of Physiological Chem. Yale University, vol. ii.

20 Chitten(le7i and Hart — Elastin and the Elastose Bodies.

2400 grams. The mass was boiled for four days in water frequently
changed, after which it was soaked for 45 hours in 8 litres of 1 per
cent, potassium hydroxide, and then boiled with the solution for 4
hours. The tissue was freed from alkali by thorough washing with
water, and was then boiled with water, frequently changed, for 16
hours. The potassium hydroxide solution on neutralization, gave a
heavy precipitate slightly soluble in excess of 0-4 per cent, hydro-
chloric acid, easily soluble in strong acid. The addition of acid in
excess caused a strong odor of hydrogen sulphide to be given off.

After boiling the tissue a second time with water, it was placed in 8
litres of 10 per cent, acetic acid and warmed for an hour and a half,
after which it was allowed to stand in the cold for 16 hours. It was
then boiled for 4 hours in the same acetic acid solution, and after-
wards washed thoroughly with water. The tissue was then placed
in 10 litres of 5 per cent, hydrochloric acid and allowed to soak for
some time in the cold, after which it was washed in running water
and macerated in water, frequently changed, for 24 hours to take out
all traces of the acid. After this treatment, the tissue was boiled
with successive portions of water for about 45 hours, when it was
placed in a large volume of 95 per cent, alcohol and allowed to re-
main for several days. It was afterwards boiled in alcohol for about
15 hours and then placed in an excess of ether, after which it was ex-
tracted with warm ether for two days.

Analysis of Elastin A.

Sub-
stance
used,
gram.

H2O
fouud.
gram.

H

N found.

N

Ash
found,
gram.

No.

CO...
found. d
gram.

c. c.

T.

°C.

Pres-
sure,
mm.

Ash

%

I

11

III

IV

V

VI

0-4250
0-3398
0-5454
0-4440
0-5125
0-2971

0-2745
0-2213

7-17
7-23

0-8381
0-6697

53-77
53-74

72-7
59-8
69-4

8-6
9-0
9-4

771-5
771-2
767-1

16-47
16-59
16-59

0-0027

....
0-9

Percentage composition of ash-free substance .

C
H

N
O

54-26
7-24

54-22
7-30

16-74

16-76

16-62

Average.

54- 24

7-^7

16-70

21-79

100-00

Gldttenden and Hart — E last In and the Elastose Sadies. 21

The tissue on being freed from water was hard, tough, and difficult
to powder. It was finally ground to a coarse powder and then re-
extracted with warm ether, until the ether on evaporation left no
appreciable residue. This took some time, for, as previously pointed
out by Horbaczewski, there apparently remains in the tissue a small
amount of fat-like matter, which dissolves very slowly in ether and
which can be completely extracted only by first grinding the purified
tissue as fine as possible. The elastin prepared in this manner we
have termed for convenience Elastin A.

A portion of the preparation dried at 110° C. gave on analysis the
results contained in the accompanying table. The methods of
analysis were the same as those previously described in former
articles on the proteoses.* By fusion with potassium hydroxide and
potassium nitrate, according to the method recommended by Ham-
marsten,f no sulphur could be found. Elastic tissue, however,
unquestionably contains sulphur, but whether it exists there as a
constituent part of the elastin molecule, or loosely united as in kera-
tin, or as a constituent of some adhering j^roteid or other substance,
it is difiicult to say. Certainly, the boiling of elastic tissue with 1
per cent, potassium hydroxide for several hours might reasonably be
expected to remove a part at least of any sulphur which might be
present, and if by this process sulphur is removed from the tissue
might it not as probably come from the elastin, as from any other
proteid substance? Treatment with acid, of the alkaline solution
obtained in the preparation of A, plainly showed the presence of
hydrogen sulphide and we therefore decided in the preparation of the
second portion of elastin to omit the treatment with alkali. Accord-
ingly, 1700 grams of carefully cleaned neck bands from oxen were
treated in the same manner as in A, except that the alkali was
omitted, and in its place, treatment of the tissue with both acetic and
hydrochloric acid was repeated twice. In subjecting elastin to the
action of 5 per cent, hydrochloric acid care must be taken, before
boiling the washed tissue with water, to see that every trace of acid
is removed, otherwise the faintly acid water formed will at 100° C.
give rise to a partial decomposition of the elastin.

After thorough extraction of the powdered elastin with ether, a
portion dried at 110° C. gave by analysis the results shown in the
following table, Elastin B.

* See Zeitschrift fiir Biologie, Band xx, p. 11, and Amer. Chem. Jour., vol. vl, p. 3.
f Zeitschrift fiir physiologische Chemie, Band ix, p. 298.

22 Chittenden and Hart — Elastin and the Elastose Bodies.

Analysis op Elastin B.

Sub-
stance
used,
gram.

H,0

found,
gram.

H

%

C0.2

found,
gram.

C

%

N found.

N
%

BaS04 after

fusion with

KOH + KNO3

gram.

Ash
found,
gram.

No.

c, c.

T.

Pres-
sure,
mm.

S

Ash

fo

I

0-5833
0-4374
0-3836

0-3744
0-2851

7-13 1-1531

53-90

----

....

----

II

7-24 0-8653 53-94

III

54-1

11-0

754-8

16-91

IV

0-4821

66-8

11-7

760-4

16-69

V

0-9095

0-0170

0-25

VI

0-7854

0-0208

0-36

VII

0-4235

0'0015

0-35

VIII

0-4583

---

0-0013

0-28

Percentage composition of ash-free substance .

c

54-06

54-10

H

7-15

7-26

N
S

----

■---

16-96

16-75

0-34

0-39

Average.
54-08

7-20
16-85

0-30
21-57

100-00

Comparison of the two products shows a fairly close agreement in
composition. In B, however, the carbon is slightly lower and the
nitrogen a trifle higher than in A. Further, Elastin B contains 0-3
per cent, of sulphur, as determined by the modified Liebig's process.*
Miiller originally reported elastin, purified, however, by the use of an
alkali, as containing 0-08 per cent, of sulphur, but this he considered
merely as an imj^urity.

Morochowetz, who appears to have made a hydration product of
elastin by the simple action of heat and water, states that it contains
0-617 per cent, of sulphur, but the details of his process we do not
know.f If the figures are correct, his elastin must have probably con-
tained double the percentage of sulphur present in elastin B-
Whether pure elastin does contain sulphur, or whether the 0-3 per
cent, present in i^reparation B is simply a constituent of some adhering
proteid, removable by alkali, we are not at present prepared to say,
but deem it probable that elastin does contain a small amount of
sulphur.

■ See studies from Laboratory of Physiological Chemistry, Yale University, vol. ii, p.

163.

\ We have seen only an abstract in Jahresberieht fiir Thierchemie fiir 1886, p. 271.

Chittenden and Hart — Elastm and the Elastose Bodies. 23

Elastin A., prepared by the usual process, shows a close agreement
in composition with Horbaczewski's product, but is quite different
from the product obtained by Miiller and Tilanus, as seen from the
following table.

Elastin A. Horhaczeivski. Miiller* Tilanus*

C 54-24 54-32 55-09-55-72 54-90-55-65

H 7-27 6-99 7-11-7-67 7-25-7-41

N 16-70 16-75 15-71-16-52 17-52-17-74

The liigher content of carbon in the preparations of Miiller and
Tilanus are doubtless due to the presence of more or less fat, not
completely extracted by ether.

Decomposition of Elastin by acid loater at 100" C.

On heating moist elastin with water containing a trace of hydro-
chloric acid, for some hours at or near 100° C. it soon commences to
swell, and after a time becomes converted into a semi-gelatinous
mass, a portion of which, as the boiling continues, becomes soluble and
passes into the acid fluid. The liquid gives no reaction with tannic
acid for gelatin, but does give a strong biuret reaction and yields a
heavy turbidity when excess of strong potassium hydroxide is added
to the solution. Evidently, under these conditions, some soluble
cleavage product or products are formed, percipitable by excess of
caustic alkali.

These products we prepared in quantity by taking a portion of elas-
tin ^, soaking it for some time in 5 per cent, hydrochloric acid, until
the softened elastin was thoroughly impregnated with the acid, then
washing out the excess of acid by soaking it for 20 hours in a laro-e
volume of water. The elastin, still quite strongly acid to test papers,
was then boiled in water for about 10 hours, when the acid liquid
was strained off from the gelatinous elastin. The boiling was then
continued with a fresh volume of water, the gelatinous mass still
containing sufficient acid to render the whole fluid distinctly acid
to test papers. Tnis process was repeated until the elastin had
been heated for 45 hours with water gradually containing less
and less acid, until finally the elastin remaining had entirely lost
its former gelatinous appearance and taken on its original look.
All of the acid fluids were united and concentrated somewhat,

* Gorup-Besanez, Physiologische Cliemie, Dritte Auflage, p. 148

24 Chittenden and Hart — Elastin and the Elastose Bodies.

then neutralized with sodium carbonate. No neutralization pre-
cipitate was observed. By further evaporation of the neutral fluid
on a water-bath, a gummy mass separated on the bottom of the
dish. It was quite tenacious, something like rubber, soluble in cold
water, biit not so readily in hot. The cold water solution of the
gummy mass, and the mother-liquor showed the following reactions:

1 . Heated in a test-tube, the sohition quickly became turbid, giving finally,
if concentrated, a sticky precipitate. The turbidity, however, immediately
disappeared as tlie fluid cooled, returning again when heat was applied
and disappearing wlien cold. When the solution was so concentrated as to
yield a heavy precipitate by heat, it naturally dissolved more slowly.

2. With chloroplatinic acid, a heavy yellow precipitate was formed,
readily soluble in alcohol.

3. Heated with acetic acid there was no change, but addition of potas-
sium ferrocyanide to the acid fluid caused a heavy precipitate.

4. Millon's reagent gave a strong reaction similar to the albumin reaction.
.5. Strong potassium hydroxide gave a fine flocculent precipitate, insoluble

in excess of the alkali.

6. Pure dilute nitric acid gave a heavy precipitate, only slowly soluble in
excess of the acid. Heated, it yielded a clear yellow fluid and with ammo-
nia gave the orange yellow color of the xanthoprotein reaction.

7. Pure dilute hydrochloric acid yielded a slight precipitate, soluble in
large excess.

8. Saturation of the sohition with ammonium sulphate gave a heavy,
gummy precij^itate.

The gummy mass and the original filtrate giving approximately
the same reactions, the solution of the former was united witii the
latter, the fluid made slightly acid with acetic acid and then satu-
rated in the cold with ammonium sulphate. The saturated fluid, on
bcino- boiled, gave a second gummy precipitate which was added to
the first.

From analogy with the reactions of the proteoses, ammonium sul-
phate should precipitate any elastose bodies formed, leaving in solu-
tion the true peptone. The precipitates formed in the manner de-
scribed were washed by being rubbed up in a mortar with hot satu-
rated ammonium sulphate solution, and the washings added to the
filtrate. A portion of the ammonium sulphate was then removed
from the solution by concentration and crystallization, and the re-
mainder by dialysis in running water. On testing the solution
for peptone, however, none could be found; the biuret test failed to
give any reaction, as did also phosphotungstic acid.

Chittenden and Hart — Elastiu and the Elastose Bodies. 25

Protoelastose.

The ammonium sulphate precipitate of elastoses, after being washed
as described above, was dissolved in water, the solution made quite
neutral by addition of a little sodium carbonate and then saturated
with crystals of rock salt. A heavy gummy precipitate resulted,
which by analogy should consist of what we may call protoelastose.

This precipitate, after being washed with saturated salt solution,
was dissolved in water and reprecipitated by salt, this process being
repeated three times. The final sodium chloride precipitate was dis-
solved in water, dialyzed until all chlorine was removed from
the solution, concentrated and precipitated with 95 per cent, alcohol.
Apparently, only a portion of the substance was precipitated by
alcohol, so the alcoholic filtrate was evaporated and the residue care-
fully dried. Further examination, however, showed that both pro-
ducts were identical, alcohol simply precij^itating a portion of the
substance.

After being dried at 110° C. until of constant weight, having then
the form of a brown powder, both ])roducts were analyzed Avith th^
results shown in the accompanying tables.

Analysis of Protoelastose (A1) Precipitated by Alcohol.

Sub-
stance

used,
gram .'

0-6852

H..0
found .
gram.

PI

%

6-92

CO.
foimd.
gram.

C

r/

N found.

N

Ash
found,
gram.

No.

c. c.

T.

°0.

Pres-
sure,
ram.

Ash
%

I

0-4271

1-3190

52-49

II

0-3926

0-2437

6-89

0-7526

52-27

....

III

0-5400

.- - .

72-9

13-7

759-4

16-13

IV

0-3883

51-9

15-4

762-9

15-95

..

V

0-7048

0-0259

3-67

VI

0-5362

----

----

----

----

....

....

0-0202

3-76

Percentage composition of ash-free substance.

c

54-51

54-28

H

7-19

7-16

N

16-75

16-56

Average.

54-39

7-17

16-65

21-79

Trans. Conn. Acad., Vul. VIII.

100-00

Nov., 1888.

26 Chittenden and Hart — Elastin and the Elastose Bodies.

Analysis of Protoelastose (Ai) from the Alcoholic Filtrate.

Sub-

HoO

CO.

N found.

Ash

No.

stance

found.

H

found.

C

Pres-

N

found.

Ash

used.

gram.

%

gram.

%

c. c.

T.

°C.

sure.

%

gram.

%

gram.

mm.

I

0-4936

0-3113

7-00

0-9626

53-18

II

0-4096

0-2592

7-03

0-8079

53-78

....

III

0-4588

... -

63-2

13-4

759-8

16-49

IV

0-4367

....

62-5

21-6

760-8

17-06

V

0-3655

0-0054

1-47

VI

0-5096

....

...

....

....

.....

0-0075

1-47

Percentage composition of ash-free substance.

c

53-96

54-58

H

7-11

7-13

N

O

16-73

17-31

Average.

7-12
17-02
21-59

100-00

The reactions of the two preparations were identical and were as
follows; both were readily soluble in cold water, but only slightly
soluble in hot water; in neutral solution, heat produced a turbidity
which disa^ipeared on cooling; the concentrated mineral acids gave
precipitates soluble in excess ; phosphotungstic acid, picric acid, tan-
nic acid, 30 per cent, acetic acid saturated wnth salt, acetic acid and
potassium ferrocyanide, and alcohol all produced precipitates; mer-
curic chloride and mercuric nitrate gave precipitates insoluble in
excess ; lead acetate and cupric sulphate both failed to give any
precipitate ; potassium mercuric iodide produced a precipitate in a
hydrochloric acid solution ; sodium carbonate and sodium hydroxide
gave heavy precipitates and both the biuret and xanthoprotein test
gave positive results. Further, the elastose is but slightly, if at all,
diffusible. Long continued concentration of an aqueous solution
leads to a separation of more or less of the substance as a gummy
mass.

In composition, the protoelastose formed by action of the dilute
acid and heat is almost identical with that of elastin itself.

Morochowetz,* in a recent paper on the laws of digestion, states
that elastin by the action of heat and water passes into a new form,

*Loc. cit.

Chittenden and Hart — Elastin and the Elastose Bodies. 21

elastose, which by further action passes into peptone. The elastose
is described as a body completely soluble in water, not jjrecipitable
by mineral or acetic acids, but its solution is rendered turbid by heat.
In its other reactions it resembles albumin. In composition, how-
ever, it differs decidedly from elastin and equally from our protoelas-
tose. Morochowetz ascribes to it 55 '9 per cent, carbon, 7*29 per cent,
hydrogen, 16*68 per cent, nitrogen and 0*617 per cent, sulphur. As
to the method employed in its preparation and purification we do
not know, but as described it evidently is not akin to pure elastose.

Deiiferoelastose.

In the salt-saturated filtrate from the first sodium chloride precipi-
tate of protoelastose, there was present a substance precipitable as a
gummy mass, by the addition of a little 30 per cent, acetic acid sat-
urated with salt. Past experience with other proteoses has shown
that, as a rule, all of the proto body is not precipitated by saturation
of its neutral solution with salt, but that there usually remains a certain
amount which on addition of salt-saturated acetic acid is precipitated,
together with more or less of the deutero body, and that from this
filtrate pure deutero can be separated by saturation with ammonium
sulphate. In the present instance, however, it appeared that the
acetic acid precipitate was of quite a different nature from the sodium
chloride precipitate and that if it was not pure deuteroelastose, it at
least contained only a ti-ace of the proto body. Further, saturation
of the acetic acid filtrate with ammonium sulphate gave only a slight
precipitate, even when the mixture was heated, showing that if the
salt-saturated acetic acid had not precipitated the deuteroelastose,
little could have been formed.

The gummy precipitate separated by acetic acid and salt was
purified somewhat by solution in water, and reprecipitation by satura-
tion with salt and addition of a little salt-saturated acetic acid. It
was then dissolved in water, made exactly neutral to test papers, and
dialyzed until all chlorine was removed from the solution. The
filtered fluid was then evaporated to dryness on a water-bath, and the
powdered residue dried at 110" C. until of constant weight.

The composition of the substance is shown in the accompanying
table, from which it is seen that the deutero body contains a notice-
ably smaller percentage of carbon than protoelastose, and a corres-
pondingly higher content of oxygen. The difference between the
sodium chloride precipitate and the acetic acid precipitate, however,
becomes far more marked when the reactions of the two bodies are

28 Chittenden and Hart — Elastin and the Elastose Bodies.

compared. Deuteroelastose was readily soluble both in cold and hot
water, and on heating a cold saturated solution only a slight turbidity
was seen which, as in the proto reaction, disappeared when the solu-
tion cooled. This turbidity was no doubt due to the presence of a
small amount of protoelastose, for later on we obtained a purer deu-
tero body having nearly all of the reactions of the present preparation
in which, however, the heat reaction was absent.

Analysis of Deuteroelastose. A 1.

Sub-
stance
used.
gram.

N found.

Hi,0
found,
griim.

CO2
found,
gram.

C

N
t

Ash
found,
gram.

No.

c. c.

T.

Pres-
sure
mm.

Ash

%

I

0-5164

0-9894

52-24

II

0-4003

0-2499

6-93

0-7708

52-51

.. .

III

0-4648

0-2924

6-98

0-8885

52-12

IV

0-4278

0-2704

7-02

0-8149

51-94

V

0-3624

.

49-8

13-8

759-8

16-43

VI

0-4818

65-8

13-9

759-3

16-30

VII

0-5370

0-0105

1-95

VIII

0-4371

----

----

....

....

....

....

....

0-0091

2-08

C

53-30

53-57

53-19

52-99

H

7-07

7-13

7-13

N

16-76

16-64

Percentage composition of ash-free substance.

Average.

53-26

T12

16-70

22-92

100-00

The deutero body, likewise, gave no precipitate whatever with
the concentrated mineral acids, nor with acetic acid and potassium
ferrocyanide. Alcohol gave no precipitate and salt-saturated 30 per
cent, acetic acid, when added to an aqueous solution of the substance,
also gave no precipitate.

Ferric chloride gave a slight turbidity, but no precipitate. With
the other reactions of protoelastose, the deutero body showed close
agreement. Pure deuteroelastose, formed later on by the action of
pepsin -hydrochloric acid, gave no precipitate whatever with concen-
trated potassium hydroxide. This reaction, Avhich is quite character-
istic of protoelastose, was fairly distinct in the deutero body first
described, which fact, coupled with the turbidity produced by heat,
may be considered good evidence that the deuteroelastose was not
perfectly pure.

Chittenden and Hart — Elastin and the Elastose Bodies. 29

Digestion of elastin by pepsin-hydrochloric acid.

An artificial gastric juice, free from both albumose and peptone,
was prejDared as follows ; the cardiac portions from the mucous mem-
branes of ten pigs' stomachs (820 grams) were placed in three litres
of 0*4 per cent, hydrochloric acid and warmed at 40° C. for two
weeks. The clear fluid filtered from the small residue of nuclein, an-
tialbumid, etc, was then saturated with ammoniiim sulphate to pre-
cipitate the pepsin. This was filtered ofi", washed with saturated
ammonium sulphate solution to remove any adherent peptone, dis-
solved in two litres of 0*2 per cent, hydrochloric acid and dialyzed
until the sulphate was entirely removed. The resultant solution
was mixed with an equal volume of 0*4 per cent, hydrochloric acid,
making a pure and active pepsin-acid mixture.

In the first digestion of elastin, 150 grams of powdered elastin A,
1500 c.c. of the pepsin-acid mixture, and an equal volume of 0-4 per
cent, hydrochloric acid, were warmed at 40° C. for 75 hours. At the
end of this time the elastin was almost entirely dissolved. The acid
fluid was filtered from the small residue remaining, and neutralized
with potassium hydroxide, without giving any neutralization precipi-
tate. The reactions of the fluid showed plainly the presence of elas-
toses similar to those formed by the action of acid. Saturation of
the fluid with ammonium sulphate gave a heavy gummy precipitate,
in the filtrate from which, nothing having the reactions of peptone
could be discovered.

Protoelastose.

The ammonium sulphate precipitate was dissolved in water and the
neutral fluid saturated with sodium chloride, by which a heavy gummy
precipitate was formed, having all of the reactions described as char-
acteristic of protoelastose. The product was purified by several re-
precipitations with salt, then dialyzed and the solution evaporated to
dryness on a water-bath, the residue powdered and dried at 110° C.
until of constant weight.

The composition of the substance (protoelastose A 2) is shown in
the accompanying table.

In reactions, the product agrees exactly with the protoelastose
already described, being precipitated by the concentrated miner'al
acids, by acetic acid and potassium ferrocyanide, by 30 per cent,
acetic acid and sodium chloride, by strong potassium and sodium
hydroxide as well as by sodium carbonate. Its a<pieous solution,
likewise, shows tlu; peculiar action towards heat, already described ;

30 Chittenden and Hart — Elastin and the Elastose Bodies.

becoming turbid when heated, clear again as the solution cools. The
product also gives the other reactions common to both proto- and
deuteroelastose.

Analysis of Protoelastose. A 2.

Sub-
stance

used,
gram.

found,
gram.

H

CO2
fouud.
gram.

C

N found.

N
%

Ash
found,
gram.

No.

c. c.

T.

Pres-
sure
mm.

Ash

%

I

II

III

IV

V

VI

0-4607
0-4952
0-5546
0-3109
0-4603
0-5303

0-2880
0-8079

6-94
6-90

0-9103
0-9763

53-88
53.76

76-2

45-8

13-9
21-0

759-4

754-8

16-41

17-06

0-0061
0-0073

1-32
1-37

Percentage composition of ash-free substance.

c

54-58

54-46

H

7-03

6-99

N

16-64

17-29

Average.

7-01
16-96
21-51

100-00

This body, which we prefer to call protoelastose, is apparently iden-
tical in composition and reactions with the hemielastin previously de-
scribed by Horbaczewski,* and separated by him from a pepsin-acid
digestion of elastin, by a method somewhat similar to the one em-
ployed by us.

At first glance, it w'ould appear from Horbaczewski's method of
separation, that his hemielastin would consist of a mixture of proto-
and deuteroelastose, instead of being identical with the proto body.
But the fact that he strongly acidified his digestive mixture with
acetic acid, prior to saturating it with salt, explains the matter.
Deuteroelastose, which is precipitable from a salt-saturated solution
by a little acetic acid, is more or less soluble when excess of the acid
is added, and hence by acidifying the mixture sufficiently, the deutero
might remain dissolved while the proto would be precipitated by
salt from an acid solution, equally as well as from a neutral fluid,

* Ueber das Verhalten des Elastins bei der Pepsinverdanung. Zeitschrift fiir
physiologische Ghemie, Band vi, p. 3,30.

Chittenden and Hart — Elastin and the Elastose Bodies. 31

Deuteroelastose.

In the salt-saturated filtrates from protoelastose, deutero was sepa-
rated as a gummy, sticky precipitate by addition of a little 30 per
cent, salt-saturated acetic acid. It was purified by reprecipitation
and dialysis, and the final neutral solution was evaporated to dryness
and the residue dried at 110° C. until of constant weight. The
amount was small and only the nitrogen was determined, 16-88 per
cent.

In reactions, the body closely resembled the deutero already de-
scribed ; its aqueous solution giving no precipitate or turbidity what-
ever on the application of heat, no precipitate with alcohol, or with
strong potassium hydroxide solution. It was likewise not precipi-
tated by acetic acid and potassium ferrocyanide, by the concentrated
or dilute mineral acids, nor by 30 per cent, acetic acid. In reactions,
the body resembles the elastin peptone of Horbaczewski, like it being-
soluble in cold and warm water and dilute alcohol, and giving the
same precipitations with phosphotungstic acid, picric acid, tannic
acid, and with potassium mercuric iodide. It also gives the biuret
and xanthoprotein reaction. It dilfuses slowly.

With our present knowledge regarding peptones and proteoses,
this body can hardly be considered as belonging to the former class,
since it is precipitable both by ammonium sulphate, and by acetic
acid when added to a salt-saturated solution ; reactions not common
to true peptones.

A second digestion of elastin A was made with the same quantities
of pepsin-hydrochloric acid, and the same amount of powdered elastin,
as in the first digestion. The products formed were separated in the
same manner as the preceding, the elastoses l)eing first precipitated
collectively by saturation of the digestive fluid with ammonium sul-
phate.

On boiling the filtrate from this ammonium sulphate precipitate, a
second gummy mass separated. This was collected on a cloth filter
and washed with saturated ammonium sulphate solution. In the fil-
trate from this precipitate, no peptone could be detected. This
second ammonium sulphate precipitate was purified by dialysis, etc.,
and the final solution evaporated to dryness and the substance dried
at 110° C. Its composition is shown in the accompanying table,
from which it is seen to be nearly identical with that of protoelastose.
The reactions, however, indicate that it is a mixture of the two elas-
toses. Thus, while the concentrated mineral acids and potassium
hydroxide give no precipitate, acetic acid and potassium ferrocyanide

32 Chittenden and Hart — Elastin and the Elastose Bodies.

produce a noticeable turbidity, as does also alcohol, and acetic acid
and sodium chloride. Moreover, the aqueous solution becomes some-
what clouded on heating, clearing up again as the solution cools.
Hence, the reactions would indicate a preponderance of deuteroelas-
tose, while the composition points to an excess of the proto body.

Analysis of the Second Ammonium Sulphate Precipitate.

Sub-
stance
used,
gram.

H.,0
found,
gram.

CO.,
found,
gram.

C

N found.

N
%

Ash
found,
gram.

No.

c. c.

57-7
57-6

T.
° C.

14-2
22-6

Pres-
sure
mm.

758-0
753-9

Ash

I

II

III

IV

V

VI

0-4783
0-3781
0-4100
0-3963
0-4756
0-5016

0-3115
0-2433

7-23

7-14

0-9404
0-7405

53-61
53-40

16-76
16-72

0-0048
0-0054

1-00
1-07

c

H

N
O

Percentage composition of ash-free substance.

Average.

54-14 53-93 5^-03

7-30 7-22 .... -... 7-26
16-92 16-89 lG-90

S2-S1

100-00

We must conclude, then, that this second gummy precipitate is sim-
ply a residue of the mixed elastoscs not at first precipitated, perhaps
from a lack of complete saturation with ammonium sulphate. Past
experience, however, has shown that deuteroproteose is not as quickly
or completely precipitated witli ammonium sulphate as the other
proteoses.

From the first ammonium sulphate precipitate, a large amount of
protoelastose (A 3) was separated, having all of the reactions charac-
teristic of this body, and the composition, when dried at 110° C,
shown in the accompanying table.

A larger amount of deuteroelastose was separated from this diges-
tion than in the preceding one, and after purification by the use of
methods already described, a portion was dried at 110° C. for analy-
sis. Its composition is shown in the accompanying table. Like the
deuteroelastose formed by dilute hydrochloric acid, it contains a
lower percentage of carbon than elastin or the proto body, while its
content of nitrogen is higher than that of elastin. Its reactions were
the same as the deutero previously obtained.

Chittenden and Hart — Elastin and the Elastose bodies. 33

Denteroelastose appears to be the same as the elastin peptone of
Horbaczewski, as already mentioned ; and the composition of the
present product resembles it, in that it contains a similar percentage
of carbon, but is unlike it in containing one per cent, more nitrogen.

Analysis op Protoelastose. A 3.

No

Sub-
stance

used,
gram.

H2O
found,
gram.

H

%

CO2
found,
gram.

I

% c. c.

vT foun

T.
° C.

d.

Pres-
sure
mm.

X

16-99
16-89

Ash
found,
gram.

Ash

I

II

III

IV

V

VI

0-2990
0-4383
0-3938
0-3233
0-5562
0-4866

0-1906
0-2750

7-07
6-97

0-5841
0-8606

53-27
53-54

57-1

47-1

19-8
21-6

760-8

757-2

0-0081
0-0079

1-45
1-62

Percentage coviposition of ash-free substance.

c

54-08

54-37

H

7-19

7-08

N

17-26

17-16

100-00

Analysis of Deuteroelastose. A 3.

No.

Sub-
stance

used,
gram.

H,0
found,
gram.

H

%

CO2
found,
gram.

C

%

1
c. c.

'? foun

T.
° C.

d.

Pres-
sure
mm.

N

Ash
found
gram.

Ash

I

II

III

IV

V

VI

VII

0-5067
0-6021
0-4550
0-6886
0-3570
0-6638
0-7748

0-3125
0-3752
0-2705

6-85
6-92
6-60

0-9681

1-1587

0-8685

52-10
52-47
52-05

96-3
51-4

13-8
22-4

759-4

754-0

16-70
16-58

0-0201
0-0247

3-02
2-91

Percentage composition of ash-free substance.

c

53-68

54-07

53-63

H

XT

7-06

7-13

6-60

;;;;

;;;;

17-21

Average,

53-79

6'9'J

-31 17-26

21-96

Trans. Conn. Acad., Vol. VIII.

100-00

Nov., U

34 Chittenden and Hart — Elastin and the Elastose Sodies.

Horbaczevv ski's elastin peptone is described as not giving a precipi-
tate with acetic acid and neutral salt. Deuteroelastose, likewise,
gives no precipitate with acetic acid and salt, except when the solu-
tion is saturated or nearly saturated with the salt.

Another preparation of deuteroelastose was separated from a
pepsin-hydrochloric acid digestion of elastin B. Its chemical com-
position is shown in the accompanying table, from which it is seen
to differ somewhat from deutero A, but like It has a lower content of
carbon than the proto body.

Analysis of Deuteroelastose B.

Sub-

H2O

CO2

N found.

Ash

I

No.

stance

found.

H

found.

C

' T.

C. C. (.

Pres-

N

found.

Ash

used.

gram.

%

gram.

%

sure

%

gram.

%

gram.

0-3560

6-68

mm.

I

0-5919

1-0934

50-37

1

II

0-4201

0-3573

6-80

0-7796

50-60

III

0-4853

64-9 140

756-8

15-91

IV

0-4719

...

66-0 33-4

754-3

16-11

V

0-7960

0-0398

5-00

vt

0-5883

....

....

....

....

.... ....

....

....

0-0397

5-04

Percentage composition of ash-free substance.

c

53-01

53-31

H

7-03

7-14

N

16-81

16-90

Average.

53-11

7-08

16-85

22-96

100-00

Digestion of dastin u'lth pancreatic juice.

A strong solution of pure trypsin was prepared, according to
Kiihne's method,* from 100 grams of dried ox pancreas and added to
150 grams of powdered elastin B in 2 litres of 0-5 per cent, sodium
carbonate. The mixture M'as warmed at 40° C. for four or five days,
a little thymol being added to prevent putrefaction. At the end of
this time, the elastin was nearly all dissolved and the filtered fluid
on being tested gave the ordinary elastose reactions. On saturation
of a portion of the neutralized fluid with ammonium sulphate, a
heavy gummy precipitate was obtained, and in the filtrate no trace
of peptone I'eaction could be found.

* Kiihoe and Chittenden, Zeitschrift fiir Biologie, Band xii, p. 196.

Chittenden and Hart — Elastin and the Elastose Bodies. 35

The entire digestive fluid was therefore neutralized with dilute
hydrochloric acid (no neutralization precipitate), and the protoelas-
tose at ouce separated by saturation with sodium chloride. The
product was purified by repeated precipitation with salt, and after
dialysis, the resultant fluid was evaporated to dryness and the
residue dried at 110° C for analysis. The product contained con-
siderable ash (7'4 per cent.), consisting mainly of calcium phosphate
and sulphate, with some oxide of iron. Its composition, seen in the
accompanying table, is somewhat different from the protoelastose
formed by pepsin-hydrochloric acid, containing as it does a notice-
ably lower percentage of carbon. In its reactions, however, it
resembles closely the preceding preparations. It is readily soluble
in cold water and the solution when heated gives the characteristic
turbidity, which disappears as the solution cools. With acetic acid
and potassium ferrocyanide, it gives the usual precipitate and also
with the concentrated mineral acids, the latter dissolving in an
excess of the acid. Unlike the protoelastose previously described,
however, it gives with cupric sulphate a precipitate soluble in excess
of the copper salt, and with sodium hydroxide no precipitate. In
all other respects, its reactions resemble those of the proto body
formed in pepsin digestion.

Analysis of the SooroM Chloride Precipitate— Trypsin Digestion of B.

Sub-

Ho.O

OO2

N found.

Ash

No.

stance

found.

H

found.

C

T.

Q

Pres-

N

found.

Ash

used.

gram.

%

gram.

%

c. c.

sure

%

gram.

%

gram.

mm.

I

0-4455

0-2635

6-57

0-8045

49-24

....

II

0-4650

0-2696

6-44

0-8360

49-03

...

III

0-3765

. - - .

. - . -

49-7

16-0

765-4

15-77

....

IV

0-6754

87-3

15-1

765-4

15-49

... -

...

V

0-6473

0-0480

7-41

VI

0-8286

....

----

----

....

----

....

0-0610

7-36

Percentage composition of ash-free substance.

c

53-15

52-95

H

7-09

6-95

17-02

16-74

Average.

53-05

7-02

16-88

SS-05

100-00

In the salt-saturated filtrate from protoelastose, the addition of
a little 30 per cent, acetic acid saturated with salt gave a second

36 Chittenden and Hart — Elastin and the Elastose Bodies.

gnramy precipitate which was dissolved in water, the solution neu-
tralized and dialyzed until all chlorine was removed. The solution,
on evaporation, left a brownish residue, which was powdered and
dried at 110° C. It was readily soluble in cold and hot water, its
solution showing no turbidity whatever when heated. It likewise
gave no precipitate with alcohol. Unlike the ordinary deutero-
elastose, it did give noticeable precipitates with the mineral acids,
soluble in excess, and also with acetic acid and potassium ferro-
cyanide. Lead acetate and cupric sulphate also gave precipitates,
soluble in excess of the metallic salt. Sodium or potassium hydrox-
ide failed to give any precipitate. In composition too, the product
showed an approach to protoelastose formed by j^epsin digestion,
but unfortunately it contained over 5 per cent, of ash, and hence the
quantitative results may perhaps be questionable.

Analysis of the precipitate produced by Salt-saturated Acetic
Acid— Trypsin Digestion of B.

Sub-

HoO

CO2

N found.

Ash

No.

stance

found.

H

found.

C

T.

° C

Pres-

N

found.

Ash

lised.

gram.

%

gram.

c. c.

sure

i

gram.

%

gram.

t

mm.

I

0-4120

0-2466

6-65

0-7780

51-49

....

....

....

...

II

0-3625

0-2185

6-69

0-6899

51-89

III 0-3431

45-6

14-7

756-5

15-81

IVi 0-3436

45-8

19-4

754-8

15-52

V

0-6973

-- - -

0-0370

5-30

VI

0-5663

----

----

----

----

----

....

----

----

0-0337

5-60

Preeentage composition of ash-free substance.

c

54-46

54-85

H

7-01

7-07

N

16-71

16-40

Average.

54-6.5

7-04

16-55

21-76

100-00

Of anything corresponding to heteroalbumose, we have found no
trace. Further, under the conditions of our experiments, no appre-
ciable amount of true peptone was formed in any of the digestions ;
at least, nothing approaching a peptone in reactions was to be found
in any of the digestive fluids, after saturation with ammonium sul-
phate. We propose, later, to attempt a study of the elastin peptone,

Chittenden and Hart—Elastin and the ElaMom Bodies. 37

Eh
O

Q
O
«

m
P
O

t— (

>

o

o
o

o

'A

o

Cd <B

1

H^

\ t~-

in

m

4ti

t-

cb

^

iC

s

0?

>

oa.

6

S c

CD

r-H C*

10

«

00

, »c

00

^

^•■3

00

CO

1

CS ^l^

CO

10

c-

S

; CO

*;-

C^

CU

_

_o

rt C/"-^ A-

so

50

1—1

CO

^^ § A.^

c«

05

GO

^

-^

t-

cb

' ^— (

T^

1—1

CJ

- — ^

,

'o

1—1

00

iO

50

W

CO

<D

.»■

'XI

05

o

2

CQ

10

t-

T-l

0?

lb

_o

03

OS

2
o
p

13

CD

OS

50

«5

CD

ro

t-

05

?*

CJ

05

a

P

<1

03

«b

1-1

o?

C7 ■

'm

p<

<+-(

O

t-

CO

1—1

C5

CO

a

^

oi

0^

o?

CO

10

o

•<

-#

^-

t-

' 1—1

1-H

■-3

10

' 01

o

<^

"3

1

1

^Ph

o W !z; Gc o

38 Chittenden and Hart — JElastin and the Elastose Bodies.

using for this purpose the elastoses just described as well as elastin
itself, and more vigorous digestive fluids, both peptic and tryptic.

In conclusion, then, we may say that elastin by the action of veiy
dilute acid at 100° C, and by the proteolytic action of pepsin-
hydrochloric acid, and of trypsin yields two primary cleavage pro-
ducts, which bear essentially the same relation to elastin that proto-
and deuteroalbumose do to albumin. Further, that protoelastose is
practically identical in reactions and composition with Horbaczew-
ski's hemielastin.

The close relationship in composition of all the bodies analyzed is
shown in the accompanying table. In considering these results, how-
ever, it is well to remember that the pancreatic preparations contain
an over large amount of ash, and hence the figures may not be
altogether trustworthy.

III. — The Imfluence of Urethax, Paraldehyde, Antipyrin,

AND ANTIFEBRl>f ON PROTEID METABOLISM. By R. H. ChIT-
TFNDEN.

I. The influence of urethan ; — from experiments made by N. P. Washburn,

Ph.B.

Ethyl-urethan, C0(NHJ0.C2H^, which has recently come into
use as a hypnotic, is claimed by R. v, Jaksch* to be a sleep-produc-
ing agent, free from the usual disagreeable after effects common to
most hypnotics. According to this observer, urethan does not ap-
pear to exert any poisonous action on the heart, nor to have any
depressing influence on the arterial system. Tt is further assumed
that the drag is without influence on digestion. Urethan acts
mainly on the brain, the peripheral nervous system being unaffected,
and according to v. Jaksch the hypnotic effect is always produced
when the drug is taken in doses of 1 gram.

Smaller doses (0'25-0*5 gram) are uncertain in their action.

In view of the somewhat peculiar action of urethan as a hypnotic,
we have undertaken to study the influence of the drug on the proteid
metabolism of the healthy organism, in order to compare its action
in this respect with that of other well known hypnotics.

Garnier,f alone, appears to have experimented in this direction.
He states that, in the case of a man, a dose of 6 grams of urethan was
followed by giddiness, etc., which condition, however, soon passed
away. In this experiment, the excretion of urea was appreciably in-
creased. In another experiment, the same observer found that a
dose of 2 grams of urethan was also followed by an increased excre-
tion of urea, in the case of a dog in a condition of hunger. A portion
of the urethan was appai-ently converted into urea and excreted as
such, but the greater part of the drug appeared in the urine unchanged.
Garnier further states that large, nearly fatal doses, of the drug check

* Abstract in Jahresbericht fiir Thierchemie, 1885, p. 70.

f Influence de I'urethane sur I'excretioQ des elements azotes de I'urine. Coiupt.
rend. soc. Biolog. 1886, p. 230.

40 R. II. Chittenden — Influence of Urethan, Paraldehyde,

metabolism, diminishing the quantity of urine excreted, as well as the
amount of urea, uric acid, etc.

The experiments about to be described were conducted wholly upon
the person of a healthy man of 66 kilos, body weight. A definite
amount of food, of known composition, was taken daily and uniform
habits of sleep, exercise, etc., were kept up during the whole time of
the experiment, which extended over a period of six weeks. In this
manner, body equilibrium was established and the daily excretions
brought to a constancy of composition, as preliminary to studying
the action of the drug. The daily diet was as follows :

312 grams fresh beef, free from fat and tendons.

368

potatoes.

227

wheat bread.

149

boiled rice.

35

butter.

28

sugar.

6

salt.

200

water.

The body-weight was ascertained each morning, and the 24 hours'
urine collected and analyzed each day as follows : nitrogen was de-
termined by the Kjeldahl method;* sulphur by fusing a given vol-
ume of the urine with pure potassium hydroxide and potassium nitrate
in a silver crucible, and ultimately precipitating and weighing the
sulphur as barium sulphate ;f phosphorus by fusion of a portion of
the urine in a like manner with potassium hydroxide and nitrate,
precipitation of the phosphoric acid from a nitric acid solution by
molybdic solution, solution of this precipitate in ammonia and repre-
cipitatiou with magnesia mixture, as ammonio-magnesium phosphate;
chlorine by ignition Avith potassium nitrate, and titration of the pre-
pared sobition with a standard solution of silver nitrate.

On April 19th, analysis of the urine was commenced and con-
tinued for fourteen days before the first dose of urethan was admin-
istered. The drug was then taken for five consecutive days, a total
of 73 grains or 4*73 grams, after which the urethan was discontinued
for seven days. A second trial was then made by giving 89 grains

* Neue metbode zur Bestimmung des Stickstoifs in organischen Korpern. Zcit-
schrift fiir Analytische Chemie, xxii, 366.

•j- For the details of the processes used, see Studies from Laboratory of Physiological
Chemistry, Yale University, vol. ii, p. 88.

Antipyrin, and Antifebrin on Proteid Metabolism.

41

Amount

of urethan

takeu.

o a5

6'S

2S oiooiT-Hosoioot-coo oioioeo

OS

CO

. CO

.§0

2 Qococot-iooojt-oooio ooiooo

t-l

^1

10

00
tH

2 a

d 000005COCO 1-^ ■ 1 «1C500»

CO

1— 1

i« S

° B

Ho

>

,• C> '77 to <?"» 0000

OSOOiOOSTHi— iO^OtH 0050SO
l-H r-l t-Ht-H— l-rHi-l -rH t-C

10
CO

T-i

t-«OCX)00C-iMIOC>?C01C 00COJ>CO

cp

lb
07

d

"0 "■*■'■'-■'■'"•■' ^"::::

Body
weight.

d 000000000 , 0000

a 000000000 0000

ca ^oooDwooooco . -5i<«ooi»

u

ji 0»0^:v)lOCOt-Xi010o3T-IC>>SOTt(

Trans. Conn, Acad., Vol. VIII.

Nov., IJ

42 Ji. JT. Chittenden — Influence of Urethan, Paraldehyde,

a s'

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Antipyrin, and Antifebrin on Proteid MetahoUsm. 43

or 5 "76 grams of urethan in three consecutive days, followed by a
period of six days in which the drug was not taken.

The tables, giving the amounts in grams for the 24 hours' urine
of the several elements determined, show plainly that urethan has a
decided action on the metabolism of the body.

On examining the results in detail, it is to be seen that urethan
has a very decided diuretic action, most noticeable on the second day the
drug was taken. In fact, this may be called the initial action of the
drug, since in both trials the amount of water excreted, after the first
increase, rapidly diminished as the dose of urethan was increased, and
indeed, the volume remained far below the average amount for two
or three days after the drug had been discontinued, or until its elimi-
nation from the system was fairly complete, when the volume of
fluid quickly rose to normal.

The excretion of nitrogen is at once affected by urethan, even a
dose of five or ten grains bringing the nitrogen noticeably below the
normal amount. In both series, the excretion of nitrogen was greatly
diminished. On discontinuing the drug, the nitrogen excreted rapidly
increased in amount, and on the third or fourth day after its discon-
tinuance, the daily excretion of nitrogen passed considerably above
the normal.

As regards the excretion of phosphorus, it would appear from the
experiments that the administration of small doses of urethan gives
rise to an increased excretion of this element, as seen from the results
obtained on May 5-9th. With larger doses of the drug, however,
the excretion of phosphorus is diminished, as seen from the results of
May 17-1 9th. As the excretion of sulphur runs parallel with the
excretion of nitrogen, both coming from the metabolism of proteid
matter, it follows that urethan when taken in small quantities
must exert an inhibitory influence on proteid metabolism, while it
stimulates the decomposition of certain phosphorized matters. In
larger doses, the inhibitory action of the drug on proteid metabolism
is still more pronounced, while at the same time the excretion of
phosphorus is also retarded.

In no case was any hypnotic action noticeable.

44 H. H. Chittenden — The Influence of Urethan, Paraldehyde^

II. The influence of Paraldehyde ; — from experiments made by J. E. Docken-

dorff, Ph.B.

In the following experiment, a full-blooded coach dog of 25 kilos,
weight was employed. The animal was confined in a suitable cage,
lined with galvanized iron and furnished with a bottom of wire net-
ting, under which was a funnel-shaped tray, the whole so arranged as
to allow all of the fluid excreta to pass into a collecting bottle under-
neath.

The animal was fed daily on a weighed diet consisting of dessicated
beef, soda crackers and water. The beef was prepared by removing
as thoroughly as possible all fat, fascise, tendons, etc., passing it
through a sausage cutter and then drying it at a temperature below
60° C, until it had lost 75 per cent, of its weight. The dried and
sampled beef was then preserved in tightly stoppered jars until
needed. The crackers were ordinary soda crackers, containing about
0-7 per cent, of nitrogen. The daily rations consisted of 60 grams of
crackers and 125 grams of the dessicated beef, soaked in 600 c. c. of
water. This diet was commenced sometime before the urine was
collected, and was continued throughout the experiment. Ulti-
mately, the 24 hours' urine was analyzed each day, according to the
methods described in the preceding experiment. Owing to irregular-
ity of urination, and the difticulty of using a catheter, the quantity
of urine obtained each day was necessarily quite variable, hence
the composition of the normal urine was determined daily for
three weeks, so that a sufficiently large number of results might
be obtained to yield an accurate average for the normal period.
Paraldehyde was then administered in gelatin capsules, about six
hours after the dog had been fed, so that the drug might not inter-
fere with digestion. Its administration was continued for eighteen
days.

The results, expressed in grams for each 24 hours' urine, are shown
in the accompanying tables.

Antipyynn, and Antlfehrin on Proteid 3Ietaholistn.

45

Normal Urine. Without Paraldehyde.

Date.

Total
Volume.

Sp. Gr

1 Reaction.
acid.

Nitrogen.

grams
12-540

Sulphur.

Phosphorus.

Amoinit of
Paraldehyde.

Api'il
21

c. c.
395

1036

grams

0-883

grams
0-862

22

840

1025

"

19-315

1-529

1-288

23

660

1031

((

18-276

1-176

1-442

24

490

1035

<(

14-622

1-038

1-096

25

610

1036

"

21-050

1-388

1-340

26

410

1036

(<

13-568

27

550

1037

"

18-550

1-396

1-202

28

415

1036

"

13-580

1-322

0-730

29

420

1037

"

14-916

0-940

0-934

30

630

1038

"

18-071

1-394

1-386

May
1

490

1040

..

18-415

1-172

1.214

2

390

1038

<«

12-896

0-826

0-80G

3

630

1038

"

23-425

1-499

1-410

4

525

1036

"

17-244

1-235

1-098

5

430

1035

"

13-849

1-138

0-862

6

375

1035

-

11-672

0-810

0-784

7

740

1032

-

23191

1-648

1-538

8

465

1031

"

13-682

1-085

0-898

9

545

1032

"

10-239

1-340

1-166

10

535

1032

"

15-756

1-119

1-134

11

495

526

1032

"

14-406

1-145

0-904

Av'age

1035

acid.

16-440

1-204

1-105

46 R. H. Chittenden — The Influence of Urethan, Paraldehyde,

With Paraldehyde (C2H40)3

Date.

Total
Volume.

Sp. Gr.

Reaction.

Nitrogen.

Sulphur.

Phosphorus.

Amount of
Paraldehyde.

May

c. c.

grams

grams

grams

grams

12

540

1031

acid.

15-032

1-346

1-044

0-424

13

...

Urine

lost.

0-740

14

540

1028

acid.

13-624

1-000

0-776

0-715

15

310

1024

6-539

0-587

0-442

0-794

16

810

1027

20-191

1-593

1-339

0-755

17

460

1031

12-793

0-874

0-932

0-784

18

690

1033

21-091

1-444

1-296

0-702

19

525

1033

16-863

1-214

1-106

1-071

20

505

1032

14-523

1-016

0-978

1-536

21

735

1029

20-844

1-475

1-292

1-499

22

545

1031

16-756

1-122

0-462

1-811

23

555

1031

17-063

1-195

1-620

2-268

24

580

1032

18-202

1-245

1-080

2-675

25

360

1030

9-935

0-734

0-590

3-111

26

800

1033

25-695

1-924

1-476

3-632

27

460

1030

12-992

0-935

0-766

4-063

28

585

1033

19-174

1-368

0-990

4-958

29

.695
570

1032

21-756

1-391

1-342

5-941

Av'age

1030

acid.

16-060

1-203

1-031

87-479

Antipyrin, and Antifebrin on Proteid Metabolism.
Without Paraldehyde.

47

Date.

Total
Volume.

Sp. Gr.

Reaction.

Nitrogen.

Sulphur.

Phosphorus.

Amount of
Paraldehyde.

May
30

31

June
1

2

3
4

c. c.
610

570

440
645
640

445

558

1028
1031

1028
1031
1030
1026

acid.

a

acid.

grams

17-672

17-080

12-201
19-550
17-798
11-112

grams
1-309

1-311

0-765
1-376
1-237
0-979

grams
0-992

1154

0-780
1-299
1-608

Av 'age

1029

15-902

1-163

1-166

Throughout the experiment, the dog appeared perfectly well, and
at no time showed any symptoms of nausea. Neither was there any
special hypnotic action noticeable.

The average of the results shows plainly that the drug has little,
if any, action on proteid metabolism. Under the influence of the
paraldehyde there was a slight increase in the amount of water
excreted. Bokai * has stated that the urinary secretion is slightly
increased by paraldehyde. In our experiment, however, the diuretic
action is not great. As regards the excretion of nitrogen, there is a
slight diminution to be seen in the paraldehyde period. There is
also a corresponding decrease in the excretion of phosphorus. The
three periods, however, show such close agreement in results, it is
obvious that, under the conditions of this experiment, paraldehyde
has not exerted any special influence on proteid metabolism.

According to the experiments of Quinquad and A. Henocque,f
paraldehyde causes a diminution of body temperature, and at the
same time a very noticeable falling off* in the excretion of carbonic
acid ; thus, according to one of Quinquad's experiments, a dog after
receiving by hypodermic injection 8 c. c, of paraldehyde gave off" 5-5
grams of carbonic acid, while it expired during the same time, three-
fourths of an hour after the injection, only 1*96 grams of carbonic
acid.

* Ueber die physiologische Wirkung des Paraldehyds. Centralblatt fur die medicin-
ische Wissenschaften. 1887, p. 412.

f Abstract in Jahresbericht fiir Thierchemie. 1884, p. 374.

48 R. H. Ghittende7i — The Influence of JJrethan, Paraldehyde,

III. Influence of Antipyrin ; — from experiments made by H. F. Adams,

Ph.B.

Previous experiments* with antipyrin, made in this laboratory,
have shown that this drug, when introduced into the stomach of
healthy rabbits, has little, if any, noticeable influence on the excre-
tion of carbonic acid or on the body temperature, except in the
case of toxic doses. Coppola,f likewise, found that the subcu-
taneous introduction of 0-1 to 0-3 gram of antipyrin in the case of
dogs led to a reduction of body temperature of only 0*25 to 0-6 of a
degree, while 0*3-0*4 gram of the di'ug was without any noticeable
influence upon the excretion of urea. In fever patients, F. Muller,J
however, had previously noticed a diminution in the excretion of
nitrogen under the influence of antipyrin, while in the case of healthy
men the excretion was afiected but very slightly, if at all. Jacubo-
witsch,§ likewise, had noticed in experiments on healtliy and fevered
children that the use of antipyrin led to a diminution both in the
quantity and specific gravity of the urine, and also a diminution in
the quantity of uric acid, phosphoric acid, sulphuric acid and of
chlorides. L. Riess,|| by carefully conducted experiments on nine
typhus fever patients, found as a j)rincipal result that antipyrin in
doses up to 12 grams per day diminished considerably the excretion
of nitrogen, the diminution ranging in six series of experiments from
2*5 to 24'7 per cent. Umbach,^ likewise, has studied the influence
of antipyrin on the excretion of nitrogen, both on a dog and on him-
self, and he found that with a definite diet the excretion of nitrogen
sank, under the influence of 4 grams of antipyrin, in two days about
2 grams, equal to 4 grams of urea. The uric acid excretion, however,
was not materially afiected.

In spite of these manifold experiments, we have deemed the matter
of sufiicient importance to warrant further study, especially with a
view to the action of the drug on the metamorphism of nitrogenous
matter in the healthy organism. Tlie experiments were therefore
tried upon a healthy man with a body weight of 77 kilograms, and

* Chittenden and Cummins. Studies, vol. ii., p. 231.

f Abstract in Jahresbericht fiir Thierchemie. 1885, p. 98.

X Abstract iu Jahresbericht fiir Thierchemie. 1884, p. 242.

§ Abstract iu Jahresbericht lur Thierchemie. 1885, p. 444.

II Archiv fiir experim. Pathol, u. Pharm. xxii, 127 ; also Abstract in Jahresbericht
fiir Thierchemie. 1886, p. 417. Ueber stickstoffausscheiduug bei antipyretischer
Fieberbehandlung.

^ Ueber den Einfluss der Antipyrins auf die stickstoffausscheidung. Abstract in
Jahresbericht fiir Thierchemie. 1886, p. 418.

Antipyrinj and Antifehrin on Proteid Metabolism. 49

under definite conditions of diet, exercise, etc. Nitrogenous equi-
librium was established prior to the experiment, and the following
daily diet was strictly adhered to throughout the entire period.

386 grams meat (beef).

340 " potatoes.

227 " wheat bread.

163 " oat meal (steamed).

28 " sugar.

42 " butter.

120 " milk.

1040 " water.

In this experiment, nitrogenous metabolism was measured by deter-
mining the urea and uric acid contained in the daily excretion, instead
of determining the total nitrogen. This was done in order to ascer-
tain whether the drug has any special action on the excretion of uric
acid. Urea was determined by Liebig's method, as modified by
Pfliiger.* Chlorine was previously determined by fusion with potas-
sium nitrate and titration with silver nitrate in the usual manner, and
then removed from the solution to be tested for urea, by a standard
silver solution. Uric acid was determined by Salkowski's method,
and phosphoric acid by titration with a standard uranium solution.

The accompanying tables give the results of the daily analyses.
From these it is evident that under the conditions of this experiment,
anti pyrin has a decided inhibitory action on the proteid metabolism
of the healthy human organism, as shown by the diminished excre-
tion of urea and uric acid when the drug is taken. Antipyrin also
tends to diminish the volume of the urinary secretion, this action
being very marked in the second series, where comparativelv laro-e
amounts of antipyrin were administered. As regards the excretion
of phosphoric acid and of chlorine, nothing definite can be said. The
raoi*e important changes produced by the antipyrin are shown in the
following table of average daily results.

Urea. Uric acid. Total P2O5. Volume. Sp.Grr.

grams. gram. grams. c. c.

Normal period 41-806 0-586 3-185 951 1028

First antipyrin period 38-375 0-556 3-026 848 1029

First after period- 42-030 0-575 2-929 929 1028

Second antipyrin period.. 40*854 0-472 2-941 822 1031

Second after period 44-220 0-537 2-923 957 1028

* Pfliiger's Archlv fiir Physiologie, vol. xxi, p. 248.
TuANs. Conn. Acad., Vol. VIII. 7 Nov., 1888

50 B. H. Chittenden — TJie Infl^ience of Urethmi, Paraldehyde,

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Antipyrin, and Antifebrm on Proteid Metabolism.

51

Amount of Aniipyrin
taken.

■J!

•5 o o o Lo ^
5 so CO so ^ ira

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Urea.

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52 R. H. Chittenden — The Influence of Urethan, Paraldehyde,

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Antipyrin, and Antifebrin on Proteid Metabolism. 53

Since this work was finished we have seen an interesting paper
by Dr. Kiimagawa* on the action of certain antipyretics on proteid
metabolism, in which is given the results of an experiment with
antipyriu on the excretion of nitrogen and uric acid, in the case of a
dog of 26 kilos, weight in a condition of nitrogenous equilibrium.

In this experiment, Dr. Kumagawa found that even large doses of
antipyrin (51 grams in 16 days) produced no change whatever in the
excretion of nitrogen (determined by the Kjeldahl method), but that
there was a very noticeable increase in the excretion of uric acid
(determined by Salkowski's method), amounting on an average to 65
per cent, above the noi'mal excretion. These results stand in direct
opposition to what we have found with somewhat smaller doses, in
experimenting on the human organism. Whether the explanation
of this ditference is to be found in the different nature of the two
organisms experimented with we cannot now say, but we hope at a
later date to explain this apparently divergent action.

IV. The influence of antifebrin ; — from experiments made by H. C.

Taylor, Ph.B.

Antifebrin or acetanilide, which has recently come into use as an
antipyretic, as a nervine and antiseptic, has been the subject of many
clinical observations but has not as yet, so far as we know, been
experimented with to ascertain its influence on proteid metabolism.
We have endeavored, therefore, to ascertain the influence of this new
antipyretic on the nutrition of the healthy human organism, believing
that such results may possibly be of greater value than those ob-
tained by experimenting on animals. At the same time it is to be
borne in mind, that an antipyretic especially may produce an eftect
upon the healthy organism quite different from that which the same
doses would produce on an organism rendered perhaps more sus-
ceptible by disease, as in fever. The experiment was therefore con-
ducted upon the person of a young man of 64 kilos, body weight,
brought into a condition of nitrogenous equilibrium and maintained
throughout the experiment upon a weighed diet of known composi-
tion. For reasons already given, the excretion of nitrogenous matter
was measured by determining in the 24 hours' urine the amount of
urea and uric acid, using the methods employed in one of the pre-
ceding experiments. Sulphur, phosphorus and chlorine Avere also

*Ueber die Wirkung eiaiger antipyretisclier Mittel auf den Eiweissumsatz im
Orgauismus. Virchow's Arcbiv, Band cxiii, p. 192.

54 R. H. Chittenden — The Influence of TTrethan, Paraldehyde,

determined each day by methods already indicated. After nitrogen-
ous equilibrium had been established, and the urine analyzed for ten
consecutive days, antifebrin was administered daily in divided doses,
at a time not to interfere with digestion, for a period of nine days.
The daily dose was gradually increased until at last there was a
slight approach to cyanosis. In all, ]3'3 grams or 205 grains of the
antipyretic were taken. No disagreeable symptoms were experienced,
but there was a very noticeable lividity of countenance with a de-
cided blueness of the lips, and a slight darkening of the skin near the
cheek bones.

The initial daily dose was 0*4 of a gram or a little over 6 grains,
and was rapidly increased to 2*6 grams or 40 grains per day, given in
thi'ee doses. There are, to be sure, many cases recorded where
apparently smaller doses have led to serious results, but careful
watching failed to show any symptoms whatever even suggestive of
any disagreeable action on the digestive system, the circulation or
respiration. Weinstein, indeed, has said that persons not suf-
fering from fever may take antifebrin for weeks together without
any effect on the circulation, while according to Herczel the long-
continued administration of antifebrin, thirty to forty-five grains
daily for six weeks, may lead to what he terms aniline anaemia with
solution and decomposition of the haemoglobin of the blood. In
fact, the latter observer considers that aniline is set free from the
acetanilide and that the decomposition of the blood-coloring matter
is due to this cause. Whether this is the cause of the cyanosis so
often spoken of in connection Avith this drug is uncertain.

In the second antifebrin period, 13*9 grams or 214 grains of the
drug were taken in seven days, accompanied at the close with the
same approach to cyanosis as before.

Examination of the analytical results shows plainly that under
the conditions of this experiment the excretion of urea is not very
greatly affected. There is, however, in both antifebrin periods a
slight increase, indicating increased proteid metabolism under the
influence of the di'ug. This increased excretion of urea is more
apparent in the individual results than in the average of the series.
Thus, in the normal urine it is to be noticed that the daily excretion
of urea never exceeded 34'5 grams, while in the first antifebrin period,
on the days when the largest doses of acetanilide were taken, the
excretion of urea amounted to 35-37 grams, and in the after period
quickly fell to about 33 grams per day. The same peculiarity is
also noticeable, to a less extent, in the second antifebrin period.

Antipyrin, and Antifehrin on Proteid 3fetaholism,.

55

<4-( ••

o n
a ^
o S3

<<

o

Chlorine.

grams

2-967

4-404

3-603

3-520

8-221

4-027

4-377

4-073

6-366

4-078 ■

4-063

o

en
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grams
0-966
0-864
1-227
0-859
1-041
0-881
0-862
0.842

0-821
1-122

00

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O

Sulphur.

grams
1-380
1-154
1-207
1-254
1-220
1-296
1-222
1-289

1-350
1-124

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gram

0-749
0-708
0-739
0-753
0-779
0-723
0-741

0-694

0-777

o
t-

o

Urea.

grams
33-331
33-280
33-486
33-947
33-058
34-502
32-619
33-440

34-370
33-334

33-536

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1280
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1340
1030
1150
1250

880

1590
1250

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1—1

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Body

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64200
64200
64000
64000
64000
64000
63800
64200

64000
64000

Date.

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56 R. 11. Chittenden — lyifuence q/ Urethan, Paraldehyde,

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Antipyrin, and Antifebrin on Proteid Metabolism.

57

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Nov., ij

58 R. H. Chittenden — Influence of Z^rethm}, Paraldehyde,

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Ahtipyrin, and Antifehrln on. Proteid Metabolism. 59

We must conclude, therefore, that acetanilide tends to increase some-
what the excretion of urea, but that with such doses as we have era-
ployed the increase in proteid metabolism cannot be great. This is
further indicated by the lack of any corresponding change in the
excretion of sulphur.

The excretion of phosphorus is also unaffected by antifebrin.

On the excretion of uric acid, however, our results indicate a
special inliibitory influence. This is quite apparent both in the
averages of the different series and in the individual results, and, if
correct, would appear to be the most marked characteristic of
antifebrin, so far as its influence on proteid metabolism is concerned.
Various observers have stated that antifebrin acts as a diuretic,
others that it decreases the secretion of water, and while doubtless
both results have been seen to follow its administration in diseased
conditions of the system, our experiment on a healthy man gives no
evidence of any action of this kind.

Since the foregoing was written we have seen the results of
Kumagawa's* experiment with antifebrin on a dog, from which he
concludes that acetanilide taken in small doses (2-3 grams per day)
does not give rise to any apprecial)le increase in the decomposition of
proteid matter, but that larger doses (4-5 grams per day) cause a very
marked increase, as indicated by the increased excretion of nitrogen.
Such doses are, however, as Kumagawa liimself admits, too large,
especially when given to a moderate sized dog, to have the results of
any practical value. Lepine,f too, experimenting on a hungry dog,
thought he found an increase in the excretion of nitrogen after giv-
ing two doses of one and two grams of antifebrin. Further, Ademski];
is quoted as considering that urea is increased, but the total quan-
tity of nitrogen decreased by antifebrin. Bokai, that the amount of
nitrogen is diminished and Berezooski that the urea decreases with
the fall of temperature. Whether these latter views are founded on
actual experiments or are mere conjecture I do not know.

According to Jaffe and Hilbert,§ rabbits fed upon antifebrin excrete
it mainly as paramidophenol-sulphuric acid, and Kumagawa has like-
wise found, in the case of a dog, that neither acetanilide nor aniline
appear in the urine, but that the antifebrin is excreted mainly as
paramidophenol united to sulphuric acid.

* Virchow's Archiv. Band cxiii, p. 171.

f See Salkowski's Bemerkung in Virchow's Archiv, Band cxiii, p. 394.
X See Report on Antifebrin in the Therapeutic Gazette, vol. xii, p. 571.
§ Zeitschrift fur phyaiologisohe chemie, xii, p. 307.

fV. — The influence of several new therapeutic agents on

AMTLOLYTIC AND PROTEOLYTIC ACTION. By R. H. ChITTEN-

DEN AND C. W. Stewart, Ph.B.

In view of the pronounced action of a number of newly discovered
therapeutic agents on metabolism, we have deemed it of importancfe
to widen our knowledge regarding their physiological action by
attempting a study of their behavior towards the amylolytic and
proteolytic ferments, with the hope of gaining some insight into
their influence on normal digestion.

The methods employed were similar to those used in previous ex-
periments of this kind,* in which the action of varying percentages
of the drug were determined quantitatively.

Influence on amylolytic action.

As amylolytic ferment, human mixed saliva was employed, filtered
and carefully neutralized, and then diluted with distilled water in the
proportion of 1 to 5. The experiments were made in series, in which
one digestion of each series served as a control for comparison. The
volume of each digestive mixture was 100 c. c, in which was present
1 gram of perfectly neutral potato starch previously boiled with a
portion of the water, 10 c. c. of diluted neutral saliva and a given
quantity of the substance to be experimented with. The mixtures
were warmed at 40° C. for thirty minutes, after which further action
of the ferment was stopped by heating the solution to boiling. The
extent of amylolytic action was then ascertained, by determining in
one-fourth of the solution the amount of reducing substances by
Allihn's gravimetric method. f For the sake of convenience, the
total amount of reducing substance was calculated as dextrose, from
which in turn was calculated the percentage of starch converted.

Antipyrin.

With this new antipyretic, several series of experiments with small
percentages were made which show clearly that the substance is

* Studies from the Laboratory of Pbysiological Chemistry, Yale University, vols, i
and ii.

f Zeitschrift fiir aualytische chemie, xxii, p. 448.

Therapeutic Agents on Amylolytic and Proteolytic Action. 6 1

without any appreciable influence. When present in larger percent-
ages, the drug was found to have a slight inhibitory influence on
amylolytic action, as the following series of experiments show :

Per cent, of
Antlpyrln.

Total amount of
reducing substances.

Starch
converted.

Relative
amylolytic action

0-3446 gram.

31-01 per

cent.

100-0

0-5

0-3446

31-01

100-0

10

0-3424

30-82

99-4

30

0-3278

2950

95-1

5-0

0-3112

28-01

93-6

Coppola,* in studying the physiological action of antipyrin, found
that three per cent, of the substance did not hinder the inversion of
cane sugar by yeast, but did prevent alcoholic fermentation. Fur-
ther, that the same percentage hindered slightly the action of malt
diastase on starch, and had a decided inhibitory influence on the
alkaline fermentation of urine.

Antifebrin.

Owing to the comparative insolubility of antifebrin or acetanilide
in water, large percentages could not be employed. Such as were
used, however, clearly show that this antipyretic has little influence
on amylolytic action.

Per cent, of
Antifebrin.

Total amouDt of
reducing substances.

starch
converted.

Relative
amylolytic action.

0-3604

gram.

32-44 per

cent.

100(1

0-10

0-3600

32-40

99-9

0-25

0-3638

32-70

100-8

0-50

0-3550

3200

98-6

0-3382

30-44

100-0

2-0

3298

29-68

97-5

In this cormection, it is interesting to notice that Kumagawa* has
found antifebrin to have a strong antiseptic action on the putrefac-
tive processes of the intestinal canal, and Van Seer has observed that
milk does not undergo fermentation if saturated with it, and also
that it will prevent albumin becoming ])utrid.f

Urethan.

With ethyl-urethan, two series of experiments were tried, with the
following results :

* Jahresbericht fur Thierchemie. 1885, p. 98.

f Virchow's Archiv, Band cxiii, p. 184.

\ See The Therapeutic Gazette, vol. xii, p. 566.

62 Chittenden and Stevmrt — Injhience of several new

Per cent, of
Urethan.

Total amount of
reducing substances.

Starch
converted.

Relative
amylolytlc action.

0-3382 gram.

30-44 per cent.

100-0

0-5

0-3550

31-95

104-6

0-3402

30-62

100-0

2-0

0-3446

31-01

101-3

3-0

0-3352

30-n

98-5

5-0

0-3268

29-40

96-0

Here, there is noticeable, with the smaller percentages, a slight
stimulation of amylolytic action, but it is not sufficiently large to be
very marked.

Paraldehyde.

This substance shows very strong inhibitory action, even 0*5 percent,
diminishing the amount of starch converted by 30 per cent. It must
be remembered, however, that the aldehyde is only slightly soluble
in water, and that it is more or less volatile. The experiments were
conducted in .small flasks and the aldehyde kept more or less emul-
sionized with the aqueous solution by shaking the mixtures, but ob-
viously the percentages given can only approximately represent the
amount actually taking part in the reaction.

Following are the results of two series of experiments :

Per cent, of
Paraldehyde.

Total amount of
reducing substances.

Starch
converted.

Relative
amylolytic action.

0-3554 gram.

31-98 per cent.

100-0

0-1

0-3534

31-81

99-5

0-2

0-3528

31-75

99-3

0-5

0-2468

22-21

69-5

1-0

0-1066

9-59

30-0

0-3554

3 1 -98

100-0

0-8

0-1392

1-2-52

39-1

10

0-0948

8-53

26-6

1-5

00620

5-58

14-3

2-0

0-0446

4-01

12-5

T/i<iIIin Sulphate.

This salt has a more marked influence on the amylolytic action of
saliva than any of the preceding substances. Very small percentages
have a noticeable stimulating action, while the presence of 0*2 per
cent, of the salt almost entirely stops the action of the ferment. The
results of the following two series show one or two small points of
diflerence, but in the main they jioint to the same general action.

Therapeutic Agents on Amylolytic and Proteolytic Action. 63

Per cent, of
ThalUn Sulphate.

Total amount of
reducing substances.

Starcli
couverteri.

Relative
amylolytic action.

0-3534 gram.

31-81 per cent.

100-0

0-025

0-3648

32-83

103 2

0-050

0-3810

34-36

108-0

0-100

0-3540

31-86

1001

0-200

0-0690

6-21

19-5

0-300

trace.

0-3652

32-86

100-0

0-025

0-3798

3418

104-0

0-050

0-3872

34-89

1061

0-080

0-3812

34-81

105-9

O'lO

0-3148

28-33

86-2

Starch
converted.

Kelative
amylolytic action

32-57 per

cent.

1000

30-08

92-4

32-97

100-0

30-96

93-9

Caffein and Thein.

These two closely related alkaloids were found to have only a
slight action on the amylolytic ferment, producing when present in
considerable amount a slight diminution in the amount of starch con-
verted. With two per cent, of the alkaloids, the following results
were obtained :

Per cent, of Total amount of

alkaloid. reducing substances.

0-3620 gram.

20 CafEein. 0-3342

0-3664

0-2 Thein. 03440

Influence on proteolytic action.

The influence of the above therapeutic agents on the proteolytic
action of pepsin-hydrochloric acid was determined, as in preceding
work of this kind,* by ascertaining the amount of fibrin digested or
dissolved in a given time, by a definite volume of standard, artificial
gastric juice, in the presence of varying amounts of the substances to
be tested. The gastric juice was made by dissolving 10 c. c. of a
glycerin extract of pepsin in one litre of 0*2 per cent, hydrochloric
acid. The volume of each digestive mixture was 50 c. c, composed
of 25 c. c. of the above mentioned artificial gastric juice, and 25 c. c.
of 0-2 per cent, hydrochloric acid containing the desired amounts of
the substances to be tested. The proteid material consisted of puri-
fied fibrin, coarsely powdered and dried at 110° C. One gram of
fibrin was used in each experiment. The digestive mixtures were
warmed at 40° C. for a certain length of time, usually two hours,

* Studies from the Laboratory of Physiological Chemistry of Yale University, vol?,
i and ii.

64

Chittenden and Stewart — Influence of several new

after which further proteolytic action was stopped by heating the
mixtures to boiling. The undissolved residues were then collected
on dried, weighed filtei's, washed thoroughly with boiling water, and
finally dried at 110° C. until of constant weight. The amount of
fibrin dissolved is taken as a measure of proteolytic action.

Following are the results obtained with the various substances
tested :

Fer cent, of
Antipyrin.

0-2

0-5

10

0-5
30
5-0

Per cent, of
Antlfebrln.

0-1

0-2

0-5

1-5

Antipyrin.

Undigested
residue.

Fibrin
digested.

RelHtlve
proteolytic action

0-507 gram

49-3 per cent.

100-0

0-622

47-8

96-9

0-573

42-7

86-6

0-688

31-2

63-3

Longer

time at 40

°C.

0-143

85-7

100-0

0-213

78-7

91-8

0-964

3-6

4-2

1-008

A)

tifehrin

Undigested
residue.

Fibrin
digested.

Kelative
proteolytic aciion

0-129 gram.

87-1 per cent.

100-0

0-145

85-5

98-1

0-166

83-4

95-8

0-212

78-8

90-5

0-371

62-9

72-2

Urethan.

'er cent.
Urethan

of

02

10

30

5-0

Per cent, of
Paraldetiyde.

0-05

0-10

0-30

1-00

2-00

Undigested
residue.

Fibrin
digested.

Relative
proteolytic action

0-148

gram.

85-2 per cent.

1000

0138

86-2

101-2

0-171

82-9

97-2

0-224

77-6

90-9

0-240

76-0

89-4

Paraldehyde.

Undigested
residue.

Fibrin
digested.

Relative
proteolytic action.

0-263

gram.

73-7 per cent.

100-0

0-219

78-1

105-9

0-246

75-4

102-3

0-255

74-5

101-1

0-264

73-6

99-9

0-269

73-1

99-2

Therapeutic Agents on Amylolytic and Proteolytic Action. 65
Thallin sulphate.

Per cent, of
lallin sulphate..

Undigested
residue.

Fibrin
digested.

Relative
proteolytic action.

0-379 gram.

62-1 2)ez- cent.

100-0

0-05

0-248

75-7

121-9

0-10

0-327

67-3

108-4

0-30

0-395

60-5

97-4

Thein and Cafein.

'er cent, of
alkaloid.

Undigested
residue.

0-105 gram.

1 -0 Thein.

0-252

4-0 "

0-179

4-0 Caffein.

0-584

Fibrin
digested.

89-5 per cent.

74-8
82-1
41-6

Relative
proteolytic action.

100-0

84-7 ■

91-8

46-5

These results show that autipyrin has a decided inhibitory influ-
ence on the action of the proteolytic ferment, and that when present
to the extent of 3'0 per cent, it practically stops all digestive action.
Antifebrin has also an inhibitory action, perhaps equal to that of anti-
pyrin, but owing to its greater insolubility in acid fluids large per-
centages of the substance could not be tested.

Uretban has only a slight retarding action, even when present to
the extent of 5 per cent.

Paraldehyde and thallin sulphate both show a very decided stimu-
lating action when small fractions of one per cent, are present, the lat-
ter, particularly, causing a much larger amount of fibrin to be di-
gested than in the control experiment.

Thein and cafFein both show an inhibitory influence on the ferment,
that of cafFein being much greater than that of thein.

On the proteolytic action of trypsin in an alkaline solution, two
substances only were tested, antifebrin and paraldehyde. The ex-
periments were conducted in the same manner as with pepsin-hydro-
chloric acid, except that a solution of trypsin in 0*3 per cent, sodium
cai'bonate was employed in place of the pepsin-acid, and the mixture
warmed for a longer time at 40° C. Following are the results ob-
tained, showing a much more pronounced inhibitory action on this
ferment than on pepsin.

Per cent, of
substance.

Undigested
residue.

0-2 antifebrin.
1*0

0-2 paraldehyde
20

Trans. Conn. A.cad., Vol. VIII

0-159 gram.

0-368

0-741

0-368
0-959

Fibrin
digested.

84-1 per cent.

63-2

25-9

63-2

4-1

Relative
proteolytic action.

1000

75-1

30-8

75-1

4-9

Nov., 1888.

V. — Caseoses, Casein Dtspeptone, and Casein Peptone. By
R. PI. Chittenden.

In a previous paper on " Casein and its Primary Cleavage Pro-
ducts,"* the writer expressed the intention of continuing the study of
the caseoses formed in pepsin digestion, and also of studying pure
casein peptone and the so-called casein dyspeptone. In the fulfillment
of this intention, experiments upon these subjects have been carried
on in this laboratory during the past two years, with the aid of sev-
eral co-workers, and the results are here presented collectively.

I. — Casein Dyspeptone ; — from experiments by L. ^4. Conner, Ph. B., axiA

C. A. Tuttle, Pli.B.

When casein is exposed to the action of pepsin-hydrochloric acid
at 40° C, or even at ordinary temperatures,-it is decomposed, as is
well known, into soluble caseoses and peptone. In every such diges-
tion, however, there always remains a certain amount of an insoluble
pasty, grayish white substance, which apparently is not susceptible
to the further action of gastric juice, no matter how long continued.
This insoluble substance, which is noticeable to a greater or less extent
in the pepsin digestion of all proteids, but particularly so with casein,
and which received from Meissner the name of dyspeptone, was
examined somewhat carefully by Lubavin in 1876,f who described
some of its properties. lie considered this casein dyspeptone as a
mixture of two distinct bodies, separable from each other by the
action of sodium carbonate. Substance A^ that portion of the dys-
peptone soluble in sodium carbonate, was described by Lubavin as a
body containing 4-6 per cent, of phosphorus, 13*3 per cent, of nitro-
gen, 48*5 per cent, of carbon and no sulphur, corresponding to the
formula C^^H^^N^POj^. Substance J5, insoluble in sodium carbonate,
was slowly soluble in sodium hydroxide, contained sulphur, but only
a small trace of phosphorus and evidently contained more or less

* Chittenden and Painter. Studies from Laboratory of Physiological Chemistry
Yale University, vol. ii., p. 156.

f Hoppe-Seyler. Med. Chem. Untersuchungeu, p. 463.

Chittenden — Caseoses, Casein Di/speptone, and Casein Peptone. 67

admixture of fat. Judging from tiie description, substance A must
have been an acid compound of the body studied, mixed witli more
or less undigested casein ; while substance J^ was doubtless a mixture
of fat and the body -1. However this may be, our results show
conclusively that no body having the formula ascribed by Lubavin
to his substance A, can be separated IVom the undigestible residue
of casein in pepsin-hydrochloric acid.

In all of the experiments to be described, the casein employed was
freshly prepared from skim milk by precipitating the greatly diluted
fluid with dilute acetic acid, washing thoroughly with water, redis-
solving the precipitate in water containing a trace of ammonia and
reprecipitating, repeating this operation three or four times.

In subjecting casein to the action of artificial gastric juice the con-
ditions were varied more or less in the individual experiments, so
that if the so-called dyspeptone be a mixture of two or more sub-
stances, the varying conditions under which the digestions were
made might so change the nature of the mixture, that on analysis, it
would become apparent.

Digestion A.

The casein from five gallons of milk was placed in four litres of
0"4 per cent, hydrochloric acid and warmed to 40° C. To this was
added 200 c. c. of a dialyzed pepsin solution, prepared from a glyc-
erin extract of the ferment, and the mixture kept at 40-45° C. for
forty-eight hours. At the end of this time there was still a compara-
tively large mass of gelatinous matter undissolved, composed in part
no doubt, of swollen casein. The entire mixture was then dihited con-
siderably with water and treated with dilute alkali to near neutraliza-
tion, leaving the fluid, however, distinctly acid. The undio-ested
matter was then filtered ofl" and washed thoroughly with water. This
partial neutralization of the digestive mixture was found necessary,
owing to the extreme slowness with which the acid fluid filtered.
The undigested matter was again warmed at 40° C. for forty-eight
hours, with four litres of a much more vigorous pepsin mixture con-
taining 0*4 per cent, hydrochloric acid. The residue still undissolved
was filtered off and washed with water. The acid filtrate gave no
precipitate whatever on neutralization. A third time the undigested
matter was warmed at 40° C. for sixty hours, with four litres of a still
stronger artificial gastric juice. The cpiantity of insoluble matter did
not appear to be diminished at all by this third treatment with pep-
sin and acid. The substance was thereupon filtered from the acid

68 (Jhittexden — Caseoses, Casern Di/sjyeptone, and Casein Peptone.

fluid, and washed with water uutil the washings gave no reaction for
chlorides. It was then treated in the cold with one litre of one per
cent, sodium carbonate, in which it appeared to dissolve completely.
On filtration there appeared a small whitish residue, which on treat-
ment with ether, dissolved in great part, thus showing its fatty
nature. There remained, however, a very small residue of a heavy,
browiiish substance too small in quantity to be considered other than
as an impurity. On adding dilute hydrochloric acid to the alkaline
fluid, no precipitate was obtained until the fluid was made distinctly
acid, when the dyspeptone was thrown down as a heavy flocculent
precipitate. In the filtrate, the biuret and Millon's test showed only
a faint trace of an albuminous body. The precipitate of dyspeptone
was washed with water until the washings gave no reaction with
silver nitrate, after which it was dissolved in one per cent, sodium
carbonate, the fluid made exactly neutral with dilute hydrochloric
acid, thymolized, and then dialyzed in running water until all chlo-
ride was removed from the fluid.

The neutral fluid of dyspeptone so obtained, was concentrated to a
thick syrup on the water-bath, and then while still warm, it was
treated with 95 per cent, alcohol and a little absolute alcohol.
A moderately heavy precipitate of dyspeptone resulted, but appar-
ently not all of the substance was precipitated. On standing for
forty-eight hours, the fluid was found in a thick, gelatinous condition.
The coagulum was insoluble in 95 per cent, alcohol, but readily and
completely soluble in water. It was therefore washed thoroughly
with alcohol, allowed to stand under absolute alcohol for several
days, then treated with cold ether, after which it was dried, pow-
dered and placed in a fat extractor and extracted with boiling ether
as long as any fatty matter Avas dissolved, a process which took sev-
eral days. About nine grams of the pure, dry substance were
obtained.

A portion was then dried at 110° C. until of constant weight,
for analysis. Its composition is shown in the accompanying table.

The methods of analysis employed were the same as those previ-
ously described. Phosphorus was determined by fusing the sub-
stance in a silver crucible with potassium hydroxide and potassium
nitrate, acidifying the mixture with nitric acid, evaporating to dry-
ness, dissolving the residue in water acidified with nitric acid and
precipitation of the phosphoric acid, first with raolybdic solution, and
lastly with magnesia mixture and final weighing of the phosphorus
as magnesium pyrophosphate.

Z 32 Cli O

VO Chittenden — Caseoses, Casein Dyspeptone, and Casein Peptone.

It is to be seen from the table of aualytical results tliat the total
phosphorus is exactly equal to the phosphorus of the ash. The ash,
as examination showed, was composed almost entirely of calcium
phosphate with a trace of iron. There was no calcium sulphate.
Taking the percentage of phosphorus at the highest figure, viz: 2*67,
and calculating it to calcium phosphate Gdi^(Vo^„, it would be equal
to 13'3 per cent, of calcium phosphate, or within 0*3 per cent, of the
ash found. Hence, it would appear that the phosphorus present in
the substance probably existed there wholly as calcium phosphate.

Digestion B.

A quantity of pure casein, equal in amount to that used in digestion
A, was warmed at 40° C. with seven litres of 0*4 per cent. hydi'O-
chloric acid, to which a quantity of purified and vigorous pepsin solu-
tion was added. After being kept at 40° C. for fifty hours, two
litres more of 0*4 per cent, hydrochloric acid, together with some
pepsin solution, were added and the mixture warmed at 40° C. for
two days more, after which it was diluted with water and the undi-
gested residue allowed to settle out. The supernatant fluid was
syphoned off, the residue washed by decantation and then again
treated at 40° C. with five litres of an active pepsin-hydrochloric acid
solution for four days. The residue still undigested was filtered ofl',
washed with water, dissolved in one per cent, sodium carbonate solu-
tion and the alkaline fluid filtered from the small amount of undis-
solved matter. From this fluid, the dyspeptone was precipitated by
hydrochloric acid, the acid compound washed thoroughly with water,
after which it was warmed at 40° C. for forty-eight hours with 1200
c. c. of 0*2 per cent, hydrochloric acid, and 50 c. c. of a strong pepsin
solution. The undigested residue, after being thoroughly washed,
was dissolved in 800 c. c. of one per cent, sodium carbonate, the solu-
tion exactly neutralized with hydrochloric a-cid and dialyzed until
chlorides were entirely removed. The clear aqueous solution was
evaporated to a syrup and the dyspeptone j^recipitated with alcohol,
after which it was treated exactly as preparation A. 9*5 grams
of pure, dried substance were obtained.

For analysis, the dyspeptone was dried at 110° C. until of constant
weight. The analytical results are shown in the accompanying
table.

The ash which was larger than in the first preparation, contained
no sulphate whatever, but was composed in great part of calcium
phosphate.

P after
deducting
P of Ash.

;;;;;;;;;;

CU t5.

1 1 1 . t , ' C.-J (?}

CO «5

; ! CO X-

;;;:;;;;;.§§

^r^ ...iiilliO,,

from the
Ash.
gram.

0-0484

, , , , 1 . . 00 Ol , , 1

» -6^ O cp , , ,

1 ' ' ' ' ' o C ' ' '

^ B + CO

oQ-sin M

1

^ ^ . • .

60 ' '

1 ^

00 m

CO -* ' 1 ■ ' i

Ash
found,
gram.

1 ■ . . OS 1 • 1 ' '
c- t. , . , ■

00

(N -"IH !0

^^ ^ . . O 00 Cv}

•^ ^ ; ; eb o> CO ; ; ; 1 ; ; ;

,-H rH ,-H

O

oil

4h 3D x> ', '

10 10 ' ' '
1 ; t- t- I- ; ! 1 1 1 1 ;

Hp

,,<J»T--|-*,

''ooos'''''''

'' 0} « T-H ''''■' '

, i;H TjH ,,,,,, ,

' iO CO ^ '■'''' '

o ■&?.

so t- .

T^ Ct 1

CO CO ','.','.',',' I ' '.

OU2
found,
gram.

0-6680
0-4465

1 GO lO

a x^ 1 ?» "?'

1 loo ' '

H.,0
fouud.
gram.

10 OJ Ill

00 3D •

CO

i 1 ! ! ' ! ! 1 I 1

Substance
used,
gram.

OTtiQO-rHiOT-ieo-rf<oeOT(<i:o
o}Q03scQ-^i-HOOcooobeoo

-^ 01 -^ CO ^ iO ^ IC H^ 50 «p

000000000000

d

"-I ?^ «5 ^

1^ t-

o W ;?; .7: Hh o

72 Chittenden — Caseoses, Casein Dyspeptone, and Casein Peptone.

Digestions C and D.

These digestions were conducted in much the same manner as the
preceding. In C, the casein was first subjected to the action of four
litres of vigorous, but purified, artificial gastric juice containing 0-4
per cent, acid, for four consecutive days. The undigested residue
was then filtered off, washed, and again treated with a vigorous pep-
sin-acid mixture for three days longer, in both cases at 40° C, The
residue still undigested was washed thoroughly with water and then
dissolved in one per cent, sodium carbonate, after which it was treated
exactly as pi-eparation A,

In 2>, the casein was warmed for three days with six litres of a
similar pepsin-acid mixture, when the undissolved residue, after being
filtered and thoroughly washed, was dissolved in diluted sodium
carbonate and reprecipitated by dilute hydrochloric acid. Then, as
in B^ the washed precipitate was redigested with a vigorous gastric
juice for several days and the residue again dissolved, after thorough
washing, in one per cent, sodium carbonate and treated exactly as
the preceding preparation.

For analysis, both products were freed entirely from fat, and ulti-
mately dried at 110° C, until of constant weight. Their composition
is shown in the accompanying tables.

Three other distinct preparations of dyspeptone were made in man-
ner similar to the preceding, except that in all, larger quantities of
pepsin-hydrochloric acid were employed and the mixtures warmed
for a longer time at 40° C. Tims in digestion E, 1562 grams of
moist casein were warmed with 9-5 litres of 0*4 per cent, hydrochloric
acid and pepsin for two days, and the undigested residue again treated
at 40° C. with 3 litres of a like pepsin-acid for seven days, and finally
treated a third time with pepsin and acid for four days, before solu-
tion in sodium carbonate, etc. Likewise in digestion 6^, 2000 grams
of moist casein were warmed at 40° C. with 11 litres of 0*4 per cent,
acid and pepsin for 21 days and the residue warmed again at 40° C.
for several days, with a fresh pepsin-acid mixture. Ultimately, all of
the three products Avere treated as previously described, and finally
dried at 110° C. prior to analysis. The analytical results are shown
in the accompanying tables.

P after
deducting
P of ash.

\ \ \ \ \

i

i

r-t
O

CO

6

pLl-fcR

\ \ \ \ \

'.

:

;

;

bos O

■;;:;;

i

\

i

1

o

p
o

•

;

\

T- 1

so

!

1

•^ g 03 ?3
. boo ■* 5b

05

O 1 ■

1 1 1 I 1 ■ 1 -^ 1 1

' ' ' ; ' ' ; I ■?

' ' ' ' o ' '

M "teS.

i

o

o
6

\

;

;

c3 ^M S

o 9 + S

; ; ; ; :

;

o
o

CO
Ol

o
o

•

:

;

1^

CO

\ \ \ \ ^
1 i' i i <ri

1-1

CO

J

;

[

;

;

£ a

^

05 =

=2 be

trj^^

9, fl a

; 13 £

hH I— I I— I ►> K. I— I

I ^ a ^ ^ >

Trans. Conn. Acad., Vol. VIII.

s a X «

t> HH M

10

M

V

'^ i^ ^O ':::i

O K Iz; M PL^ O

Nov., 1888,

r- ^ -^ I

o _

+ cs I

&D3

^5 ^- a

^ S a= 2

TiO

cs &M a

,

,

,

,

O

OS

,

,

,

sli&

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;

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o

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Ash
fouad.
gram.

-*

to

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1

;

o

CO

'

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;

'

1

'

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'

o
o

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6

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Chittenden — Caseoses, Casein Dyspeptone, and Casein Peptone. 75

Dyspeptone E.

Sub-
stance
used,
gram.

found,
gram.

H

N found.

N
%

Ash
found,
gram.

No.

COo
found,
gram.

C

c. c.

T.
°C.

Pres-
sure,
mm.

Ash
%

I

II

III

IV

V

VI

0-7272
0-3746
0-3177
0-5570
0-3319
0-3330

0-3758
0-1927

5-74
5-71

1-1528
0-5967

43-23

43-17

34-06
58-33

13-0
12-2

759-6
760-3

12-84
12-61

0-0496
0-0499

14-94
14-98

Percentage composition of ash-free substance.

C
H

N

50-84
6-75

50-75
6-72

15-10

15-15

Average.

50-80

6-73

15-12

Dyspeptone F.

Sub-
stance
used,
gram.

found,
gram.

CO2
found,
gram.

C

%

N found.

N
%

Ash
found,
gram.

No.

c. c.

T.

° G.

Pres-
sure
mm.

Ash

fo

I

0-5375

0-2837

5-86

0-8772

44-50

....

'

II

0-5686

0-3039

5-93

0-9317

44-68

... -

III

0-4649

50-3

12-4

764-0

13-11

IV

0-4100

44-7

13-0

765-5

13-16

.-..

V

VI

0-3938
0-4347

-- —

0-0536
0-0590

13-61
13-59

Percentage composition of ash-free substance.

c

51-50

51-70

H

6-78

6-87

N

15-17
Dyspeptone G

15-21

Average.

51-60

6-83

15-19

Sub-
stance

used,
gram.

H.,0
found,
gram.

H

%

CO2
found,
gram.

fo

N found.

N

%

Ash
found,
gram.

No.

c. c.

T.
" 0.

Pres-
sure
mm.

Ash

%

I

0-7545

0-3915

5-76

1-2148

43-90

....

n

0-4094

0-2137

5-79

0-6604

43-98

III

0-6268

- . - -

....

70-6

13-8

760-5

13-56

IV

0-7803

88-8

14-0

760-3

13-61

....

V

0-4356

0-0552

12-66

VI

0-5654

----

----

----

----

,----

0-0722

12-76

Preeentage composition of ash-free substance.

Average.
C 50-35 50-44 .... .... 50-39

H 6-61 6-65 .... .... 6-63

N 15-55 15-61 15-58

76

R. H. Chittenden — Caseoses, Casein Dyspeptone^

Table Showing the Average Composition of the Several Dyspep-

TONES.

A.

B.

C.

D.

E.

P.

G.

Casein.*

C -

51-15

51-07

51-29

51-82

50-80

51-60

50-39

53-30

H

7-18

7-41

7-26

7-44

6-73

6-83

6-63

7-07

N ._.

15-16

15-41

15-23

15-48

15-13

15-19

15-58

15-91

S - ------

0-71

0-71

0-68

0-78

0-83

P

0-87

25-80

25-40

25-54

24-48

....

----

....

23-03

Ash.-.. -

13-67

15-41

12-43

14-19

14-96

13-60

12-71

0-98

It is evident from the more complete analyses of the first four
products, that the dyspeptone as prepared by us contains essentially
the same percentage of sulphur as the original casein ; further, that
instead of being a phosphorized compound, it apparently contains no
phosphorus whatever, other than that combined with calcium. Very
noticeable, is the large percentage of ash in all of the preparations.
This we were not able to materially reduce by any process of purifica-
tion, and as the ash of the original casein, like that of the dys-
peptone, was composed almost wholly of calcium phosphate, it would
appear as if all of the phosphate from the mother substance had
attached itself to the dyspeptone.

In their content of carbon, all of the seven preparations show a very
close agreement, while they differ from casein by containing two per
cent, less carbon. The nitrogen of the dyspeptone is, likewise, a little
less tlian that of casein, while the individual preparations show
throughout a very close agreement in their content of this element.
In composition, therefore, all of the seven preparations, although rep-
resenting considerable variation in the method of production, show a
sufficiently close agreement to indicate their identity. Compared
with casein, the lower percentage of carbon would point to their
production by hydration, and it would appear from the analytical
data that the so-called casein dysjjeptone, formed by gastric digestion,
is a mixture of calcium phosphate with a hydration product of casein,

*CbitteDden and Painter. Studies from Laboratory, vol. ii. p. 113.

and Casein. Peptone. 77

the hydrochloric acid compound of which is insoluble in water and
dilute acid. The dyspeptone itself is quite readily soluble in cold
water, the solution remaining unchanged on boiling.

Following are some of the reactions of dyspeptone.

Addition of acetic acid to an aqueous solution of the substance
produces a heavy white precipitate insoluble in moderate excess, but
partially soluble in a large excess of the acid. On heating the strongly
acid fluid, the precipitate dissolves completely and the fluid remains
clear on cooling. Addition of potassium ferrocyanide to the clear
acid fluid gives only a slight turbidity.

Dilute hydrochloric and sulphuric acid both give a heavy white
precipitate, insoluble in slight excess of acid, but entirely soluble
in a large excess on application of heat. Even 0-2 per cent, hydro-
chloric acid precipitates the dyspeptone completely.

Dilute nitric acid, likewise, precipitates the dyspeptone, but the
precipitate is far more soluble in excess of the acid. On w^arming the"
acid solution, it quickly turns yellow, and with ammonia gives the
orange yellow color of the xanthoprotein reaction.

Cupric sulphate and potassium hydroxide give the violet color of
the biuret reaction.

Cupric sulphate and ferric chloride both give heavy precipitates,
insoluble in excess.

Potassium hydroxide and lead acetate give, on boiling, a distinct
reaction for sulphur.

Mercuric chloride, added in small quantity to a cold aqueous solu-
tion of the dyspeptone, gives no precipitate, but when added in
excess and the mixture is heated, a heavy white i^recipitate is formed,
insoluble on cooling.

The dyspeptone is precipitated by saturation of its aqueous solution
with ammonium sulphate, but not by sodium chloride, even on heat-
ing. Addition of acetic acid, however, to the salt-saturated fluid
gives the usual precipitate of dyspeptone.

Casein antialbumid.

On heating casein with sulphuric acid and water at 100° C, it is
decomposed, as is well known, into soluble products and an insoluble
antialbumid. We have found, however, that the antialbumid pre-
pared in this manner is quite difierent in composition from the dys-
peptone formed in gastric digestion. In one experiment, where about
two kilograms of pure, moist casein were heated with two litres of
water and 100 grams of concentrated sulphuric acid for seven hours

'78 i?. H. ChitteMflen — Gaseoses, Casein Dyspeptone,

at 100° C, and the residue so obtained treated again in a like man-
ner with the same strength of acid, a comparatively large amount of
casein antialburaid was obtained, which unlike the dyspeptone from
a gastric digestion, was only slowly soluble in dilute sodium carbon-
ate. Freed from any adhering soluble products by treatment with
several litres of a vigorous gastric juice for two days at 40° C, it still
dissolved slowly in dilute sodium carbonate. By long contact with
a one per cent, solution of the alkaline carbonate it finally dissolved,
leaving but a small residue. From this solution, the antialbumid
was reprecipitated by hydrochloric acid, and after thorough washing
with water, it was again dissolved in sodium carbonate, the fluid
made exactly neutral and then dialyzed until all chloride was re-
moved. After concentration of the fluid and precipitation with alco-
hol, etc., the antialbumid was dried and analyzed. It contained 18
percent, of ash. The ash-free substance contained 54*4 per cent, of
carbon, 68 per cent, of hydrogen, and 14"8 per cent, of nitrogen;
showing thus a much higher percentage of carbon, and a lower per-
centage of nitrogen than the dyspeptone formed by pepsin-hydro-
chloric acid.

Both casein antialbumid and dyspeptone are dissolved with more
or less readiness by alkaline solution of trypsin and are converted
by long warming at 40° C. into a peptone-like body, presumably
antipeptone.

II. Caseoses ;— from experiments by Charles Norris, Jr., Plt.B., and

C. A. Tuttle, Ph.B.

In a previous study of the caseoses formed in pepsin digestion,* we
were much impressed with the peculiar behavior of protocaseose
towards acids. Unlike the proto bodies from other proteids pre-
viously studied, aqueous solutions of the substance gave heavy pre-
cipitates with dilute acids. Protocaseose, as then separated, was
readily soluble in 0-4 per cent, hydrochloric acid, but addition of
stronger acid invariably produced a decided precipitale, soluble,
however, in a still larger excess of acid. This peculiarity rendered
the protocaseose an object of some interest to us, and further study
of the conditions favoring its formation in gastric digestion has
shown us that, apparently, the nature of the body precipitated by
saturation with salt, as well as the body precipitated by salt-satu-
rated acetic acid is modified by the strength of the pepsin solution,

* Chittenden and Painter. Studies from Laboratory of Physiological Chemistry,
vol. ii, p. 195.

a7id Casein Peptone. 79

and the length of time the casein is subjected to the action of the
ferment. In our earlier work with the proteoses, we deemed it es-
sential in attempting a study of the primary products of proteolytic
action to use as weak a ferment solution as possible, and to discon-
tinue its action as soon as solution of the proteid was complete, in
order that there might not be too great a loss through formation of
peptone. Further study, however, especially by use of the am-
monium sulphate reaction, has shown us that the formation of pep-
tone is a far less rapid process than generally supposed. Indeed, in the
majority of artificial digestions with pepsin-hydrochloric acid, as or-
dinarily conducted, the ammonium sulphate reaction will show the
entire absence of peptone. True peptone appears to be formed only
by the action of a very vigorous pepsin mixture and that long con-
tinued.

In all of our previous experiments, the casein was either subjected
to the action of a very weak pepsin mixture or else, in the use of a
stronger ferment, exposed to its action for a few hours only. We
now find that by using a far more vigorous pepsin mixture and by
continuing its action for several days instead of hours, there is still
not a trace of [jeptone to be found in the filtrate from the ammonium
sulphate precipitate of the caseoses, but that the caseoses them-
selves, particularly the proto and deuterocaseose, differ somewhat,
both in composition and reactions from the products previously sep-
arated. The discovery of this fact has led us to a further study of
the caseoses formed in pepsin digestion, by which we have been able
in many ways to verify our former observations and at the same
time extend our knowledge of these interesting primary cleavage
products of casein. We have also extended our work by studying
the caseoses formed through the action of try|)sin, and dilute sul-
phuric acid.

A. Caseoses formed by pepshi-hydrochloric acid.

In all of these experiments the pepsin mixture was very power-
ful, and was especially prepared to insure freedom from both albu-
moses and peptone.* The casein was, likewise, thoroughly pure,
having been freshly prepared from skim milk by precipitation
with 0'2 per cent, hydrochloric acid, and reprecipilation three or four
times after ■solution in ammoniacal water.

* Studies from the Laboratory of Physiological Chemistry, vol. ii, p. 133.

80 R. H. Chittenden — Caseoses, Casein Dyspeptone,

Digestion A.

Nearly 2 kilos, of moist casein were warmed at 40° C. for a
little more than two days, with 10 litres of 0*4 per cent, hydro-
chloric acid containing sufficient of the pepsin mixture to insure vig-
orous action. After partial neutralization of the acid, the clear fluid
was filtered from the semi-gelatinous dyspeptone, made exactly neu-
tral with sodium hydroxide and then evaporated until moderately
concentrated. On filtering the concentrated fluid through paper, a
small residue remained, somewhat gummy, insoluble in dilute acid,
but readily soluble in dilute sodium carbonate, from which it was
precipitated by either hydrochloric or acetic acid. The amount was
too small for study, but it seemed to resemble in reactions casein
dyspeptone.

The neutral fluid containing the caseoses gave no precipitate
whatever on addition of 0*4 per cent, hydrochloric acid or even
stronger acid, and in this respect differs from the earlier diges-
tions in which the ferment action was continued for a short time
only. With dilute acetic acid, however, a slight turbidity was pro-
duced, the amount of which was too small to admit of any study of
its character.

The caseoses were precipitated collectively in the form of a heavy
gummy precipitate, by saturation of the neutral fluid with ammo-
nium sulphate. On boiling the filtrate from this ammonium sulphate
precipitate, a small quantity of a second gummy precipitate was
obtained. In the filtrate from this second precipitate, no trace of a
peptone-like body could be discovered by any of the ordinary tests.
Apparently only caseose bodies had been formed.

The first and main ammonium sulphate precipitate, after being
washed by trituration with a saturated solution of ammonium sul-
phate, was dissolved in water and the perfectly neutral fluid satu-
rated in the cold with sodium chloride. By this means a heavy
gummy precipitate was formed, which after being washed with a
saturated solution of sodium chloride was redissolved in water, and
reprecipitated by saturation of the neutral fluid with salt. After
three or four reprecipitations, the protocaseose was considered suffi-
ciently pure.

In this digestion, there appeared to be present more heterocaseose
and dyscaseose than in our former experiments, as was evidenced by
the small insoluble residues remaining each time the precipitated
protocaseose was redissolved in water. These residues of hetero- and

and Casein Peptone. 81

dyscaseose were very small, but still sufficiently laro-e to enable us
to make out their general characters. Similarly, Dr. Thierfelder*
found in the purification of his protocaseose, or " propeptone I" as
he terms it, a small, insoluble residue each time he dissolved the
sodium chloride precipitate in water. It would appear, therefore,
that in a vigorous or long continued digestion there is a much greater
probability of the hetero body being formed than when the ferment
is allowed to act only a short time on the casein. This, however, is
contrary to Neumeister's views regarding the order of formation of
the proteoses.

In order to free the precipitated protocaseose from salt and any
adhering heterocaseose, it was dissolved in water and dialyzed until
all chloride was removed. The neutral solution was then concen-
trated to a syrup, and the proto body precipitated by alcohol.
During the concentration of the fluid, a gummy-like mass separated,
similar tothe separation of protoelastose. This, however, dissolved
more or less completely as the mixture cooled. The precipitated
caseose, after being extracted several times with alcohol and with
warm ether, was partially dried, ground to a fine powder, re-
extracted with ether in a fat extractor and finally dried at 110° C.
until of constant weight.

On analysis it gave the following results :

Protocaseose A.

I. 0-4947 gram substance gave 0-9794 gram CO„ = 53-98 per cent. C.

II. 0-380-2 gram substance gave 0-2397 gram H.,O = 7-00 per cent. H.

III. 0-3862 gram substance gave 0-2467 gram H.,O = 7-09 per cent.
H and 0'7670gram CO„i=54-15 per cent. C.

IV. 0-4953 gram substance gave 64-8 c. c. N at 20-8° C. and
760-3 mm pressure = 15-72 per cent. N.

V. 0-3764 gram substance gave 50-1 c. c. N at 22-5° C. and 760-6
mm pressure = 15-95 per cent. N.

VI. 0-4599 gram substance gave 0-0046 gram ash = 1-00 per cent.

VII. 0-4103 gram substance gave 0-0041 gram ash = 0-99 per cent.

Percentage composition of ash-free substance.

Average.

C 54-53 54-69 ..- --.. 54-61

H .... 7-1] 7-07 ' 7-11

N .... 15-88 16-11 15-99

* Zur Kenntniss der Cfiseinpeptone, Zeitsclirifit fur ph3'siologiselie Chemie, x, p. 577.
Tkans. Conn. Acad., Vol. VIII. 11 Nov., 1888.

82 R. H. Chittenden — Caseoses, Casein D^speptone,

This protocaseose is thus seen to have a higher percentage of car-
bon than the proto bodies previously* studied, although one prod-
uct was then obtained with 53-93 per cent. C, 7' 1 7 per cent. H, and
16-05 per cent, N.

In reactions, likewise, this protocaseose differs somewhat from the
protocaseoses previously obtained, and as the characters of the pres-
ent body have been verified by the reactions of several other prod-
ucts similarly prodiiced, we are led to believe in their constancy.
Moistened with water, the powdered protocaseose becomes immedi-
ately gummy and soon dissolves to a perfectly clear fluid, which on
addition of considerable water becomes decidedly cloudy or tur-
bid. Treated with a large amount of water at the outset, the pro-
tocaseose dissolves more slowly, giving a more or less turbid fluid.
In dilute acid and in dilute sodium carbonate it dissolves to a per-
fectly clear fluid.

Towards heat, aqueous solutions of protocaseose act exactly like
protoelastose. Even when warmed very gently, the solution be-
comes quickly turbid and if concentrated gives more or less of a floccu-
lent precipitate. On cooling, the turbidity disappears, reappearing
as the fluid is heated. Like protoelastose also, a solution of the
caseose body on being rapidly concentrated deposits more or
less of the substance as a gummy mass, which, however, will dis-
solve in cold water, or if the fluid is not too concentrated will dis-
solve in the mother liquid as it cools. Dilute acetic acid added to
an aqueous solution of protocaseose gives no precipitate whatever,
but potassium ferrocyanide added to the acid fluid gives a heavy
precipitate.

Dilute hydrochloric acid produces no precipitate.

Dilute nitric acid added to an aqueous solution of the caseose
gives a heavy white precipitate, which on gently warming, quickly
dissolves while the fluid takes on a faint pink or rose color, wliich
on further warming changes to a bright yellow or reddish yellow
color. If the nitric acid solution is not warmed too long, the pre-
cipitate reappears as the mixture cools.

Solution of cupric sulphate gives a heavy, greenish white precipi-
tate when added to an aqueous solution of protocaseose.

As already stated, the first precipitates of protocaseose obtained
by saturating the neutralized digestive fluid with salt, were not en-
tirely soluble in water; a small residue remained, apparently

* See Chittenden and Painter. Studies from Laboratory of Physiological Chem-
istry, Yale University, vol. ii, p. 197.

and Casein Pejitone. 83

wholly insoluble. This residue, whicli by analogy should consist of
hetero and dyscaseose was somewhat soluble in salt solution and
wholly soluble in dilute sodium carbonate and dilute acid, both acetic
and hydrochloric, even dissolving in 0-2 per cent, hydrochloric acid.
It was dissolved in sodium carbonate, the fluid neutralized without
giving any precipitate, and dialyzed until the sodium chloride was
entirely removed. The neutral and clear fluid was then concentrated
to a syrup, precipitated by alcohol and the precipitate dried at 110° C.
On treatment with cold water, it was now found soluble to a large
extent, though a certain amount of gummy matter was still insolu-
ble. The soluble portion showed all of the reactions of the proto
body ; the solution being rendered turbid by heat, clear again on
cooling, and giving in the cold a heavy precipitate with nitric acid,
soluble when heated. This behavior of the insoluble heterocaseose
towards dilute sodium carbonate would seem to imply a conversion
of this substance into protocaseose, or perhaps a reconversion of the
coagulated heterocaseose dyscaseose, into heterocaseose proper.

Acetic acid precipitate.

On adding a little 30 per cent, acetic acid to the oi'iginal salt-
saturated filtrate from the first sodium chloride precipitate of proto-
caseose, etc., a heavy, flocculent precipitate settled out, which in
amount far exceeded the protocaseose and which on standing, soon
became gummy. Excess of acid was avoided, as the precipitate
was somewhat soluble in a large amount of the reagent. The
gummy mass, after being washed as thoroughly as possible with
saturated salt solution, was treated with cold water, in which the
greater portion of the substance dissolved, the solution made neu-
tral, dialyzed, concentrated to a syrup, and the substance precipitated
with alcohol. It was then thoroughly extracted with ether and
finally dried at 110° C.

In reactions, it diftered decidedly from protocaseose; in water
it was quickly and completely soluble and the solution when heated
gave no coagulum whatever, or at the most only the slightest ap-
proach to a turbidity.

Dilute nitric acid, in the cold, gave no precipitate ; when heated, the
acid fluid changed to a reddish yellow color, which quickly turned
yellow.

With acetic acid, an aqueous solution of the substance remained
perfectly clear. Potassium ferrocyanide, however, when added to
the acid fluid gave a heavy precipitate.

84 R. H. Chittenden. — Caseoses, Caaein Dyspeptone^

Saturation of the aqueous solution with salt gave a slight turbidity.
Cupric sulphate gave a heavy precipitate, soluble in excess.
On analysis, the substance gave the following results, which show
a composition quite different from that of protocaseose.

Acetic Acid Precipitate A.

I. 0*4629 gram substance gave 0-2812 gram H„0=r6'74 per cent.
H and 0-8608 gram 00^=50-71 per cent. C.

II. 0-3277 gram substance gave 0-1977 gram H2O = 6'70 ])er cent.
H and 0-GOoO gram CO3=50-34 per cent. C.

III. 0-4253 gram substance gave 53-1 c. c. N at 130" C. and 761-8
mm pressui-e=: 15-01 per cent. N.

IV. 0-4843 gram substance gave 60-8 c. c. N at 13-0° C. and
760-8 mm pressure= 15-07 per cent. N.

V. 0-3649 gram substance gave 0-0108 gram ash = 2-96 per cent.

VI. 0-3947 gram substance gave 0*0124 gram ash = 3-14 per cent.

Percentage composition of ash-free substance.

Average.

C 52-30 51-92 .-.- 5S-10

H 6-96 6-90 6-93

N 15-48 15-54 .15-51

As previously stated, the above original precipitate produced by
acetic acid was not entirely soluble in water. A small residue
remained, which after being washed with water was dissolved in
dilute sodium carbonate, and the solution neutralized with dilute
hydrochloric acid, without yielding any neutralization precipitate.
The solution was then dialyzed, concentrated to a syrup and the
substance precipitated by alcohol. It was now found, to a great
extent, soluble in water, the solution showing no turbidity by heat
and giving no precipitate with nitric acid. Cupric sulphate gave a
heavy precipitate, and dilute acetic acid added to the aqueous solu-
tion produced quite a heavy precipitate, not readily soluble in excess
of the acid. The amount of substance was too small to admit of
analysis, and the reactions are hardly sufficient to identify it. It is
evidently not heterocaseose, for it is only the acetic acid compound
that is insoluble in water, not the caseose substance itself. In many
respects it appears like casein dyspeptone, and as this substance
is precipitated by saturation of its aqueous solution with ammonium
sulphate and not by sodium chloride, the presence of a trace of this
body might not be impossible.

and Casein Peptone. 85

Amnwniiun sulphate precipitate.

On adding ammonium sulphate in substance to the above salt-
saturated acetic acid fluid, a slight gummy precipitate was obtained,
readily and completely soluble in water, and which, after removal of
the salts by dialysis and concentration of the fluid, was precipitated
by alcohol. In reactions it did not differ materially from the body
obtained by precipitation with acetic acid, except that with cupric
sulphate only a slight precipitate was produced. With acetic acid
and potassium ferrocyanide, on the other hand, a distinct precipitate
Avas obtained, while with nitric acid in the cold no turbidity whatever
was produced.

In composition, however, it differed decidedly from the preceding
preparations, although as it contained considerable ash, nearly 10 per
cent., the result perhaps can be considered only as an approximation
to the truth.

Following is the percentage composition of the ash-free substance :

Average.

48-23

6-94.

15-63 15-75 15-69

This body, which by analogy should be nearly pure deutero-
caseose, evidently approaches much nearer to our conception of a
true casein peptone than any of the preceding preparations. Another
body, however, has been obtained with a still lower content of
carbon and with reactions still more closely approximating to true
peptone.

In the first precipitation of the caseoses fi-om this digestion by
saturation of the original fluid with ammonium sulphate, it will be
remembered that a small amount of a second gummy precipitate was
obtained on heating the cold saturated ammonium sulphate filtrate.
This gummy precipitate of a caseose body, after purification by
dialysis and precipitation with alcohol, was found to consist of a
substance extremely soluble in water, the solution giving no precipi-
tate with acetic acid and potassium ferrocyanide, neither with nitric
acid nor with cupric sulphate. After being dried at 110° C. it gave
on analysis the following results :

I. 0*7280 gram substance gave 0-4239 gram 11.^0=0-46 per cent.
H and 1*2160 grams 00^ = 45 -54 per cent. C.

II. 0*5020 gram substance gave 0-2905 gram 1120 = 6*40 per cent.
H and 0*8442 gram C0„ = 45'85 per cent. C.

c

48.33

48-13

H

6*88

7-01

N

86 R. H. Chittenden — Caseoses, Casein Dyspe^'itone,

III. 0-3291 gram substance gave 42-1 c. c. N at 13-9° C. and
756 mm pressure = 15-33 per cent. N.

IV. 0-6835 gram substance gave 87*0 c. c. N at 14-5° C. and
756 mm pressure= 15*30 per cent. N.

V. 0-5370 gram substance gave 0*0230 gram asli=4*28 per cent.

VI. 0-3525 gram substance gave 0-0154 gram ash = 4*36 per cent.

Percentage composition of ash-free substance.

Average.

C 47-57 47-87 .... 47-7^

H 6-75 6-70 .... 6-73

N .... 15-99 15-96 15-97

This substance, since it is precipitable by ammonium sulpliate,
cannot be considered a true peptone, yet in composition it closely
approaches both the ampho- and antipeptone from fibrin,* which it
also resembles somewhat in reactions, except in its behavior towards
ammonium sulphate.

It is thus evident from the foregoing, that in this active and com-
paratively long continued digestion there is a much smaller amount
of protocaseose present than was found in our former experiments.
Indeed, deuterocaseose appears to predominate, while at the same
time the protocaseose is modified both in composition and reactions,
due in part without doubt, to adhering heterocaseose. Further,
we are inclined to consider the presence of at least two forms of deu-
terocaseose. We have generally considered that a proto body is never
completely precipitated by saturation of its aqueous solution with
salt, and that consequently the precipitate produced by acid in the
salt-saturated fluid must be a mixture of proto and deuteroproteose, and
this we have usually found to be the case. In the present diges-
tion, however, the acetic acid precipitate contained only a very small
amount of protocaseose, for as previously stated this precipitate when
purified gave only a slight turbidity on saturation of its aqueous solu-
tion with salt and no precipitate whatever with nitric acid ; both of
which reactions would indicate freedom from any large amount of pro-
tocaseose. This view is further substantiated by the great difference
in the percentage of carbon of the two bodies. To be sure we have,
with Nenmeister, looked on the cupric sulphate reaction as a means
of distinguishing between proto and deuteroalbumose, but it does
not necessarily follow that the same reaction will hold good for all
proteoses. The acetic acid precipitate does indeed give a strong

* Kviline and Cbittenden, Studies, vol. ii, p. 40.

and Casein Peptone. 87

reaction with cupric sulphate, but the striking differences in composi-
tion and reactions between the purified sodium cliloride precipitate
and the acetic acid precipitate (when freed from acid) point to a
totally different nature, and we are inclined to consider the latter as
a deutero body, probably contaminated with a little protocaseose,
and for convenience we propose to call it a deuterocaseose.

It is unquestionably very difficult, if not almost impossible, to isolate
the individual caseoses in a state of perfect purity. Whenever one
is precipitated, it usually brings down with it more or less of any
other caseose present and such admixtures are very hard to remove.
It is, we think, owing to this fact that we have not been able to
obtain a deuterocaseose sufficiently free from protocaseose as not to
give any precipitate on saturation with sodium chloride.

That form of caseose in this digestion which was not precipitated
by salt, or by salt and acetic acid, but which appeared on addition of
ammonium sulphate in the cold, is probably a mixture of a deutero and
and what we term (3 deuterocaseose, with possibly an intermediate
body.

The name (3 deuterocaseose, we apply to that caseose not readily
precipitable by saturation with ammonium sulphate in the cold, and
which is generally found in a greater or less quantity in the filtrate
from the precipitate produced by saturation with ammonium sulphate.
It is precipitated fairly pure, as a sticky gum, by simply boiling the
saturated ammonium sulphate filtrate and is especially characterized
by its low content of carbon, and by its non-precipitation with acetic
acid and potassium ferrocyanide, with nitric acid, and with cupric sul-
phate. It stands, unquestionably, nearer to peptone than a deu-
terocaseose and is doubtless formed from the latter by the continued
action of the ferment.

Digestion B.

In this digestion, 2 kilos, of moist casein were warmed at 40° C.
for eight days, with about 5 litres of 0*4 per cent, hydrochloric acid
containing an active pepsin solution, after which the mixture was
partially neutralized with sodium carbonate and filtered from the
dyspeptone. The clear fluid was then made exactly neutral (no neu-
tralization precipitate) and concentrated to a thin syrup. When cold,
proto and heterocaseose were directly precipitated by satm-ation
of the solution with salt. Protocaseose was purified by repeated
precipitation with salt, etc., as described under A. In this process,
the same insoluble residues of hetero and dyscaseose were met witii

88 H. H. Chittenden — Caseoses, Casein Dyspeptone,

as in the previous digestion. The purified protocaseose showed the
same peculiar reactions as protocaseose A., viz: with nitric acid a
heavy white precipitate, soluble on warming; with heat alone, a dis-
tinct turbidity or coagulum, disappearing as the solution cooled ; and
giving with a large amount of water a more or less turbid fluid, from
whiich on standing a little gummy matter separated.

Dried at 110° C. and analyzed, the following results were obtained.

Protocaseose B.

I. 0-3470 gram substance gave 0-2192 gram H.,O = 7'02 per cent. H.

II. 0-2935 gram substance gave 0"1854 gram II„O = 7-01 per cent.
H and 0-5779 gram 002 = 53-69 per cent. C.

III. 0-2952 gram substance gave 0-5853 gram CO^ = bA-0l jDer
cent. C.

lY. 0-2989 gram substance gave 38-6 c. c. N at 13-8° C. and
760-1 mm pressure =15*52 per cent. N.

V. 0-5564 gram substance gave 72-8 c.c. N at 13-1° C. and
760*1 mm pressure=15'69 per cent. N.

VI. 0*3594 gram substance gave 0*0047 gram ash=r30 per cent.

VII. .0*3523 gram substance gave 0*0045 gram ash = l*27 per
cent.

Percentage composition of ash-free substance.

Average.

54-39 54-58

7-10
15*71 15*90 15-80

In composition, therefore, as in reactions, this body is apparently
identical with protocaseose A.

In the oi'iginal salt-saturated filtrate from protocaseose, acetic
acid produced a heavy, gummy precipitate, which was dissolved in
water and purified in the same manner as the corresponding body in
digestion A. Like the latter, it was readily and completely soluble
in water, the solution giving no turbidity whatever by heat, nor on
the addition of either nitric or acetic acid. With cupric sulphate, a
heavy precipitate was formed, as also with acetic acid and potassium
ferrocyanide.

Dried at 110" C it gave the following results on analysis:

Acetic acid precipitate B.

I. 0*3429 gram substance gave 0-2057 gram H.,0 = 6*66 ])er cent.
H and 0*6315 gram CO.,=50*22 per cent. C.

c

54-77

H

7-10

7-09

N

and Casein Peptone. 89

II. 0-4345 gram substance gave 0-2734 gram H^O = 6*72 per cent.
H and 0-8358 gram CO„ = 50-48 per cent. C.

III. 0-3316 gram substance gave 43-5 c. c. N at 13-7" C. and
756-3 mm pressures 15-70 per cent. N.

IV. 0-3258 gram substance gave 43*1 c. c. N at 13 2° C. and 759-1
mm pressure= 15*86 per cent. N.

V. 0-3306 gram substance gave 0-0126 gram ash=3-81 per cent.

VI. 0-2262 gram substance gave 0-0085 gram ash = 3-76 per cent.

Percentage composition of ash -free substance.

Average.
C 53-17 52-43 .... .... 5^-30

H 6-93 6-99 .... .... 6-95

N .... .... 16-32 16-49 16-40

This body appears to differ from a deuterocaseose obtained in
digestion A by nearly 1 per cent, of nitrogen, but in all other respects
is practically identical with it.

As in digestion A, the precipitate first obtained by the addition of
acetic acid was not entirely soluble in water, a residue remained
soluble in dilute sodium carbonate, and which comported itself
exactly like the insoluble residue obtained in the preceding diges-
tion, apparently being a trace of casein dyspeptone.

The original filtrate from the above acetic acid precipitate, on
saturation with ammonium sulphate, gave an additional precipitate,
hardly sufficient for analysis, but which when purified proved to be
identical with the corresponding body fi'om A and like it giving
with cupric sulphate only a very slight precipitate.

It is thus seen that in a vigorous pepsin digestion of casein there
are formed, in addition to dyspeptone and heterocaseose, at least three
distinct caseoses all soluble and differing from each other both in
composition and reactions. As compared with casein, protocaseose
is somewhat peculiar in containing a higher percentage of carbon
than the mother substance. All of the other products show a very
much smaller content of carbon. The relative composition of the
products is shown in the following table :

Casein.

Proto-
caseose
A.

Proto-
caseose
B.

a Deutero-
caseose
A.

a Deutero-
caseose
B.

/? Deutero-
caseose
A.

c

53-30

54-61

54-58

52-10

52-30

47-72

H

7-07

7-11

7-10

6-93

6-95

6-73

N

15-91

15-99

15-80

15-51

16-40

15-97

Trans. Conn. Acad., Vol. VI FF. 12 Nov., 18SS.

90 It. H. Chittenden — Caseoses, Casein Dyspeptone,

Protocaseose agrees closely in its content of carbon with the
" propeptone I " of Thicrfelder.* This investigator has separated
from a pepsin digestion of casein, by a process similar to our method
of separating the proto body, a substance to which he gives the
above name and which contained 54'63 per cent, of carbon and 7-45
per cent, of hydrogen. In reactions it was similar to protocaseose,
except that aqueous solutions of the substance remained perfectly
clear on warming. Nitrogen was not determined. Thierfelder also
separated from the filtrate from his " propeptone I," a second body,
by addition of hydrochloric acid, to which he gives the name " pro-
peptone II." This substance, which corresponds to our a deutero-
caseose, he found to contain 49-8 per cent, carbon, 7"1S per cent,
hydrogen and 14-23 per cent, nitrogen. Judging from the method
of separation, however, the body analyzed must liave been an acid
compound of the caseose and not the caseose body itself. In reac-
tions, so far as they are given, the substance was not different from
a deuterocaseose.

b. Caseoses from WeyVs casehi 23eptone.

This commercial product, sent to lis from Germany, we have
examined according to the foregoing methods and have found it, as
might be expected, composed almost entirely of caseoses. It was
completely soluble in water and gave with acetic acid a slight tur-
bidity, somewhat increased by addition of potassium ferrocyanide.
By saturation of its aqueous solution with sodium chloride, only a
comparatively small precipitate was obtained, greatly increased, how-
ever, by addition of acetic acid.

200 grams of the powder were dissolved in water and the caseoses
precipitated collectively by saturation of the fluid with ammonium
sulphate, in the form of a heavy gummy precipitate. On heating
the filtrate from this precipitate of caseoses until a crust of ammo-
nium sulphate formed on the surface of the hot fluid, a second
gummy precipitate gradually separated, which after purification by
dialysis, etc., was finally precipitated by alcohol, and a portion dried
at 110° C. for analysis.

This substance, representing a form of caseose not readily precipi-
table by ammonium sulphate and thus indicating its close approach
to true peptone, is apparently identical with the ft deuterocaseose
similarly obtained in our own digestion, but present here in much

* Zeitschrift fiir physiologische Chemie, Band x, p. 585.

and Casein Peptone. 91

larger quantity. In water it was readily and completely soluble,
the solution giving no turbidity whatever by heat, nor with dilute
nitric acid. With cupric sulphate only a very slight turbidity was
produced, and with acetic acid and potassium ferrocyanide a corre-
spondingly slight turbidity. Its aqueous solution on being saturated
with salt and then made slightly acid with acetic acid showed a small
flocculent precipitate, doubtless representing the substance which
gave the slight turbidity with cupric sulphate and potassium ferro-
cyanide, viz : a deuterocaseose. On analysis the following results
were obtained :

ft deuterocaseose from WeyVs casein peptone.

I. 0-3641 gram substance gave 0*2121 gram H„0 = r47 per cent.
II and 0-6087 gram CO^=45-58 per cent. C.

II. 0-4600 gram substance gave 0-2708 gram H„0 = 6-54 per cent.
H and 0-7652 gram CO.^ = 45-36 per cent. C.

III. 0-3298 gram substance gave 42-2 c. c. N at 13-9° C. and
755*4 mm pressure =15-31 per cent. N.

IV. 0-3418 gram substance gave 43-5 c. c. N at 14-4° C. and
756-1 mm pressure =15-31 per cent. N.

V. 0-3514 gram substance gave 0-0153 gram ash =4-35 per cent.

VI. 0-3599 gram substance gave 0-0155 gram ash =4-30 per cent.

Percentage composition of ash- free substance.

Average.
47-50

_6'79

16-00 15-84 15-92

In composition, therefore, as well as in reactions, this body re-
sembles the ft deutero described under A, and like it is especially
characterized by its exceedingly low percentage of carbon.

F'rom the first ammonium sulphate precipitate of caseoses, proto-
caseose Avas separated by saturation of the aqueous solution of the
above precipitate with salt. As in similar precipitates from the
preceding digestions, there was considerable heterocaseose present,
showing itself as an insoluble residue when the sodium chloride pre-
cipitate was dissolved in water for reprecipitation. Purified after
the methods previously described, the protocaseose showed the usual
reactions characteristic of this body, its aqueous solution growing
turbid when heated, giving a precipitate with nitric acid, etc.

Dried at 110° C. it gave on analysis the following results :

c

47-61

47-40

H

6-76

6-83

N

92 M. H. Chittenden — Caseoses, Casein Dyspeptone^

Protocaseose, from WeyVs casein 2)eptone.

I. 0-4291 gram substance gave 0-259'7 gram Iljd^%-12 percent.
H and 0-8140 gram CO.,=51-'70 per cent. C.

II. 0-364'7 gram substance gave 0-2155 gram H„Or=6'56 per cent.
H and 0-6935 gram 00^=51-85 per cent. C.

III. 0-4115 gram substance gave 52-5 c. c. N at 136° C. and
760-8 ram pressure= 15-11 per cent. N.

IV. 0-7730 gram substance gave 96-9 c. c. N at 13-8° C. and 761-2
mm pressure:z= 15-10 per cent N.

V. 0-2901 gram substance gave 0-0121 gram ash=:4-16 per cent.

VI. 0-2959 gram substance gave 0*0120 gram ash=:4-05 per cent.

Percentage composition of ash-free substance.

Average.

c

53-93

54-08

1

54-01

H

7-01

6-84

6-92

N

15-75

15-70

15-72

Addition of salt-saturated acetic acid to the filtrate from proto-
caseose, gave a moderately heavy, gummy precipitate which, after
purification and removal of the acid, was completely soluble in
water, the solution showing no turbidity by heat, but giving a slight
turbidity with dilute nitric acid. It gave all of the reactions men-
tioned as characteristic of this precipitate. It w^as not analyzed.

The original filtrate from the acetic acid precipitate gave on sat-
uration with ammonium sulphate, a small gummy precipitate which
after purification was found wholly soluble in water, the solution
giving no turbidity by heat and no precipitate with nitric or acetic
acid. With acetic acid and potassium ferrocyanide, however, a dis-
tinct turbidity was produced and with cupric sulphate a heavy pre-
cipitate. The substance was not analyzed.

It is thus evident that Weyl's so-called casein peptone, however it
may be prepared, contains essentially the same kind of caseoses
found in our own digestions with pepsin-hydrochloric acid. In addi-
tion, a small amount of a substance was found not precipitable by
saturation with ammonium sulphate and Avhich gave no precipitate
with nitric or acetic acid, nor with acetic acid and potassium ferro-
cyanide, and with cupric sulphate only a slight turbidity. This
body, which doubtless was amphopeptone mixed with a little
caseose, was obtained in too small quantity for analysis.

and Casein Peptone. 93

c. Caseoses formed hy dilute sulpJmric acid at 100° C.

Two kilos, of pure moist casein were heated in a flask with 2
litres of water and 100 grams of pure sulphuric acid at 100° C. for
seven hours. The residue of casein and antialbumid was again
warmed with a like amount of fresh acid and water for six hours.
The acid fluids were united, neutralized with sodium carbonate with-
out giving any noticeable precipitate, and then evaporated until
moderately concentrated. On cooling, considerable tyrosin and
leucin crystallized from the fluid. With cupric sulphate, the solution
gave a heavy precipitate which dissolved in sodium hydroxide with
a reddish color. Sodium chloride and ammonium sulphate both pro-
duced heavy ])recipitates when added to saturation.

The caseoses were separated from the fluid by saturation in the
cold with ammonium sulphate, and from this precipitate, protocaseose
was separated by solution in water and precipitation with sodium
chloride. The sodium chloride precipitate was dissolved in water,
the substance roprecipitated by saturation with salt, and again dis-
solved in water to which a trace of sodium carbonate was added
to make the mixture quite neutral. On dialysis, a small amount of
gummy heterocaseose separated, mixed with a little tyrosin. The
clear fluid, now free from salts, was concentrated on a water-bath.
As the evaporation advanced, a brown gummy mass settled out,
which was, however, readily soluble in cold water, for as the concen-
trated fluid cooled at night the gummy matter entirely disappeared,
reappearing as the fluid was again heated. The final concentrated
fluid was treated with alcohol, the gummy precipitate boiled with
alcohol repeatedly to free it from any adhering tyrosin, and finally
extracted with ether and dried at 110° C.

Analyzed, it gave the following results :

Frotocaseose, formed by sulphuric acid.

I. 0"4405 gram substance gave 0.2*718 gram H^OrzG'SS per cent.
H and 0-8756 gram CO„=54"20 per cent. C.

II. 0*;3783 gram substance gave 0'2341 gram H„0 = 6'87 per cent.
H and 0.7604 gram C02=ro4-81 per cent. C.

III. 0*3786 gram substance gave 48-3 c. c. N at 17-5° C. and 753*4
mm pressure= 14*93 per cent. N.

IV. 0*3780 gram substance gave 47*8 c. c. N at 17*0° C. and 754*0
mm pressure=:14*84 per cent. N.

V. 0*3701 gram substance gave 0*0113 gram ash = 3*05 per cent.

VI. 0*5112 gram substance gave 0*0154 gram ash = 3*02 per cent.

94 M. II. Chittenden — Caseoses, Casein Di/speptone,

Percentage composition of ash-free substance.

Average.

C 55-90 56-50 56-20

H 7-07 7-09 .... .... 7-08

N --.- -..- 15-40 15-31 15-S6

Alter being dried at "110° C. the substance dissolved with diffi-
culty in water, leaving a large residue, but was readily and com-
pletely soluble in 0-2 per cent, hydrochloric acid and in 0*2 per cent,
sodium carbonate. From the alkaline solution it was reprecipitated
by hydrochloric acid, and not readily dissolved by an excess of the
acid. Aqueous solution of the substance was rendered turbid by heat,
the turbidity disappearing as the solution cooled. With dilute nitric
acid, a white precipitate was formed, readily soluble on warming and
I'eappearing as the solution cooled. Acetic acid and potassium ferro-
cyanide gave a heavy precipitate.

Thus in many respects this body resembles protocaseose formed
by pepsin-hydrochloric acid, but is apparently characterized by a
somewhat higher percentage of carbon.

The filtrate from the original sodium chloride precipitate, treated
with salt-saturated 30 per cent, acetic acid, gave a flocculent, chang-
ing to gummy precipitate which was readily and completely soluble
in water. The solution made exactly neutral was dialyzed without
showing any evidence of a hetero-like body, finally concentrated and
precipitated by alcohol. After being dried at 110° C, the substance
was only partially soluble in water. The aqueous solution was ren-
dered turbid by heat, clear again on cooling, and gave with nitric
acid a heavy white precipitate as also with acetic acid and potassium
ferrocyanide.

Analyzed, it gave the following results :

Acetic acid precqntate of easeose (a deuterocaseose ?) formed hy
dilute sidphnric acid.

I. 0-4056 gram substance gave 0-2383 gram H„0 = 6-52 per cent.
li and 0-7733 gram CO^=5l-99 per cent. C.

II. 0-4697 gram substance gave 0*2740 gram H,^0 = 6-48 per cent.
H and 0'8910 gram CO„ = 51-73 per cent. C.

III. 0-5379 gram substance gave 66-5 c. c. N at 14-9° C. and 752-1
mm pressure= 14-58 per cent. N.

IV. 0*4150 gram substance gave 0-0210 gram ash = 5'06 per cent.

V. 0*5136 gram substance gave 0*0247 gram ash=4*80 per cent.

mid Casein Peptone. 95

Percentage composition of ash-free substance.

Average.

C S4-68 54-42 54-55

H 6-86 6-81 .... 6-84.

N ---- 15-33 15-33

This caseose, instead of being a pure cleutero boclj^, appears to be
a mixture of proto and a deutero as indicated by its behavior towards
heat and nitric acid ; indeed, the behavior of this acetic acid precipi-
tate resembles the acetic acid precipitate of caseose obtained in our
previous work, where the two bodies were plainly precipitated to-
gether. In composition, while it agrees closely with protocaseose
formed by pepsin-hydrochloric acid, it has a higher content of carbon
than the corresponding a deutero, but bears about the same relation
to the sulphuric acid protocaseose as the a deutero formed by pepsin
to its corresponding protocaseose.

On boiling the original ammonium sulphate-saturated filtrate, a
second gummy precipitate gradually separated from the hot fluid.
This caseose, after purification by the usual methods and drying
at 110° C, was entirely soluble in hot and cold water, and
was especially characterized by yielding with acetic acid a heavy
white precipitate, soluble in excess of the acid. With nitric acid it
also gave a white precipitate, soluble in excess of acid. Cupric
sulphate likewise gave a heavy precipitate. The cold water solution
was not rendered turbid by heat.

Analyzed it gave the following results:

yS deuterocaseose, formed hy dilute sulplmrlc acid.

I. 0*3952 gram substance gave 0-2308 gram H.,0 = 6-49 per cent,
H and 0*7282 gram GO^=bO-2o per cent. C.

II. 0*3518 gram substance gave 0*2064 gram H.,0 = 6*51 per cent.
H and 0*6440 gram CO„ = 49-92 per cent. C.

III. 0*4727 gram substance gave 59*2 c. c. N at 15-0° C. and
754*9 mm pressure = 14*81 per cent. N.

IV. 0-4279 gram substance gave 0*0233 gram ash = 5*44 per cent.

V. 0*5184 gram substance gave 0*0219 gram ash = 5*38 per cent.

Percentage composition of ash-free substance.

Average.
C 53*10 52-77 .... 52-93

H 6-86 6-89 .... 6-87

N 15.66 15-66

96 i?. H. Chittenden — Caseoses, Casein Dyspeptone,

Thus, this body, which corresponds to the ji deuterocaseose
formed by pepsin-acid, has a relatively higher content of carbon and
also varies in certain of its reactions, which resemble rather those
of a deutero, and even the protocaseose of a pepsin digestion, than
those of a genuine ji deutero.

Altogether, the three caseoses separated from the sulphuric acid
solution of casein, while showing a certain general relationship to
the caseoses formed by pepsin-hydrochloric acid, are sufficiently
diflferent in their individual reactions to suggest at least some differ-
ence in their nature.

d. Caseoses formed hy the action of trypsin.

In subjecting casein to the action of trypsin, care was taken that
the ferment solution should be as free as possible from all products
of the self-digestion of the pancreatic tissue. Dried pancreas from
the ox, prepared according to Kiihne's method, was warmed with
0-1 per cent, salicylic acid at 40° C for 24 hours, after which the acid
extract was neutralized and made alkaline with sodium carbonate to
the extent 0-3 per cent. The alkaline solution, well thyniolized, was
warmed at 40° C. for several days in order to convert the albuminous
matters present into easily diffusible products, after which it was
dialyzed in running water for some time, and the solution ultimately
evaporated to dryness at 40° C. This residue, being extracted with
a small volume of water, gives a fairly pure solution of trypsin, free
from objectionable impurities.

In the formation of the caseoses, 2200 grams of pure, moist casein
were soaked in 3 litres of 1 per cent, sodium carbonate for several
days, and the mixture well thymolized to prevent putrefaction. The
trypsin solution was then added, together with some water, and the
whole warmed at 40° C. At first, the mixture was quite limpid, the
casein being dissolved in the alkaline fluid, but after two or three
days, gelatinous lumps began to appear on the bottom of the dish
and finally a soft coagulum appeared on the surface and all through
the mixture, resembling the separation of antialbumid. This coagu-
lum gradually disappeared and at the end of five days the digestion
was stopped, and the alkaline fluid filtered from the undigested resi-
due. The latter was thoroughly washed with thymolized water and
the washings added to the filtrate. This residue of undigested mat-
ter was found to be insoluble in 0-5 per cent, sodium carbonate and
also in 0*2 per cent, hydrochloric acid, but on being warmed with
pepsin-hydrochloric acid it was in time almost completely dissolved.

and Casein Peptone. 97

Foi; separation of the caseoses the digestive fluid was neutralized
with dilute hydrochloric acid, giving only a slight neutralization
precipitate, and the neutral fluid concentrated. When moderately
concentrated, the solution was placed in a cool place for several
days to allow as much of the leucin and tyrosin to crystallize as
possible. The caseoses were then separated from the filtrate, in
the form of a gummy precipitate, by saturation of the fluid with
ammonium sulphate. On boiling the filtrate from the first precipi-
tate, and adding still more ammonium sulphate, a second gummy
precipitate gradually settled out of the hot saturated fluid. This
precipitate was separated from the fluid, the latter being saved for
the detection of any peptone formed, washed with hot saturated
ammonium sulphate solution, then dissolved in water and dialyzed
until all traces of sulphate were removed. In the dialysis, no signs
of any gummy heterocaseose or other like body was noticed. The
caseose was precipitated from the suitably concentrated fluid with
alcohol, boiled repeatedly with alcohol to free it from any adhering
tyrosin, and finally dried at 110° C.

The substance was extremely soluble in water, the fluid remaining
perfectly clear when heated. Acetic acid produced a heavy precipi-
tate, soluble in excess, and in the acid fluid potassium ferrocyanide
gave a heavy precipitate. Dilute nitric acid, added to the aqueous
solution produced a white precipitate readily soluble in excess of acid,
and when heated showed the xanthoprotein reaction.

Cupric sulphate also gave a heavy white precipitate. On analysis,
the following results were obtained :

fi deiiterocaseose, formed by trypsin.

I. 0"3355 gram substance gave 0*1847 gram H30 = 6*ll per cent.
H and 0-5990 gram C02 = 48-68 per cent. C.

II. 0-3459 gram substance gave 0*1871 gram H„O = 6*01 per cent.
H and 0*6120 gram 00^ = 48*24 per cent. C.

III. 0*5399 gram substance gave 63*0 c.c. Nat 13*8° C. and
762*6 mm pressure = 13*63 per cent. N.

IV. 0*4166 gram substance gave by fusion with koh + kno,
0*0300 gram BaSO^=:0*98 per cent. S ; after deducting sulphur of
ash = 0*95 per cent.

V. 0*5496 gram substance gave by fusion with koh + kno,
0*0380 gram BaSO_=0'95 per cent. S ; after deducting sulphur of ash
= 0*91 per cent.

Teans. Conn. Acad., Vol. VIII. 13 Nov., 1888

c

53-81

53-31

H

6-76

6.64

N

....

S

o

98 M. H. Chittenden — Caseoses, Casein Dyspeptone,

VI. 0-4804 gram substance gave 0-0365 gram = 9-59 per cent.

VII. 0-4272 gram substance gave 0-0405 gram ash=9-43 per cent.

VIII. Ash from 0-8070 gram substance gave 0-0181 gram BaSO^
=0-03 per cent. S,

Percentage composition of ash-free substance.

Average.

53-36

6-70

15-07 15-07

0-95 0-91 0-93

23-74

100-00

This sample of /5 deutero shows very close agreement in composition
and reactions with the like body formed by dilute sulphuric acid ;
both being characterized by the reaction with acetic acid and show-
ing, by their reactions and composition, a closer relationship to the
proto-like bodies than their non-precipitation by ammonium sulphate
would appear to warrant. In composition, however, this body
shows a much smaller percentage of cai'bon than the caseose precipi-
tated by acetic acid.

For separation of the other caseoses formed in this digestion,
the first ammonium sulphate precipitate was dissolved in water
and protocaseose precipitated by saturation of the solution with
sodium chloride. The precipitate, which was not very heavy, was
purified by reprecipitation and dialysis. On dissolving the first salt
precipitate in water quite a little residue was noticed, soluble in
dilute sodium carbonate, but readily precipitated by the least trace
of acid. The final neutral solution was concentrated, giving when
heated a heavy coagulum which finally came together as a gummy
mass. The clear fluid, separated from the gum, gave still another
coagulum as the heating was continued. On cooling, the gummy
matter readily dissolved. As the evaporation continued and the
fluid became concentrated the gummy matter dissolved even in the
hot fluid, and the caseose was finally precipitated while hot with
alcohol, and the precipitate boiled repeatedly with alcohol for the
complete removal of leucin and tyrosin.

In reactions, this body was apparently identical with protocaseose
formed by pepsin-hydrochloric acid, except that with acetic acid and
also with hydrochloric acid it yielded a heavy white pi-ecipitate, sol-
uble in excess of acid. In water it dissolved almost completely,
the solution, however, becoming turbid when heated and if sufficiently

and Casein Peptone, 99

concentrated giving a gummy , deposit as the heating continued.
Further, the reaction with heat was the only respect in which this
body differed from the preceding ammonium sulphate pi-ecipitate.
Owing to insufficient quantity it was not analyzed.

In the filtrate from the first sodium chloride precipitate, a second
caseose was precipitated by addition of a little 30 per cent, acetic
acid. At first it separated as a flocculent precipitate, but on stand-
ing changed to a gummy mass, which dissolved more or less readily
in water, and completely so when a little sodium carbonate was added
to neutral reaction. After reprecipitation, the aqueous solution of the
substance was neutralized, dialyzed, the solution concentrated without
separation of any gummy matter, and the substance finally precipitated
with alcohol. The precipitate was boiled repeatedly with alcohol and
finally dried at 110° C. The dried substance was readily soluble in
water and also in dilute acetic acid, potassium ferrocyanide producing
in the latter solution a slight turbidity only. Added to an aqueous
solution of the substance, acetic acid produced a heavy white pre-
cipitate readily soluble in excess of the acid. Nitric acid, likewise,
produced a heavy white precipitate not so readily soluble in excess.
Cupric sulphate also gave a heavy white precipitate. The aqueous
solution of the caseose gave no turbidity whatever, when heated.

Analyzed it yielded the following results :

Acetic acid precipitate of caseose {a deuterocaseose f) formed by

trypsin.

I. 0-5594 gram substance gave 0-3300 gram H.p = 6-55 per cent.
H and 1-0924 grams CO^=53-25 per cent. C.

II. 0-3670 gram substance gave 0-2157 gram H„0 = 6-53 per cent.
II and 0-7158 gram CO^=53-19 per cent. C.

III. 0-3811 gram substance gave 44-4 c. c. N at 12-8° C. and
765-1 mm pressure =14-07 per cent. N.

IV. 0-2848 gram substance gave 33-0 c. c. ISl at 13-4° C. and
765-5 mm pressure =13-98 per cent. N.

V. 0-4351 gram substance gave 0-0230 gram ash =5-28 per cent.

VI. 0-4033 gram substance gave 0-0210 gram ash =5-26 per cent.

Percentage conqwsition of ash-free substance.

Average.
C 56-20 56-14 .-. ..-- 56-17

H 6-91 6-89 .... 6-90

N .... .... 14-85 14-75 U'SO

100 M. II. ChUtenden — Caseoses, Casein. Dyspeptone,

It is thus obvious from the foregoing that by the action of trypsin,
caseoses are formed of the same general nature as those formed by
the action of pepsin-hydrochloric acid, and by hot dilute sulphuric
acid, but with higher contents of carbon.

In other digestions with trypsin, made especially for the prepara-
tion of casein peptone, these caseoses were again separated and the
foregoing reactions verified. The quantities, however, were too
small to admit of their analysis.

III. Casein peptone ; — from experiments by Charles Norris, Jr., Ph.B.

Supersaturation of a digestive fluid with ammonium sulphate,
under proper conditions, suffices to entirely remove the preliminary
products of proteolytic action. Proper conditions, however, are not
obtained by simply adding the ammonivim salt to a cold fluid, for as
has been already pointed out an additional precipitate of proteose
can nearly always be obtained, by heating the cold saturated solution
until a thick crust of the ammonium salt forms on the hot fluid. In
the present state of knowledge, we assume as peptone any amorphous
product of proteolytic action precipitable by alcohol, and not pre-
cipitable by heating with ammonium sulphate added to saturation.
Unquestionably, the albumose precipitated only by long boiling of a
saturated ammonium sulphate solution, is much nearer to true pep-
tone than those bodies more easily precipitated, but at present we
are not inclined to accept as true peptone any body precipitable by
ammonium sulphate under any conditions whatever. In the present
series of experiments we have aimed to prepare a casein peptone,
by the action of trypsin, entirely free from albumose in order to
study its composition and reactions.

In the digestion of casein with trypsin, described in the preced-
ing section, the filtrate from the ammonium sulphate saturation was
heated for some time and the slight gummy film of caseose sep-
arated, after which as much of the ammonium sulphate as possible
was removed by alternate crystallization, treatment with alcohol, etc.
The last traces of the ammonium salt were removed by long con-
tinued dialysis in running water, and when finally the fluid gave no
reaction with barium chloride it was evaporated to a syrup and pre-
cipitated with alcohol. The precipitated peptone was freed from
any adhering tyrosin and leucin by repeated treatment with boiling
alcohol, and finally dried at 110° C. until of constant weight. This
proved a long operation. The peptone was so exceedingly Hygro-
scopic and held on so tenaciously to the water, that it was only after

and Casein Peptone. 101

long-continued drying that a constant weight was reached and then
the odor was strongly suggestive of partial dissociation. In a pre-
vious article* on peptone, attention was called to the peculiar odor of
valerianic acid almost invariably noticed when the fibrin peptone was
dried at 110° C, but in that case it was found impossible to bring
the product to a constant w^eight. With casein peptone, the same
odor was noticeable on drying the product for analysis, but after a
few days heating at 110° C, the weight of the pi-oduct remained
fairly constant.

On analysis, the casein peptone yielded the following results :

Casein antipeptone A.

I. 5792 gram substance gave 0*3185 gram H^O — 6'11 per cent.
H and 0-9951 gram CO,,=46'85 per cent. C.

II. 0'4692 gram substance gave 0-2562 gram H„O = 6-07 per cent.
H and 0-8060 gram C0.^ = 46-84 per cent C.

III. 0-4357 gram substance gave 55-5 c. c. N at 14-0° C. and 762*0
mm pressure = 15-27 per cent. N.

IV. 0-6534 gram substance gave 83*0 c. c. N at 14*0° C. and 761*0
mm pressure^ 15*18 per cent. N.

V. 0*9703 gram substance gave 0656 gram ash=6*76 per cent.

VI. 0*4939 gram substance gave 0-0326 gram ash = 6*60 per cent.

VII. 0*7032 gram substance gave by fusion with koh + knOj
0*0386 gram BaSO^=0-75 per cent. S.

VIII. 0-6526 gram substance gave by fusion with koh + knGj
0-0337 gram BaSO^=0-71 per cent. S.f

Percentage composition of ash-free substance.

Average.

C 49-94 49-93 .... .... j^9-94

H 6-51 6-50 .--. 6-51

N .... .... 16*35 16*26 .... ... 16-30

S .... 0*70 0-66 0-68

O - ^6-57

100-00

Somewhat to our surprise, on testing the purified and dried pep-
tone, we found that its aqueous solution gave a heavy wiiite precipi-
tate with nitric acid, soluble in excess of acid ; likewise, a heavy
precipitate with acetic acid, also soluble in excess ; with cupric sul-

* Kuhne and Chittenden, Peptone. Studies, vol. ii.

f The ash contained only a slight iinweighable trace of sulphate.

102 B. H. Chittenden — Caseoses, Casein Dyspeptone,

phate a heavy precipitate; and on saturation with ammoninm sul-
phate in the cold an abundant gummy precipitate, the latter plainly
indicating the presence of caseose. As the product originally
gave no precipitate with ammonium sulphate, it would apparently
follow that the peptone hy long drying at 110° C. had been, in
part, reconverted into caseose, and as the solution gave no turbid-
ity by heat it would imply that the caseose formed by the recon-
version of the peptone was the deutero body. Results similar to
these were obtained by Kiihne and Chittenden with fibrin peptone.

In a second digestion of trypsin, 1 kilo, of moist casein was
warmed at 40° C. with 4 litres of 0*6 per cent, sodium carbonate
solution containing trypsin, well thymolized, for one week. On
the third day, the gelatinous coagulum already described made its
appearance, but gradually disappeared and at the end of the week
there was only a very small residue remaining. The neutralized and
concentrated digestive fluid, freed from more or less tyrosin by cool-
ing and crystallization, was in this case treated with rock salt to sat-
uration, yielding, however, only a small precipitate of proto- and
heterocaseose, which on purification agreed in reactions with the
caseoses previously described. Addition of salt-saturated acetic
acid to the sodium chloride filtrate from the foregoing caseoses, failed
to give any precipitate whatever, and as the saturation of the fluid
with ammonium sulphate gave only a slight gummy precipitate it is
evident that in this digestion the casein was almost completely con-
verted into peptone. In order to be quite sure of the complete re-
moval of everything precipitable by the ammonium salt the mixture
was boiled for some time with an excess of ammonium sulphate, and
the filtrate treated as described under peptone A for the complete
removal of tyrosin and ammonium sulphate.

The final product was exceedingly gummy and parted with the
last traces of adhering alcohol very slowly. In fact, we found it best
to dissolve the final alcoholic precipitate of peptone in a little water,
and to drive ofi" the alcohol from the solution by heat, after which the
fluid was evaporated and the gummy peptone finally transformed
into a friable mass by drying on a water-bath, and at last completely
dried at 110° C. After its final precipitation with alcohol, an aque-
ous solution of the peptone gave no precipitate whatever, with nitric
or acetic acid, neither with acetic acid and potassium ferrocyanide
nor with cupric sulphate, or at the most nothing more than a faint
turbidity. After being dried at 110° C. until of constant weight,
the product then gave a decided gummy precipitate by saturation

and Casein Peptone. 103

of its aqueous solution with ammonium sulphate, and like peptone A
gave precipitates with nitric and acetic acid and with cupric sul-
phate.

On analysis the following results were obtained :

Casein antipeptone £.

I. 0-3770 gram substance gave 0-2023 gram lrLfi = b-QQ per cent.
H and 0-6535 gram CO„=47-25 per cent. C.

II. 0-2687 gram substance gave 0-1442 gram H„0=5-96 per cent.
H and 0-4667 gram 00^=47-36 per cent. C.

III. 0-3732 gram substance gave 44-9 c. c. N at 14-4° C. and 757-0
mm pressures 14-31 per cent. N.

IV. 0-3801 gram substance gave 46-0 c. c. N at 14-0° C. and 756-0
mm pressure =14 -3 8 per cent. N.

V. 0-4760 gram substance gave 0-0374 gram ash=:7-86 per cent.

VI. 0-5396 gram substance gave 0-0429 gram ash = 7-95 per cent.

Percentage composition of ash-free substance.

Average.

C 51-35 51-42 51-38

H 6-47 6-47 .... ..- 6-47

N 15-54 15-61 15-57

In a third digestion with trypsin, 2 kilos, of casein were warmed at
40° C. for five days with 3 litres of 0-5 per cent, sodium carbonate
containing an active trypsin solution, well thymolized. At the end of
the second day, considerable casein antialbumid separated from the
solution, this time more as a gummy precipitate than as a gelatinous
coagulum. On the sixth day, the alkaline fluid was filtered from
the small undigested residue, neutralized, concentrated, and the
caseoses precipitated by saturation with ammonium sulphate. In
this digestion, there was present only a very small trace of caseose
precipitable by saturation with salt, but considerable precipitable by
salt-saturated acetic acid. After repeated boiling of the ammonium
sulphate-saturated fluid, for complete removal of caseoses, the peptone
remaining was separated, purified and dried as already described.
The prodiict dried at 110° C. gave on analysis the following results.

Casein antipepto7ie C.

I. 0-3104 gram substance gave 0-1720 gram H„0 = 6-15 per cent.
H and 0*5252 gram CO., = 4614 per cent. C.

II. 0-2756 gram substance gave 0-1513 gram H^O = 6-10 per cent.
II and 0-4638 gram CO, = 45 -89 per cent. C.

c

49-64

49-40

H

6-63

6-57

N

104 R. IT. Chittenden — Caseoses, Casein Dyspeptone,

III. 0-5432 gram substance gave 68-0 c. c. N at 14-6° C. and
756-4 mm pressure= 14-88 per cent. N.

IV. 0-6280 gram substance gave 0*0449 gram ash = 7-15 per cent.

V. 0*4060 gram substance gave 0-0288 gram ash = 7 -09 per cent.

Percentage composition of ash-free substance.

Average.
49-52

6-60

15-99 15-99

This peptone, like the preceding, after being dried at 110° C, gave
a small gummy precipitate on saturation of its solution with ammonium
sulphate, and also gave a precipitate with nitric and acetic acid, and
a slight turbidity with cupric sulphate.

In composition, all three of the peptones show a smaller
percentage of carbon than the caseoses formed by trypsin, but
somewhat to our surprise the percentage of carbon is higher
than in some of the caseoses formed by pepsin-hydrochloric acid.
The nature of the substance, however, which at present affords
but little proof that we have to deal with a single body, the
extreme difficulty of obtaining it in a condition of dryness suit-
able for analysis, and the almost utter impossibility of freeing
it from adhering inorganic salts, all tend to throw doubt on the
analytical data as expressing the composition of pure casein pep-
tone. As already stated, there is unquestionably more or less of a
decomposition or change attending the drying of the peptone. So
pronounced is the hygroscopic character of these bodies, that when
partially dried they will gain weight over strong sulphuric acid, and on
being taken from the air bath Avhile drying at 110° C. we often no-
ticed on damp days a hissing noise as if from the rapid absorption of
water. Further, as the drying progressed the odor of valerianic acid
became quite pronounced and on testing the dried product, it was
often found to have an acid reaction so pronounced in many cases as
to give a sour taste to the peptone, in addition to the characteristic
bitter. Before drying, the peptones were usually found to have a
neutral reaction. These facts coupled with the changed behavior of
the product towards ammonium sulphate, point to a change in the
nature of the peptone, which may well be assumed to affect its com-
position, and hence we would have our analytical figures taken with
some allowance.

and Casein Peptone. 105

We also prepared a peptone, entirely non-precipitable by saturation
with ammoniuni sulphate, by boiling 20 grams of pure deuterocaseose
with 500 c, c. of 3 per cent, sulphuric acid for 14 hours. This prepara-
tion, after purification, was too small in quantity for analysis, but in
reactions it showed close agreement with the antipeptones prior to
their long drying at 110° C, viz: non-precipitation by acetic acid and
potassium ferrocyanide, by nitric acid, by cupric sulphate, and by
saturation with ammonium sulphate both in neutral and in acid
solutions.

Trans. Conn. Acad., Vol. VIII. 14 Nov., If

VI. — Some Expekiments on the Influence op Arsenic and
Antimony on Glycogenic Function and Fatty Degenera-
tion OF THE Liver. By R, H. Chittenden, and J. A.
Blake, B.A., Ph.B.

Saikowsky's* oft-quoted experiments on rabbits with antimonic
and arsenious acids have made clear that in both arsenical and anti-
monial poisoning there is pronounced fatty degeneration of the liver,
with a lessening of the hepatic glycogen and in some cases even a total
disappearance of it. With antimonic acid, Saikowsky found in his
original experiments, that one-half to one gram of antimonic acid or
other preparation of antimony per day, for fourteen or nineteen days
in succession, gave rise to a fatty degeneration embracing the liver,
kidneys, and even the heart. This has been verified by the experi-
ments of Grohe and Hosier, who also state that in the duchy of
Brunswick the peasantry give to the geese, when producing the
famous fatty livers, a certain quantity of the white oxide of antimony
every day.f With arsenic, Saikowsky likewise found that when
rabbits are poisoned by a small dose so as to live from three to six
days, the liver becomes much enlarged and very fatty and the glyco-
genic function nearly or quite abolished.

It is very evident, therefore, that in large quantities both arsenic
and antimony have a special action on tissue changes, particularly
on the liver. In the experiments referred to above, the quantities of
poison given were quite large and with arsenic, particularly, their ad-
ministration was soon followed by death. As neither of these sub-
stances are ordinarily used in medicine for an acute eifect, it seemed to
us of interest to study the action of small doses on the tissue changes of
the liver, with a view to ascertaining whether non-toxic doses of these
two poisons would produce a similar effect. It is ordinarily stated
that in poisoning with antimony, phosphorus, and arsenic the nitro-
genous products of tissue waste appear in the urine in much larger
quantity than normally, owing to the increased decomposition which
is going on. I Experiments of our own, however, have shown that

*Virchow's Archives, Band xxxiv, p. 78.

•)• Quoted from H. C.Wood's Therapeutics, p. 161.

Ij. Brunton's Pharmacology, Therapeutics and Materia Medica, p. :^60.

Chittenden and Slake — Influence of Arsenic and Antimony/. 107

small repeated doses of antimonious oxide are without influence on
the excretion of nitrogen, sulphur, and phosphorus, and that hence
when taken in non-toxic doses it has no noticeable action on proteid
metabolism.* Without doubt, toxic doses do materially aifect the
nutrition of the body, but with a dog of 13 kilos, weight the admin-
istration of repeated doses of antimonious oxide, to the extent of 17
grains in 13 days, led to no apparent change in the amount of nitrogen
etc. excreted, although the presence in the 24 hours' urine of 13-23
milligrams of antimony (Sb) gave evidence of decided absorption.

We have therefore tried a few experiments on rabbits and fowls
to see what effect small Repeated doses of arsenic and antimony would
have on the liver, as indicated by its content of fat, glycogen, and
sugar. The experiments were made in pairs, in which one animal of
each pair served as a control for comparison, while the other, kept
under the same conditions of diet, etc. so far as possible, was fed
each day with arsenic or antimony as the case might be. At the end
of the period both animals were killed and the livers analyzed.
Naturally, the animals of each pair were of the same age, from the
same brood, and so far as possible of the same body weight. During
the experiment they were kept on a weighed diet of cracked corn,
meal, etc., and were confined in suitable cages.

The methods of analysis were as follows : after determining the
body weight, the liver was quickly removed, weighed and sampled
by chopping, 10 grams or thereabouts were then weighed out accu-
rately, dried on a water bath, ground to a fine powder and extracted
in a fat extractor with warm ether until the fat was entirely removed.

For glycogen and sugar, 20-40 grams of the sampled liver were
thoroughly extracted with hot water, (continuous extraction with
water, frequently renewed, for several days) the extracts united, con-
centrated to a very small volume and precipitated with a large excess
of 95 per cent, alcohol. The precipitate of glycogen, etc., was washed
with alcohol, dissolved in a small volume of cold water to 100 c. c,
sufficient hydrochloric acid added to make the fluid contain 2 per
cent. HCl and heated on a water-bath for 15 hours. After neutrali-
zation, the volume was made up to 200 c. c. and the sugar determined in
25 c. c. of the fluid by Allihn's gravimetric method, and the glycogen
calculated therefrom. The liver sugar was determined by evaporating
the alcoholic fluid from the glycogen precipitate, dissolving the x'esi-
due in a little water, adding sufficient sulphuric acid to make the mix-

* Ghitteaden and Blake. Studies from Laboratory of Physiological Chemistry, Yale
University, vol. ii, p. 94.

108 Chittenden and Slake — Influence of Arsenic

ture contain 2 per cent. H^SO^, boiling for two hours to convert the
sugar wholly into dextrose, and then, after neutralization of the acid
fluid and diluting to 200 c. c, testing its reducing power by Allihn's
method.

Experiment I.

Action of arsenic on a fowl.

Period of dosing.

May 20-24, 0-1 grain AsoOa daily.

" 25-31, 0-2 " " "

June 1-15, 0-3 " " "

6*4 grains.

A, control fowl. B, arsenic foivl.

Body weight May 20, 1814 grams. 1814 grams.

" " June 16, 1871 " 1686 "

+ 57 -128

Weight of liver, June 16, 28-378 grams. 34-635 grams.

The liver of B, showed unmislakedble signs of fatty degeneration.

Determination of fat in liver.

A. control, 9210 grams liver gave 0-3347 gram fat, = 3-63 per cent.

B. arsenic, 13848 " " " 1-8561 " " = 13-40 "

+ 9-77 per cent.
Determination of glycogen and sugar.

A. B.

Weight of liver used, 20-167 grams. 20-787 grams.

Glycogen A, control.

Volume Equivalent Equivalent Total Per

used. Weight Cu. in dextrose. in glycogen. amount. cent.

25 c. c. 00744 gram. 0-0380 gram. 0-0342 gram. 02736 gram. 1-35

25 00727 0-0371 0-0334 0-2672 1-32

Glycogen B, arsenic.
25 c. c. 0-1720 gram. 0-0879 gram. 0-0791 gram. 0-6328 gram. 3-04

25 0-1713 0-0875 00787 0-6296 3-02

Sugar A, control.

25 c. c. 0-0299 gram. 0-0159 gram. 0-1272 gram. 063

25 0-0332 0-0176 0-1408 0-69

Sugar B, arsenic.

25 c. c. 0-0185 gram. 0-0102 gram. 0-0816gram. 039

25 0-0162 0-0091 0-0728 0-35

100 grams of breast muscle from £ gave 04 milligram of ^s.

In this experiment, then, we have as the apparent result of the long-
continued feeding of arsenic a loss of body weight, a decided increase

and Antiinony on Glycogenic function^ etc.

109

in the weight of the liver, a large increase in the liver fat, a gain in
the amount of liver glycogen, and a loss in liver sugar.

Experiment II.
Action of arsenic on a fowl.

Period of dosing.
May 20-24, 0-1 grain AsjOa daily.

" 25-31, 0-2 " "

June 1-15, 0-3 " " "

'' 16-18, 0-4 " " "

T'G grains.

Towards the end of the experiment the arsenic fowl appeared to
be suffering somewhat from the toxic action of the poison, consequently
the arsenic was discontinued after the 18th, but the fowl died on the
21st, and was immediately analyzed.

A, control fowl.

B, arsenic foiul.

Body weight May 20, 1644 grams.

1531 grams.

" June 21, 1757

1247

+ 113

-284

Weight of liver, June 21, 55830 grams.

33-729 grams,

Determination of fat in liver.

A, control, 13740 grams liver gave 0-7462 gram fat.

= 5-40 per cent.

B, arsenic, 10-179 " " " 0-4925 "

= 4-83

—0-57 per cent.

Determination of glycogen and sugar.

A.

B.

Weight of liver used, 42-090 grams.

23-550 grams.

Glycogen A, control.

Volume Equivalent Equivalent
used. Weight Cu. in dextrose. In glycogen.

Total
amount.

Per
cent.

25 c. c. 0-5213 gram. 0-2500 gram. 0-2250 gram.

1-8000 gram.

4-27

25 0-5228 0-2504 0-2253

1-8024

4-28

Sugar A, control.

25 c. c. 0-0359 gram. 0-0188 gram.

0-1504 gram.

0-35

25 0-0379 00198

0-1584

0-37

In J5, no trace of a reduction could be obtained for glycogen and
only an unweighable trace for sugar.

In this experiment, therefore, we find a decided loss in body weight
under the influence of the arsenic, a loss in the weight of the liver, a
slight diminution in the liver fat and neai'ly a complete disappear-
ance of both glycogen and sugar.

110

Chittenden and Blake — Influence of Arsenic

Experiment III,

Action of arsenic on a rabbit.

Period of dosing.
May 25-31, 02 grain AsoOs daily.

June 1-15, 0-3 " " "

" 16-23, 0-4 " " "

9'1 grains.

A, control rabbit. B, arsenic rabbit.

Body weight May 25, 1644 grams. 1502 grams.

" " June 24, 16l6 " 1586

— 28 +84

Weight of liver June 24, 61'360 grams. 46'760 grams.

Determination of fat in liver.

A, control, 15'320 grams liver gave 0*4126 gram fat, := 2"69 per cent.

B, arsenic, 14-435 " " " 0-2888 " " = 2-00 "

—0*69 per cent.
Determination of glycogen and sugar.

A. B.

Weight of liver used, 45'940 grams. 32-325 grams.

Glycogen A,* control.

Volume used Equivaleut Equivalent Total Per

lor reduction. Weight Cu. in dextrose. in glycogen. amount. cent.

25 c. c. 0-2036 gram. 0-1045 gram. 0-0940 gram. 1-8800 grams. 4-13

25 0-2027 01041 0-0937 1-8740 4-07

Glycogen B, arsenic.

25 c. c. 0-5230 gram. 0-2503 gram. 0*2252 gram. 1-8016 grams. 5-57

25 0-5223 0-2500 0'2250 1-8000 5-54

Sugar A, control.

25 0. c. 0-0177 gram. 0-0098 gram. 0-0784 gram. 0-lt

25 0-0173 0-0097 0-0776 0-16

Sugar B, arsenic.

25 c. c. 0-0254 gram. 0-0137 gram. 0-1096 gram. 0-34

25 0-0232 0-0126 0-1008 0-31

100 grams of muscle tissue yielded 0-3 milligram of J.5.

Here, we find under the influence of the arsenic, an apparent gain,
though slight, in body weight, an evident diminution in the weight
of the liver, together with a diminished percentage of liver fat. Ap-
parently, however, the amount of liver glycogen and sugar are some-
what increased. The liver showed no approach to fatty degeneration
on microscopic examination.

* The glycogen solution after boiling with acid, etc., was made up to 500 c. c. instead
of 200 c. c.

and Antimony on Glycogenic function., etc. Ill

Experiment IV.
Action of antimony on a fowl.

Period of dosing.
May 20-24, 0-2 grain SbjOa daily.

." 25-31. 0-4 "

June 1-7, 0-6 " " "

" 8-14, 0-8 " " "

13-6 grains.

A, control fowl. B, antimony fowl.

Body weight May 20, 1559 grams, 1616 grams,

" " June 15, 1531 " 1828 "

— 28 +212

"Weight of liver June 15, 22'995 grams. 31-799 grams.

Determination of fat in liver.

A, control, 7*985 grams liver gave 0-4600 gram fat, = 5-76 per cent.

B, antimony, 11-409 " " " 0-6741 " " = 5-91 "

+ 015 percent.
Determination of glycogen and Sugar.

A. B.

Weight of liver used, 15-010 grams. 20390 grams.

Glycogen A, control.

Volume used Equivalent Equivalent Total Per

for reduction. Weight Cu. in dextrose. in glycogen. amount. cent.

25 c. c. 0-0577 gram. 0-029G gram. 0-02C6 gram. 0-2128 gram. 1-41

25 0-0563 0-0289 0-0260 0-2080 1-38

Glycogen B, antimony.
25 c. c. 0-1011 gram. 0-0514 gram. 0-0462 gram. 0-3696 gram. 1-81

25 0-1015 00516 0-0464 0-3712 1-82

Sugar A, control.

25 c. c. 0-0198 gram, 0-0109 gram. 0-0872 gram. 0-58

25 00211 0-0115 0-0920 0-61

Sugar B, antimony.

25 c. c. 0-0300 gram. 0160 gram. 0-1280 gram. 0-62

26 0-0277 0-0148 0-1184 0-58

100 grams of breast muscle gave 1-2 milligrams Sb.

These results would seem to indicate that antiraonious oxide tends
to increase the body weight, and apparently also that of the liver.
The liver fat is obviously not much affected in this experiment, while
the glycogen shows a slight increase in the antimonial fowl.

112 Chittenden and Blake — Influence of Arsenic

Experiment V.
Action of antimony on a fowl.

Period of dosing.
May 20-24, 0-2 grain Sb^Os daily.

" 25-31, 0-4 "

June 1- 7, 0-6 " " " .

" 8-15, 0-8 " " "

" 16-19, 1-0 " " "

18*4 grains.

^, control fowl. B, antimony fowl.

Body weight May 20, 1701 grams. 1757 grams.

" " June 20, 1644 " 1814

-57 +57

Weight of liver June 20, 34'177 grams. 35'440 grams.

Determination of fat in liver.

A, control, 9*477 grams liver gave 0-3890 gram fat, ^=. 4'10 per cent.

B, antimony, 9-579 " " " 0-5088 " " = 5-31 "

+ 1-21 per cent.
The appearance of the liver in B indicated fatty degeneration.

Determination of glycogen and sugar.

A. B.

Weight of liver used, 24-700 grams. • 25-871 grams.

Glycogen A, control.

Volume used Equivalent Equivalent Total Per

for redaction. Weight Cu. in dextrose. in glycogen. amount. cent.

25 c. c. 0-2253 gram. 0-1160 gram. 0-1044 gram. 0-8352 gram. 3-38

25 0-2253 01160 01044 0-8352 3-38

Glycogen B, antimony.
25 c. c. 0-2480 gram. 0-1281 gram. 0-1153 gram. 0-9224 gram. 3-56

25 0-2479 0-1280 0-1150 0-9200 3*55

Sugar A^ control.

25 c. c. 0-0360 gram. 0'0189 gram 0-1512 gram. 0-61

25 00360 00189 0-1512 0-61

Sugar B, antimony.

25 c. c. 0-0383 gram. 0-0200 gram. 0-1600 gram. 0-62

25 0-0372 00195 0-1560 0-60

100 grams of breast muscle gave I'S miUigrams Sb.

In this experiment, there was pronounced fatty degeneration of the
liver in the antimonial fowl. Further, as in the last experiment, there
was apparently a slight increase both in body weight and in the
weisrht of the liver.

and Anthnony on Glycogenic function, etc.

113

Corresponding with the fatty degeneration, there was found 1 -2 per
cent, more fat in the antimonial liver than in the control.

The liver glycogen was also slightly increased under the influence
of the antimony.

Experiment VI.

Action of anti'mony on a rahhit.

Period of dosing.

May 25-31, 0-4 grain SboOa daily.

June

" 1

1- 7, 0-6 "
8-15, 0-8 " ''• ''
16-21, 1-0 "

19-4 grains.

A, control rabbit. B*

antimony rabbit.

Body weight May

25, 2012 grams.

1814 grams.

" June 23, 1998 "

1701 "

— 14

-113

Weight of liver June 23, 38-219 grams.

42-741 grams.

Determination of fat in liver.

A, control, 9-375 grams liver gave 0-3575 gram fat,

= 3-81 percent

B, antimony, 10-971

" 0-3422 " "

= 3-12

— 0-69 per cent

Determination of glycogen and sugar.

A.

B.

Weight of liver used,

28-844 grams.
Glycogen A, control.

31-770 grams.

Volume used

for reduction. Weight Cu.

Equivalent Equivalent
in (le.xtrose. in glycogen.

Total
amount.

Per
cent.

25 c. c. 0431 gram.
25 0-0441

0-0224 gram. 0-0201 gram.
00229 0-0206

Sugar A, control.

0-1608 gram.
0-1648

0-55
0-57

25 c. 0. 0-0445 gram.

0-0231 gram

0-1848 gram.

0-64

25 0-0440

0-0229

0-1832

0-63

In B, there was no reduction for eitlier glycogen or sugar.
100 grams of muscle gave TO milligram Sb.

In this experiment, towards the end, the animal was evidently suf-
fering from the toxic action of the antimony, and the body weight is
seen to be noticeably diminished. The weight of the liver, however,
was apparently increased by the antimony, although there was a

*The last two days the antimony rabbit ate very little. Hence, the o-tide was not
given after the 21st. The animal was evidently much affected by the antimony and
its general nutrition was at the last very poor.

Trans. Con.v. Acau., Vol. VIII. 15 Dec, 1888.

114 Chittenden and Slake — Influence of Arsenic and Antimony.

diminution in the liver fat, and a complete disappearance of both
glycogen and sugar.

While these experiments are far too few in number to generalize
from, yet it would appear that small doses of antimonious oxide long
continued tend to increase body weight, and particularly the weight
of the liver. Further, the increase in liver weight is accompanied
by an increased percentage of fat and a slight increase of liver
glycogen.

With corresponding doses of arsenious oxide, on the other hand,
the results would indicate a diminution in body weight, likewise a
diminution in the weight of the liver and also in the amount of liver
fat. As regards glycogen, two of the results show an increased
amount in the arsenical livers, while in one experiment there was a
total disappearance of glycogen. On the other hand, as Experiment
No. I indicates, there may be, with small doses of arsenic, a very
pronounced fatty degeneration of the liver, accompanied by an in-
creased liver weight and an increase in the percentage of liver
glycogen.

VII. — The Nature and Chemical Composition of the Myosin
OF Muscle Tissue. By R. H. Chittenden and G. Wyckoff
Cummins, Ph.D.

In spite of the interest attached to this peculiar proteid substance,
little attention has been directed to its chemical nature since the
time of its discovery by Kiihne.* In fact, Danilewskyf is the only
investigator who appears to have studied its chemical relations to
any extent, and so far as we are aware no attempt has ever been
made to ascertain its chemical composition. With this fact in mind,
it has been the main object of the present investigation to study the
chemical composition of pure myosin, and to determine the diffei'-
ences that may exist in the chemical nature, or in the properties, of
myosin prepared from various animal sources. Unfortunately, our
work was completed before Halliburton's]; recent paper on muscle-
plasma was published, otherwise we should have attempted to verify
some of his interesting discoveries i-egarding the muscle clot, and
possibly have modified somewhat the character of our work.

The researches of Kiihne, IIoppe-Seyler,§ Weyl,|| and Danilewsky
have shown that myosin, both from the animal and vegetable king-
doms, is a globulin body, soluble in dilute sodium and ammonium
chloride solutions, as well as in sulphate of magnesia and sulphates
of the alkalies, and precipitable therefrom either by the addition of
salt in substance or by dilution with much water. As stated by
Danilewsky, myosin is most advantageously extracted from muscle
tissue by ammonium chloride, in solutions of from 7-20 per cent.
We have, likewise, found this to be the case by comparative tests,
and have, therefore, in all of our preparations of myosin for analysis,
used ammonium chloride as the extractive, preferably of 15 percent,
strength. For separation of myosin from the ammonium chloride

* Kiihne, Protoplasma, Leipzig, 18G4. Also Lehrbuch der physiologische Chemie.

\ Myosin, seine Darstellung, Eigenschaften, Umwancllimg in Sjmtonin und Riickbil-
dung aus demselben. Zeitschrift fiir physiologische chemie. Band v, p. 158.

:]: Journal of Physiology, vol. viii, p. 133.

§ Handbuch der Chem. Analyse, 4 Auflage, p. 236.

H Beitrage zur kenntniss thierischei* und pflanzlicher eiweisskorper, Zeitschrift fiir
physiologische Chemie, Band i, p. 72.

116 Chittenden and Cummins — Nature and Chemical

solution we have mainly employed two methods, either dilution with
a large volume of distilled water, or dialysis of the ammonium
chloride solution until the salt is entirely removed. In this latter
process, we have often observed that the sepai'ation of myosin
partakes more of the nature of a coagulation than of an ordinary
precipitation. A moderately strong ammonium chloride solution
of myosin, on being placed in a parchment bag and suspended in
running watei", will ordinarily at the end of two or three days be
converted into a semi-solid, jelly-like mass, which later on contracts
more or less, but still shows all the characters of a genuine clot.
In fact, we are inclined, with Halliburton, to consider this a genuine
re-coagulation rather than a j^recipitation. We are inclined, how-
ever, to believe that separation of myosin by dialysis is hardly as
satisfactory in the preparation of small quantities, as precipitation by
water, since on dialysis the jellying of the myosin naturally tends to
enclose some of the salt and also any other proteids possibly present
in the solution, while by precipitation with water the myosin is floc-
culent, easily washed and thus more surely freed from both salt and
albumin.

Myosin A, from ox muscle.

The first sample of myosin was prepared from a freshly killed ox.
The finely chopped muscle from the thigh was freed, so far as possi-
ble, from all traces of blood and soluble albumin by long soaking
and frequent kneading with water. The water was frequently
changed and kept thoroughly thymolized so as to prevent any ap-
proach to putrefaction. When the washings failed to give any tur-
bidity by heat, or by acetic acid and potassium ferrocyanide, the
washed tissue was placed in 10 litres of a 15 per cent, solution of am-
monium chloride for extraction of the myosin. The resultant opales-
cent fluid was filtered through paper and the myosin precipitated
by saturation with sodium chloride. The precipitate was quickly
strained off, dried somewhat between folds of filter paper, dissolved
in a small amount of water and reprecipitated by treatment of the
fluid witli a large volume of water. The precipitate so obtained
was washed with water until the washings gave no reaction for
chlorides, when it was treated with weak alcohol and finally with
95 per cent, and absolute alcohol, and ether.

When partially dried, it was ground tine and further dried at 110°
C. until of constant weight, for analysis.

Its composition is shown in the accompanying table.

Cotnposition of the Myosin of Muscle Tissue.

117

^

CO ^ bd £

_r (3 4- 03

CO «t3 'SR m

g 02 K

^

o»

00 , , ,

(C

"e^

1 1 1

■*

■^ , , 1

<1

1 1 1

'

1-1

T-l ' ' '

y—i

tH

^

T3

S

CS

bO

1 1 1

•

O

C<J , 1 ,

't»

□

1 1 1

1

■r-l

<

<2

,

'

o
o

0-0

tH

OS

1 , CD

?o

1

1 1 1 1

1^

"^

! i o

•

1 I 1 1

<D

3 _•

1 I ■*

! 1 <x>

^- ! 1 ! I I
^11111

(» g

' lO

lO ' ' ' '

t^

! ; c-

^ 1 1 i 1 I

PL|

1 1 t-

o , , , , .

H^

I I '^

<=> ; I ' : !

°

1—1'''''

d

1 1 i>

■7^ 1 1 1 > '

1 * ■,_<

"

' 00

00

S be

'^ o bb

> >

I— I It-I K<(

?^ 1^ >H >~t

-si -

s

o

CO

o

a,

O K 1^ 03 O

118 Chittenden and Oronmins — N'ature and Chemical

Myosin B^froin ox muscle.

A solution of myosin in 15 per cent, ammonium chloride was pre-
pared from 4 kilos, of fi-eshly killed beef, as described under A.
The myosin was then separated from the solution by dialysis, con-
tinued until nearly all of the chloride was removed. The last traces
of the ammonium salt were separated by filtration of the gelatinous
myosin through chamois skin, and washing with water. The prepa-
ration was then treated exactly as A, and dried at 110° C. for analysis.

By long continued contact with water, as in dialysis, the precipi-
tated or coagulated myosin is rendered insoluble in dilute salt solu-
tions, as noticed by Weyl and others, hence in this preparation it was
not possible to purify the substance by reprecipitation.

The composition of the product (see the accompanying table) is
essentially the same as that of the preceding preparation.

Myosin C, from sheep's muscle.

This sample of myosin was obtained from fresh mutton in essen-
tially the same manner as tlie preceding preparation, viz : by extrac-
tion of the thoroughly washed tissue with 15 per cent, ammonium
chloride, and separation of the myosin by dialysis.

On analysis, it was found to possess a somewhat higher percentage
of carbon than the two myosins from ox muscle, but in other respects
it was identical with them.

Myosin D, from calfs muscle.

A solution of myosin was prepared from 3 kilos, of fresh, lean mus-
cle from a young calf, by extraction of the thoroughly washed tissue
with 15 per cent, ammonium chloride solution.

From one-half of the filtered fluid, myosin was precipitated by
dilution with water and purified by washing with water until chlo-
rides were entirely removed. It was then washed with weak alcohol,
finally with absolute alcohol and ether, and then dried at 110° C. for
analysis (D').

From the other half of the ammonium chloride solution, myosin
was separated by addition of ammonium chloride in substance. The
precipitate was freed from excess of ammonium chloride by addi.
tion of just enough water to dissolve the salt, after which the floc-
culent myosin was strained off, dissolved in a little water, and precip-
itated by pouring the solution into a large volume of water. It was

Composition of the Myosin of Muscle Tissue. 119

S after
deducting
S of Ash.

1-26
1-24

aj-^

1-31
1-29

SZ '^ ^

o g + 2

0-0597
0-0594

Sof
Ash.

sub.
0-05

BaSOi

from the

Ash.

gram.

0-0046

j2 1 , , ' , 05 OS , , ,
OJ-^ , , , , cp O , , ,
< ' i ' ' o o ' ' '

Ash
found,
gram.

00072
0-0046

iz;^

16-34

16-48

'6

a

a
t5

CL,

764-6
762-6

hP

1 1 00 J> 1 1 ■ 1 1

I ! OS OS ! 1 1 ! !

ci

67-8
81-3

C)"6S.

52-16
52-12

CO2

found,
gram.

0-8237
0-7515

^ CO

fr|-^ <? 9

'^ ^

q^p Si;;;;':;

"^ &c 06;;:;;;;

Substance
used,
gram.

0-4306
0-3932
0.5061
0-6005
1-0425
0-6690
1-7115
0-6277
0-6358

6

!2h

1— II— lh-|i-~,i-i.l— II— II— 1^^

1— (1— ii^r*K>i— II— ii-<(

^ - > ^ ^ ^

«3^

CO

"^ t^

CO

■M

?^

iS ^ T-l

O ffi » CO O

120

Chittenden and Cummins — Nature and Chemical

S after
deducting
S of asli.

1-20
1-25

1

1-24
1-29

0^ + 2

0-0699
0-0667

CD

"o-go-l ; ; ; ; ; ; g ; ;

BaSOi

from the

ash.

gram.

0-0037

Si , , , , o t- , , ,

00 -^ , , , , <p »p , , ,

<j 1 , 1 1 <^ J^ 1 . ,

Ash
found,
gram.

0-0031
0-0051

Jz;-^

16-29
16-41

p

0)

i e
£ s

761-1
762-1

10-2
10-2

c3

61-8
101-3

o-^

52-95

52-88

o g £

0-9603
0-6863

W^

7-08
7-08

....

0-3150
0-2255

Substance
used,
gram.

0-4945
0-3539
0-4600

0-7477
0-5170
0-8969
1-4139
0-7761
0-7059

i^ to >~l >-H

o tri ^

cc O

Composition of the Myosin of Mtiscle Tissue. 121

then washed with water, alcohol, and ether, and dried at 110° C.
(D").

The composition of both products is shown in the accompanying
tables.

From the analytical data it is seen that the two preparations show
close agreement, although there are minor differences ; D", for exam,
pie, containing a slightly higher percentage of cai-bon and a corres-
pondingly lower percentage of nitrogen than D\ Further, D" con-
tains less than half as much ash as the other pi-eparation. Both show
fairly close agreement with the my(tsins from ox muscle, with per-
haps a slightly higher average percentage of nitrogen.

Myosin E, from fish.

So far as we are aware, little attention has been paid to the my-
osin from fish flesh. Myosin is assumed to be present and is supposed
to be of the same general nature as the myosin from other forms of
muscle tissue.

We first ti'ied the separation of myosin from fresh cod (Gadiis
callarias or G. morrhna), using 2 kilos, of the fresh, lean tissue ob-
tained in market. The final ammonium chloride extract (15 per
cent.) failed to give anything more than an insignificant precipitate,
either by dilution with water or by saturation of the fluid with
sodium chloride.

A second preparation was attempted from the flesh of the halibut
[Hippoglossus vulgaris) ; 2*5 kilos, of fresh tissue, free from fasciae, fat
and integument were thoroughly extracted with thymolized water
for several days, the chopped tissue being well rubbed up with the
water to insure complete removal of soluble albumins, etc. The
thoroughly washed fibre was then triturated with a 15 per cent,
ammonium chloride solution and allowed to stand in contact with it
for 24 hours. The filtered fluid gave a decided precipitate on boiling,
and also on addition of salt to saturation.

Myosin was separated from the ammonium chloride solution by
dialysis as a more or less gelatinous precipitate, and was prepared
for analysis by washing with thymolized water, and treatment with
alcohol and ether.

Dried at 110° C. until of constant weight, it yielded the following
results :

I. 0-3335 gram substance gave 0*2043 gram H„0 = 6 08 per cent.
H and 0-6267 gram C0,= 51-16 per cent. C.

Trans. Conn. a.cad., Vol. VIII. lu Dec, 1888.

122

Chittenden and Cummins — Natxire and Chemical

S after
deducting
S of ash.

1-29
1-29

M-eR

1-39
1-39

S50

rT O + 2

0-0679
0-0575

Sof
Ash.
% of
sub.

•

0-10

BaSOi

from the

ash.

gram.

0-0130

<1

2-95
3-91

Ash
found,
gram.

0-0291
0-0214

1 , «> «) 1 1 1 . 1
^^ 1 I i «b 1 ! i ! I

tH T-H

Pi
O

S-l

!2i

Pressure
mm.

749-8
768-1

c. c.

87-4
82-6

00 CO .... 11 .
tH CO 1 1 1 1 •' •

»o o '■'''' ■

CO2

found,
gram.

0-6646
0-7395

W^

6-91
6-93

found,
gram.

0-2301
0-3439

Substance
used,
gram.

0-3541
0-3925
0-6283
0-6065
0-9821
0-7361
1-7182
0-6707
0-5690

d

>

e^ t^ *N '~H

I Tj<

O K ^ cc O

Composition of the Myosin of Muscle Tissue.

123

5>0 .

t« a j3

1

St:3 M

J

,

,

,

1

,

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CO

[ ;

;

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T3 03

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ta
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M-SM t*)

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found
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found
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124

(Jhittenden and Cummins — Nature and Chemical

S after
deducting
S of ash.

1-25
1-35

1 ■ 1 1 1 1 1 o o t-
OQ-^ ,iiiiiiCp-rt<<?4

' ' ' ' ' ' ' -1-1 T-l 1-1

og + 2

OS Ol JO

C- OT -*

' ' ' ' 1 ' iO O IC

! 1 I I 1 I 1 "^ ■? 9^

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OQ < ^ m

lO , 1 1

1 , 1 I . O . 1 1

' ' ' ' ' o ' ' '

* 2 . o

'>< -^ -c2 P , 1 , 1 1 • o ' 1 '

eg a S 2 o ■

A 11 , CO 35 , 11 1
M -^ , , 1 1 OJ (M , , , ,
<1 iiii^t1|"'i'

Ash
found,
gram.

00 o
' o o '

l2;\R.

lO t-

1 -* ip ' ; ; ; ;
! ! «b <£> 1 ! 1 ! ! i

t3

a
o

2
i a

oa "

£ a

Oh

! 1 '^p t- ; ; 1 ! ; 1

> ' lo LO ; ; ' ;
I ; t- c- ; ; ; ; ; I

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, 1 CO ^ 1 1 1 . . .

' ' xH M I ! ! 1 ! ;

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o> «o . , , , , 1 , ,
e* ep

CO,
found,
gram.

0-8109
0-7019

w ■««.

05 CO Ill

OS o 1 . 1 ■ 1 1 1 '
i) t- ''''■'' '

H,0
found,
gram.

0-3661
0-2315

Substance
used,
gram.

0-4339
0-3655
0-6159
0-6100
0-7144
0-7000
1-4144
0-6112
0-5189
0-5882

6

'^ B ^ "^ > tt

5B 3i '--'

I* €» t^ li)

V.

S? Oi ?o

*> "S'i "iP

!* CO t-

o W !z; oj o

Composition of the Myosin of Muscle Tissue.

125

bo •

^^ 3 'S

® '^ en

1

1

'

'

o

^

•

•

CO

o*

^-o °

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T-H

T-H

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<M

CC1^

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:

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CO

T-H

,

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o

^ fl + 03

1

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'

'

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^^ . •

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126 Chittenden and Gtmxmins — Nature and Chemical

II. 0-4895 gram substance gave 67-8'='= N at 15-4* C. and 761-6'"'"
pressure = 16-50 per cent. N.

III. 0-4939 gram substance gave 0-0116 gram ash = 2-35 per cent.

Percentage composition of ash-free substance,
C 52-39, H 6-97, N 16-74.

The amount of myosin obtained from the fish muscle was not at
all commensurate with the amount of flesh experimented with.
Danilewsky,* however, found in the muscle tissue of the perch 8-56
per cent, of myosin as contrasted with 7-5 per cent, in ox muscle.
Whether our inability to separate a corresponding amount from the
tissue of the cod and halibut was due to the character of the tissue,
to its possible staleness, or to inferior methods of separation we
cannot say. Certainly, the yield of myosin in oar case was not as
great as from the muscle tissue of the ox, calf, lamb or sheep.

In composition, and in such reactions as we were able to try, the
fish myosin was not unlike the myosin from other preparations.

Myosin F^ from ox muscle.

A solution in 15 per cent, ammonium chloride was prepared as in
preceding cases and the myosin precipitated by addition of a large
amount of water.

One half of the precipitated myosin was washed with water until
all ammonium chloride was removed, then with alcohol and ether,
and dried for analysis (F').

The otlier half of the myosin was washed somewhat with water,
not enough to render it insoluble, then dissolved in 10 per cent,
sodium chloride solution, reprecipitated by dilution with water, thor-
oughly washed and dried (F").

The composition of the two products is shown in the accompanying
tables. The i-esults show close agreement with each other, and with
the preceding preparations.

Myosin G^ from ox muscle.

According to Danilejvsky, a 15 per cent, solution of ammonium
chloride is stronger than necessary for the extraction of ^myosin from

* Ueber die Abhanghigkeit der coDtractionsart der muskeln von der mengen ver-
haltnissen einiger ihrer Bestandtheile. Zeitschrift fiir physiologische Cliemie. Band
vii, p. 124.

Composition of the Myosiyi of Muscle Tissue. 127

muscle tissue. In fact, microscopic examination showed that 15 per
cent, solutions of this salt generally cause destruction of the muscle
fibres, while weaker solutions of the ammonium salt, as 5 per cent.,
extract the myosin equally well, without injuring the structural
elements in the least.

Thinking that possibly a purer myosin might be obtained by the
use of a weaker salt solution, several kilos, of thoroughly washed
muscle tissue were rubbed up with an excess of 5 per cent, ammo-
nium chloride and kept in contact with it for 48 hours. From the
filtered solution, myosin was separated by addition of a large volume
of water and the product washed and dried for analysis (G').

The tissue remaining after extraction as described, was treated
with a fresh 5 per cent, solution of ammonium chloride, and from
this fluid a second precipitate of myosin was obtained by addition of
water. This also was washed and dried for analysis (Gr").

The residue of tissue still i-emaining was then extracted with a 15
per cent, solution of ammonium chloride. The extract so obtained
gave a flocculent precipitate on saturation with sodium chloride, and
a coagulum appeared on boiling the solution, but no precipitate could
be obtained on addition of a large volume of water. It is thus evi-
dent that there was an almost complete extraction of myosin by the 5
per cent, solution of the ammonium salt.

On precipitating the small amount of myosin present in the 15 per
cent, ammonium chloride solution by saturation of the fluid with
salt, a clear filtrate was obtained which gave a turbidity on boiling
and also on addition of acetic acid and potassium ferrocyanide, thus
showing the presence of a small amount of soluble albumin.

The composition of the two myosins, as seen from the accompany-
ing tables, does not differ at all from that of other preparations.

Myosin H., from muscle of sheep.

In this preparation, the thoroughly washed and chopped muscle
was pai'tially extracted with a 5 per cent, ammonium chloride solu-
tion, and the myosin separated by addition of water. It was then
washed and dried for analysis (H^).

The residue of tissue was extracted with a 15 per cent, solution of
ammonium chloride and the myosin separated from this solution,
likewise, by addition of water (H").

128

Chittenden a^id Cummins — Nature and Chemical

S after
deducting
S of ash.

1-18
1-14

m\K

1-26

1-22

BaS04 after

fusion with

KOH + KNO,

gram.

0-0484
0-0506

S of
Ash.
% of
sub.

0-08

o^^" a

Mi a CO ^

05 5 C3 £

0-0071

■

1-47
1-51

Ash
found,
gram.

0-0071
0-0109

J^J-fis.

16-24
16-32

a

Pressure,
mm.

761-8
765-6

12-0
13-6

d

85-5
77-6

0-fe%,

52-31
52-21

o § §

0-7437
0-8979

M^

7-11

7-05

H2O
found,
gram.

0-2482
0-2974

Substance
used,
gram.

0-3877
0-4690
0-6343
0.5718
0-4814
0-7234
1-2048
0-5281
0-5719

6

>5

1— 11— (1— l»>t>l— II— II— (M

»Ci

o^

^

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f^i

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,

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s

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T-H

1

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■^

eo

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S

V

u K !z; 02 o

Composition of the Myosin of Muscle Tissue.

129

S after
deducting

S of Ash.

1-14
1-13

ca m

1 , , , ■ . oj oi

' ' ' ' ' ' ' 1-1 T-l

d § + 2

§8 S

6 6

BaSOi
from the

Ash.
gram.

0-0055

-^ , , , , T-l CO , , ,
^•^ ,,,, -rH OS ,, .

Ash
found,
gram.

1-1 lO
. . « O '

1 1 ! ! o o i ■ I
X o o

1 CO OS

^ . . ^ -*

^^^ ; ; cb «b ; , ; : :

g

£ , , 1

^ ^ , , GO >*

S 9 ' ' '^'^ ^ ' ! ! 1 !

|g , ; ^ g ; ; ; ; :

Cu

, , CO 1— 1 , , , , 1

CO »o

00 00

CO t- • I 1

lO Id '

CO,
found,
gram.

r- c« '

O CQ '

OS t- ' 1 ■ ' ■ ■ •

o o 1 I I I 1 ! '

td -^

^ o , , , 1 , , ,

O OS 1 , , 1 1 1 1

j^ 50 '''''' '

oi^ a

OCJ CO ' ' 1 ' ' ' '

O CO

CO CQ 1 1 1 1 ■ 1 I

o o ! ! ' ' 1 1 1

Substance
used,
gram

0-4788
0-3802
0-8286
0-4869
0-5740
0-5348
1-0088
0-5490
0-5918

d

h— < 1— I ^"^ f^ ^-i. HH HH '^■^

- ^ ^ > ^ -

>-H !i) Co 1--I
1-^ Cci -^1 '~H

't^.

=

o

o

CO

lO

<»

00

o

Ol

o>

r-

h?

»o

s

li)

o

!~

t-

CO

n^

t-

O M ^ cc O

Trans. Conn. Acad., Vol. VIU.

17

Dec, 1888.

130

Chittenden and Cummins — Nature and Chemical

^

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•^

1-1

-^

11^^^

;

;

:

1

1

'•

1—1

T-l

CO

CO

1—1

m'%°

'o m

00

ts

00

(ji>^

;

;

1

:

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1-1
1^

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op

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■«1<

CO

CO

"S &M g

;

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j

in

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lO

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1

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f

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§•1^5.

'

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pqv^g

;

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;

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t-

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,

,

hC "^

o

\

:

''

■

1

'

'

T-l

-<*

,

,

,

,

,

,

,

o'B s

:^

00

l

',

1

I

1

_'

;

CQ

c«

t

1

1

1

t

•

t

Wo ^

o

o

•

'

'

'

'

o

o

-*

t-

o

ic

o

IC

CO

OS

CO

S-^' a

o

so

CD

o

OS

OS

CJ

00

CO

35

o

'^

o

-5t<

m

OJ

« » g

!>3

-*

O-t

iC

®

CJ

IC

m

to

^ 3 fcc

O

o

o

o

o

1-1

o

o

o

CO

d

!2i

HH

t— 1

1— 1
1— t

>

HH

>

>

1— 1
1— 1

>

HH
HH
»— 1

>

X

^

1^

OS

OS

§

»^

»-<

^

"-H

^i^

S 00 r^

o W t<; a: C

Composition oj the Myosin of Muscle Tissiie.

131

deducting
Sof Ash.

1-17
1-11

, OJ o

' 1 ' ' ' ' ' I-l T-l

0^ + 2
g.9g^

S8 §

■* 1*

1 . 1 1 1 p p

'■'''' o 6

BaSOi

from the

Ash.

gram.

1 t • 1 1 1 •'^ 1 1

1 1 1 1 O ' 1
1 , 1 1 , 1 O 1 ,

' ' ' o ' '

<

1 CO CO , 1 1

1 -jtH TlJ , ,
' ' ' ' 1^ rH ' ■ '

Ash.
found,
gram.

' ' ' o 6 ' ' '

l-H «D 1
, 1 «0 JO ' ' 1 1 >

^^ ; ; i cb : ; ; ; ;

1 l-H l-H

n

.2

S a I 1 t- lo ■ 1 • . 1

aj a . 1 o CD " ' 1 1 ■
^ II * t 1 1 1

^"

"* Oi . 1 1 1 1

: ' -^ -^ ; ; ' ; '
' ' 1-1 1— 1 ' 1 1 1 1

1 op O 1 , , , ,

: : ■^ <© 1 1 ! I '

00 CO ' ' ' ' '

o-^

e« OS

T-t 1^ i ! 1 1 i I 1

COj
found,
gram.

0-7716
0-4526

M>^

i> -rH 1 1 1 1 1 1 1
P P 1 • . I 1 1 ,
<0 t- '■'''' '

OS J> 1 . . 1 . . 1

»o OS 1 1 1 ' I I 1

lO TfH

o o 1 1 I I 1 I I

Substance
used,
gram.

0-4075
0-2374
0-5898
0-2675
0-6164
0-5958
1-3133
0-5413
0-4759

d

'^

13

i^

^

a<

<» l— tH
S^l CJ £^

O S !2; CO o

1.S2

Chittenden and Cummins — Nature and Chemical

deducting
S of Ash.

1-23
1-25

co-^

1-26
1-28

0-0458
0-0552

»o

o-g o^ ; ; ; ; ; ; ; ; 9 ; ;

CO ^ ts, 55 . •' 1 1 1 , ' . 1 o \

BaSOi

from the

Ash.

gram

0-0020

Ash.

0-65
0-68

Ash
found,
gram.

0-0043
0-0046

^zj-^

16-73
16-72
16-77

a
.2

6

CO Q

CO a

— —

759-8

757-8
760-7

15-4
14-7
15-0

d

109-1

70-6

105-9

0-6R,

52-48
52-48
52-38

c-vs a

OSS

0-7053
0-9404
0-5596

K^

7-01
7-13

HoO
found,
gram.

0-2318
0-1868

Substance
used
gram.

0-3665
0-4900
0-2913
0-7746
0-5013
0-7524
0-6629
0-6741
1-0959
0-4967
0-5992

No.

HHI— IHHK.^1— II— II— ItxjMl— 1

i-it-iEL'^t>'-ii-iLJ^t<i

,_| 1— t I-' [^ )_H 1— 1 '■^

>

V

o td £5 cC o

Composition of the Myosin of Muscle Tissue.

133

Resume of the Analytical Results.

My

osin A, from ox._.
B, " '•_.

C,

D",

D",

E,

F',

F",

G',

G",

H',

H",

I,

Average -

Fibrinogen* - _ .
Fibrin* ......

Paraglobulin*
Egg albuminf

sheep
calf . -

halibut

sheep

c

H

N

S

52-84

7-12

16-89

1-49

52-51

7-09

16-53

1-25

53-24

7-12

16-45

1-23

52-84

7-11

17-14

1-33

52-97

7-13

16-96

1-25

52-39

6-97

16-74

52-99

7-11

16-73

1-29

52-83

7-10

16-74

1-31

53-05

7-19

16-52

1-18

52-82

7-11

16-80

1-16

52-84

7-10

16-91

1-28

52-57,

7-10

16-84

116

52-79

7-12

16-86

1-26

52-82

7-11

16-77

1-27

52-93

6-90

16-66

1-25

52-68

6-88

16-91

1-10

52-71

7-01

15-85

1-11

52-18

6-93

15-81

1-87

21-66
22-62
21-96
21-58
21-69

21-88
22-02
22-06
22-11
21-87
22-33
21-97

21-90

22-26
22-48
23-32
23-21

* Hammarsteu, Jahresbericht der Thierchemie, vol. x, p. U.

f Chittenden and Bolton. Studies from Laboratory of Physiological Chenfiistry,
Yale University, vol. ii, p. 134.

134 Chittenden and Gurmnins — Nature and Gheudcal

The two products are seen to be pi'actically identical in compo-
sition, and show close agreement with the other preparations of
myosin.

Myosin I, from mitscle of ox.

This, the last product analyzed, was obtained from an ammonium
chloride extract of the washed muscle tissue from a freshly killed
ox, by dialysis. After being washed and dried at 110° C. it was ana-
lyzed with the results shown in the accompanying table.

On comparing now the composition of the thirteen preparations of
myosin analyzed, there is seen to be a very close agreement through-
out. Further, on comparing the average of the analytical results
with Hammarsten's figures for fibrinogen, the composition of the
two bodies is seen to be almost identical. Compared with egg
albumin, the most striking difference in composition is the low
content of sulphur, and even if we assume with Danilewsky* that
the sulphur of the ash comes from the sulphur of the myosin the
correction therefor would not materially lessen the difference.
Myosin also contains nearly one per cent, more nitrogen than egg
albumin.

Goagxdation of niyosi^i solutions.

According to Kuhne,f the coagulation of myosin in a 10 per cent,
sodium chloride solution occurs at 55-60° C. WeylJ obtained similar
results. Danilewsky§ states that a 12-15 per cent, ammonium chloride
solution of myosin becomes somewhat turbid at 42-43° C, decidedly
turbid at 45-50° C, while at 55° C. a flocculent precipitate sep-
arates. The same investigator also found that the more concentrated
the ammonium chloride solution was, the lower the temperature at
which the turbidity and separation of a precipitate occurred, although
never below 40° C. In common witli other proteid bodies a slight
degree of acidity lowers the temperature of coagulation, while a cor-
responding degree of alkalinity raises it. According to the recent

* Zeitschrift fiir Physiologische Chemie, Band v, p. 161.
f Lehrbuch der Physiologischen Chemie, p. 275.
\ Zeitschrift fiir Physiologische Chemie, Band i, p. 77.
§ Zeitschrift fiir Physiologische Chemie, Band v, p. 160.

Composition of the Myosin of Muscle Tissue. 135

work of Halliburton,* muscle-clot or myosin is formed from a sub-
stance (myosinogen) in the muscle-plasma which coagulates by heat
at 47° C. and 56° C, thus indicating that it probably consists of two
distinct proteids which coagulate at these temperatures respectively.
Further, Halliburton considers that whenever myosin is dissolved in
a suitable saline solution, it is at once re-converted into myosinogen,
or rather into two proteids which resemble myosinogen in that they
have the same heat coagulation temperatures and that they are con-
vertible into myosin by dilution of their saline solutions. In salted
muscle-plasma, Halliburton recognizes five distinct proteid bodies,
distinguishable by fractional heat coagulation, viz :

47° C, a flocculent, somewhat sticky precipitate.
56° C, a more abundant and very sticky precipitate.
63° C, a finely flocculent precipitate, not sticky.
73° C, a finely flocculent precipitate, not sticky.
A non-coagulable albumose.

Of these, the two proteids coagulating at 47° C. and 56° C, make
up the muscle-clot or myosin.

With these preliminary statements, we proceed to the results ob-
tained in our study of the coagulation points of the diff'erent myo-
sins, simply prefacing it with the explanation that our experi-
ments were performed previous to reading Halliburton's paper.
Naturally, the temperature of coagulation offers the simplest and
surest means, in conjunction with the determination of composition,
of detecting any difierence in the character of the myosins from
different sources. And with this as the main object the following
experiments were tried. In every experiment, the muscle tissue was
chopped quite fine and very thoroughly extracted with water, well
thymolized, prior to solution of the myosin with the saline fluid.

We first demonstrated to our satisfaction that a 5 per cent, ammo-
nium chloride solution of myosin invariably coagulates at a lower
temperature than a 5 per cent, sodium chloride solution, and further,
that the original saline extract of washed muscle shows approxi-
mately the same temperature of coagulation as the salt solution of
precipitated myosin.

* Journal of Pli}'siology, vol. viil, p. 14S.

136 Chittenden and Cummins — N'ature and Chemical

Flocculent
Turbidity. precipitate.

A. 5 per cent. NH^Cl extract of sheep's muscle. 43° C. 47° C.

B. 5 per cent. NH^Cl solution of II„0 precipi-

tate of myosin, from A. 44° 48°

C. 5 per cent. NaCl solution of II^O precipitate

of myosin, from A.

D. 5 per cent. NH^Cl extract of ox muscle.

E. 5 per cent. NaCl solution of H^O precipitate

of myosin, from D.

F. 5 per cent. NH^Cl extract of lamb's muscle,

G. 5 per cent. NaCl solution of H^O precipitate

of myosin, from F.
H, 5 per cent. NH^Cl extract of calf's muscle. 41°
I. 5 per cent. NaCI extract of same.

J. 5 per cent. NH^Cl extract of breast muscle

from chicken.
K. 5 per cent. NaCl extract of same.

L. 5 per cent. NH^Cl extract of leg muscle from

chicken.
M. 5 per cent. NaCl extract of same.

In all of these trials, the filtrate from the flocculent precipitate
failed to show any further coagulation, although the temperature
was raised to above 75" C.

The results show a varying difference in the coagialation points of
the ammonium and sodium chloride solutions, but on an average the
difference amounts to eleven degrees. It further appears that the
myosin sohitions (in NaCl) from lamb, calf, and chicken muscle have
a somewhat lower coagulating point than the corresponding extracts
from ox and sheep muscle.

In another series of results, shown in the accompanying table, the
same difference in coagulation point shows itself, and it is further seen
that the extracts from rabbit's and halibut's muscle have a still lower
coagulating point. Further, in this series of experiments, the filtrate
from the first coagulum gave a second precipitate on raising the tem-
perature, and in the case of the muscle from ox and lamb, a third
coaffulum was obtained at 74^ C.

57°

62°

40°

44°

57°

62°

44°

47°

51°

57°

41°

44°

53°

56°

43°

46°

53°

57°

45°

48°

56°

61°

(Jomposition of the Myosin of Muscle Tissue.

137

Coagulation of Myosin Solutions.

Nature of the Solutiou.

5 per cent. NaCl.
Sheep's muscle.
Ox

Rabbit's "
Pig's

Lamb's ' '
Calf's

Halibut's "
Ox

Lamb's "
Calf's
Chicken

(breast)
(leg)

Coagulation of the Solution.

Lobster*

Plocculent

Turbidity.

precipitate.

57" C.

63 C.

58^

63°

52°

57°

56°

60

53°

60°

53°

57°

47"

53°

57°

62°

55°

62°

53°

59°

53°

57°

56°

61°

65°

Coagulation of the filtrate.

Turbidity.

Flocculent
precipitate.

68° C.

73° C.

66°

72°

62°

66°

65"

70°

66°

70°

55°

60°

64°

66°

63°

68°

63°

66°

61°

67°

The results collectively fail to show the presence of a proteid
coagulating at 47° C, given by Halliburton as characteristic of myo-
sin, but they do show a coagulutn at 56° C. or thereabouts, correspond-
ing to Halliburton's second proteid. This diflference is possibly due
to the character of the saline solution, Halliburton having used a
magnesium or ammonium sulphate solution. The coagulums obtained
at higher temperatures show an approach, at least, to the figures found
by Halliburton in his fractional coagulation of salted muscle-plasma.

* Lobster did not yield enough myosin to permit of an exact determination, still a
decided turbidity was present at 65° C. The addition of water to the 5 per cent. NaCj
solution, gave a flocculent precipitate which changed to a glairy mass resembling much
the white of an egg. On washing with water it became insoluble, but differed in
general appearance from similar precipitates from other sources.

Trans. Conn. Acad., Vol. VIII.

18

Deo., 1888.

138 Chittenden and Cummins — My om,n of Muscle Tissue.

A careful study of the preceding results, combined with what has
been known concerning the chemical properties of myosin, seems to
justify the assumption that myosin, as it occurs throughout the ani-
mal kingdom, is a single chemical compound, doubtless formed, as
suggested by Halliburton, by the interaction of one or more myosin-
ogens and a ferment body. That myosin is a single body, is sup-
ported by the observed agreement in chemical composition and the
general uniformity in the temperature of coagulation of myosin from
various animal sources, and is furthermore to be inferred from the
similarity in function of the tissue in which it occurs most abun-
dantly.

VIII. — Myosinoses, By W. Kuhne and R. H. Chittenden.

Of the primary digestion jDroducts of the various proteids, the
albumoses from fibrin * and egg albumin f have been more or less
carefully examined and analyzed, as also the globuloses,J the case-
oses,§ and elastinoses. || Further, the vitelloses ^ have likewise been
prepared from crystalline phyto-vitellin and their properties ascer-
tained. With the hope of gradually completing the list we have
undertaken a study of the primary digestion products of myosin, the
results of which we now present.

For the preparation of myosin, large quantities of finely divided
ox muscle were extracted with cold water until the fluid no longer
gave reaction for albumin, after which the tissue was placed in an
excess of a 15 per cent, ammonium chloride solution, and the myosin
ultimately precipitated from the filtered fluid by dialysis.** The ex-
traction of myosin with ammonium chloride, after the method of
Danilewsky,ff is far better in every way than the older method of
extraction with 10 per cent, sodium chloride, since myosin is dis-
solved more rapidly and completely by the ammonium salt, even
from coarsely divided muscle, and at the same time forms a more
easily filterable fluid. Further, the advantage of precipitating the
myosin by dialysis in running water, instead of pouring the ammo-
nium chloride solution into a large excess of water, consists in the
avoidance of the large volumes of fluid necessary in the preparation
of such a large quantity of myosin, while at the same time the my-
osin is obtained equally free from soluble salts. At the best, how-
ever, the preparation of such a quantity of myosin as was needed in
the present investigation involved a large amount of labor and a
comparatively low temperature, hence compelling us to take the
winter season for the work. The united products obtained in this
manner were treated ultimately with dilute alcohol, whereby the
semi-gelatinous mass was naturally more or less shrunken, and finally
with ether.

* Zeitschrift fiir Biologic, Band xix, p. 159 and Band xx, p. 11.

f Studies from Laboratory of Physiological Ciiemistry, Yale University, vol. ii, p. 126.

J Zeitschrift fiir Biologic, Band xxii, p. 409.

§ This volume, p. 66. ||This volume, p. 19.

^ Ueber vitellogen, by Dr. R. Neumcister. Zeitschrift fiir Biologic. Baad xxiii, p. 2.

** See the preceding article.

ff Zeitschrift fiir Physiologische Chemie, Band v, p. 158.

140 W. Kiihne and R. H. Chittenden — Myosinoses.

A sample of the product, carefully dried at 110° C. until of con-
stant weight, yielded the following results on analysis.

Myosm.

I. 0-3665 gram substance gave 0'2313 gram H„0 = 7*01 percent.
H and 0-7053 gram C0^= 52-48 per cent. C.

II. 0-4900 gram substance gave 0*9404 gram 00.^= 52-48 per cent.
C.

III. 0-2913 gram substance gave 0-1868 gram \{JJ=z1-Vi per
cent. H and 0-5596 gram CO^ = 52-38 per cent. C.

IV. 0-7740 gram substance gave 109-1 c.c. N at 15-4° C. and 759-8
mm pressure = 16-73 per cent. N.

V. 0-5013 gram substance gave 70-6 c.c. N at 14-7° C. and 757-8
mm pressure = 16-72 per cent. N.

VI. 0-7524 gram substance gave 105-9 c.c. N- at 15-0° C. and 760-7
mm pressure = 16-77 per cent. N.

VII. 0-6629 gram substance gave 0-0043 gram ash =r 0-65 per
cent.

VIII. 0*6741 gram substance gave 0-0046 gram ash = 0-68 per
cent.

IX. The ash from 1-3370 grams substance gave 0-002 gram BaSO^
= 0-02 per cent. S.

X. 0-4967 gram substance gave by fusion with koh + KNO3, after
Hammarsten's method, 0-0458 gram BaSO^:=r26 per cent. S; de-
ducting 0-02 per cent. S of ash = 1*24 per cent.

XI. 0-5992 gram substance gave after fusion with koii + knOj
0-0552 gram BaSO^=l-28 percent. S; deducting 0-02 per cent.=i 1-26
per cent.

Percentage composition of ash-free myosin.

Average.

- 52-79

.... 712

16-86 16-83 16-88 16-86

- 1-35 1-27 1-26

21-97

c

52-82

53-83

52-73

H

7-06

7-17

N

s

....

....

....

100-00

The ash consisted almost entirely of calcium phosphate.

Digestion of Myosin.

The myosin purified as described with alcohol and ether, proved
so resistant to the action of pepsin-hydrochloric acid, tliat its diges-

TT^. Kilhne mid R. H. Chittenden — Myosinoses. 141

tioii could be accomplished only by repeated treatment with the fer-
ment. The gastric j nice at first emyloyed was prepared by warming
120 grams of mucous membrane from a pig's stomach with 1200 c. c.
0-4 per cent, hydrochloric acid for 24 hours at 41° C, filtration
through paper, dilution of the acid fluid with an equal volume of
water and further exposure to a temperature of 40° C. for four days,
in order to convert any dissolved albumoses into peptone.

The digestive fluid so obtained contained 0"2 per cent. HCl and
0-5 per cent, solid matter.

200 grams of finely powdered myosin were placed in 2 litres of
this gastric juice, 2 litres of 0*2 per cent, hydrochloric acid added
and the whole warmed at 40° C. for two days. As only a little of
the myosin appeared to be dissolved, the acidity was increased to
0*4 per cent, and the mixture continued at 40° C. for 24 hours longer.
Although a large residue still remained undissolved, the entire
mixture was made neutral with sodium hydroxide and strained
through a cloth filter. The undigested residue, together with the
abundant neutralization precipitate, was again warmed for several
days at 40° C. with 2 litres of 0*4 per cent, hydrochloric acid con-
taining 7 "5 grams of scrapings from a stomach mucous membrane.
After stopping the action of the pepsin by neutralization, this second
digestive fluid was added to the first. In spite of the energetic
action of the pepsin (tested by allowing a little of the solution to
act on boiled fibrin), there still remained considerable undissolved
substance together with considerable neutralization precipitate, the
whole apparently very resistant to the action of the ferment. In the
united solutions there was present less than 100 grams of organic
matter, of which it is fair to presume about 10-5 grams consisted of
impurity in the form of substances from the stomach membrane.
When it is remembered, however, that well prepared gastric juice
contains only a very small amount of substances precipitable by the
salts used in separation of the proteoses, it is fair to assume that this
impurity in the digestive fluid is unimportant in the study of the
myosinoses. In all, nearly 60 grams of myosinoses were obtained.

For separation of the myosinoses, the united neutral filtrates were
concentrated to about one-sixth of their volume and saturated with
crystals of rock salt, by which the fluid was converted into a gelatinous
mass. On adding saturated salt solution to a portion of the filtered
fluid it was rendered decidedly turbid ; consequently, three volumes
of a saturated sodium chloride solution were added to the mixture,
after which it was found that neither salt in substance or in solution

142 W. KiXhiie and R. H. Chittenden — Myosinoses.

would give any further turbidity. From this it is evident that the
neutral digestive fluid is not completely precipitated by simple satu-
ration with sodium chloride, but that complete precipitation is
reached only when the absolute quantity of salt stands in a certain
proportion to the proteoses present.

After this separation of the first portion of the myosinoses, which
would naturally consist of proto, hetero, and dysmyosinose, the re-
mainder were precipitated first with salt-saturated 30 per cent,
acetic acid and then, after removal of the sodium chloride by dialy-
sis, with neutral ammonium sulphate. In the solution i-emaining
from this last precipitation, peptone was detected by the biuret re-
action.

Protomyosinose.

The sodium chloride precipitate, after thorough washing with
saturated salt solution, dissolved almost entirely on being rubbed up
with water.

What did remain undissolved, showed the reactions of dysalbu-
mose. it was insoluble in water and in salt solutions of all strengths,
but easily soluble in 0*1 per cent, hydrochloric acid, much more diffi-
cultly soluble in 0*5 per cent, sodium carbonate, by neutralization
only partially precipitated and gave the biuret reaction. From the
filtrate of the neutralized hydrochloric acid solution, some hetero-
myosinosc in the form of a flocculent precipitate was obtained by
dialysis. This hetero body was soluble in dilute sodium chloride
solutions, insoluble in water, precipitable by salt in substance, and
after the manner of the albumoses gave a precipitate with nitric acid
in tlie cold, which disappeared as the mixture was warmed, reappear-
ing as the solution cooled. This heteromyosinose, formed from dys-
myosinose by a process of retrogression, amounted to considerable ;
about 1'5 grams.

Reactions of Protomyosinose.

In order to purify protomyosinose the aqueous solution of the
substance was freed from sodium chloride by dialysis, by which only
traces of heteromyosinose separated, and the solution evaporated to a
thin syrup. The fluid was filtered from a slight flocculent albumin-
like prccii)itate insoluble in hot water, somewhat further concen-
trated and the pure myosinose separated by alcohol. After washing
with alcohol and ether it appeared as a light, white powder. The
yield amounted to about ten grams.

Wl Kilhne and R. H. Chittenden — Myosinoses. 143

The reactions of protomyosinose agree in general with those of
protoalbumose, but with one important difference, viz : that an
aqueous solution of the former, free from salt, of whatever concen-
tration, is not rendered turbid by nitric acid. Addition of even a
little sodium chloride to the acid fluid, however, is sufficient to cause
a heavy precipitate, soluble as the mixture is warmed but reappear-
ing as the solution cools. The precipitate is also soluble in an ex-
cess of the acid in the cold.

Protomyosinose is readily soluble in distilled w^ater, the solution
showing a weak, but unquestionably alkaline reaction. In this solu-
tion, as well as in a solution rendered acid by acetic acid, cupric sul-
phate produces a heavy turbidity, which on boiling almost entirely
disappears. Acetic acid and potassium ferrocyanide produce a heavy
precipitate, insoluble in glacial acetic acid. Neutral lead acetate
gives no precipitate. Basic lead acetate and mercuric chloride
both produce a heavy turbidity. An aqueous solution of the
myosinose boiled with sodium hydroxide and lead acetate is colored
deep bi'own or black; with sodium hydroxide and cupric sulphate a
beautiful red. Concentrated sodium hydroxide, as in protoelastose,
produces a heavy flocculent, gelatinous precipitate. By saturation
of an aqueous solution of protomyosinose with sodium chloride, only
a portion of the substance is precipitated ; the portion remaining dis-
solved is precipitable by acid.

The composition of the substance, dried at 110° C. until of con-
stant weight, is shown by the following analysis.

Protomyosinose.

I. 0-3'722 gram substance gave 0-2387 gram H^O = 7"12 per cent.
H and 0-7090 gram C0^= 51-95 per cent. C.

II. 0-4239 gram substance gave 0-2688 gram H^O = 7-05 per cent.
H and 0-8048 gram CO^= 51-77 per cent. C.

III. 0-3319 gram substance gave 0-2138 gram IT.^0 = 7-15 per cent.
H and 0-6308 gram C0^= 51-83 per cent. C.

IV. 0-4798 gram substance gave 67-5 c. c. N at 14-8° C. and 762-1
mm pressure = 16*79 per cent. N,

V. 0-5663 gram substance gave 78-3 c. c. N at 15-1° V. and 768-2
mm pressure = 16-64 per cent. N.

VI. 0-4429 gram substance gave 62-7 c. c. N at 15-8° C. and 758-2
mm pressure = 16-77 per cent. N.

VII. (»-5494 gi-am substance gave 0-0062 gram ash = 1-13 percent.

VIII. 0-7080 gram substance gave 0-0081 gram ash=l-14 per cent.

1 44 W. JKiihne and R. S. Chittenden — Myosinoses.

IX. The ash from 1-2574 grams substance gave 0-0132 gram BaSO^
= 0-14 per cent. S,

X. 0-4183 gram substance gave after fusion with koh + kno^
0-0450 gram BaSO^=l-47 per cent. S ; after deducting 0-14 per cent.
S of the ash = 1-33 per cent.

XI. 0-3963 gram substance gave after fusion witli koh 4- kno^
0-0411 gram BaSO^=: 1-42 per cent. S; deducting 0*14 per cent.rr
1-28 per cent.

Percentage composition of ash-free protomyosinose.

Average.

--- .. .--. o3-4a

-... 7-17

16-98 16-83 16-96 16-93

1-35 1-80 1-33
32-16

c

52-53

53-35

52-41

H

N

7-19

7-13

7-32

S

o

100-00

The ash contained only calcium phosphate and sulphate, with a
little ferric oxide.

Deuteromyosinose.

This myosinose, as we have before mentioned, was obtained mainly
by saturation with ammonium sulphate. After removal of the
greater portion of protomyosinose by saturation with salt as de-
scribed, deuteromyosinose was in part precipitated by salt-saturated
acetic acid, but this precipitate could not be used on account of the
large amount of proto body precipitated with it, as shown by the
cupric sulphate reaction. This method of treatment, however,
although necessitating the loss of considerable deuteromyosinose,
enabled us to remove the proto body completely and thus ensure a
pure specimen of deuteromyosinose on treatment of the filtrate with
ammonium sulphate. Naturally, before saturating the fluid with the
ammonium salt, the sodium chloride was removed by dialysis. The
myosinose thus precipitated by saturation with ammonium sulphate
was dissolved in water and dialyzed until the sulphate was entirely
removed, or to such an extent that the fluid gave only the slightest
turbidity with barium chloride even on long standing. We hastened
the removal of the sulphate by repeated evaporation and renewed
dialysis of the concentrated fluid. The substance was finally pre-
cipitated from the suitably concentrated fluid, as a white powder, by
alcohol and washed with alcohol and ether. It weighed forty-five
grams.

W. Kuhne and It. H. Chittenden — Myosinoses. 145

Reactions of deuteromyosinose.

Like the preceding myosinose, this body also reacts alkaline in an
aqueous solution and this property exercises more or less of an influ-
ence on certain of its reactions. In order to convince ourselves of the
absence of traces of either proto or heteromyosinose, we paid particu-
lar attention to the behavior of the deutero body towards the cupric
sulphate reaction, which according to Neumeister's observations is a
decisive test on this point. A fairly concentrated solution of our
preparation was not rendered turbid by cupric sulj^hate in the cold,
but after boiling and then cooling the mixture, a slight turbidity
appeared. Solutions of the substance so concentrated as to be
almost syrupy, gave a slight turbidity at once, the turbidity disap-
pearing when the solution was heated and reappearing as the fluid
cooled. By partial neutralization of the alkalinity of the myosinose
solution, leaving the fluid, however, still alkaline to delicate test
papers, precipitation by cupric sulphate was entirely prevented. As
protomyosinose is precipitated by the copper salt equally well in an
acid fluid, we are led to consider our deuteromyosinose entirely free
from this impurity.

In general, deuteromyosinose shows much the same reactions as
deuteroalbumose, but is somewhat different from the latter in that it
is more difficultly precipitable. Acetic acid and nitric acid produce
a precipitate only after addition of sodium chloride to saturation.
Acetic acid and potassium ferrocyanide give a decided turbidity, not
soluble in glacial aectic acid. Basic lead acetate and mercuric
chloride both produce a precipitate, insoluble in excess of the rea-
gent. Cold nitric acid quickly produces an intense yellow color.
The biuret reaction comes out distinctly, but on boiling the my-
osinose with sodium hydroxide and lead acetate only a faint brown-
ing of the fluid is obtained.

The composition of the substance, dried at 110° C, is shown by
the following analysis :

Deuteromyosinose.

I. 0-2121 gram substance gave 0-1394 gram 11^0= 7-30 per cent.
H and 0-3896 gram C0„=: 50*09 per cent. C.

II. 0-2115 gram substance gave 0-1390 gram H^O = Y'30 per cent.
H and 0-3891 gram 00^= 50-12 per cent. C.

III. 0-3830 gram substance gave 53-6 c. c. N at 15*0° C. and
762-5 mm pressure = 16-71 per cent. N.

Trans. Conn. Acad., Vol. VIII. 19 Dec, 1888.

146 W. Kilhne and It. H. Chittenden — Myosinoses.

ly. 0-2940 gram substance gave 41*4 c. c. N at 15-7° C. and 759-V
mm pressure = 16*'72 per cent. N.

V. 0-4672 gram substance gave 0-0080 gram ash =1-77 per cent.

VI. 0-5462 gram substance gave 0-0094 gram ash = 1-72 per
cent.

VII. The ash from r0034 grams substance gave 0-0116 gram
BaSO^=0-15 per cent. S.

VIII. 0-3343 gram substance gave after fusion with koh + KXOg
0-0315 gram BaSO^= 1-29 per cent. S ; after deducting 0-15 percent.
S of the ash = 1-14 per cent.

IX. 0-4050 gram substance gave after fusion with koh + kno
0-0420 gram BaSO^rr: 1-42 per cent. S; deducting 0-15 per cent.:=
1-27 per cent.

Percentage composition of ash-free deuteromyosinose.

Average.

C 50-95 50-98 .... ..-. 5097

H 7-42 7-42 • .... 7-43

N 17-00 17-01 17-00

S .... -. .... 1-16 1-28 1-22

O 23-.39

100-00
The ash consisted only of calcium pliospiiate and sulphate, with
some oxide of iron.

This is as far as we have been able at present to carry our study of
the myosinoses, since hetero and dysmyosinose appear to have been
oresent in the digestions only in very small quantity. Myosin
purified by alcohol, as was the preparation employed by us, is so diffi-
cultly digestible that it is attacked only by the most energetic pepsin
mixture, and this has the disadvantage of rapidly converting hetero
and protoproteose, which according to Neumeister's * investigations
are formed in the beginning of digestion, into the deutero body ; con-
sequently in the present instance we could expect a large amount only
of deuteromyosinose. Corresponding with this view, we obtained for
45 grams of deuteromyosinose, only 10 grams of protomyosinose and
but 3 grams of hetero and dysmyosinose. Further, the unavoidable
loss attending the separation of these bodies was probably greater
with deuteromyosinose than with the others. Portions of the fluid
from the first and second digestion, tested before they were united,
showed also a difference in that the fluid from the first digestion con-

* Loc. cit.

H

N

s

7-12

16-86

1-26

21-97

7-17

16-93

1-32

23-16

7-42

17-00

1-23 ■

33-39

W. Kuline and R. II. Chittenden — llyosinoses. 147

tained far more proto and dysmyosinose than the latter, which, on the
other hand, was particularly rich in deuteromyosinose. From this it
is evident how an albuminous body can in one sense be difficultly
digestible, in that its solution takes place slowly and its primary
cleavage products form . gradually, and yet the latter be further
transformed, under the continued action of the ferment, far more
readily and completely. Our conception of digestibility needs there-
fore to be broadened, after having for so long embraced simply
the time required for solution of the proteid, or, in the case of pep-
sin digestion, the hardly attainable extreme of complete conversion
into peptone.

On comparing now the results of the analyses of myosin and the
two myosinoses in the following table,

C

Myosin 52-79

Protomyosinose. - _ 52-43
Deuteromyosinose .50-79

there is seen to be only a small ditference in composition between
myosin and protomyosinose. The content of cai-bon in the latter is
only 0-36 per cent, less than in the former, that of nitrogen 0-06 per
cent, greater and oxygen only 0-17 per cent, greater. Between deu-
teromyosinose and the undigested proteid, on the other hand, there
is a far greater difference in composition, the content of carbon being
1-82 per cent, less, while nitrogen is 0-14 per cent, greater and oxy-
gen 1-42 per cent, greater. In hydrogen, both myosinoses show a
small increase over myosin, while the content of sulphur is practi-
cally unaltered.

That portion of the myosin, which was apparently not further
alterable by gastric juice, together with the somewhat large neutral-
ization precipitate, we attempted to digest by the action of trypsin.
This was only partially successful, for although the trypsin solution
consisted of an extract from 20 grams of dry pancreas in 2 litres of
0*4 per cent, sodium carbonate, and the proteid matter was warmed
with it at 40*^ C. for six days, we were not able to bring more than
half of the material into solution. The digestive fluid behaved some-
what peculiarly, in that with a certain excess of acetic acid it gave
a fine pulverulent white precipitate. After removal of this substance,
the solution gave no turbidity whatever with sodium chloride, or
with sodium chloride and nitric acid, and only a very slight one with
ammonium sulpliate ; hence it contained no myosinoses. Peptone,
however, was formed in considerable quantity.

IX. — The Relative Absorption of Nickel and Cobalt, By
R. H. Chittenden and Charles Norris, Jr., Ph.B.

When nickel and cobalt were first discovered they were supposed
to be possessed of decided toxic properties, and nickel particularly
was looked on as more poisonous than copper. Examination, how-
ever, of many of the supposed cases of nickel poisoning led to the
view that toxic action was due, mainly at least, to the presence of
arsenical impurities, with which German nickel particularly was
known to be contaminated. Gradually, therefore, the view has be-
come widespread that nickel and cobalt are no more poisonous than
iron, with which chemically they are so closely related. We have
not, however, been able to find many very definite statements re-
garding their physiological or toxic action. Blake * in his study of
the relation between isomorphism, molecular weight, and physiolog-
ical action, places the sulphates of nickel and cobalt in the same
group with copper, zinc, iron, etc., and further arranges cobalt, cop-
per and zinc together in a sub-group, on account of their arresting
the action of the heart and preventing the coagulation of the blood.
Nickel, liowever, is placed with manganese on account of its exert-
ing a marked influence on the nervous system. As to the intensity
of their physiological action, Blake apparently considers nickel and
copper of the same strength, while cobalt is figured as one-twentieth
stronger. Both salts, liowever, kill by arresting the action of the
heart and in lethal action cobalt stands first.f Woodman and Tidy J
state that 30 grains of the oxide of cobalt given to a dog proved
fatal in a few hours, whilst 3 grains of the sulphate injected into a
vein proved fatal in four days. With nickel, the same writers state
that vomiting is freely induced in a dog by a dose of 20 grains of
the sulphate, whilst 10 grains injected into the jugular vein will de-
stroy life instantly. Finally, Brunton and Cash have found that
nickel and cobalt, like most other metallic salts, cause slight contrac-
tion of the blood vessels. §

* American Jouraal of Science and Arts, vol. vii, p. 194.
f See Brunton's Pharmacology and Therapeutics, p. 51.
X Forensic Medicine and Toxicology, p. 171 and p. 214.
gBruDton's Pharmacology, p. 246.

Chittenden and JVbrris — Absorption of Nickel and Cobalt. 149

Our experiments have been conducted wholly upon rabbits, the
main object being to study the distribution of the absorbed poison.
The salts used were chemically pure cobalt and nickel nitrates, dried
over sulphuric acid. They were administered by mouth in gelatin
capsules.

We first endeavored to gain some idea of the relative toxic action
of the two salts. For this purpose two rabbits, weigliing 2 kilos,
each, were dosed as follows :

Experiment I.

Kabbit A.

Rabbit B.

Oct.

1,

10 a.

m.

0-150 gram

Ni(N03).

0-150

gram

CO(N03)..

1,

4 p.

m.

0-102

0-100

2,

10 a.

ni.

0-150

0-150

2,

5 p.

m.

0-101

0-100

3,

9 a.

m.

0-200

0-200

3,

4 p.

m.

0-200

0-200

4,

9 a,

,m.

0-251

0-250

4,

5 p.

in.

0-250

0-250

5,

10 a.m.

0-350

0-350

5,

4 p.

m.

0-350

0-350

6,

9 a.

m.

0-500

0-500

6,

3p,

.ni.

0-500

rams

0-500

grams

3-104 g:

3-100

Both rabbits died on the morning of the Yth, apparently from
heart failure. The urine was examined each day, but in both cases
was entirely free from either sugar or albumin. On the 3d, the
cobalt rabbit appeared troubled with involuntary micturition and
defecation, and on the 5th there was quite pronounced partial paral-
ysis of the hind quarters. The appetite remained good up to the
6th. On the 6th instant, after administration of 500 milligrams of
the cobalt salt, there was a loose diarrhoea continuous till death, de-
cided pai-alysis of the hind legs, total loss of appetite with a decided
lowering of the body temperature. After death, the body weight
was found to have diminished three-fifths of a kilo. With the nickel
rabbit, there was no diarrhoea whatever and the paralysis of the
hind legs, visible on the 6th, was not as pronounced as with cobalt.
The loss of body weight was the same as in the cobalt rabbit.

On post-mortem, the only noticeable abnormal feature with either
cobalt or nickel was a slight congestion of the lining membrane of
the stomach and intestines. The stomach, however, was found full
of undigested food, as if the salt had interfered with the digestive
process.

150 R. n. Chittenden and C. JSTorris — 77ie Relative

It is thus seen that neither the cobalt or nickel salt can be called a
violent poison, since comparatively large amounts are required to
produce a toxic eiFect, and even then the action is somewhat slow.
This is still more clearly seen in the next experiment.

Mxperiment II, with cobalt.

A vigorous black doe, weighing 2 kilos., was dosed as follows :

Oct. 8, --- 0-200 gram cobalt nitrate.

9, 0-400 "

" 10, 0-600 "

" 11, 0-700 "

" 12, 0-250 "

" 13, 0-250 "

" 14, 0-250 " " " •

" 15, ...0-550 "

" 16,. .1-000 "

" 17, 1-500 "

" 18, ...2-000 "

7-700 grams.

The animal died on the 19th, of heart failure. On the 11th, when
7 decigrams of the salt were given, the animal appeared sickly, with
loss of appetite, high rectal temperature, etc., but by diminishing
the dose of cobalt the animal rapidly recovered. On the 1 7th, how-
ever, with increase in the dose of salt there was diarrhoea, with a
slight indication of paralysis of the extremities. At no time did the
urine contain either sugar or albumin. There was a decided loss of
body weight, nearly one-third. On post-mortem, the stomach and
small intestines were found somewhat inflamed, and the liver showed
signs of a slight fatty degeneration.

Immediately on the death of the animal the internal organs were
removed 'and the absorbed cobalt determined.

The method of analysis, both for cobalt and nickel, was as follows :
The finely divided tissue was oxidized with dilute hydrochloric acid
and potassium chlorate, after the usual method. From the solution
so obtained, chlorine was removed by evaporation, the fluid made
alkaline with ammonia and the cobalt or nickel precipitated by a
stream of hydrogen sulphide gas. The washed sulphide, after igni-
tion, was then dissolved in nitro-hydrochloric acid, the free acid en-
tirely removed by heat and the chloride converted into sulphate by
addition of concentrated sulphuric acid. Ultimately, the sulphuric
acid solution of sulphate was diluted somewhat with water, made
strongly alkaline with ammonia and the metal separated by elec-
trolysis.

Absorption of Nickel and Cobalt. 151

Following are the amounts of absorbed cobalt found in experi-
ment II :

Total weight Weight Co per 100

of organ. of Co. grams of tissue,

grams. milligrams, milligrams.

Stomach and contents. 81-75 57-5 70-34

Small intestines - 43-90 13-8 31-44

Large intestines... 26-40 40-3 15210

Cgecum .-- 154-00 302-5 196-43

Liver.. .-.. 45-75 8-9 19-43

Kidneys 10-30 lost

Heart.. 8-80 0-7 7-95

Lungs 5-82 0-9 15-46

Muscleof legs- 100-00 1-9 1-90

Muscleofback 27-45 1-5 5-46

Brain 8-75

a • 1 ^ Q «Q k 0-8 6-45

Spmal cord - 3-63 )

Considering the large amount of cobalt nitrate administered, the
extreme solubility oi the salt, and the length of time intervening
between the iirst and last dose, it is somewhat surprising that the
amount absorbed was not greater. Evidently a large portion of the
cobalt passes directly through the alimentary canal, pi'obably com-
bining with the proteid matter of the food to form an insoluble and
indigestible compound.

Experiment III, with nickel.

A black and white doe of 4 kilos, body weight was dosed as fol-
lows :

Oct. 15, 0-200 gram nickel nitrate.

" 16, 0-400 " "

" 17, 0-600 "

-' 18, 0-800 "

" 19, 0-500 " " "

2-500 grams.

On the morning of the '20th, the animal was found dead, the only
noticeable symptoms having been general weakness, loss of appetite
and diarrhoea. The loss of body weight was quite pronounced,
amounting in the six days to 1-4 kilos. The urine was entirely free
from albumin and sugar. There was a little inflammation of the
stomach. In this experiment, the toxic action would appear to have
been greater than that of the cobalt in the preceding experiment.
The distribution of the absorbed poison was as follows ;

152 R. H. Chittenden and C. Norris — The Relative

Total weight Nl per 100 grams

of organ. Weight of Ni. of tisaue.

grams. milligrams. milligrams.

Stomach and contents 115-00 12-2 10-68

Small intestines _.. 67-87 11-4 16-88

Large intestines 37-45 3-9 10-41

Cajcum 190-00 9-4 4-95

Liver - 87-50 5-1 5-83

Kidneys 18-85 0-8 4-24

Heart .-. • 8-90 1-3 14-60

Lungs 14-15 0-9 6-36

Muscle of legs 292-00 1-2 0-41

Muscle of back-.. 129-00 3-8 2-95

Brain... 875 21 2400

Spinal cord 500 1-0 20-00

Spleen-..- 2-00 0-6 30-00

The amount of nickel found in the alimentai-y tract is naturally
not so large as in the case of cobalt, where the final doses were larger
and the diarrhoea not so bad. Of the absorbed nickel, the distribu-
tion is essentially the same as with cobalt. The amount in the kidneys
and liver is not as large as would be expected from the size of the
doses and the soluble character of the salt. It suggests that only
a small portion of the salt given is absorbed, and that elimina-
tion goes on with comparative slowness. Quite striking is the
peculiar distribution of the nickel in the muscle tissue, the amount
in the muscles from the back being seven times as large as in the leg
muscles. The same peculiarity is likewise noticeable with cobalt.
Also noticeable is the comparatively large amount in the brain and
spinal cord, more of the poison in proportion to the weight of the
organ being found here than in either the liver or kidneys.

Experiment lY^ with cobalt and nickel.

In this experiment, two rabbits of approximately the same body
weight were dosed with nickel and cobalt respectively as follows :

Nov. 1,

RabUt A.

Rabbit B.

1,

0-200

gram C0(N03)...

0-200

gram Ni(N03

2

0-150

0-350

3,

0-250

0-200

4,

0-400

0-300

5,

0-250

0-500

6,

0-800

0-300

7,

0-500

0-800

8,

0-500

2-550

grams.

3-150

grams.

Absorption of Nickel and Cobalt.

163

Both animals were found dead on the 9th, In the cobalt rabbit,
the stomach, lungs, kidneys and brain were found more or less
congested and there was considerable diarrhoea before death. In the
nickel rabbit, there was no diarrhoea and but little congestion. Rec-
tum was found full of hard faeces.

Following is the distribution of the poison in the two rabbits.

Rabbit A, cobalt.

Total weight
of organ.

grams.

Stomach and contents 76-65

Small intestines 58'70

Large intestines - 20'10

Caecum -. 108-00

Liver... ... 91-08

Kidneys 13-55

Heart ... 13-37

Lungs - - 9-62

Muscle of legs 196-00

Muscle of back 95-85

Brain 8-87

Spinal cord 4-95

Veight of Co.

Co per 100 grams
ol tissue.

milligrams.

milligrams.

10-8

14-09

7-5

13-77

JO-9

54-23

50-5

46-76

3-3

3-62

0-8

5-90

0-9

6-73

1-8

13-50

1-2

0-61

1-7

1-77

1-2

18-53

0-9

18-18

Rabbit B, nickel.

Total weight
of organ.

grams.

Stomach and contents 64 80

Small intestines 53-30

Large intestines 13-10

Liver 70-00

Kidneys 11-30

Heart.-.. 6-72

Lungs 7-93

Muscle of leg 157-00

Muscle of back.... 82-00

Brain ...:.. 9-00

Spinal cord 3-98

Very noticeable in both of these results, as in the preceding ex-
periments, is the comparatively large amount of poison in the brain
and spinal cord, also the same relative distribution in the muscle
tissue of the legs and back previously commented upon. The large
amount of j^oison in the lungs and heart, as contrasted with the liver
and kidneys, is also quite noticeable.

Trans. Conn. Acad., Vol. VIII. 20 Dec, 1888.

Weight of Ni.
milligrams.

18-4

Niper 100 grams
of tissue.

milligrams.

38-39

2-0

3-83

2-0

16-53

5-6

8-00

0-8

7-05

0-8

11-90

2-1

26-51

1-1

0-70

2-0

3-44

1-2

13-33

1-7

42-71

154

a. H. Chittenden and C. JVbrris — The Relative

Experiment V, with nickel.
In this experiment, 1-8 grams of nickel nitrate were grven by
mouth during five days to a rabbit ofr5 kilos, weight, the individual
doses being of about the same size as in preceding experiments. On
the sixth day, the animal died of heart failure. Following is the
distribution of the poison.

Total weight Ni per 100 grams

of organ. Weiglit of Ni. of tissue.

grams. milligrams. milligrams.

Stomach and contents 73-00 3-2 4-38

Small intestines 59-10 1-8 3-04

Large intestines 36-40 9-9 27-20

Caecum 176-00 36-5+ 20-74 +

Liver.-. 58-00 8-1 13-96

Kidneys '. 10-27 0-8 7-79

Heart 7-42 1-3 16-63

Lungs. 6-52 0-2 3-06

Muscle of legs 97-30 2-6 2-67

Muscle of back 35-00 3-7 10-'57

Experiment VJ, with cobalt.

In this experiment, an attempt was made to ascertain something-
regarding the elimination of the cobalt through the kidneys. For
this purpose, the urine was collected each day and the cobalt deter-
mined by the same method as used in the analysis of the organs.
The rabbit (body weight 2-5 kilos.) was dosed as follows :

Nov,

Dec.

27,

0-100 gram

CO(N03)2

28,

0-200 "

"

29,

"

30,

0-300 "

a

1,

0-200 "

ii

2,

a

3,

0-400 "

"

4,

0-400 '•

"

5,

0-400 "

"

6,

0-400 "

"

7.

0-400 "

"

8,

0-500 "

"

9,

10,

0-500 -

11,

0-500 "

ns.

••

4-300 grai

Examination of the 2^ hours' urine.
30 c.c. contained 1-3 milligrams Co.
50 " " 6-1

73

120

6-

23-3

Absorption of Nickel and Cobalt. 155

The animal died on the 12th. On the last day, 15 c. c. of thick, dark
brown, viscid urine were voided which contained 1'8 milligrams of
cobalt. 7-15 grams of fseces excreted during the last 24 hours were
found to contain about 100 milligrams of cobalt. Obviously a large
amount of the cobalt passes directly through the alimentary canal,
while of the absorbed portion considerable is eliminated through' the
urine, much more indeed than the small amounts found in the kidneys
would appear to warrant.

On analysis of the organs of this rabbit, the folloAving results
were obtained :

Total weight Co per 100 grams

of organ. Weight of Co. of tissue,

grams. milligrams. milligrams.

Liver-.... 66-00 2-6 3-94

Spleen.-.. 0-40 0-5 125-00

Kidneys 15-30 0-5 8-27

Heart.. 10-73 0-1 0-93

Lungs 7-02 1-0 14-24

Muscle of leg 140-00 9-8 7-00

Muscle of back 130-00 4-7 3-61

Brain.. 8-35 1-5 17-96

Spinal cord 4-10 2-0 48-78

Here, as in many of the preceding cases, the lungs contain a
higher percentage of the poison than either the liver or kidneys,
while the brain and spinal cord contain a still higher percentage.

With this rabbit, convulsions were noticed shortly before death
and the breathing was very labored, as if the respiratory muscles
were aftected.

An experiment, similar to the preceding, tried with nickel, led to
a like result as regards the elimination of the poison. The first 24
hours' urine (25 c. c.) contained 1-6 milligrams of nickel, while the
portion (70 c. c.) passed during the 24 hours preceding death con-
tained 8-1 milligrams of the metal. Further, 3-5 grams of faeces
excreted during the last 24 hours contained 17*5 milligrams of nickel.

It is obvious from the foregoing that soluble nickel and cobalt
salts are possessed of decided toxic properties, but that their poison-
ous action is somewhat slow and manifested only when compara-
tively large amounts of the salts are administered. Further, so far
as our experiments show, the two salts act very much alike. Both
apparently cause death by stopping the action of the heart and also
produce more or less disturbance in the alimentary tract, interfering
with digestion, producing more or less inflammation of the mucous

156 Chittenden and JSTorris — The Relative

membrane of the stomach and intestines, and causing a more or less
persistent diarrhoea. Unlike ui-anium, these salts have no apparent
action on the kidneys or liver, sections of hardened tissue from these
organs showing no change of structure. Further, in every case the
urine of the poisoned animal was entirely free from sugar and albu-
min throughout the experiment. Both salts tend to produce a par-
tial paralysis of the extremities, more pronounced possibly with
cobalt than with nickel. They enter the circulation quickly, are
rapidly distributed to all parts of the body and are in turn more or
less rapidly eliminated by the kidneys. Considerable, however, evi-
dently passes directly through the alimentary tract and is excreted
through the faeces.

Both salts appear to raise the internal body temperature quite
decidedly, the rectal temperature rising even two or three degrees
centigrade. The blood vessels of the eai's, on the contrary, quickly
become constricted under the influence of the salts, and the ears ap-
pear white and quite cold.

The storage power of the individual organs is somewhat peculiar.
The spinal cord and brain, in the majority of the experiments, stand
first in their power of picking up and retaining the nickel and cobalt.
This is in close accord with what has already been found with solu-
ble forms of arsenic,* and more recently with strychnine sulphate.f
It would have been interesting in this connection to have seen
whether, as with arsenic, the form of the poison modifies the
relatiA'e amount absorbed by the brain and spinal cord, but this we
did not have time to try. Again, as with soluble forms of arsenic,
the muscle tissue shows in several of the experiments, as Nos. V and
VI, a marked affinity for nickel and cobalt, retaining a larger percent-
age of the metals than either the liver or kidneys. Still more notice-
able, in all of the experiments but one, is the much larger amount of
poison in the muscles of the back than in the muscles of the leg; a
constant difference occurring in a tissue of the same kind, and hardly
explainable on the ground of difference in vascularity. A somewdiat
similar distribution of arsenic in the muscle tissue was found by
one of us a few years ago in an arsenical poison case.J

* Chittenden, Amer. Chem. Journal, vol. v, p. 8, also Studies from Laboratory of
Physiological Chemistry, vol. i, p. 141. Scolosuboff, Bulletin de la Societe Chiraique
de Paris, vol. xxiv, p. 125.

f R. W. Lovett, Journal of Physiology, vol. ix, p. 99.

\ Chittenden, Amer. Chem. Journal, vol. v, p. 12.

Absorption of Nickel and Cobalt. 157

Another striking featiu-e in the storage of nickel and cobalt by
the tissues, is the comparatively large amount retained by the lungs
and heart, the amount found in these organs generally exceeding the
amounts stored up in the liver and kidneys.

Trans. Conn. Acad., Vol. VIII. 21 March, 1889.

X. — Results obtained by Etching a Sphere and Crystals op
Quartz with Hydrofluoric Acid. By Dr. Otto Meyer and
Samuel L. Penfield.

A few years ago one of us * published the results of an experiment
of etching a sphere of calcite with acetic acid in which the symmetry
of a calcite crystal was brought out by the character of the etchings
on the sphere and the final result of eating away the greater part of
the calcite was a crystalline figure with rounded faces, but with a
decided steep scalenohedral habit with truncations at the extremities
of the vertical axis. This suggested to us the idea of trying similar
experiments on spheres cut from other crystals. The difficulty of
course lies in obtaining spheres of perfectly pure homogeneous ma-
terial ; the results furnish, however, an interesting and instructive
means of studying the symmetry of any crystalline substance and as
parts of the sphere are parallel to all possible faces of a crystal, as
soon as the relation of the sphere to the axes of the crystal is made
out the character of the etchings in any particular part of the sphere
will determine the character of the etching produced by the solvent
on any crystal face parallel to that particular part of the sphere.
The ease with which spheres of Japanese quartz can be obtained and
the readiness with which quartz yields in certain directions, to the
action of hydrofluoric acid, made the following experiments quite
easy, while the results as will be seen are far more striking than one
would at first suj^pose.

The results of our experiments will be better understood by re-
viewing some experiments made in 1855 by F. Leydolt f on quartz
crystals in which he showed that hydrofluoric acid acts very un-
equally on the diflerent kinds of faces, so that not only the right and
left-handed character of the crystals, but also all the complexity of
twinning can be made to appear by etching. The experiments were
repeated by us by placing simple quartz cx'ystals from Herkimer,
N. Y., in strong hydrofluoric acid and leaving them till sufficiently
distinct etchings were produced. In these experiments, some of
which were carried on in cold and some in hot acid, the character of

* Meyer, Jahrb. Min., 1883, i, 74.

f Sitz-ber. der Wiener Akad., 1855, xv, p. 59.

Etching a Sphere of Quartz with hydroflnoria acid. 159

the etching was in all cases the same, and as qiiartz is dissolved by
the acid very slowly it is not probable that slight changes in the tem-
perature or strength of the acid would have made any appreciable
difference. On the ordinary quartz combination of prism, ni, I, lolo,
positive rhombohedron r, 1, lOii and negative rhombohedron 2,-1,
0111 the following etchings are very easily developed. The positive
rhombohedron r yields most readily to the action of the acid becom-
ing covered with elongated unsymraetrical depressions having a hori-
zontal direction, the heaviest part being to the right in a right
handed crystal, fig. 1, plate I, and to the left in a left handed crystal
fig. 2, plate I. The top and middle edges of these depressions are
nearly straight, the bottom slightly curved, the widest end is ter-
minated by a straight edge having the direction of the zonal edge
between r and the adjacent z face. These etchings are distributed
thickly over the r faces, and although they are not all exactly alike,
their general character is well represented in figs. 1 and 2. The
effect of this action is also to eat away and replace all of the edges of
the crystal toward which the heaviest ends of the etchings are turned ;
thus in a right-handed crystal between r and r (lOll and ilOl), r and
z (lOll and OlTl) and r and m (lOll and Ollo) all to the right, while
the corresponding edges to the left toward which the points of the
depressions on r are turned, are left perfectly sharp, except of course
the upper parts where r, loli forms a short edge with the adjacent
r, on 1 face to the left. In a left-handed crystal, this same phenomena
can be observed only with the corresponding edges eaten away to
the left instead of to the right. This replacement of the edges is not
shown in figs. 1 and 2, but is shown in the original figures of Leydolt,
who also determined the symbols of the faces replacing the different
edges. According to our experience the replacement of the edges
appears more like an accumulation of little facets, all reflecting the
light simultaneously, than a replacement made by a single face and
for a discussion of the symbols of the faces and the determination of
the twinning structure of quartz as shown by the etchings we refer
our readers to the original paper of Leydolt. If the crystals are left
in the acid for a sufl&ciently long time the edges between the rhombo-
hedron faces become so far eaten away that nothing is left of the
original rhombohedron faces and the prism is left terminated by the
etching faces alone, which flatten out the crystal very much in the di-
rection of the vertical axis.

On the negative rhombohedron z^ the etchings are of an entirely
different character, composed of a system of shallow depressions

160 Meyer and Penfield — Residts obtained by Etching a Sphere

with curved contours, giving a sort of feather-like marking with the
direction of greatest action turned toward the heaviest etching on
the positive rliombohedron, figs. 1 and 2, plate I.*

The prismatic faces are much less acted upon than the rhombo-
hedron faces, the etchings varying somewhat in character but con-
sisting essentially of four sided depressions with long and short
vertical edges parallel to the edges of the prism, one straight steep
edge on the side of the positive rhombohedron r and parallel to the
zonal edge between m, and 2, and a shorter slightly curved edge on
the side of the negative rhombohedron z. These etchings have
definite relations to the symmetry of the crystals and are of reverse
character on right, fig. 1, plate I, and left, fig. 2, plate I, handed
crystals. On adjacent prismatic faces, the longer or shorter vertical
edges are turned toward each other, and by prolonged etching the
alternating prismatic edges, toward which the shorter vertical edges
of the etchings are directed, are slightly eaten away while the other
prismatic edges remain sharp and perfect.

From a consideration of the above we can now more readily under-
stand the action of hydrofluoric acid on a sphere cut from a simple
quartz crystal. A sphere of about 2'tt4 c. m. diameter was purchased in
New York, and etched by placing it in a lead crucible containing rather
a strong commercial hydrofluoric acid, such as can be bought in rubber
bottles from dealers in chemicals. The exact strength of the acid
was not determined. No special care was taken to place the sphere
in any particular position in the acid, its position being accidentally
changed nearly every day when the acid was removed, and the solu-
tion of the quartz going on so slowly that the acid had a chance to
act apparently equally on all similar parts of the sphere. During
the progress of the etching, which was carried on slowly in the cold,
photographs of the etched sphere were obtained at three stages, which
seemed well suited for illustration.

* According to my experience these etchings on the rhombohedron faces furnish
one of the best methods of showing to a beginner in crystallography that the six faces
which usually terminate a quartz crystal, are not the faces of an hexagonal pyramid,
and all alike, but are those of a positive and negative rhombohedron. To prepare
sections for showing this with a microscope, crystals should be etched till the mark-
ings are sufficiently distinct, tlien by cementing the crystal, with the etched face down,
to a glass plate with Canada balsam and cementing glass plates on either side, the
quartz can be ground away witli emery till the glass plates form a large wearing sur-
face and the quartz is ground to just tlie thickness of the glass plates ; tiien after re-
moving the slice of quartz and cleaning it, it can be cemented to an object glass with
the etched surface up and is ready for examination with the microscope. — Penfield.

and Crystals of Quartz loith hydrofluoric acid. 161

After leaving the sphere in the acid for a few hours, the etchings
were distinctly observed and their arrangement on the sphere was
such that its crystalline nature and relation to hexagonal axes could
be determined. The location of the extremities of the vertical axis
was marked by the centers of two triangular patches on opposite
sides of the sphere, while tlie character and arrangement of the
prominent etchings on the positive rhombohedron indicated the right
handed character of the crystal from which the sphere was cut as
well as tlie location of the extremities of the lateral axes. After
being in the acid for about four days some of the etchings were very
prominent, and the spliere had the appearance represented in figures
1 and 2, plate II. In figure 1 we are looking down upon the sphere
in the direction of the vertical axis. In the centre there is a distinct,
somewhat hexagonal field, the center of whicli marks the extremity
of the vertical axis. This whole portion is one where the etching
has gone on very vigorously, and with the microscope it can be seen
that the surface is composed of minute triangular pyramids grouped
closely together. About this, three prominent parts, which are ar-
ranged in the alternating sections of the hexagon, indicate the
position of the positive rhombohedron by the greater extent of the
etching, leaving very distinct prominences with their steep sides
turned to the right. A distinct ridge or marking, from which the
lines of etching go ofl" very distinctly, can also be seen about in the
center of each negative rhombohedron. In fig. 2, we are looking at
the sphere about at right angles to a prismatic face. A little above
the center of the figure and trending to the right, the prominent
etchings, indicating the position of the positive rhombohedron, can
be seen, while below and to the right they can also be seen in the
position of the lower positive rhombohedron. On what may be
called the equator of the sphere, midway between the above men-
tioned prominent etchings on the positive rhombohedrons above and
below, the extremity of one of the lateral axes can be located a little
to the right of the center of the figure. On much of the surface near
the equator of the sphere, the original polish has not been destroyed.
The vigorous action of the acid at the extremities of the vertical axis
is plainly seen accompanied already by a slight flattening of the
sphere.

After exposing the sphere again to the action of the acid for about
two weeks it had the appearance represented in figures '6 and 4, plate
11, In figure 3, where we are looking down upon the sphere in the
direction of the vertical axis, three parts on the equator, located by

162 Meyer and Penfield — Results obtained hy Etching a Sphere

the right hand and upper and lower left hand angles of the hexagon,
indicate one extremity of each of the three lateral axes and from
these parts the lines of etching run out very beautifully toward the
center and the prominent marking on the rhombohedron faces. In
figure 4, where we are looking at right angles to the vertical axis,
besides the decided flattening, a rhombic portion, about in the center
of the field, is conspicuous, the center of which locates the extremity
of one of the lateral axes. On this portion not only could the origi-
nal curved surface of the sphere be detected but also the original
polish. The acid having had apparently no action on this portion
of the sphere, while the etched portions come up to meet this with
sharp and distinct angles. Owing to a slight misunderstanding a
mistake was made in photographing figure 4, which was not discov-
ered till it was too late to correct it. If we imagine the sphere
turned 90° so that the unattacked portion would appear at the right
and seen at a tangent, while one of the two similar portions which
are now behind and out of sight would appear in the front and a
little to the left, the quartz would appear in just the right position
to compare with figures 2 and 6. As it is we are looking at the
crystal not at right angles to a prism in but at right angles to a prism
of the second order I2l0.

By exposing the quartz for about one month longer to the action
of the acid it appeared as represented in figures 5 and 6, plate II. In
figure 5, which is again a vertical view, we can readily locate the ex-
tremities of the three lateral axes by the right hand and upper and
lower left hand angles of the hexagon. At these parts the curved
contour of the sphere is preserved for a short distance, but between
them there is a decided tendency toward a triangular cross section.
The sphere as will be seen from figure 6 has become extremely flat-
tened and the upper and lower portions meet along a very sharply
defined line. The etchings seem to arrange themselves along parallel
lines or ridges and some idea of their beautiful arrangement can be
obtained from the larger reproduction shown in figure 3, plate I. In
figure 6 we notice, in addition to the extreme flattening, two of the
three portions where the acid has had very little action, one taken at
a tangent to the right, the other a little to the left of the center ;
these appear as very conspicuous parallelograms ; they have a
curved surface similar to that of the original sphere, and although
the original polish has disappeared from them only the finest etchings
can be detected with the microscope. It can almost be said that the
acid has had no action on these three surfaces, at least not enough to

and Crystals of Quartz with hydrofluoric acid. 163

destroy the original polish of the sphere till toward the very end of
the experiment and not enough to appreciably diminish the diameter
of the sphere. Although the original diameter was not accurately
measured care was taken soon after commencing the etching to cut a
hole in a card board very carefully just large enough to allow the
sphere to pass and at the conclusion of the etching the quartz just
touched at these three points when passed through the same hole.

The quartz was still exposed to the action of this acid for about a
week, but the general eftect was not different from that shown in
figures 5 and 6. Of course the sphere was still further flattened in
the direction of the vertical axis and the three parts at the extremi-
ties of the lateral axes where the acid had acted least, became con-
siderably changed, being flattened out more in a vertical direction
and therefore appearing as parallelograms, relatively much more
elongated horizontally. Figure 3, plate I, is from a photograph
taken at this stage in which the relation of these parts to the longer
sharp edge between them is less than in figure 5, plate II. At this
stage also the etching of the sphere was stopped and the specimen
deposited in the collection of Professor George J. Brush, New
Haven, Conn.

In review it will be noticed, as is the case in all etching, that the
acid acts very unequally on different faces of a crystal and therefore
on different parts of the sphere, equally, however, on those similar
parts of the sphere which are similarly situated with reference to
hexagonal axes. The action is greatest at the two extremities of the
vertical axis. The action seems to be, especially toward the end, to
lift off" or dissolve away layers of molecules from above and below
while there are three parts, which are each one of the ends of three
lateral axes, where the acid exerts practically no solvent action.
These parts diminish in size as the action of the acid continues but
not by any action of the acid upon them directly, except as the
molecules were taken away from above, below and at the sides.
This is one of the most striking features of the experiment that in
these three directions quartz is almost absolutely insoluble in hydro-
fluoric acid. As a study in symmetry the experiment all along was a
very interesting one. The etched sphere could never be divided by
a plane into symmetrical halves and showed throughout all of the
experiment the tetratohedral character of a right-handed quartz
crystal. The sphere was cut from a crystal which would have shown
etchings like those in figure 1, plate I. The accompanying illustra-
tions give only a faint idea of the beauty of the etched sphere, it

164 Meyer and Penfield — Mesults ohtahied by Etching a Sphere

being impossible to reproduce the delicacy and beauty of the mark-
ings as they appeared on the perfectly transparent material of the
quartz.

Pyro-electrrical experiments. — To further test the relation of the
sphere to crystallographic axes, it was heated for some hours in an
air bath to 100° C, and on cooling dusted with a mixture of red oxide
of lead and sulphur, according to the excellent method described by
Professor A. Kundt of Strassburg.* The red oxide of lead and
yellow sulphur arranged themselves in six alternating vertical bands
about the equator of the sphere, the red bands being located midway
between the heaviest parts of the etchings on the positive rhombohe-
drons above and below. On the etched sphere represented in figure
2, plate II, a red band ran vertically a little to the right of the cen-
ter, a yellow one a little to the left of the center and so on, three red
alternating with three yellow about the equator of the sphere. On
the etched sphere represented in figures 5 and 6, plate II, the slightly
attacked parallelogram parts, representing the ends of the lateral
axes, were yellow while the center of the sharp edges midway be-
tween them were red. These pyro-electrical phenomena, according
to B. von Kolenko,! indicate that the crystals from which our sphere
was cut was a right-handed one ; that part of the sphere where neg-
ative electricity develops on cooling and where the positively elec-
trified red oxide of lead deposits, indicating the position of the pris-
matic edge, where the right-handed trapezohedral faces above and
below would occur. The even distribution of the pyro-electricity
into six alternating positive and negative sections prove the simple
character of the sphere and the absence of twinning. This latter
was very important for the success of our experiment, lor if the
sphere had been cut from a complicated twin crystal the etchings
would have arranged themselves in a very confused manner, and the
shapes produced by the etching would have been very much modified.

In closing we wish to express our thanks to Mr. J. M. Blake of
New Haven, for the care which he took in photographing such a
difficult, transparent object, and to Mr. E. Bierstadt of New York,
for the pains which he took in the preparation of the plates,

Mineralogical Laboratory, Sheffield Scientific School, Feb. 4th, 1889.

* Ann. d. Phys. u. Chera., 1 883, xx, p. 592.
t Zeitsch. f. Kryst, ix, 1884, p. 1.

and Crystals of Quartz toith hydrofluoric acid. 165

EXPLANATION OF PLATES.

Plate I.

1. Etchings produced ou the faces of a right handed quartz crystal by hydrofluoric
acid.

2. Etchings produced on the faces of a left handed quartz crystal by hydrofluoric
acid.

3. Final result of eating away the greater part of a sphere (cut from a right handed
quartz crystal) with hydrofluoric acid during a period of about eight weeks. Seen in
the direction of the vertical axis. Tiie angles of the hexagon mark the extremities of
the lateral axes.

Plate fl.

'1 and 2. Appearance of the etched sphere after being in the acid about four days.
1. Seen in the direction of the vertical axis. 2. Seen in the direction at right angles
to the vertical axis and a prism of the first order.

3 and 4. Appearance of the etched sphere after being in the acid about three
weeks. 3. Seen as in 1. 4. Owing to a mistake in taking the photograph seen about
at right angles to a prism of the second order.

5 and 6. Appearance of the etched sphere after being in the acid about seven
weeks. 5. Seen as in 1 and 3. 6. Seen as in 2.

Trans. Conn. Acad., Vol. VIIT. 22 Dec, 1889.

XI. — New England Spiders of the Families Dbassid^,
Agalenid^e and Dysderid^e. By J. H. Emerton.

Drassidse.

The Drassidm have long bodies like the Agalenidce and Lycosidce,
but most of them are a little flattened above and walk with their
bodies near the ground ; the first and second feet are directed for-
ward and the third and fourth backward. The feet have but two
claws, under which is usually a cluster of hairs sometimes so thick
as to conceal the claws. The under sides of the tarsus and metatarsus
are sometimes covered with hairs, especially on the first and second
legs, and these hairs are often flattened or thickened at the end.
The ce})halothorax is low in front, the highest part being in the
middle or farther back. The eyes are in two nearh^ straight rows.
The spinnerets are cylindrical with the tubes on the end, and the
upper and under pairs are nearly equal in length.

None of the Drassidm make webs for catching insects, though
many of them make nests, usually flattened tubes, in which they
hide in winter or while moulting or laying eggs. Most of them live
on the ground and hide under leaves and stones. A few, as Gluhiona,
Ghiracanthium, and Anyphmna live in summer on plants several
feet above the ground.

Many species are found adult at all seasons and probably live
several years.

The cocoons are flat, some are attached by one side, but most of
them lie loose in the nest or hiding place.

A large number of American species were described and figured
by Hentz, most of them under the generic name Herpyllus. Of
these I have identified nine : II. descriptus., crocatus, alarms, ecclesi-
asticus, ater, h'dineatus, variegatiis, and Cluhiona saltahunda, pallens.
Several others have been described by Thorell, and of these I have
identified Gnaphosa hrumalis Thor., Proc. Boston Soc. Nat. Hist.,
vol. xvii, 1875, and Gnaphosa conspersa, G. scudderi, and Prosthe-
sima melancholia Thor., Bulletin Hayden's U. S. Geol. Survey, vol.
iii, No. 2, 1877. The specimens of this family in the Museum of
Comparative Zoology in Cambridge, Mass., have been named by
Keyserling, and I have adopted his names for several species though
they are not yet published.

Neto England Brass khe, Agalenidm and Dysderidm. 167

Micaria Westdug.

This genus was first separated from Drassus by C. Koch under
the name of Macaria, which had before been used for a genus of Lepi-
doptera, and was therefore changed by Westring to Micaria in 1851.
They are all small and slender spiders with the cephalothorax arched
upward in the middle, without any dorsal groove or o\^\y a thick
opaque spot in place of it. The abdomen and usually the cephalo-
thorax are covered with flattened scales sometimes brightly colored
and ii-idescent. The tarsus and metatarsus of the first and second
feet have a double row of flattened hairs on the under side.

Plentz's Herpyllus auratas, found farther south, belongs to this
genus and is nearly related to 31. longipes.

Micaria longipes, uew sp.

Plate III, figures la, 1&, Ic, \d, It;, ]/; 1//.

Largest specimen 5"'"^ long. PI, iii, fig. 1. The cephalothorax
is twice as long as wide, widest across the middle. Head not much
narrowed. The eyes occupy half the width of the head. The front
row is nearly straight, the upper row with the middle eyes highest.
Eyes all nearly of the same size. The cephalothorax is highest in
the middle, curving downward toward both ends. The abdomen is
one-half longer than the cephalothorax and about as wide, blunt at
both ends and drawn in a little at the sides and above about one-
third its length from the front. The legs are long and slender, the
fourth pair longest. The colors are light yellowish brown with gray
hairs and scales which have green and red metallic reflections on the
abdomen. The legs are darker from before backward, the front
pair all light yellow except the femur, and the fourth and third pairs
with longitudinal brown stri})es that nearly cover the tarsal joints.
The cephalothorax is without markings. The abdomen has a pair of
white stripes at the constricted spot and a less distinct pair near the
front end. At the hinder end it is almost black. The scales of the
abdomen are of various forms, those of the white spot are long with
several branches at the base figs. U, le, those of the front of the
abdomen are more simple, fig, Ic, and those behind the white mark
ings are half as Avide as long \vith two rows of short branches fig. \f
at some distance from their edges. The under side of the abdomen
is as dark as the upper side. The white markings extend under half
way to the middle line. The sternum is nearly twice as long as
wide, widest at the second pair of legs and narrowed to a point
behind.

168 J. H. Emerton — Kew England Drassidm,

The maxilhe are a little notched on the oiiter sides and straight
on the ends except at the inner corners. The labium is two-thirds
as long as the maxillte. It is narrowed toward the tip, where it is
about half as wide as at the base, fig. \c(.

The male palpi are small, the patella and tibia are about of the
same length ; at the base of the tibia on the upper side is a large
tooth nearly as long as the diameter of the palpus, and curved
strongly forward, fig. \h. The tarsus is as long as the tibia and
patella together, and pointed at the end. The palj^al organ is small,
fig. \h.

The epigynum of a female from Salem, Mass., appears as in tig. lA
with two oblique openings near the posterior edge.

Salem and Medford, Mass., under stones and leaves. Adult male
in August and adult female in June.

Micaria montana, new sp.

Plate III, figures 2, 2a.

This is smaller than the common species. A female measures 4™™
long. The cephalothorax is not twice as long as wide and the widest
part is behind the middle. The abdomen is twice as long as wide,
not constricted or truncated at either end. The cephalothorax and
legs are light yellow-brown, the legs lighter toward the ends. The
abdomen is greenish brown with iridescent scales. Across the mid -
die is a distinct narrow white line and a less distinct one crosses the
front of the abdomen. On the hinder half of the abdomen are four
or five white spots. The white markings extend a short way under
the abdomen. The epigynum, Plate in, fig. 2a, has two oblique
openings near the posterior edge turned more toward each other and
less downward than in Micaria longipes.

Mt. Washington, N. H., July 1, 1874, east side, near the Ledge.

G-eotrecha, uew genus.

This genus includes a number of American spiders described by
Hentz, under the name of Herpyllus. Besides Ilerpyllus descrijAus
and crocatus, H. ornatus, H. long'qyalpus, H. marmoratus, II. cruci-
ger^ H. conarius^ H. triUneatus, probably belong in it. II. descrip-
tus and H. crocatus were placed by Koch in the genus Agrmca,
with which they agree in the shape of the maxillae and position
of the eyes. In this, he was followed by Keyserling, who named
the specimens of that species in the Museum of Comparative
Zoology, in Cambridge, Mass., Agreeca crocata. In the same collec-

Agalenidm and Dysderidm. 169

tion another species, Geotrecha hivittata, is named by Keyserling
Gastlaneira hiinttata, the genus Castianeira having been named by
Keyserling, in 1879, for a South American spider, with a long,
slender cephalothorax and a slender abdomen with the front part
hardened and differently colored from the softer part.

In our species of Geotrecha, the cephalothorax is about two-thirds
as wide as long and narrowed in front, more in some species than in
others. The abdomen is longer and a little wider than the cephalo-
thorax. It sometimes has a small, hard patch at the front end which
is of the same color as the rest of the back and not easilj'^ seen. The
abdomen is round, not flattened above as it usually is in Prosthesima.
The legs are long and slender. The hairs on the under side of the
first and second legs are only slightly flattened and thickened, and the
claws concealed by a thick bunch of hairs. The maxillae are nearly
sti'aight as in Agroeca and the labium is as short as wide. The eyes
are close together in the middle of the front of the head, the front
row nearly straight and the hind row with the middle eyes highest.
The middle eyes of both rows are largest and farther apart than
they are from the lateral eyes. The spinnerets are very small and
close together. The colors are dark brown and black, with white or
bright colored markings.

The male palpi have the patella and tibia both short and the tarsus
long and tapering. The palpal organ is round at the base and tajjers
to a fine point. The epigynum has two simple openings dii'ected
backward, and differing in size and distance apart in different species.

G-eotrecha bivittata.

Castianeira bivitkita Keyserling, specimens in Cambridge Museum.
Plate III, figures 3a, 36, 3c, 3d.

Length, 7 or 8"'™. Legs of fourth pair, 10 or 11™'". The cepha-
lothorax is widest across the middle in front of the dorsal groove,
Plate III, fig. 3a, and is about half as wide at either the front or
hinder end. The abdomen is usually about as long as the cephalo-
thorax and widest at the hinder third. It is sometimes slightly
drawn in at the sides and above over the front white marking. This
is caused by contraction in alcohol, the front end of the abdomen
being hardest contracts less than that part just behind it. The legs
are long and tapering, the fourth pair longest.

The cephalothorax is dark brown. The abdomen is of the same
color, a little lighter, with two white cross stripes, one about the
middle of the back and the other, a less distinct one, farther for-

1*70 -T. H. Emerton — New England Drassiclw,

ward. The femora of all the legs are striped lengthwise with brown
and yellow. The hind legs are brown with a little yellow on the
upper side of the patella and tibia. The other legs are yellow, some-
times with brown stripes on the under side. The white marks on
the abdomen extend underneath half way to the middle line. The
front hard part of the under side of the abdomen is lighter than the
hinder part, and the sternum is of the same color. The coxa? are
lighter yellowish brown.

The cpigynum shows through the skin as three dai-k spots and has
two openings directed backward, fig. 8f7.

The male palpi have the patella very short, about half as long as
the tibia, fig. 2>h. The tarsus is very large and dark colored, wide at
the base and tapering toward the tip. The palpal organ is simi-
larly shaped, with a rounded bulb through which the coiled tube
can be seen, and a slender tip lying in a groove in the tarsus, fig. 3c.

Hentz's Herpyllus zonarius and trilineattis seem to be near this
species.

It lives under leaves at all seasons of the yeai", and though not so
quick in its motions as crocata is a difficult spider to catch except in
cold weather, when it is often sifted from leaves in a torpid condi-
tion.

Massachusetts, Connecticut, and in N. Pike's Long Island collection.

The color is sometimes lighter, the whole cephalothorax above
and below being light orange color, and the legs the same color, with
the longitudinal brown stripes very narrow and indistinct.

In young individuals of both varieties the sternum is wider and
more convex than in adults.

Geotrecha pinnata, new sp.

Plate III, figures 4, 4a.

The largest specimen is 7™"^ long, cephalothorax, S"*™.

The cephalothorax is shaped much as in G. crocata. The abdo-
men is oval, not so much narrowed in front as in the other species.

The cephalothorax is light brown. The abdomen is grayish
brown with several white transverse stripes. The two widest
stripes are in the same position as the two stripes of C. bivittata.
On the front of the abdomen is another stripe, and on the posterior
half are four or five others, some of them incomplete on the middle
of the back. The femora of all the legs are light brown, the first,
second and third legs are yellow, except the femora. The fourth
legs have the tarsus and the ends of the tibia yellow, the rest light
brown. Plate iii, fig. 4.

Agalenidm and Dysderidce. 171

Epigynum, like that of crocata, but with the holes larger and
nearer together, fig. ^a.

The palpal organs and male palpi resemble those of crocata, but
are a little larger.

Three specimens, of different ages, from Medford, Mass., under
leaves with C. bivittata. Three adult females from Topsfield, Mass.,
Sept. 3d, under log in woods. Males and females in N. Pike's Long-
Island collection.

Geotrecha crocata.

Agroeca crocata Keys., specimens iu Miis. Comp. Zool , Cambridge, Mass.
S Herpyllus descriptus Hentz.
S Herpyllus crocatus Hentz.

Plate III, figures 3Z;, 3c, 3d.

Length of female, 8 to 10'""^ ; cephalothorax, 4™'". The cephalo-
thorax is nearly twice as long as wide and widest across the dorsal
groove. It is not narrowed behind as much as in G. bivittata. The
abdomen is usually longer than the cephalothorax and a little wider
at the widest part.

The cephalothorax is very dark brown or black, and the femora
and coxa? of all the legs are the same color. The abdomen is black
with a bright red spot of variable shape and size at the posterior end.
The spot turns yellow in alcohol. In some specimens it is wanting.
In the males the red spot is usually larger, sometimes extending the
whole length of the abdomen. The hind legs are black or brown
their whole length, a little lighter at the ends ; the other legs are
yellow, except the femora. The under side of the body is all black.

The epigynum has two small round openings, wide apart, a little
in front of the transverse fold. Plate iii, fig. 3d.

The male palpi are much like those of C. bivittata, but the tarsus
and palpal organ are only about half as large and the patella and
tibia are nearly equal in length. The tibia has a short process on
the under side. Figs. 3b, 3c.

This spider lives among stones in dry, open places. It is easily
alarmed and moves very rapidly. The flat, parchment-like cocoons
common on stones in pastures are probably made by this species,

Massachusetts and Connecticut, and in N. Pike's Long Island col-
lection.

172 J. H. Emert07i — N'ew England Drassidm,

Prosthesima l. Koch.

Ceplialothorax widest in the middle and more than half as wide in
front. Eyes near together, occupying about half the width of the
head. The middle eyes of both rows smaller than the lateral and
nearer together than they are 'to the lateral eyes. Upper row
straight, or the lateral eyes a little farther back. Sternum large
and nearly as wide as long. Maxillae wide in the middle and but
little widened at the ends, PI. iii, fig. 6a. First and second legs with
flattened haii'S under the tarsus and part of the metatarsus.

Prosthesima atra.

Herpyllns ater Hentz.

Prosthesima funesta Keyserling, speeiraens in Mus. Comp. Zool., Cambridf^e, Mass.

Prosthesima melancholica Tliorell, Bull. Hayden's U. S. Geol. Survey, Yol. Ill, 1877.

FL.A.TE III, FIGURES 6, 6c, Gcl.

Female, 8""" long ; cephalothorax, 3™"\ Male smaller. Ceplialo-
thorax and abdomen both a little flattened above. Cephalothorax
narrow in front, about half as wide as in the middle. Plate iii, fig. 6.
Abdomen oval, the hinder half usually a little wider than the front.
Sternum very large, almost as wide as long, fig. 6a. Maxillae and
labium large and a little shorter and wider than in P. ecdesiasHca.
Feet 1 and 2 with flattened hairs under the tarsus and half the
metatarsus. Feet 3 and 4 with fine hairs in the same places.

The whole body is black in most individuals, sometimes, especially
in the young, yellowish broAvn on the ends of the feet and under the
abdomen.

Epigynum large and distinct with two small depressions in front
and large openings behind surrounded by a thick brown rim, fig. 6d.

The male palpus is short with a very large tursus, as long as the
tibia and patella together and more than half as wide. The process
on the outside of the tibia is about as long as the tibia itself and
nearly straight. The palpal organ has a small fine tube and several
small hooks and processes all at the tip end of the palpus, fig. 6c.

This spider lives under stones and leaves. The cocoon is flat on
one side, by which it is attached, and convex on the other. It is
white, or sometimes a little pink.

Mt. Washington, N. H., Eastport, Me., Massachusetts, and in
N, Pike's Long Island, N. Y., collection.

P. melancholica was found by Dr. A. S. Packard at Monitou,
Colorado, 1875.

Agalenidce and Dysderidm. 173

Prosthesima depressa, new sp.

Plate III, figures 8, 8a.

A smaller species than atra. Female, 6™™ long. The head is
much smaller than in atra and the eyes larger and closer together.
Plate III, fig. ft. The cephalothorax and abdomen are black. The
first and second legs have the tarsus and metatarsus pale yellow, the
rest of the legs black except a pale spot on the outside of each
femur. The third and fourth legs have the tarsus and metatarsus
pale, the tibia black at the distal end, the leg becoming lighter from
this point to the base. Underneath, the coxae are darker from
behind forward.

The epigynum has the openings at the sides, farther forward, and
the ridges over them thicker and shorter than in atra, and the two
little depressions in front appear to be wanting, fig, 8«.

Medford, Mass., July 23.

Prosthesima ecclesiastica.

Prosthesima profiinqua Keys.
Herpyllus ecdesiasticm Ilentz.

Plate III, figure 7, In, lb, 1c, Id.

This spider is about 8 or 10™"' long, a little smaller than Gnaphosa
conspersa which it resembles in foi'm and habits, but from which it
is easily distinguished by its colors. The cephalothorax is black at
the sides and has a whitish stripe in the middle. The abdomen is
black at the sides with a bright white stripe in the middle that
extends from the front end about two-thirds its length. At the
hind end of the abdomen, just over the spinnerets, is another white
spot. The legs are dull black turning to brown in alcohol, as does
the cephalothorax. The under side of the abdomen is dark at the
sides and light in the middle.

The eyes cover about half the width of the head. The two rows
are nearly equal in length, the hinder only a little the longer. Seen
from above, both rows appear straight. PI. iir, fig. 7.

The maxillae are widened at the end, the outer corner sharp and
the inner rounded off down to the lip. Fig. 7«.

The male is much smaller than the female but similarly marked.
The male palpi are small. The patella and tibia are short, and the
tarsus is as long as both together. The process on the tibia is slender
and about equals the tibia in length. It is on the outer side, show-
ing indistinctly from above. It is slightly forked at the tip. The

Trans. Conn. Acad, Vol. VIII. 23 Dec, 1889.

174 J. H. Emerton — N'eio England DrassidfB,

palpal oi'gan is simple, with two short processes on the outer end.
Fig, 7 c, 7 c?.

The epigynum has a small oval opening at the posterior end of a
dark area. Fig. 1h.'

Under stones. Boston, Salem, Danvers, Wood's Holl, Mass.; Al-
bany, N. Y. ; Providence, R. I.

Poecilochroa Westr., Simon.
Plate IV, figures la, 3a.

The cephalothorax is narrowed towai'd the front, as in Prosthe-
sima, and more narrowed in males than in females. The two rows of
eyes are far ai)art, the hinder row a little longer than the front row,
with the lateral eyes farther back than the middle ones. The middle
eyes are farther apart than they are from the lateral eyes. The
labium is not much longer than wide and a little narrowed toward the
end. The maxillae are about twice as long as the labium. They are
narrower at the base and widen to the insertion of the pal])i. From
the palpi the maxillte curve inward and nearly meet in front of the
lip. The outer corners are turned outward. PI. iv, figs, la, 3a.
The colors are bright and the markings distinct.

Poecilochroa variegata.

Ilerpyllus va.riegatus Hentz.

Drassiis variegatus Keyscrling, specimens in Mus. Com p. Zool , Carabfidp^e, Mass.

Plate IV, figures 1, 1&, Ic.

This is one of the most distinct and brightly colored species of the
family. PI. iv, fig. 1. The cephalothorax is bright orange, a little
darker toward the eyes. The abdomen is black with three white
transverse stripes from the middle of which a T-shaped white mark
extends half way to the front stripe. On the front half of the ab-
domen the white stripes are usually partly colored with orange. The
femora of the first and second legs are black. The distal end of the
femur and both ends of the tibia of the fourth legs are black. The
legs are otherwise orange colored. The hinder row of eyes is con-
siderably longer than the front row, the rear lateral eyes being
their diameter nearer the sides of the head than those of the front
row. The head of the male is much narrower than that of the
female. The male palpus has a process on the outer side of the tibia
half as long as the tarsus, tapering toward the end and slightly bent
inward at the tip. The tube ends near the outer end of the tarsus
and is supported by a short thick process. Figs, lb, le.

Agalenidm and DysderidoB. 175

This spider is common under leaves in dry Avoods. Eastern Massa-
chusetts; Dublin, N, H.; New Haven, Conn.

Poecilochroa montana, new sp.

Plate IV, figures 'J, 2a.

This species is a little larger and less brightly colored than
P. variegatai PI. iv, fig. 2. Female 8""" long, cephalothorax
3.5™'". The arrangement of the eyes and proportions of the body
are about the same. The cephalothorax and legs are dark brown,
the hinder ones a little the lighter. The abdomen is black with a
pair of white spots near the front end and another pair across the
middle nearly united in the middle. The sternum and coxae are
dark brown. The epigynum is dark brown with a small opening at
the hind end. Fig. 2a.

Mt. Washington, N. H., on the road to Gorham.

PcBcilochroa bilineata.

HeTpyllus hilintatus Heutz.

Plate IV, figure 3, 'da.

A very distinct species on account of its markings. Cephalothorax
white, with two black stripes and a fine black line on the edge,
each side. PL iv, fig. 3. Abdomen thickly covered with long hairs,
white in the middle and at the sides, and with two wide black stripes
that do not extend quite to the end. The under side of the abdo-
men is white with a black stripe each side. Fig. 3a. The legs are
covered with gray and white hairs.

The female is 7™™ long, cephalothorax 3""". The head is about
half as wide as the middle of the thorax. The eyes are small, the
middle pair in each row farther apart than they are from the lateral
eyes. The two rows are widely separated. Sternum oval, widest
in the middle. Spinnerets long. Epigynum small, with a single
opening directed backward just in front of the transverse fold.

Gnaphosa Latr., 1804.

GnaphOSa brumaliS Thorell, Proc. Boston Soc. Nat. Hist., vol. xvii, 1875.
Gnaphosa scudderi Thorell, Bull. Hayden's U. S. Geol. Survey, vol. iii, 1877.
Plate IV, figures 5, 5a, 56.

This species is a little smaller than G. conspersa. A female of the
usual size is 10™'" long, cephalothorax 4™™, while conspersa grows to
the length of 12 or 15""".

176 J. H. Enierton — Neio England DrassidcB,

The colors are the same as those of conspersa ; cephalothorax and
legs dark brown and abdomen rusty black.

The epigynum has the openings rather wider apart, and the front
middle appendage flat, wrinkled at the edges, and with a hard spot
in which is a small hole near the end. This apj^endage resembles
the finger in the same position in Ex>eira. PI. iv, fig. hh.

The male palpus has the tibia rather shorter and its outer process
longer than in conspersa, fig. 5. The tube of the palpal organ
is only about half as long, its base being nearer the middle of the
tarsus. The middle hooked appendage is as long as in conspersa,
but much more slender, fig. ha.

Under stones on Mt. Washington, Ni H., from the ledge upward,
with cocoons of eggs July 1.

Males and females from Ellis Bay, Anticosti, July 23, S. Hen-
shaw in collection of Boston Soc. Nat. Hist.; females with cocoons of
eggs.

The specimen named by Thorell was from Strawberi-y Harbor,
Labrador, collected by A. S. Packard in 1864.

G. scudderi was found by A. S. Packard at the Garden of the
Gods, Colorado, in 1875.

G-naphoSa conspersa Thorell, Bull. Ilayden's U. S. Geol. Survey, vol. iii,
1877.
Gnaphosa giganfea Keysorliug, spueiraeus in Mus. Comp. Zool., Cambridge, Mass.
Plate IV, figures 4, 4a, ib, 4c, 4rf, 4e.

This spider is 12 to 15'"'" long and rusty black in color. Some
specimens freshly moulted are dull yellowish or greenish gray, and
old individuals have a. brownish color. The whole body and the legs
are covered with long hairs. The cephalothorax is wide in front and
the eyes are not so close together as in Pythonissa hnhecilla, or in
Prosthesima, The hinder row of eyes is a little longer than the front
row, and the lateral eyes are larger and farther back than the middle
ones. PI. IV, fig. 4. The middle hinder ej^es are a little oval and
oblique, diverging toward the front. The mandibles are large and
strong, on the inner side under the claw they have a wide flat tooth
with irregular and serrated edge, and near the inner corner two large
pointed teeth, fig. Ah. The maxillae are very wide and curve inward
so as nearly to meet ai'ound the end of the lip. Their outer corners
are rounded, tig. 4«. The spinnerets are stout and the lower pair
are widely separated. The male differs but little from the female.
The male palpi have the patella and tarsus both short and the

AgalenidcB and Dysderidce. 177

tarsus as long as both of tliem. The tibia has a short pointed pro-
cess extending forward over the tarsus, fig. 4c. The tarsus is curved
outward at the end. The tube of the palpal organ is slender and
extends along the outer edge of the tarsus for its whole length. At
the outer end of the palpal organ is a short process flat at the end
and curved downward, fig. 4c?. The epigynum has a long opening
on each side and a short transverse pit in front of them in the
middle, fig. 4e. In western specimens the opening at the hinder
part of the epigynum has the sides more nearly parallel, not diverg-
ing forward as much as in those from New England.

It lives under stones and leaves. The cocoon is white and flat,
with its diameter as great or greater than the length of the spider.
The female stays near the cocoon, but makes no nest.

All over New England, from the White Mountains, N. H., to New
Haven, Connecticut ; Adirondacks, N. Y.

Thorell's specimens were collected by A. S. Packard in 1875. A
female with cocoon of eggs on Gray's Peak, Colorado, over 11,000
feet high, on fir, Kelso's cabin, Colorado, and a small one from
Idaho, all adult females and all smaller than most eastern specimens.

Pythonissa Koch.

Pythonissa imbecilla Keyserling, speeimen fioni Kentucky iu Mus. Coinp

Zool., Cambridge, Mass.

Plate IV. figures Ga, G?;, 6c, <6d.

The male is about 4'""' long and the female 5""". The two rows
of eyes are nearly of the same length, the hindei" row almost straight,
with the lateral eyes only slightly farther back than the middle
ones. The lateral eyes of both rows are larger than the middle eyes.
The maxilhe are almost as wide as long and are curved inward so as
to touch in front of the lip. PI. iv, fig. 6«. The front edge of the
maxilhe is straight with the corners only slightly rounded. The
mandibles are small and the wide tooth under the claw, fig. 6^, can
be plainly seen just in front of the maxillae with another pointed
tooth on its inner side. The cephalothorax, both above and below,
and the legs are orange-brown with black hairs. The abdomen is
bluish black with a few v/hitish hairs at the hind end and around
the four muscular spots near the middle. The epigynum, as in fig. <od.
The patella and tibia of the male palpus are very short and not so
thick as the femur. The tarsus is as long as the patella and tibia
together. The palpal organ is large and complicated, the tube show-
ing plainly across the outer end, fig. 6c.

178 J. H. Emerton — Neio England Drassidoe,^

Under stones. IMales from Dublin, N. H., and Dedbam, Mass.
Females from Bliiehill, Milton, Mass,

Drassus.

Plate IV, figure la.

Tbe genus Drassus of Walckenaer included tbe greater part of
tbe present family DrassidiB, as well as some Agalenidai and Cini-
Jlonidm. From tbis, various genera bave been separated from time
to time, leaving tbe present Drassus an ill defined group containing
species differing greatly among tbemselves and forming several
groups, whicb f urtber study will no doubt make it possible to sepa-
rate. Tbe only two species wbicb I place in tbis genus belong, one
near tbe Eui'opean D. lapidosus and tbe otber near D. troglodytes.
In tbese species the cepbalotborax is wider in front and less flattened
tban in Gnaphosa and Prosthesima. The eyes are small and sepa-
rated by spaces at least as wide as tbeir diameter. Tbe front row is
nearly straight. Tbe posterior row is longer, and curved with the
lateral eyes lower tban tbe middle. The middle hinder eyes are oval
and turned apart toward tbe front, and are nearer together than to
the lateral eyes. The mandibles and maxillae are large and stout.
Tbe maxillje are widened on both sides bej'ond tbe insertion of tbe
palpi, the outer corners are slightly rounded and tbe inner corners
slope obliquely toward the lip. PI. iv, fig. 7 a. Tbe lip is about
half as long as the maxilhe. The colors are gray and drab with fine
short, white or gray hairs, and only faint markings on tbe abdomen.

Drassus saccatus, new sp.

Plate IV, figures 7, 7c, Id.

Tbis is one of tbe most common of our Drassidce. PI. iv, fig. 7.
The female grows to be 12""" long, with legs 10""" to I5™"\ The
color is light gray sometimes with indistinct transverse dark mark-
ings on tbe abdomen. The color of the front part of the bead is a
little darker and the feet and mandibles and maxillae are brown.
Tbe abdomen is long and slender as in Clubwna. The epigynum is
small and has two dark round depressions just in front of the fold,
fig. Id.

Tbe male is smaller and more slender. Tbe male palpi are very
long tbe patella, and tibia are together as long as the femur, and all
are as long as tbe femur of tbe first legs. The tibia has a small
process on tbe outer side, fig. 7c. Tbe tarsus is long and narrow.

Agalenidm and DysderidcB. 179

and the palpal organ is small with a short tube near the distal end,
fig. 7c.

These spiders live under stones in a large bag of silk in which the
female stays with her cocoon of eggs. In the early summer a male
and female live together in the nest, the female often being imma-
ture.

White Mountains, N. H., to Connecticut,

Drassus robustus, new sp.

Plate IV", figures 8, 8a, 86, 8c.

This is a smaller and shorter legged species than the preceding.
PI. IV, fig. 8. The female is 8'"™ long. The colors are darker and
redder, especially toward the head. The sternum, maxillje and man-
dibles are dark brown. The head is as wide as in D. saccatus, and
the eyes are a little closer. The epigynum is large, light colored in
the middle, and with a dark ridge each side, fig. 8a. The male is
much smaller than the female. The palpal organs and tarsi are large
and round. The tarsus is short and has a short curved process that
extends over the tarsus on the upper side, fig. 8^, 8c.

Medford, Mass., July.

Clubiona Latr.

Cephalothorax very wide in front. Front row of eyes straight, or
with the middle pair a little higher than the lateral, nearly equal in
size and equidistant, or the middle a little larger or farther apart
than they are from the lateral. Upper line of eyes longer and slightly
curved with the middle pair highest, the eyes all larger than those of
the front row, and the middle pair usually farthest apart. Plate v,
fig. 8c. Maxilla? long, narrow at the base, and much widened beyond
the insertion j)f the palpi. Fig. 10. The mandibles are stout and
convex at the base in the females. In the males the mandibles are
more slender, longer and tapering at the tips, sometimes with sharp
ridges along the sides. Legs slender, the fourth pair longest. Feet
with long claws, the first and second pairs with the under side of
tarsus and metatarsus covered thickly with hairs widened at the end.
The abdomen is truncated in front and tapering behind. The colors
are always pale gray and drab, usually with darker brown on the
head around the eyes, and rarely a light brown or gray pattern on the
back of the abdomen. The body is covered with short and fine hairs
which give it a soft silky appearance without concealing the color of
the skin.

180 J. n. Emerton — Keto England Drassidoe^

Most species vary greatly in size, some mature individuals being
twice as large as others of the same age and sex. They live on
plants in summer, and in winter hide under bark or stones, and have
at all seasons flat tubular nests of silk.

Clubiona crassipalpis Keyserling, specimens in Mus. Comp. Zool., Cam-
bridge, Mass.

Plate V, figures 1, la, Ih.

Male 8"^™ long, sometimes smaller. Cephalothorax two-thirds as
wide as long. Male mandibles convex in front for two-thirds their
length, with a low ridge along the inner side and around the end of
the convex portion. The thin keel on the front outer edge is sharp
but short, extending only a little over the convex part.

The tibia of the male palpus is as long as the patella. The process
on the outer side is long and slender, the end curved inward over the
tarsus. Plate v, figs. 1, \a, \h.

Abdomen marked with brown irregular veins, the rest of the body
pale. Head a little darker toward the front.

Massachusetts, Connecticut, Albany, N. Y., Providence, R. I., and
in N. Pike's Long Island collection.

Clubiona mixta, new sp.

Plate V, figures 'la, Ih.

Resembles C. crassipalpis and of about the same size. The man-
dibles of the male are similarly shaped, l)ut the convexity and the
internal ridge are less prominent. The male palpi are a little more
slender and the patella proportionally longer. The process of the
tibia has the upper tooth nearer the hook than in crassipalpis, mak-
ing the process appear wider and stouter, fig. 2. The tarsus is a little
smaller than in crassipalpis. Plate v, figs. 2rt, 2&.

Salem and Marblehead, Mass.

Clubiona tibialis, new sp.

Plate V, figures 3, .'{«, 3&.
Male 6*5""" long, another 5"'"' long. The male mandibles are slen-
der and tapering, without any distinct ridges on the front. The male
palpi are short and the tibia and tarsus both ver;y large. The tibia
is very complicated in shape, having a large hook on the outer side, a
short thick process on the inner side, and a thickened edge in front
that meets a slight elevation on the back of the tarsus. The tarsus
is long and large, and so is the palpal organ. Plate v, figs. 3, 3«.

Agalenidce and Dysderidce. 181

A female, apparently of this species, is 6""° long. The front legs
are shorter than in the male ; the mandibles are stout and convex in
front. The epigynum is large with a deep rounded notch in the
middle and a slight ridge each side. Fig. Zb.

Eastern Massachusetts, and in N. Pike's Long Island collection.

Clubiona canadensis, new sp.

Plate V, figures 4, 4a, Ah, 4c.

Male V"^'" long, others smaller. Mandibles tapering and rounded,
without ridges on the upper side. Male palpi short, tibia shorter
than patella, with a complicated process on the outer side ending in a
long sharp point with a round notch in the upper edge. Plate v
figs. 4, 4a. The tarsus is more than twice as long as wide, bent
downward at the end. The palpal organ has a large bulb with small
appendages at the end. Fig. 4J.

Female a little larger, epigynum with only two depressed spots
just in front of the transverse fold. Fig. 4c.

Abdomen dark, with brown irregular lines. Cephalothorax pale,
not darkened toward the front.

The common species on Mt. Washington, N. H., from the Glen to
the highest trees, under stones and in moss ; also from Montreal,
Canada.

Clubiona minuta, new sp.

Plate V, figures 11, lla, \]h.

This little spider is about 3'"" long and in its general appearance
resembles a pale C. rubra. The male palpi, however, show it plainly
to be a different species. The patella is longer than wide, as in
rubra, and the tibia is short and wide at the end. Its appendage
on the outer side is very simple, appearing from above like a thin
spine at the side of the tarsus and not overlapping it. PI. v, fig.
11. From the outer side it is seen to be flat, wide at the base, and
tapering from the middle to a blunt point, fig. 11a. The palpal
organ has a small hook on the inner side, fig. \\b, very different
from the large hook of C. rubra.

Male from Readville, Mass., June 15, on bushes.

Clubiona pusilla, new sp.

Plate V, figures 5, 5a, 5&.

One male 6™"^ long, another only 4"'°\ Head nearly as wide as
the thorax.
Trans. Conn. Acad.. Vol. Till. 24 Jan., 1890.

182 J. H. Emerton — N'eto England Drassidce,

Front row of eyes half as loug as the head is wide, the eyes of
equal size and equidistant; upper row, longer by the diameter of
the lateral eyes. The upper eyes are larger than those of the front
row, and all about the same size, the middle pair a little farther
apart than they are from the lateral eyes.

Mandibles with a thin keel on the front outer edge, half as long as
the mandible.

Colors, pale on the legs and palpi; cephalothorax brownish yellow,
darkest in front; abdomen covered with fine brown markings.

Male palpi with the tibia shorter than the patella, A flat wide
process on the outer side, PI. v, fig. 5 a, extends forward over the
tarsus. The bulb of the palpal organ nearly fills the under side of
the tarsus, the tube is short and curved round the end of the tarsus
so as to point backward ; over the base of the tube is a short stout
hook, instead of a large hook as in rubra, fig. 5.

Salem and Beverly, Mass.

Clubiona rubra Keyserling, specimeus in Mus. Com p. Zool., Cambridge, Mass.
Plate V, figures 6, Ga, 6&, 7, la, 1h, 8, 8ffl, Sb.

This is one of several closely similar species, the classification of
which cannot M^ell be understood without comparing large numbers
from many diiferent i)laces. Chihlona ahhottii L. Koch, is this spe-
cies or very close to it.

Males and females are 3 to 4"""' long. The eyes are large in pro-
portion to the size of the spider, and cover the whole width of the
front of the head.

The color after keeping in alcohol is redder than in most species.

The epigynum is notched at the hinder edge, the depth of the
notch varying in diflferent individuals. PI. v, figs. 6c, 7c.

The male palpi have the patella and tibia of the same length.
The relative length of these joints differs in the allied species. The
tibia is wider than long and has a large appendage on the outer side,
divided from the main part of the tibia by a deep notch on the
under side, figs. 6, 7, 8. The appendage consists of two parts, figs,
6a, 7a, 8a, the under one longest and a little notched at the end.
The size and length of this process varies in different individuals.
On the dorsal side of the tibia, on the front edge, is a small tooth,
varying in size in different spiders, figs. 6, 7, 8, The palpal organ
has a very large middle process, figs. 6^, 7^*, 8/>.

White Mountains, N. H., to Connecticut,

Agalenidm and Dysderidce. 183

Clubiona ornata, new sp.

Plate V, figures 9, 9a.

Female 8""" long. The abdomen is pointed behind and more
narrowed in front than in most species. Both abdomen and cepha-
lothorax are less flattened than in most species. The mandibles of
the female are not very stout and less swelled at the base than usual.

This is one of the few sjDecies with a colored pattern on the ab-
domen. A dark stripe runs along the middle, of a different width in
different individuals, but generally narrow and tapering behind. At
the sides of this are two white or light yellow stripes with irregular
edges, and beyond this the bi'own sides of the abdomen. PI. v, fig.
9. The body is pale underneath. The epigynum is notched at the
edge of the transverse fold, fig. 9a.

Mt. Washington, Dublin, N. H., and Massachusetts.

Clubiona excepta L. Koch.

Clubiona pallens Heutz.

Plate V, figures 10, 10«, 10^ 10c, \Qd.

Female 7'"™ long, cephalothorax 3""". The abdomen is not usually
much larger than the cephalothorax and unlike most species has a
pattern on the back similar to that of Aniaurohius and Tegenaria,
or in very light individuals consisting of three roAvs of gray spots
on a white or pale yellow ground. PI. v, fig. 10«. The cephalothorax
and legs are light yellowish brown, dai'kest on the head and mandi-
bles. The spinnerets are rather long. The epigynum has two round
brown spermathecae that show through the skin, close together just
in front of the transverse fold. In front of these are two oblicjue
openings directed forward and inward.

The males are not much smaller than the females. The male
palpi are slender, the tibia only a little longer than the patella, and
the tarsus nearly as long as both together. The tarsus is oval, about
half as wide as long, and rounded on the upper side. The papal
organ is narrow and covered by the tarsus. On the inner side is a
thin appendage, the free edge of which lies along the middle line
and covers the long straight tube. On the outer side near the end
of the tube is a straight process directed forward, and at its base a
hook directed backward, fig. 10c. At the end of the tibia on the
outer side is a short flat process with a small curved tooth on the
upper corner, tig. 1()^>.

Massachusetts, Conneaticut, and in N. Pike's Long Island collec-
tion. Under stones and leaves, sometimes in white cocoons.

184 J. H. Emerton — N'eio England Drassidce,

Chiracanthium C Koch.
Chiracanthium viride, new sp.

Plate V, figures 12, 12a, 126, \2d.

Female 8™'" long, cephalothorax: S™"". Ceplialothorax three-fourths
as wide in front as at the widest part, fig. 12a. Eyes spreading over
nearly the whole width of the head; the lateral eyes close together;
the upper row a little longer than the front row, eyes in both rows
at equal distances apart. Abdomen widest in the middle, tapering
behind. First pair of legs a third longer than the fourth. Sternum
widest just behind the first pair of legs and tapering to a point
between the fourth coxae. The mandibles and maxill?e are dark
brown. The rest of the body is pale yellow, the cephalothorax a
little darker than the rest, and a gray stripe covers the middle of
the front of the abdomen. The epigynum has a large oval opening
covered by a hard dark brown lump. PI. v, fig. \2d.

In the male the mandibles and legs are longer and the difference
in length between the first and fourth legs is greater. The palpi
are as long as the second femur. The tibia is twice as long as the
patella, and has on the outer side a tliin black process, a little curved
toward the tarsus, and on the upper side a thick blunt process ex-
tending a little way over the back of the tarsus. Between the tw^o
processes of the tibia a sharp process of the tarsus extends backward,
a little curved down at the end, figs. Via, 126.

Female, Dedham, Mass., July 26. Male, Saugus, Mass., June 12.

Trachelas L. Koch.
Trachelas ruber Kejseriing.

Plate V, figures 13, 13a, 13c, 13c?.

Female 10'"™ long, cephalothorax 4'""' long and 3'"'" wide. The
cephalothorax is widest in the middle opposite the second pair of
legs and narrows to 2'"'" at the hinder end, the sides of the hinder
half being nearly straight. The head is very wide and high, the
highest part half way between the eyes and the dorsal groove.

The eyes are all about the same size and far apart. The front
row is nearly straight, the middle eyes a little higher than the
lateral, this row is half as long as the head is wide. The hinder
row is much longer, the middle eyes are about as far from the front
middle pair as they are from each other, the lateral eyes are about
the same distance from the middle ones, but much farther back on
the head, figure 13. The mandibles and maxillte are large and resem-

Agalenidm and Dysderidm. 185

ble those of Cluhiona. The abdomen is oval and very regnlar in
shape. PI. V, fig. 13. The eephalothorax is very thick and hard,
and dark brown. The abdomen is light yellow with no markings,
except four small brown spots near the middle, and a gray streak
over the dorsal vessel. The hairs are very short and scattered so
that the skin appears soft and smooth.

The first pair of legs is a little the longest insteacl of the fourth
pair, as in the European species, and both the first and second pairs
are much stouter than the third and fourth. The palpi are slender,
the tarsal joint thickened at the tip. The legs are darker fi*om back
to fi'ont, the front pair reddish brown, not so dark as the eephalo-
thorax, and the hind pair is yellow. The epigynum has two dark
brown round depressions close together.

Pale individuals are sometimes found with all the legs yelloAvish
white, eephalothorax light brown with white eyes, and the abdomen
light gray.

The males are smaller than the females, sometimes not more than
half as large. The tibia of the palpus is shorter than the patella,
and has a short hook on the outer side. The tarsus is small and the
bulb of the palpal organ is so large that it extends beyond the tar-
sus on both sides. The bulb is round and has a distinct tube which
rests in a groove of the end of the tarsus, figs. 13c, 13f7.

Under stones and leaves and sometimes on fences in autumn. In
general appearance and color it resembles Dysdera. Massachusetts
and Connecticut, and in N. Pike's Long Island collection.

Anyphaena Sundeviiii.

Plate Yl, figures 1, la.

eephalothorax highest behind. Eyes of the front row equal in
size and equidistant, the lateral eyes a little the highest. Upper row
of eyes longer than the front row, the middle eyes highest, all of the
same size and larger than those of the front row and at equal dis-
tances apart. Abdomen widest in the middle and a little pointed
behind.

Maxillae long and widened at the tij^s but not so much widened as
in Cluhiona.

The opening of the trachea? is farther forward than in other
genera, in some species approaching nearly to the epigynum. PI.
VI, fig. la.

The colors are pale. The male palpi are large and complicated.

186 J. H. Einertou — JVeio Etigland Drassidm^

Anyphaena rubra, new sp.

Plate VT, figures 1, la, 16.

Female 8 or 9"^"^ long, cephalotliorax 3"'"\ Abdomen half longer
than the ceplialotliorax and about as wide, tapering backward from
the middle to the spinnerets, PI. vi, iig. 1. The cephalothorax and
legs are pale yellowish brown. The cephalothorax has two darker
longitudinal bands. The abdomen is white or light yellow with two
stripes made up of brown or red spots. The mandibles are dark
brown. This is the largest and stoutest species.

The epigynum has two large curved openings, turned toward each
other, between which is a long depression widened at the front end.
The long spermatheca^ show through the skin just behind the open-
ings, fig. 1 h. I have not seen the adult male.

Massachusetts and Connecticut, and in N. Pike's Long Island col-
lection. On plants and under stones.

Anyphaena incerta Keys., specimens in Mus. Comp. Zool., Cambridge, Mass.
Plate VI, figures 2, 2a, 26, 2c, 2d.

Female 5™™ long, cephalothorax 2™"'. The cephalothorax is about
a quarter longer than wide, rounded at the sides, and highest in the
middle. The front of the head is very low, so that the front eyes are
not their diameter from the base of the mandibles. The front row
of eyes is nearl^^ straight. The upper row is longer and more
curved, with the middle eyes highest, and the eyes of this row are
all larger than those of the front row. The abdomen is large in the
female, as in all the species of this genus, widest just behind the
middle and a little pointed behind, PI. vi, fig. 2.

The color is light brownish yellow with gray markings. The
cephalothorax has two indistinct longitudinal stripes and a fine black
line over the leg« on each side. On the abdomen are two rows of
faint spots and oblique lines. The legs have a few faint markings
across the joints.

The maxillje are straight at the sides and rounded at the ends on
the inner side. The labium is small and not half as long as the
maxillae.

The epigynum has a large dark brown process in the middle at
the front end, fig. 2d.

The tibia of the male palpus has a large double process on the
outer side, the upper branch of which is pointed, and the lower blunt
with a rounded tooth on the upper side, figs. 2a, 2b, 2c.

Under leaves in winter, Salem and Swampscot^, Mass.

Agalenidm and Dysderidce. 187

Anyphsena calcarata, new sp.

Plate VI, figures 3, 3a, 3&, 3c, 3d

The same size as A. hicerta, but lighter colored and with longer legs
and longer spines. The front legs are longer than the fourth in both
sexes. The markings are the same as in the other species and the
spots on the front of the abdomen are more distinct than in the
others.

The epigynum has a thin edge extending backward a little over
the transverse fold and reaching from one respiratory opening to the
other. In the middle is a small hole with a short tooth-like ridge
directed backward on each side. PI. vi, fig. '3d.

The male palpi have the outer half of the femur twice as thick as
the base with a few large spines on the upper side near the end.
The patella is as wide as long and shorter than the tibia. The
appendage on the outer side of the tibia is very small and does not
extend forward beyond the base of the tarsus, fig. Zb, 3c ; near the
base of the tibia on the under side is a blunt tooth, fig. 3«, 3c. The
tarsus resembles that of A. incerta. The palpal organ has the
middle process very stout and curved inward at the end, fig. 3a.

The coxa3 of the fourth pair of legs have on the under side a small
pointed process directed outward. The coxge of the third pair have
on th^ under side a curved process directed inward with a short
tooth on the hinder side near the middle, and in front of this a short
blunt tooth directed backward, fig. 3.

West Haven, Conn., July, on plants, and in N. Pike's Long Island
collection.

Anyphsena saltabunda.

Cluhiona saltabunda Hentz.

Plate VI, figures 4, 4a, 4b, 4c, 4d.

This is a veiy long-legged and slender species. The female is 4™""
long, the abdomen but little longer than the cephalothorax. The
front leg is 10"'™ long, fourth leg 7"'"'. The palpi are slender and as
long as the femora of the first legs. The whole body is white with
two broken gray bands on the cephalothorax and two rows of gray
spots on the abdomen.

The male is about as large as the female. The male palpi are lono-,
the tibia of very complicated shape. It is curved outward and has
near the base on the outer side a long, thin forked process. PI. vi,
fig. 4. The tarsus is of the usual shape. The palpal organ has a

188 J. H. Emerton — ISTew England Drassidm,

short slender tube resting against the tip of the tarsus. Behind the
tube is a thin hooked process, and on the inner side a long process
with small black teeth at the end, fig. 4, 4a, \h, 4c.

The epigynum has a long transverse opening a little in front of
the fold, fig. 4f?.

Massachusetts, and Meriden, Conn.

Phrurolithus Koch and Westring.

Micariosoma Simon.

Small spiders sometimes with bright markings and iridescent
scales. The legs of the first and second pairs have a double row of
strong spines under the tibia and metatarsus. PI. vi, fig. bh. The
maxillae are short and wide. The palpi of the males are very large
compared with the size of the spider, and have a long stout process
on the outer side of the tibia. The arrangement of the eyes and the
pattern of the dorsal markings resemble those of jLgroeca.

Phrurolithus pugnatus, new sp.

Plate VI, figfres 6, &a, 6&, 6c.

2™"^ to 3'"™ long. Cephalothorax round, narrowed at the head as
in alarius. Abdomen usually shorter and rounder than in alarms.
PI. VI, fig. 6. Cephalothorax and legs bright yellowish brown.
Abdomen dark brown with transverse light markings which vaiy in
different individuals. Light yellowish beneath, except around the
spinnerets and epigynum.

Epigynum with two oblique openings at the front end fai-thest
from the transverse fold. Parts of the })alpal organ are sometimes
found in the openings of the epigynum, fig. 6e.

The male palpi are large in proportion to the size of the spider.
The femur has a short process near the base on the inner side. The
patella is as short as wide, but the tibia is nearly as large as the
tarsus and wider at the distal end. On its inner side is a long stout
tooth projecting forward, and on the outer side a longer curved one
as in P. alarius. The tibia is oval and the palpal organ short and
round, not extending backward at the base as in alarms, figs. 6a, 6b.

Herpylliis parens Hentz resembles this species.

Massachusetts and Connecticut.

Agalenidce and Dysderidce. 189

Phrurolithus alarius.

Herpyllus alcmus Hentz.

Plate VI, figures 5, 5a, 5&, 5(Z, 5/, 5^^, 5/;.
Full grown female 4'"'" long, cephalothorax 1.5'""\ PI. vi, fig. 5.
The cephalothorax is nearly as wide as long, rounded at the sides.
The head is about half as wide as the thorax and the eyes are close
together and all about the same distance apart, fig. 5a. The middle
eyes of the upper row are oval and turned obliquely, nearest together
towards the front.

The abdomen is oval, widest behind, and a little flattened on top.
The legs are long and slender, except the tibi?e and metatarsi of the
first and second pairs which are twice as thick as the same joints of
the other legs, fig. 5. The legs are light yellow or white with gray
hairs, except the tibia and patella of the first pair, which are black
or dark gray with the tip of the tibia white. The tibia and patella
of the second pair are marked with lighter gray in the same way.
The tibia and metatarsus of the first and second pairs have two rows
of strong black spines on the under side, fig. bb.

The cephalothorax is light yellowish with a black edge each side
and a few irregular radiating gray marks forming two indistinct
longitudinal stripes. The abdomen is gray with transverse white
markings which vary greatly in shape and size in diflrerent individuals,
figs. 5, 5a. The abdomen is covered with fiat branched hairs that
are iridescent, changing from light grayish-green to pink with the
motions of the spider, fig. 5d. The under side of the body is pale
with a dark mark in front of the spinnerets, and in some individuals
a few irregular marks along the sides.

The male palpi are large. The femur is thickened on the under
side near the outer end, forming a short black pi'ocess covered with
short stiff hairs. The patella and tibia are both short. The tibia
has on the outer side a long process slightly curved downward that
extends along the side of the tarsus for half its length, fig. bg. The
palpal organ is so long that its base extends over the end of the
tarsus, fig. 5f.

The epigynum has two lai'ge openings turned toward the sides a
little in front of the transverse fold, fig. 5h.

It lives on and under stones in dry open ground and runs with
great swiftness short distances at a time. When still it lies close to
the stone with the tibiae drawn up over the back, as in fig. 5, the
thickened and colored legs of the first pair are then the parts of the
spider most easily seen.

Massachusetts and Connecticut.

Trans.. Conx. Acad., Vol. VITI. 25 Jan., 1890.

190 J. H. Emerton — IVew England Drassidm,

Agroeca Westriug.

AgrcBca pratensis, new sp.

Plate VI, figures 7, la. lb, 1c, Id, 1e.

Female T'"'" long, cephalothorax S"^-". The cephalothorax is widest
and highest behind the middle, the head a little more than half as
wide as the thorax. The abdomen is widest across the hinder third
and not much pointed behind. PI. vi, fig 7. The front roAV of eyes
are close together, the middle ones half their diameter highest. The
upper eyes are a little larger and about their diameter apart, the raid-
die pair much the highest, fig. 7.

The mandibles are very convex in front and flat at the sides. The
maxillae are straight on both sides and a little roimded on the inner
corners. The labium is half as long as the maxillae and as Avide as
long. The sternum is large and as wide as long, fig. 7a. The legs
are stout, the fourth pair longest. Under each metatarsus are three
pairs of slender spines, under the first and second tibiae two pairs, and
under the third and fourth tibia? three pairs. The cephalothorax,
legs, and mouth parts are light brownish yellow. The cephalothorax
has a fine dark edge on each side and a row of radiating dark lines
each side forming two broken dark longitudinal bands. The abdo-
men has two rows of gray oblique markings on a light ground, fig 7.
Epigynum with a long brown piece in the middle, fig 7e.

Male about the same size with the abdomen a little smaller. The
male palpi are large and stout. The patella and tibiae are the same
length, the latter a little bent and with a short pointed spine on the
outer side. The tarsus and palpal organ are short and wide. The
palpal organ has a short blunt process on the outer side that projects
over the edge of the tarsus, figs. 76, 7c, Id.

This spider lives under leaves and in short grass and resembles a
Lycosa in its gait and genei'al appearance, and also the common
Anyphmna incerta.

Eastern Massachusetts ; Providence, Rhode I. ; Albany, New York.

Agalenidse.

The Agalenidce have the cephalothorax longer than wide, with the
cephalic part higher than the thoracic, and distinctly separated from
it by grooves or marks at the sides. The head is usually higher
than in the DrassidcB and the body less flattened. The upper spinner-
ets are two jointed, the terminal joint pointed and provided with
si)inning tubes along the inner side. In most species these are longer

Agalenidm and JDysderidce. 191

than the other spinnerets. The feet have three claws. The Agalen-
idm make large flat or irregular webs with a tube or hiding place at
one side from which they run out and seize the insects that alight on
the web. The Agalenidm run on the upper side of the web with their
back upward, while LinypMa, which makes similar flat webs, runs on
the under surface, back downwards.

CcBloteS Blackwall.

The difference between Coilotes and Tegenaria is not a very dis-
tinct one. I have placed in Ca?lotes those stouter and shorter legged
species with the mandibles prominent in front, and in which the palpi
of the males have processes on both patella and tibia.

The eyes are in two rows, nearly straight, and differ but little in
size and distance apart. The mandibles are stout and convex. The
maxilh^ are wide at the ends, rounded on the outer corners, and
obliquely truncated on the inner. The labium is about half as long
as the maxilla;, a little narrowed and truncated at the tip. The
colors are dark gray and brown.

Coelotes medicinalis.

Tegenaria medicinalis Hentz.

Plate VII, figures la, \h.

Female 12"^'" long ; cephalothorax 5'"'", fourth leg 15'""', Front
row of eyes straight, the middle eyes largest, eyes of upper row all
of the same size and about equal distances apart, the middle eyes
highest. The lateral eyes of both rows are close together, those of
the upper row farthest toward the sides. Head high and wide, dis-
tinctly separated from the thoi'ax by grooves each side. Abdomen
oval, widest behind. Legs moderately stout.

Cephalothorax yellowish brown, darkest in front, marked with
radiating gray lines forming two longitudinal dark liands. Abdo-
men gray Avith irregular pale spots. A double row of oblique pale
spots in the middle, in most specimens broken and irregular, tig. 1.
The legs are light yellowish brown Avith light indistinct gray rings.

The spinnerets are short.

The epigynum has a large piece in the middle with a branch each
side that extends outward and forward ending in a point. In front
are two small rounded teeth directed inward. PI. vii, fig. la.

Male palpus short, with a large and wide tarsus. The patella and
tibia are both short and of about the same length. The outer pro-

192 J. H. JEmerton — New England Drassidm,

cess of the patella is half as long as the patella and blunt and crooked
at the end. The tibia has on the uppei- side a short pointed process
near the base and a blunt one near the end on the outer side, both
concealed by a curved ridge. On the under side of the tibia is a
process directed forward. The tube of the palpal organ is slender
and supported at the end by two large processes, hg. \h.

Chateaugay Lake, Adirondacks, N. Y., from F. A. Bowditch, 18 7 8 ;
Swampscott, Mass., May 8.

Coelotes longitarsus, new sp.

Plate Vir, figures 2, la.

Male 7™"" ; cephalothorax .3*5'""\ Head wide and high, highest
half way between the eyes and tlie dorsal groove. First leg almost
as long as the fourth. • Legs stout. Abdomeft oval, the hinder half
a little the wider. Cephalothorax and legs yellowish brown, a few
radiating darker lines on the thorax, and the front of the head a little
darker. Legs darker toward the tips. Abdomen dark gray with a
median lighter stripe in front and a double row of lighter oblique
marks on the hinder half, much as in medicinalis.

The patella and tibia of the male })alpi are both as sliort as wide.
The patella has a long tooth, widest and truncated at the tip, directed
forward on the outer side. The tarsus is widest at the base and
pointed at the tip. At the base on the outer side it has a stout
process extending backward and inward as far as the patella. Plate
VII, fig. 2.

A small female found at the same time and probably the same
species has a small, simple oval opening at the posterior part of the
epigynum. Fig. 2a.

Mt. Carmel, Conn.

Coelotes montanus, new sp.

Plate VI I, figures 3, 3a.

12"^°^ long ; cephalothorax S™"" ; 4th leg of female 18'"'", 4th leg of
male 20""".

Epigynum with the holes open, and oblique turned nearly forward.
Plate VII, fig. 3.

Male palpus with short patella and tibia, the tibia shortest. The
patella has a short conical, black spine on the outer side near the
end, directed forward, under this is a smaller spine. On the outer
side of the tibia, which is shorter than the inner side, is a short
process turned forward a little at the tip. Fig. 3a.

Agalenidm and Dysderidm. 193

The colors and markings are like those of the two preceding
species. The legs, especially of the male, are a little longer in pro-
portion to the size of the body.

Chateaugay Lake, Adirondacks, N. Y., from F. A. Bowditch. A
small male from New Haven, Conn., is apparently of this species.
Its palpi differ slightly from those of the Adirondack males as shown
in figs. 4, 4a.

Ccelotes hybridus, new sp.

Plate VII, figures 4, Aa.

This species is only distinguished from C longitarsus by small
differences in the shape of the male palpi. The spur at the base of
the tarsus is very short and extends backward to a short process on
the middle of the tibia. The posterior inner corner of the tarsus is
differently shaped from this part in longitarsus, having a deep notch
shown in figs. 4, Aa. The tibia is a little longer than that of
longitarsus and shaped much like that of niedlcinalis. The patella
resembles that of longitarsus. In size, markings, and colors this is
like both rnedicinalis and longitarsus. The palpal organ is almost
exactly like that of longitarsus.

One male from Chateaugay Lake, Adirondacks, N. Y., from F. A.
Bowditch.

Tegenaria Latr.

These spiders differ from Cmlotes in having the legs longer and
more slender and the abdomen generally rounder and shorter. The
mandibles are less convex in front, the palpi of the males have no
processes on the patella, and the palpal organ is proportionally smaller
than in Coelotes.

Tegenaria derhamii Scopoii, 17G3; Thoreii, i87?..

Tegenaria civilis Blackwall, 1861.
Tegenaria domeslica Simon, 1875.

Plate VII, figures 6, 6a, Gb, 6c.
This is a house spider found all over the world. Female 10"™
long ; cephalothorax S'"'". Plate vii, fig. 6. The legs are long and
tapering, 4th leg 18™'" long. Tarsi and metatarsi slender. The head
is high and wider in front than it is opposite the first pair of legs.
The abdomen is short, only a little longer than the cephalothorax.
The front row of eyes is straight, the middle ones smallest. iTpper
row longer, lateral eyes close to those of the front row, the middle
ones much higher, fig. 6.

194 J. H. Emerton — New England Drassidm,

Cephalothorax and legs light yellowish brown, the legs with some
indistinct gray rings. Abdomen pale with gray markings which are
usually plainest on the hinder half. The upper spinnerets are twice
as long as the lower and the terminal joint is nearly as long as the
basal.

Epigynum with small oblique openings at the sides. Fig. 6c.

The male palpi have the patella and tibia long and both about the
same length. The tibia has on the outer side, about one-fourth its
length from the end, a short blunt process, fig. 6a, under this is a
shorter process lighter colored and directed forward, fig. Qh. The
tarsus is slender and pointed and the appendages of the i:)alpal organ
are small. Figs. 6a, 6^.

Tegenaria brevis, uew sp.

Plate VII, figures 5, 5a, 5/*, 5c.

A small species, 5 to 6""^ long. Cephalothorax two-thirds as wide
as long. Abdomen short and widest behind. The mandibles are
small and but little convex in front. The cephalothorax and legs are
pale yellowish brown with black hairs, the legs and palpi are lightest
at the base and darker toward the ends. The spines are very long
and slender. The abdomen is in some individuals pale with dark
hairs ; in others there is a gray herring-bone marking, and gray
marks along the sides.

The male palpi are slender and without any appendages on the
patella. The tibia has a short stout tooth on the outer side a little
behind the end. The tarsus is small and pointed. The palpal organ
is round and too large to be covered by the tarsus. Plate vii, fig. 5.
The head of the male is narrower than that of the female, and the
thorax wider, figs, bh, 5c.

The epigynum appears to the naked eye like two parallel dark
brown marks. It has a large posterior opening, widest behind, and
partly divided into two at the front edge, fig. 5a.

The short round abdomen and gray markings make this S2)ider
resemble Steatoda marmorata.

Mt. Washington. N. H. ; Massachusetts ; New Haven, Conn.

Cicurina Menge, ]»71. Simon, J875.
This genus differs but little from Coelotes except in the palpi of
the males which have the patella without processes, the tarsus long
and narrow, and the tibia short with a large appendage on the outer
side. The tube of the palpal oi'gan is long and supported in various
complicated ways. .

Agalenidm and Dysderidce, 195

Cicurina complicata, now sp.

Plate VII, figures 7, 7«, 76.

This is a small, stout species, the largest individuals iiieasurinsf
7'"™ in length, and the cephalothorax 3'""\ The cephalothorax, legs,
and palpi are uniform yellowish brown. The abdomen is pale with
scattered gray markings both above and below, in some individuals
forming an indistinct herring-bone pattern on the dorsal side. The
sternum is as wide as long and nearly as wide jn front as in the mid-
dle. The mandibles are stout, and in the females very convex in
front. In the males the head is narrower and the mandibles smaller.

The palpi of the males have the patella simple and about as long
as wide. The tibia has a short tooth near the base on the outer side
the middle part is turned inward, and on the outer side at the end is
a large flat and crooked appendage that in its natural position
appears to be part of the palpal organ. Plate vii, fig. 7. In fig. 7a
the process is shown from the side partly tiirned away from the pal-
pal organ. The tarsus is long and narrow and rounded at the tip,
fig. 76. The tube of the palpal organ is very long, beginning at the
base of the tai'sus it runs along the inner side around the tip, whei-e
it is supported by the edge of a flattened appendage, and backward
along the outer side, the end being under the flat tibial process.

The epigynum has a simple oval opening behind, and the tubes
show through the skin in light-colored individuals.

•Blue hill, Milton, Mass., and Salem, Mass., under leaves, in winter.

Hahnia Koch.

Upper eyes all about the same size, the middle pair, highest and
farthest back and farther from each other than they are from the
lateral eyes. Front eyes nearer together, the middle ones highest.
The lateral eyes of both rows close together.

The spinnerets are in a single transverse row, the upj^er pair being
outside the others. The outer pair has two nearly equal joints and
the next pair have a short second joint at the tip.

The opening of the tracheje is in the middle of the abdomen
instead of dii*ectly in front of the spinnerets, as in most of the
family.

Small spiders resembling Tegenaria and delotes, except in the
spinnerets.

196 J. IS. Emerton — New England Drassidm,

Hahnia bimaculata, new sp.

Plate VII, figures 8, 8a, to 8/.

Length, 2-5""°. Eyes lai-ge, both rows with the lateral eyes low-
est. Lateral eyes much nearer together than the middle pairs.
Front eyes a little the larger, both rows strongly curved, the middle
eyes highest. Maxillae short and wide, the front edge straight
except on the inner corner. Sternum as wide as long, widest oppo-
site the second legs. The spinnerets are long, the terminal joints of
the outer pair nearly as long as the basal joint. The tracheal open-
ing is nearer the epigynum than the spinnerets. Plate vii, fig. 8.

The cephalothorax, sternum, and mouth parts are reddish brown.
The abdomen is light gray with many irregular pale spots and a
double row of pale oblique markings in the middle. In the middle
of the front half of the abdomen are two orange colored spots. The
under side of the abdomen is pale with a few gray spots. The legs
are pale with gray rings, two rings on the femur, tibia, and metatar-
sus. The skin over the epigynum is very transparent and shows
two convoluted tubes almost always unsymmetrical, figs. 8c, c?, e, _/'.
The male palpus has on the outside of the tibia, near the end, a
pointed process as long as the diameter of the tibia. At the base of
the patella on the outer side is a small pointed black process curved
a little forward, fig. 8^. The tarsus is nearly as wide as long but
slightly pointed at the tip. The palpal organ is flat with a long
thin tube extending along the inner side around the end, fig. 8a.

Common under dead leaves. Massachusetts, Connecticut, and Mt.
Washington, N. H.

Hahnia radula,

new sp.

This spider resembles II. bimaculata but is nearly twice as large.
On the under side of the first and second legs and pal])i of the male
the hairs are raised on short transverse ridges so that the leg ajjpears
serrated when seen from the side. Plate vii, figs. 10, 10a. The gray
markings of the abdomen and rings around the legs are less distinct
than in hiinaculata. The little spine at the base of the patella of
the male palpi is shorter than in bimaculata and sharply turned for-
ward. The appendage of the tibia is the same as in bimaculata, and
the tarsi and palpal organs are very similar.

One male, Jaffrey, N. H., Aug. I.

Agalenidce and Dysderidce. 197

Hahnia cinerea, new sp.

Plate VII, figures 9, 9a, 96.

Length, 1-5 to 2™°^.

Cephalothorax light, with dark radiating markings. Abdomen
dark gray with scattered small white spots and a double median
row of oblique light markings somewhat like Cmlotes. Plate vii,
fig. 9. The legs are light yellowish brown Avith patella?, coxae, and
the ends of the longer joints paler than the rest. The basal joints
of the s])innerets are light yellowish brown like the legs. Terminal
joint of outer spinnerets shorter than basal. The tracheal openings
are nearer the spinnerets than the epigynum. The skin over the
epigynum is rather opaque and but little of it can be seen. The
palpi of the male have the patella and tibia both short and each has
a long, slender process on the outer side which is flexible and
variously curved at the end, fig. 9/>. The tarsus is short and oval.
The palpal organ has at the base a short feather-like appendage.
The tube is slender and curved around the distal end of the tarsus.
It has near the end a short soft appendage of the bulb, fig. ^a.

Salem, Beverly, Swampscott, Cambridge, Roxbury, Mt. Tom,
Mass., and New Haven, Conn.

Agalena Waick.

Large hairy spiders with long legs and very long upper spinnerets.
The head is high and the middle eyes of both rows are much higher
than the others. The web is flat and more regular and closely woven
than in Tegenaria.

Agalena naevia Walck. and Bosc, 1841; Heutz, 1848.

^grafena j30<fen Blackwall, Ann. and Mag. Nat. Hist., vol. xvii, 1846.
AgaUnopsis alhipilis G-iebel, Zeitsch. Gesammt. Nat., 1869.

Agalena americana Keys., Zool. botan. Gesellsch., Wien, 18*77, male witli short-
tubed palpal organs, from Illinois.

Plate VIII, figures 1, la, Ic, Id, If, Ig, Ih, li, IJ, Ik, U, Im, bi.

This is the common grass spider all over the United States. It
varies greatly in size. A lajrge male measures 14™'" long, 4th leg
35""". A large female, 18""" long, 4th leg 30""", while a small adult
male is only 7""" long, and the 4th legs 15™"*. Plate viii, fig. 1.

The cephalothorax is long and the cephalic part separated dis-
tinctly from the thoracic by grooves radiating from the dorsal
depression. The head is high and wide in front and contracted a
little just in front of the first pair of legs. The two rows of eyes

Trans. Conn. Acad., Vol. VIII. 26 Jan., 1890.

] 98 J. H. Emerton — Neio England Drassidm,

are strongly curved, tlie lateral much lower than the middle pairs, so
that the middle f I'ont eyes and the lateral hinder eyes form together
a nearly horizontal line. The thorax is marked by radiating grooves
between the legs. The abdomen is about twice as long as wide,
widest in the front half, a little truncated in front where it overlaps
the thorax, and tapering behind. The upper spinnerets are two or
three times as long as the under. The legs are long and tapering,
the fourth pair longest. The maxillae are much widened at the tips
and nearly straight on the front edges.

The cephalothorax has two wide longitudinal dark stripes. The
abdomen has a light longitudinal stripe in the middle, straight in
front, and herringbone-shaped in the hinder half, generally a little
darkened in the middle and lightest at the edges. The sides of the
abdomen are dark, or covered with dark spots close together toward
the middle stripe and more scattered toward the sides. On the ven-
tral side the abdomen has a middle dark stripe, sometimes lighter in
the middle.

Though the markings vary but little the colors vary from light
yellow, with pale gray markings, to dark reddish brown, with black
and gray spots, the colors being usually modified by long gray hairs
both in dark and light individuals. The joints of the legs are all
dark toward the end. Large individuals are, as a rule, darker col-
ored than small ones.

The palpi are long in both sexes. In the males the femur is long,
the patella not much longer than wide, and without apjDcndages ;
the tibia about as long as patella, widened at the distal end, the
outer side extending forward along the edge of the tarsus and having
a short blunt tooth, figs. \c, \g, \d. The tarsus is large, the basal
half oval and the tip narrowed into a long point. The palpal
organ, especially the tube, which is largely developed in this species,
instead of having a constant form, as in most spiders, varies
extremely. The most common form is that shown in tigs, la, 1^,
with a stout flat tube coiled in one and a half turns under the tarsus,
and with the tip turned outward away from the tarsus. On the
outer side of the palpal organ near the end of the tube is a short
thin tooth with the outer edge turned downward and the corner
usually forming a blunt tooth directed toward the end of the palpus.
This variety is found in spiders of all sizes and shades of color from
all parts of the country. Among large spiders from various locali-
ties occurs the form of palpal organ shown in fig. \h. In this the
tube is much longer and more slender, and tei'minates in a sharp

Agalenidm and Dysderidm. 199

point tui'iied inward toward the tarsns. A third variety, shown in
fig. 1(7, occurs less often but on spiders of all sizes and from differ-
ent parts of the country. The spiral here hardly makes more than
one turn and is so small as to be covered entirely by the tarsus. At
the tip the tube is twisted so as to turn the opening downward.
This is the form named Agalena americana by Keyserling, in 1877,
from Illinois ; there is one in the Cambridge museum, from Penikese
Island, Mass., named by Keyserling " var. americana,'''' and I have
seen specimens from Indiana, from Providence, R. I., New Bedford,
Mass., and Brooklyn, N. Y., in N. Pike's collection. These three
varieties seem to be distinct and I have seen no intermediate forms.
Fig. le is a palpal organ from Providence, R. I., having an unusually
large tube ; fig. l/'is the palpus of a small spider from Jaifrey, N. H.,
in which the tube is slender and the spiral unusually small.

The shape of the external opening of the epigynum is even more
variable than that of the palpal organ. The most common variety
is shown in fig. \h, taken from a female found in copulation with the
'male from which the palpus fig. \a was drawn. Figs, l^, \j, \Jc,
show a slight variation from this form by short teeth on the front
edge of the opening. Figs. \l, 1/h, Iw, have these teeth united and
extending backward across the opening nearly dividing it into two.
The three last are all fi'om large dark colored spiders like those hav-
ing palpi as in fig. Ih.

Comparison of a large number of specimens from the neighbor-
hood of Boston, Mass., showed that 69 males had palpi like fig. la,
and 5 like fig. \h ', 98 females had the oval epigynum, fig. \h, and 37
the partly divided epigynum, figs. 1/, \m, \n.

The web of this species consists of a flat sheet, shaped accord-
ing to the supports to which it is fastened, from one side of
which extends a tube at the mouth of which the spider usually
stands. The tube is open at the lower end, from which the spider
escapes if the web is entered by too large an enemy.

The webs are made in all kinds of places. In early summer great
numbers are made on short grass, but large webs are seldom made
in such situations and it is probable that spiders that do not find
more favorable places as they grow lai'ger, never live to become
adult. The largest webs and the best developed spiders are found
among stones and shrubs where there are convenient hiding places
and supports for the web, Avhich in a good situation is enlarged as
the spider grows until it becomes a foot or more wide and propor-
tionally thick and strong. The long spinnerets are used in making

200 J. H. Emerton — New l^igland Drassidm,

this flat web, the spider walking along slowly, swinging the spin-
nerets from side to side, making a band of very fine threads at each
stroke. The web does not appear to be at all adhesive, it merely
offers insects a convenient place to rest upon and the spider depends
on his quickness of movement for their capture. Large webs usu-
ally have many supporting threads running up into fences and
bushes and these 'perhaps help to trip the wings of flying insects
and cause them to fall on the web, as similar threads do in the webs
of Linyphia.

The pairing of this spider takes place on the web of the female.
The female lies still with feet drawn up as if dead. The male lays
her on one side under his thorax with her ventral side forward and
inserts one of his palpi into the epigynum at frequent intervals for a
long time, the soft parts of the palpal organ suddenly swelling and
again contracting. When tired with one palpus he turns the female
around and over so that she lies on the other side with her head in
the opposite direction and uses the other palpus. The eggs are laid
in a flat white cocoon, usually covered with a thick flat cone of silk
with which considerable dirt is often mixed. The eggs are laid
under stones or bark and on fences and buildings of all kinds, where
they are partly sheltered, from August to October, and the females
often remain and die on or near the cocoon. Adults are occasionally
found under leaves in winter, but it is doubtful if any live until the
next season. The eggs hatch early in the spring and the young spi-
ders come out in May.

It appears to be the most common spider all over the United
States.

Dysderidse.

Spiders with only six eyes and with the openings of the tracheae in
the front of the abdomen, just behind those of the air sacs, so that
they appear to have four air sacs like the Mycjalidm. The family is
a small one and the genera differ greatly in the structure of the feet
and mouth parts.

Dysdera interrita Hentz.

Plate VIII, figures 2, 2a, Ih, 2c, Id.
Female 12™"^ long. Cephalothorax 5™"> long and S'""^' wide. The
front of the head is wide and curved forward in the middle. The
eyes are small and close together. PI. viii, fig. 2. The mandibles
are half as long as the cephalothorax and inclined forward and much
narrowed toward the end. The maxillae are small, pointed at the

Agalenidoe and Dysderidm. 201

tips and widest half way to the base of the palpus. Fig. 2a. The
labium is long and widened at the base. The sternum is widest in
the middle, narrowed behind, and truncated at the front end. The
coxae are very long, fig. 2«. The first legs are longest and the fourth
next. The feet have only two claws and under them a thick brush of
flattened hairs. Fig. 'ib. The patellne are only about a fourth shorter
than the tibiae. The abdomen is long, oval and a little pointed behind.
The cephalothorax and mandibles are reddish-brown. The legs are
lighter colored and more yellow and become a little darker from behind
forward. The abdomen is dirty white or yellow without markings.

The male differs little from the female. The palpal organ is as
long as patella and tibia of the palpus. The terminal half is a
little curved inward, and on the outer side is a short blunt tooth a
little curved upward. Figs. 2c, 2d.

Swampscott, Brookline, and Roxbury, Massachusetts. This is the
only Dysdera I have seen from New England, and as Hentz's D.
mterrita came from Massachusetts, this is probably the species. It
agrees very closely with D. crocata Koch = D. ruMcunda Blk.
The palpal organ of D. mterrita is straighter, as seen from in front,
than that of crocata.

Ariadne Savigny and Aiulouin.

Ariadne bicolor.

Pylarus hicolor Hentz.

Plate VIII, figure ?,, 3a, 3h, 3c, 3d.

Female 9™"' to 10"^™ long. Plate viii, fig, 3. Cephalothorax long
and narrow, widest opposite the third pair of legs. Jn the male the
cephalothorax is proportionally much wider. The dorsal groove is
very small, and the head is not separated very distinctly from the
thorax. The abdomen is oval, widest across the middle. The first,
second, and third pairs of legs are turned forward. The first pair is
longest, the second next. The legs are all stout and the first and
fourth pairs have the patella and tibia much thickened. The color of
the cephalothorax and legs is darker from behind forward, the fourth
legs being light yellow and the first legs and front of the head dark
brown. The abdomen is pale at the sides and dark purplish-brown
above and below, darkest along the middle of the back.

The feet have three claws, fig. 8c?. The tibia and metatarsus of
the first and second legs have two rows of strong spines on the under
side, four pairs on the tibia and eight or ten pairs on the metatarsus.
The sternum is long and widest in the hinder half. The maxilla? are

202 J. H. Emerton — N'ew England Drassidce, etc.

long and narrow, widened a little half way btween the tip and the
insertion of the palpus. The palpi are short and stout, fig. 3. The
middle eyes are close together. The upper lateral eyes are about
twice their diameter from the middle pair, and the front eyes are
close to them, about half their diameter nearer the middle line.

The male is a little smaller than the female and has the thorax
wider and the legs longer and more slender, fig. 3a. The metatarsus
of the first feet is crooked at the base with a spine on each side, the
outer one nearest the base, fig. 3a. The male palpi are but little
longer or stouter than those of the female. The tibia is a little
thickened. The palpal organ is attached to the under side of the
tarsus ; it has a round bulb about as thick as the tibia is long,
which narrows on the outer side into a short finely pointed tube
tliat curves sharply inward, fig. 3a.

It lives under stones and leaves, or in long yellowish tubes only
wide enough to hold the spider under stones or in cracks of trees.
In July and August the cocoon with twenty or thirty eggs is made
in the tube with the female, and the young come out of the cocoon
and live in the tube for a short time with the female.

Massachusetts, Connecticut, and in N. Pike's Long Island collec-
tion.

INDEX.

Agalena, 107.

americana Kej^s., 197.

na3via, 197.

potteri Blk., 197.
Agaleuidae, 190.

Agalenopsis albipilis Giebel, 197.
Agrceca Wstr., 190.

crocata Keys., 168, 171.

pratensis, 190.
Anyphzena Simd., 185.

calcarata, 186.

incerta Keys., 187.

rubra, 186.

saltabimda, 187.
Ariadne bicolor, 201.
Castianeira Keys., 169.

bivittata Keys., 169.
Chiracanthium Koch, 183.

viride, 183.
Cicurina, 194.

complicata, 195.
Coelotes Blk., 191.

hybrid us, 193.

longitarsus, 192.

modicinalis, 191.

moutanus, 192.
Clnbiona Latr., 179.

abbottii L. Koch, 182.

cauadensis, 181.

crassipalpis, 180.

excepta L Koch, 183.

minuta, 181.

mixta, 180.

ornata, 182.

pallens, 183.

pusilla, 181.

rubra, 182.

saltabunda. 187.

tibialis, 180.
Drassida;, 166.
Drassus, 178.

robustus, 179.

saccatus, 178.

variegatus, 174.
Dysdera interrita, 200.
Dysderidse, 200.
Geotrecha. 168.

bivittata, 169.

crocata, 171.

pinnata, 170.
Gnaphosa Latr., 175.

brumalis, 175.

Gnaphosa consper?a, 176.

gigantea, 176.

sciidderi, 175.
Hahuia Koch, 195.

bimaculata, L96.

cmerea, 197.

radula, 196.
Herpyllus alarius, 188.

ater, 172.

auratiis, 167.

bilineatus, 175.

conarius, 168.

crocatus, 168, 171.

cruciger, 168.

descriptus, 168, 171,

ecclesiasticus, 173.

longi palpus, 168.

marnioratus, 168.

oroatus.

trilineatus, 168, 170.

variegatus, 174.

zonarius, 170.
Micaria, 167.

longipes, 167.

montana, 168.
Micariosoma Simon, 188.
Phrurolithus, 189.

alarius, 188.

pugnatus, 188.
Poeciiochroa, 174.

bilineata, 175.

montana, 175.

variegata, 174.
Prosthesima, 172.

atra, 172.

depressa, 173.

ecclesiastica, 173.

fune.sta. 172.

melancholica, 172.

propinqua, 173.
Pylarus bicolor, 201.
Pythonissa, 177.
Pythonissa imbeciUa, 177.
Trachelas, 184.

ruber, 184.
Tegenaria Latr., 193.

brevis, 194.

civilis Blk., 193.

derhamii, 193.

domestica, 193.

medicinalis, 191.

204 J. H. Mnerton — New England Drassidm^

EXPLANATION OF PLATES.

Plate III.

Fig. 1. Micaria longipes, la, maxillaj and mandibles, from below ; \b, palpus of male ;

Ic, scale from front of abdomen; Id, le, white scales from spots on abdomen;

]/, scale from hinder half of abdomen; \h, epigynum.
Fig. 2. Micaria montana ; 2a, epigynum.
Fig. 3. Geotrecha hivittata, ventral view; 3a, dorsal view ; 3&. 3c, palpi of male; Zd,

epigynum; 3e, hair of abdomen.
Fig. 4. Geotrecha, pinnata; 4a, epigj'num.

Fig. 5. Geotrecha crocaia, dorsal view; 56, 5c. palpi of male; 5(Z, epigynum.
Fig. 6. Prosthesima atra; 6a, ventral view ; 6&, (^c, palpi of male; 6rf, epigynum ; 6e,

foot of first pair; 6/, foot of fourth pair; (jg, hair of abdomen.
Fig. 7. Prosthesima ecclesiastica ; la, ventral view; 1h, epigynum: 7c, Id, palpus of

male.
Fig. 8. Prosthesima depressa; 8a, epigynum.

Plate IV.

Fig. 1. Poecilochroa variegata; la, ventral view; \h, \c, palpus of male; \d, epi-
gynum.

Fig. 2. Poecilochroa montana; 2a, epigynum.

Fig. 3. Po&cilochroa hilineata ; 3a, ventral view.

Fig. 4. Gnaphosa conspersa; 4.a, ventral view; 41), end of mandible; 4c, outer side of
male palpus; 4(^, palpal organ ; 4e, epigynum.

Fig. 5. Gnaphosa brumalis, outer side of male palpus ; 5a, palpal organ ; ^b, epi-
gynum.

Fig. 6. Pythonissa imhecilla; 6a, ventral view; 66, mandible; 6(', palpus of male;
Qd, epigj'num.

Fig. 7. Drassus saccatus; la, ventral view; 76, outer side of male palpus; 7c, tibia
and tarsus of male palpus, under side; 7d, epigynum.

Fig. 8. Drassus rohustus; 8a, epigynum; 86, c, palpus of male.

Plate V.

Fig. 1. Clubiona, crassipalpis, male palpus showing palpal organ; la, tiVna of male
palpus; 16, outer side of male palpus.

Fig. 2. Clubiona mixta, male palpus, upper side showing tibia ; 2a, under side, show-
ing palpal organ ; 26, outer side of patella and tibia.

Fig. 3. Clubiona tibialis, male palpus, upper side; 3a, outer side; 36, epigynum.

Fig. 4. Clubiona canadensis, male palpus, upper side of tibia and tarsus; Aa, outer
side of male palpus; 46, under side of tibia and tarsus, showing palpal organ;
4c, epigynum.

Fig. 5. Clubiona pusilla, under side of male palpus, showing palpal organ; 5a, upper
side, showing form of tibia ; 56, inner side.

Fig. 6. Clubiona rubra, male palpus of spider from Saugus, Mass. ; 6a, outer side ;
66, palpal organ; 6c, epigynum.

Agalenidm and Dysderidm. 205

Fig. 7. Gluhiona rubra, palpus of large male from Lynn, Mass.; 7«, outer side; 1h,
palpal organ ; 7c, epigynum.

Fig. 8. Clubiona rubra, palpus of small oiale from New Haven, Conn. ; 8a, outer
side; Sb, palpal organ.

Fig. 9. Clubiona ornata ; da, epigynum.

Fig. 10. Clubiona excepta; 10a, dorsal view; 10&, 10c, palpus of male; \0d, epi-
gynum.

Fig. 11. Clubiona minuta, male palpus, upper side; 11a, outer side; 116, under side.

Fig. 12. Chiracanthium viride; 12a, dorsal view; 126, male palpus, outer side; 12c,
upper side ; 1 2d!, epigynum.

Fig. 13. Trachelas ruber; 13a, sternum and mouth part; 136, epigynum; 13c, 13c?,
male palpus.

Plate VI.

Fig. 1. Anyphoitia rubra; la, ventral view of abdomen, showing at x the tracheal

opening; 16, epigynum.
Fig. 2. Anyphcena incerta; 2a, 26, 2c, palpus of male; 2d, epigynum.
Fig. 3. Anyphcena calcarata, under side of coxae of male, showing spurs on 3d and

4th pairs; 3a, 36. 3c, male palpi; 3d, epigynum.
Fig. 4. Anyphcena saltabunda, male palpus, under side; 4a, 46, 4c, upper and side

views; 4c?, epigynum.
Fig. 5. Phrurolithus alarius, in natural position at rest; 5a, light-colored variety ; 56,

first leg; 5c, maxillae; 5c/, irridescent scales of abdomen; 5e, 5/, bg, male palpi;

5/i, epigynum.
Fig. 6. Phrurolithus pugnatus ; 6a, upper side of male palpus ; 66, outer side ; 6c, epi-
gynum, showing part of palpal organ in one side.
Fig. 7. Agroeca pratensis ; 7a, sternum and mouth part; 76, 7c, 7ci, palpus of male ;

7e, epiigynum.

Plate VII.

Fig. 1. Ocelotes medicinalis, enlarged 4 times; let, epigynum: 16, palpus of male,

under side; Ic, upper side.
Fig. 2. Cidotes longitarsus, palpus of male, upper side ; 2a, epigynum.
Fig. 3. Ccelotes montanus, epigynum ; ?>a, palpus of male ; 36, patella and tibia of male

from New Haven, Conn.
Fig. 4. Ccelotes hybridus, male palpus, upper side of patella and tibia; 4a, under side.
Fig. 5. Tegenaria brevis ; 5a, epigynum; 56, head of female ; 5c, head of male.
Fig. 6. Tegenaria derhmnii, enlarged 4 times; 6a, palpus of male; 66, tarsus and pal-
pal organ; 6c, epigynum.
Figs. 7, 7a, 76. Gicurina complicata, male palpi; 7, under side; 7a, outer side with

large process of the tibia separated from the tarsus; 76, upper side, showing the

narrow tarsus and short curved tibia.
Fig. 8. Hahnia bimaculata, under side of abdomen, showing trachasl opening and

spinnerets ; 8a, male palpus, under side of tibia and tarsus ; 86, outer side of tibia

and tarsus ; 8c, 8c?, 8e, 8/, various forms of epigynum.
Fig. 9. Hahnia cinerea; da, male palpus, under side; 96, outer side.
Fig. 10. Hahnia radula, maxilla of male; 10a, hair of first leg of male.

Trans. Conn. Acad., Vol. Vlll. 27 Jan., 1890.

206 ./. H. Emerton — N'eio England iJrassidce, etc.

Plate VIII.

Pig. 1. Agalena Jicevia. male; la, common form of male palpus from a specimen from
Peabody, Mass. ; \h, long tubed variety of palpal organ from Woodbridge, Conn. ;
Ic, outer side of tibia of same palpus; \d, short tubed form of palpal organ of a
specimen from New Bedford, Mass. ; le, palpal organ with unusually large tube of
a specimen from Providence, R. I. ; 1/, small palpal organ from Jaftrey, N. H. ; Ig-,
outer side of small male palpus from Salem, Mass. ; \h, common form of epi-
gynum from female in copulation with the male from which Fig. 1 was drawn ;
\i and ly, epigynum from Providence, R. I. ; \k. epigynum with small process on
front edge from Salem, Mass.; IZ, Im, Iw, epigynum from several large spiders
from Massachusetts.

Fig. 2. Dysdera interriia; 2a, sternum, coxae, and mouth parts; 2b, foot; 2c, side of
male palpus; 2d, palpal organ from front.

Fig. 3. Ariadne bicolor, female; 3a, male, head, palpi, and front leg; 3b, sternum
and mouth parts of female; 3c, male palpus; 3d, front leg of female.

APR 1^1893

[Presented to the Oouuecticiit Academy of Arts and Sciences, May 20th, 1891.]

XII. — The Development of a Paleozoic Poriferous Coral.
By Charles E. Beecher. (With Plates IX-XIII.)

The origin and affinities of many groups of paleozoic corals are
still obscure. The main elements of the recognized system of
classification seem to be stable, yet so little is known of the
growth and structure of a number of important groups, that they
occupy a different place in almost every arrangement of the genera.
Each fact of development affords data which eliminate, to a degree,
the w^ant of knowledge concerning their origin and relations. Un-
less the growth of the organism is obscured by pronounced acceler-
ated or degradational features, its interpretation is simple, and
throws much light on its ancestral history. Paleozoic types in
general are least modified in their development by acceleration.
They usually show some marked expression of their jjrototype, and
also the succession of changes through which they have passed
during their evolution.

The species here discussed was originally described as Miehelinia
lenticularis. Hall,* from the Lower Helderberg group of New York.
If Miehelinia is entitled to recognition, it will exclude this form, as
it is without tabular. PleurocUctyuni, as now defined, is more in har-
mony with these features, and, therefore, the species M. lenticularis
is here referred to this genus. The large calices and their constant
origin at the basal epitheca are not, however, essential characters of
Pleurodictyimi. The structure and growth of this species indicate
that it represents one of the simpler types of poriferous corals. For
this reason, its development is without the numerous modifications
necessary in more complex forms, and its laws of growth are not
complicated.

Development of Plenrodictyum lenticulare. — The nepionic stage is
well marked. It comprises the growth of the corallum to the com-
pletion of a simple initial cell. This primitive cell or nepionic stage
(Plate IX, figure 1, V, 8) has the foi'm of an oblique inverted cone
flattened on one side. The flattened area represents the lower or
attached side, and the oblique base of the cone is occupied by the

* Twenty-sixth Kept. N. Y. State Museum of Nat. Hist., p. 113, 1874.
Tr.-vns. Conn. Acad., Vol. VTII. 28 July, 1891.

208 C. E. Beecher — Developmoit of a Paleozoic Poriferous Coral.

aperture of the corallite. The apical portion is smooth for about
one-fourth the length of the cell. Then the concentric lines of
growth become apparent, and over the distal half, radiating ribs are
also developed. The interior of the apex is granulose. At about
the middle of the cell, the granules are arranged in rows, forming
the beginnings of the septal lines.

The simple growth of the initial cell continues until the entire
procumbent portion is completed. A thickening of the margin then
takes place, and an upward growth of the corallite is initiated. At
the commencement of this upward growth, the first bud starts out
from the lateral edge of the initial calyx, either to the right or left
of the axis. This condition represents the first nealogic stage. The
bud resembles the parent cell in all particulars, and reaches consider-
able size before the second appears, as shown in Plate IX, figures 9, 10.
The visceral cavities are confluent, as the initial apex of the bud
opens into the calyx of the first cell.

The succeeding nealogic stages, to the completion of the first
circle of peripheral calices, have been observed mainly from the
epithecas of mature or nearly full grown corallums, rejjresented on
Plate X, figures 1, 2. In these examples, the lines of growth are so
perfectly shown, that all the stages are distinctly marked, and may
be satisfactorily studied.

What is here considered as the second nealogic stage is repre-
sented on Plate IX, figure 3, showing the initial corallite, with the
fii'st and second buds on opposite sides. This process of alternate
gemmation from the parent cell continues until the circlet of calices
is completed, as shown in figures 4, 5, and 6. In this species, the
normal number of peripheral calices is seven, making eight corallites
in the completed nealogic corallum. The last cells to be formed are
(1) the sixth and seventh budding from the anterior side of the first
calyx, and (2) the eighth or posterior cell. Plate IX, figure 12, repre-
sents the completed nealogic corallum, with the initial cell and six
well developed peripheral calices. The eighth has just begun to
fill up the space between the second and third. It will be noticed
that there is a direct correspondence in the size of the calices to
their relative age. The first calyx is much the largest. Then, de-
creasing serially, come the second, third, fourth, fifth, sixth, and
seventh, while the eighth is undeveloped. An inspection of the
upper surface of a mature corallum will thus usually determine the
order of successive calical additions. After the ajjpearance of the
posterior, or eighth calyx, the corallum commonly grows to double

C E. lieecher — Development of a Paleozoic Poriferous Coral. 209

the diameter of the completed nealogic stage, resulting in the
normal ephebolic or mature condition, as represented on Plate XI,
figures 1, 2. Nearly all the full grown specimens found agree in
this respect.

A coralhun rarely presents any departure from the normal number
of calices. Plate XII, figure I, is an example of a variation in the
number of peripheral corallities, for in this specimen, there are
eight in the circle, instead of the usual seven. A variation in the
opposite direction is shown in another sjiecimen having five well
develo[)ed corallites about the parent cell. Old age characters are
expressed in two ways : First, the cell walls become thickened
around the margin of the epitheca without destroying the S3anmetry
of the corallum, as shown in Plate XII, figure 2 ; Second, by the
indefinite and unequal development of the peripheral cells, together
with the addition of calices budding from the cells forming the
primary circle. One specimen, appearing at first sight as an exam-
ple of cell division or fission, is shown in Plate XIII, figure 2. It may
be explained as resulting from the abnormal growth of the second
and adjacent calices, four and eight. , This lateral impulse further
resulted in sending off the small, peripheral, tertiary corallites num-
bered in the figures 9, 10, 11, 12, and 13.

It should be understood that this arbitrary expression of normal
and abnormal growths applies only to the species P. lenticulare.
The same numerical arrangement will not hold good for genera like
Favosites, MicheUnia^ Striatopora, etc. Otherwise, it is believed,
the general laws of growth here brought out will hold good for
these and other related genera.

Some doubt may exist as to the propriety of referring the speci-
mens illustrated on Plate IX, figures 9-11, to P. lenticulare. Unfor-
tunately, material of this kind is rare and difficult to obtain. With
the exception of the position and direction of the first bud (figure
10), all the characters agree, so far as can be observed, with ordinary
specimens of P. lenticulare. The second cell of the corallum repre-
sented in Plate X, figure 1, curves rapidly backwards, although at
first the axis has an anterior direction. Taking this view of the
specimen, Plate IX, figure 1 1, it is not difficult to see how the succeed-
ing enlargement and curvature of the bud could extend backwards,
thus properly limiting the size of the eighth or last of the primary
circlet of calices.

The method of determining the relative age and succession of the
corallites can be seen in Plate X, figures 1, 2, and Plate XI, figure 2.

210 C. E. Beecher — Development of a Paleozoic Poriferous Coral.

The initial cell occiqtie.s the central position, and forms the boss or
apex of the basal ejiitheca. The first bud is nearly on a plane with
the base of the initial cell and is the one nearest the apex. The
second and successive buds are respectively more distant, and at a
higher level. Specimens having broad surfaces of attachment to
foreign objects have these distinctive features of the epitheca obliter-
ated, and the only guide to the order of the corallites then lies in
their comparative size and position on the upper surface of the
corallum.

General conclusions. — The first feature to be noted in the develop-
ment of a poriferous coral, as here described, is the simple cyathi-
form character of the initial corallite. This nepionic stage is
without mural pores, and has an epitheca over the entire exterior of
the cup. The septal lines become developed toward the end of
this stage. These features are in harmony with the young of many
paleozoic corals, such as Cladochonus, Aidoiyora, or Syringopora,
and clearly indicate a primitive, simple, and imperforate ancestry
for the Perforata. A similaf origin and development obtains in
Pavosites, as may be seen from the figure of a young colony of P.
Porbesi, var. occidentalis, given by Professor Hall.*

The first nealogic stage, represented by the primitive corallite
with one bud, is the first transition towards both a compound and a
perforate coral, Plate IX, figure 9. This stage has two calices, mak-
ing it a comj)Ound coral, and has an opening through the cell walls
or connecting channel between the corallites, forming the first mural
pore. The manner of growth and the structure of the corallum at
this stage are suggestive of Atdojwnr, and should be given consider-
able significance. The visceral cavities in Atdopora are confluent,
and rudimentary septa or lines of spinules are often present. Poni-
ingeria has a growth resembling Aulopora and Syringojiora. It is
without pores on the portion where the corallities and buds are free,
but when these are in juxtaposition at their bases, mural pores are
developed. The upward groAvth of the initial cell of P. lenticulare
proceeds but a short distance before the circlet of peripheral coral-
lites is completed. Thus at this stage there are at least seven mural
pores opening into the primary calyx. If this tendency to the
formation of numerous buds persists throughout the upward growth

* Indiana Geol. and Nat. Hist., 11th Rept. of tlie State Geologist, pi. i, figure 12,
1881.

(J. E. Beecher — iJevelopment of a Paleozoic Poriferous Coral. 211

of the corallites, the non-development of the buds consequent upon
the adjacent living corallites would naturally result in the produc-
tion of mural pores. The basal epitheca limits the fleshy portion of
the organisms, and represents an area unfavorable to the acquisition
of food or for the natural development of calices. Therefore, it
would prevent both tlie maintenance of mural pores and the growth
of basal buds.*

A Favosites in which one or more cells became inactive or dead
shows in its subsequent growth the closing over of this area by the
budding of the surrounding cells. Each cell is connected with the
parent by an apical pore, Plate XIII, figures 3, 4. Without this
opportunity to bud afforded by the death of one or more corallites, or
by their divergence, the adjacent cells would have developed only
mural pores. In the figure of P. problematicwn given on Plate XIII,
figure 2, three of the initial poi'es are indicated by dotted lines from
23. No distinction can be made between these and the ordinary pores,
except that the latter are usually not as large. Tliis difference in
size would be expected, as the primary pore represents the bud which
succeeded in producing a corallite ; -whereas the other attempts at
budding resulted no further than the production of mural pores. The
conclusion to be drawn is, that the mural pores in such genera as
Favosites, Striatopora, Pleurodictyum, Michelinia, etc., are ineffec-
tual attempts at budding, resulting only in the perforation of the
cell walls. This explanation agrees with the pronounced and per-
sistent tendency to gemmation characteristic of the genera men-
tioned. They also represent compound forms having individualized
epithecas, and this feature naturally arises from the same system of
budding obtaining in the simple corals.

Professor Verrill has shown that the presence or absence of tabulae
is of little or no importance in a natural classification.! Therefore,
the non-tabulate feature of P. lenticulare is without special conse-
quence in a discussion of the relations of this species with Favosites,
or other tabulate poriferous genera.

* The presence of basal mural pores or openings through the epitheca has been
asserted by Meek and Worthen (Pal. Illinois, vol. iii, p. 409, 1868). The specimens
from which this observation was made, are from a friable sandstone, which does not
usually preserve minute details with much distinctness. The depressions between
the spinules on the septal lines could easily be mistaken in a cast for the filliugs of
mural pores, and it is believed by the writer, that this interpretation should be
given. P. lenticulare occurs as calcareous or silicified, and in the condition of casts.
No basal mural pores are present. Also, none can be observed in the casts of P.
pvohlemalicum, from Pelm, Germany.

f Am. Jour. Sci., vol. iii, p. 187, March, 1872,

212 C. E. Beecher — Development of a Paleozoic Poriferous Coral.

If the preceding interpretations of structure and affinites are cor-
rect, a simple, conical imperforate, non-tabulate prototype, or proto-
corallum, may be assumed for the Madreporaria Perforata. The
next derived form, represented by the early nealogic stages of P.
lenticxdare^ has the structure and growth of Aulopora, and consists
of the parent cell with one or more buds. At this stage, which may
be called the Atilopora-stage, the initial eorallite has the same num-
ber of mural pores as developed buds, for each bud leads into the
parent cell by a basal opening or pore. Aulo^yora may thus be con-
sidered as representing a primitive type of a poriferous coral, in
which the number of pores in each eorallite corresponds to the num-
ber of buds given off plus one connecting it with the pai*ent cell.
Some species of this genus are free throughout most of their growth
[A. subtennis, Hall), agreeing closely with the erect growth of
Pomingeria and Syringopora. This fact removes one of the im-
portant arguments against the relations of Aulopora with these
genera. The corallites of Aulopora usually send off buds before
turning out of the common axis of the branch or colony, after which
no gemmation commonly takes places. By the explanation here
advanced, this lack of a tendency to gemmation in the distal por-
tions of the corallites in this genus accounts for the absence of
mural pores when such portions are in contiguity. The periods of
gemmation in Pomingeria are periodic. Several buds, often tonn-
ing a verticil are given off from the parent eorallite. Considerable
elongation of the tubes takes place before other series of buds are
produced. The budding is prolitic at these points, and here also
occur the mural pores. The latter are therefore developed when
the period of gemmation is in force. If pores are formed elsewhere
when the corallites happen to come into juxtaposition, it may pos-
sibly be explained as the result of a stimulus produced by the con-
tiguit}' of the animals. Further observations are necessary to show
that pores exist at other places than the bases of the verticils or
points where numerous buds are given oft' and where from crowding
the corallites are in juxtaposition.

It therefore seems, that, primarily, the development of nuiral
pores is identical or homologous with the process of gemmation.
Whether this cause is operative in such forms as Golumnopora or
Alevopora yet remains for investigation. The porous condition of
the walls in these genera may be an inherited character Avithout an
active exciting cause, or it may be teleologically different.

Tale Museum, New Haven, Conn., May 20th, 1891.

(J. E. Beecher — Development of a Paleozoic Poriferous Coral. 213

Fig.

1

Pig.

2.

Fig.

3.

Fig.

4.

Fig.

5.

Fig.

6.

Fig.

1.

Fig.

8.

Fig.

9.

EXPLANATION OF PLATES.

Plate IX.

Pkurodictyum lenticulare.
Lower side of initial cell, or nepionic stage. x ?,^.
Lower side of initial cell with one bud ; first nealogie stage, x 3i.
Initial cell with two buds ; second nealogie stage, x 3^.
Initial cell with three buds; tliird nealogie state, x 3i.
Initial cell with four buds ; fourth nealogie stage, x 3^.
Completed nealogie stage; shouing initial corallile (!) and seven peripheral

corallites (2 -8). x 3^.
Interior of nepionic corallite. x 3^.
Exterior of same specimen x 3^.

Upper side of specimen representing first nealogie or Auhpora -stage, con-
sisting of nepionic cell and one bud Apex of bud opens into visceral
cavity of parent cell, x 3i.
Fig. 10. Profile of same; showing obliqne apertures of corallites, and thickened

margin of parent cell x 3|.
Fig, 1 1. Lower side of preceding, x 3i.
Fig. 12. Upper side of completed nealogie stage; showing inception of eighth cell.

Figs. 1-G are taken from epilhecal lines of growth shown in Plate X, figure
2. Remaining figures are from actual specimens. Numbers refer to
order of calical succession.

Lower Helderberg Group. Albany County, New York.

Plate X.

Pleurodictymn lenticulare.

Fig. 1. Lower or epithecal side of specimen; showing successive alternate gemma-
tion from parent coralHte. x 3^.

Fig. 2. Similar specimen with lines of growth more strongly marked. The order of
luidding is opposite to that of preceding specimen, x 3^.
Lower Helderberg Group. Albany County, New York.

Plate XL

Pleurodidyum lenticulare.
Fig. -1. Outline of cahces of specimen Plate X, figure 2; showing central primary
cell and seven peripheral calices numbered in tiie order of their develop-
ment, x 3f .
Fig. 2. The same ; side view.

Lower Helderberg Group. Albany County, Neiv York.

214 C. E. Beecher — Development of a Paleozoic Poi-iferous Coral.

Plate XII.

Pleurodictywii lenticulare.
Fig. 1. Outline of upper side of specimen with eight peripheral calices. x 3^.
Fig. 2. Upper side of symmetrical specimen; showing general features of calices,
mural pores in central eorallite, and thickened epithecal border, x 3|.
Lower Helderberg Group. Albany County^ New York.

Plate XIII.
Pleurodictyum lenticulare.
Fig. 1. Calical diagram of geratologic specimen; showing enlargement of second,
fourth, and eighth corallites, and the addition of tertiarj^ cells, forming a
second series of peripheral calices. x 3^.
Lower Helderberg Group. Albany County, Neiv Torh.

Pleurodictyum prohlematicum.
Pig. 2. Lower side of cast of corallum with epitheca removed ; showing proximal
extremities of several corallites. Upper edge of figure represents portion
of periphery of corallum. Thus, lower angle of each eorallite represents
the point of budding from parent cell, and is connected with it by a pore,
shown for three of the corallites by dotted lines from ^). It will be
noticed that all the pores in the angles are larger than the others. Otlier-
wise, these and the initial pores cannot be distinguished from the ordinary
mural pores between the flat sides of the corallites, x 7.
Devonian. Pelm, Germany.

Favosites epidermatus ?
Fig. 3. Side view of mature eorallite with attached intermural bud. Specimen

broken from interior of a large colony, x 3^.
Fig. 4. The same front view, with bud lemoved ; showing pore or mural opening (p)
at lower point of attachment of bud, corresponding to those indicated in
figure 2. X 3|.
Corniferous limestone. Cherry Valley, Neiv York.

XIII. — Symmetrical Cell Development in the Favositid^.
By Charles E. Bebchbe. (With Plates XIV, XV.)

The majority of compound corals included in the I^avositidce are
composed of polygonal, prismatic cells or corallites in juxta2)osition.
When, however, these cells become free, their form is cylindrical.
The polygonal form of closely arranged cells is therefore explained
as the natural result of crowding.

The species Pleurodictyimi lenticulare, Hall, sp., is an example of
simple cell growth and multiplication. In the development of this
species, as shown by the writer in the previous paper, the initial
corallite is first conical. The growth of a peripheral series of buds
results in changing the sub-circular section of the parent corallite
into a polygon. The buds are angular on the sides in juxtaposition
to the parent cell and adjacent buds, but on the free portion of their
periphery they are cylindrical. The subsequent growth of peri-
pheral buds brings the first series wholly within the cbrallum, and
they are then polygonal in section like the parent corallite.

In compact corals with long cell tubes, as Michelinia and Favosites,
there is a maximum limit to the size of the corallites. Thus, the
form of the cells which have reached this limit of diametral exten-
sion is that of equal hexagonal prisms. This is of course due to the
well known fact of six equal tangent circles about a central circle of
the same size. Then from crowding, or from the elimination of the
interstitial spaces, they assume a regular hexagonal form. The
specimen of Clelstopora geometrica, illustrated by Edwards and
Haime,* represents the maximum size of the cells and their equal
development in this species. Although the tubes are not long, the
calices are nearly of the same size, and regularly hexagonal.

After the completion of a circle of calices about the parent cell of
the corallum, enlargement takes place, (1) by buds from the peri-
phery, and (2) by intermural gemination. The first is not attended
by any phenomena differing from the production of the primary
circlet of calices about the initial cell. The second takes place
under other conditions, and is the chief method of increase in the
growth of large corallums having numerous corallites.

* Monographie des Polypiers Fossiles des Terraiaes Palceozmques, p. 252, pi. 17, fig.
3. 1851.

Trans. Conn. Acad., Vol. VIII. 29 July, 1891.

216 C. E. Beecher — Symmetrical Cell Development in JFavositidce.

The radial arrangement of the tubes in a large hemispherical or
cylindrical mass tends to make the axes of the corallites diverge.
This divergence can be taken up only by an increase in the diameters
of the tubes, or by the addition of new calices between the others.
The latter mode is called intermural gemmation. In Favosites and
allied genera, the maximum size of the corallites is soon reached,
and the expansion of the coral is mainly derived from intermural
growth. The study of this method of increase, properly begins
after one or more rows of calices have been developed aboixt the
par-ent cell, and the calices have reached their full dimensions.

The following description of a symmetrical system of intermural
cell multiplication was observed in a hemispherical specimen of
Michelinia convexa, D'Orbigny, from the Corniferous limestone of
the Falls of the Ohio. It shows very clearly the stages of develop-
ment of the interstitial buds, and their modifications. Other corals
were examined to the same end, and were found to agree in all
essential particulars, whenever their growth was not irregular from
their condition of fixation, or from the excessive development or
death of a number of the corallites. An exact number of peripheral
buds is not necessary to illustrate the general laws of intermural
growth. The buds produced from any given cell cannot always
agree with symmetrical method here described, on account of the
crowding of similar sei'ies from adjacent or neighboring corallites.
After eliminating these variations, it was found that the process of
intermural gemmation in general is quite uniform, and closely con-
forms to that in 3fichelinia convexa.

Plate XIV, figure 1, represents diagrammatically the top of a coral-
lum composed of a central parent cell and six equal peripheral buds,
making seven nearly equal calices in the corallum. The upward
growth of these corallites and the divergence due to the direction of
their axes tend to separate them from the parent cell. In conse-
quence of this separation of the corallites, they would naturally
assume a cylindrical form, and there would thus appear triangular
interspaces between the tangent points of any three adjacent calices.
These angles, therefore, afford the only opportunities for the intro-
duction of a set of intermural buds, and their initial triangular form
is determined by the conditions of growth. The smallest number of
buds which can be symmetrically placed and compensate for the
divergence of the corallites is three, one from each alternate angle
of the hexagon, Plate XIV, figure 2,

C. E. Beeches — Symmetrical Cell Development ui Favoaitidce. 217

If these interstitial cells were to grow without the introduction of
others, until the original peripheral series was completely separated
from the parent or central cell, there would result a corallum con-
taining only triangular corallites. There is, however, a manifest
tendency of the organism to the production and maintenance of a
cylindrical form, or of a prism with nearly equal radial axes, as in a
hexagonal or polygonal prism. To accomplish this, and further to
take up the divergence of the corallites, three new interstitial buds
are introduced at the remaining three unmodified angles, as shown
in figure 3, At this stage, there are six symmetrically disposed
triangular buds, or intermural cells, about the central corallite,
truncating its original angles, and making it a twelve sided prism.
This stage is the third toward the formation of a series of mature
interstitial calices.

During the third stage, the intermural buds increase in size until
they completely surround the parent cell. Then further growth
truncates their adjacent angles, thus adding two more sides to each
bud, making them pentagonal in section. This marks the fourth
stage of intermural growth. At the same time, the central corallite
loses six of its sides, and returns to its early hexagonal form. The
axes have revolved 30°, and the original sides have now become the
angles of the corallite, Plate XIY, figure 4.

At this period of growth, it is necessary to consider a series of
buds on the periphery of the corallum, marked l", 2", etc., in Plate
XIV, figures 3 and 4, They are first triangular in form like the others,
and of two sizes, owing to their different ages. The growth of this
series continues until they touch and truncate the angles of the first
series (l', 2', etc.), producing the fifth condition or stage. The first
series of buds has now three hexagonal and three pentagonal coral-
lites, Plate XIV, figure 5.

In the last or sixth stage, figure 6, the further growth of all the
intermural cells results in a corallum of nineteen nearly equal hexa-
gonal corallites. The original parent cell (1) is at the center, the
first six intermural cells (1', 2', etc.) completely surround it, and the
six new peripheral corallites (1", 2", etc.) are interposed between the
members of the original circlet (1, 2, etc.). The effect of this inter-
mural growth, then, is to dissociate all the first series of corallites
from the parent cell and from each other.

The changes taking j^lace in the number and form of the cells
may be tabulated as follows :

218 C. E. Beecher — Symmetrical Cell Development in FavositidoB.

stages.

Nepionic.
First completed nea-
logic or first con-
dition requisite to
intermural gem-
mation.

2d stage.

3d stage.

■Ith stage.

5th stage.
6th stage.

I I Number of Number of

Form of primary cell. Whole No. intermural! sides of
of cells. buds. buds.

cone.

6 sided prism.

9 sided prism.

12 sided prism.

6 sided prism.

6 sided prism.
6 sided prism.

7

11
16

19

19

19

Buds are developed in Favosites and Michelinia whenever there
is a space or opporUinity for their growth, unless the corallum is
affected by some abnormal condition. If this tendency to form a
solid mass of corallites were not so strong, and if the process of
budding took place only at comparatively remote intervals, the
corallum would have the form of Romingeria. It is evident in
Michelinia convexa, that if the divergence of the corallites was
considerable and not wholly filled by intermural growths, there
would result a verticil of corallites about the parent cell which
would soon become free. The peripheral corallites, also, would be-
come separated. Then after further growth, the parent cell Avould
give off another verticil of buds, the other corallites, likewise,
develop similar verticils, and the whole form and mode of growth
be like that of Momingeria. From this point of view, Bondngeria
may represent an early form of symmetrical cell development in the
poriferous corals. The acceleration of the periods of gemmation,
and consequent approximation of the corallites carrying their ver-
ticils of buds, would produce all the conditions of cell growth and
intermural gemmation exhibited by Favosites or 3Iichelinia.

Summary : — The growth of intermural buds compensates for the
natural divergence of the corallites. New cells are introduced
whenever the old corallites have reached their maximum size, and
when their divergence approaches a separation of the cell tubes.

The form of the buds is first that of a triangular pyramid or prism,
and is due to the mechanical conditions of growth. During subse-

C E. Beecher — Symmetrical Cell Development in Favositidm. 219

quent increase, they touch and truncate each other, changing from
triangular to five- and six-sided prisms. Complete symmetrical
normal development produces a corallum with equal hexagonal
calices. The process of intermural gemmation changes the sides
of the parent cells to angles, and the older corallites, originally in
juxtaposition, become separated from each other by new series of
interstitial calices.

Yale Museum, New Haven, Conn., May 20th, 1891.

EXPLANATION OF PLATES.
Plate XIV.

Michelinia convexa.
Pig. 1. Diagrammatic representation of upper surface of corallum; consisting of

parent cell. A, and six peripheral corallites, 1, 2, 3, etc.
Fig. 2. The same; showing the introduction of three triangular intermural buds,

1', 2', 3', etc.
Fig. 3. Third condition; with six triangular buds about the parent corallite, and

three on the periphery of the corallum.
Fig. 4. Top of corallum ; showing further growth of preceding corallites, with the

addition of three peripheral calices, 4", 5", 6".
Fig. 5. The same during a succeeding stage ; showing increase in size of corallites,

and modifications produced.
Fig. 6. Completed growth of first system of intermural gemmation ; showing disso-
ciation of original series of corallites (A, 1, 2, 3, etc.), and representing condition

preparatory for new series of interstitial corallites.

All figures natural size.

Plate XV.

Michelinia convexa.
Fig. 7. Development of a group of corallites from initial conical cell to corallum
with nineteen calices. The figure represents parallel horizontal sections through
the corallum ; showing the number and form of the calices, their order of devel-
opment, and the modifications taking place during growth. The parent cell is
marked A ; first series of calices, 1, 2, 3, etc.; first series of intermural buds,
1', 2', 3', etc.; peripheral series, 1", 2", 3", etc. Notation corresponds with that
of preceding plate.
Natural size.

Trans. Conn. Acad., Vol. VIIL 30 Oct., 1891.

XIV. — New England Spiders of the family Attid^. By

J. H. Emerton.

The Attidse are distinguislied by a peculiar arrangement of the
eyes. The front of the head is wide and square and the front row
of eyes directed forward and nearly straight or with the lateral pair
a little the highest. The front middle pair are larger than the
others and often much larger, so that at first sight the spider appears
to have but two eyes. Behind the lateral eyes of the front row are
two very small eyes and still farther back, often near the middle of
the cephalothorax, are two others a little larger, sometimes as large as
the front lateral pair. See figures on Plates XVII and XVIII.

The relative length of the legs is very variable. The fourth pair
is usually the longest, but often the first, and even in some species
the third pair. The legs of the first pair are usually thickened, and
oft