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THE VITAMINES

BY

CASIMIR FUNK

ASSOCIATE IN BIOLOGICAL CHEMISTRY

College of Physicians and Surgeons

New York City

Authorized Translation

from
Second German Edition

X 'y

BY

HARRY E. DUBIN

FORMERLY ASSISTANT IN RESEARCH MEDICINE
University of Pennsylvania
Philadelphia, Pa.

BALTIMORE

WILLIAMS & WILKINS COMPANY

1922

COPYRIGHT 1922

WILLIAMS & WILKINS COMPANY
X '\^

Made in United States of America

%; /U9 ^^

The right of publication in English
is reserved

COMPOSED AND PRINTED AT THE

WAVERLY PRESS

BY THE WILLIAMS & WILKINS COMPANY

BALTIMORE, MD., U. 8. A.

CONTENTS

Preface to the first edition 9

Preface to the second edition 11

Translator's note 13

Introduction •. 15

Historical survey 19

Introductory 35

PART I
THE VITAMINE REQUIREMENTS OF PLANTS AND ANIMALS

Chapter I. The role of the vitamines in the vegetable kingdom 49

Yeast 51

Bacteria 56

Fungi 64

Higher plants 65

Chapter II. The role of the vitamines in the animal kingdom 73

Life without bacteria 76

Protozoa 79

Metazoa 81

Growth of tissue in vitro 81

Insects 83

Fish 85

Amphibia 85

Birds 87

Chickens — normal nutrition ; chicken beriberi 87

Pigeons — beriberi 95

Pathological anatomy and chemical pathology of beriberi in birds. 103

Mammals 109

Rats 110

Mice 125

Guinea pigs 127

Rabbits 134

Cats 135

Lions 136

Dogs - 136

Domestic animals — sheep, goats, reindeer, pigs, horses, cows,

oxen, (stijfziekte, lamziekte, pica) 140

Monkeys ..t^.. 153

Man.. .sri\Gi\,r.Aj>^.. . 155

4 CONTENTS

PART II
CHEMISTRY, PHYSIOLOGY AND PHARMACOLOGY OP THE VITAMINES

''///;*

Chapter I. The antiberiberi vitamine — Vitamine B 163

Chemical investigation of rice polishings '. 167

Chemical investigation of yeast 177

Other sources of supply 192

Synthetic experiments 193

Stability of the antiberiberi vitamine against heat and chemical and

physical agents 194

Demonstration and estimation of antiberiberi vitamine 195

The possible identity of vitamine B with the substance stimulating

the growth of animals and of yeast 203

Specificity of antiberiberi vitamine 205

Physiology and pharmacology of antiberiberi vitamine 207

The influence of the dietary composition on the vitamine requirement . 210

Chapter II. The antirachitic vitainine — Vitamine A 218

Chemistry of cod liver oil 218

Chemistry of vitamine A in butter and other sources 220

Nature of vitamine A 222

Demonstration of vitamine A 225

Chapter III. Relationship of the antiberiberi and antirachitic vitamines

to lipoids 228

Chapter IV. The antiscorbutic vitamine — Vitamine C 231

The chemistry and the nature of vitamine C 231

Chapter V. Vitamine content of various foodstuffs in the natural and

prepared condition 236

Influence of heating and cooking on the vitamine content 238

Vitamine content of the cooking water 241

Influence of drying 241

Influence of canning 242

Influence of ageing and storing 243

Cereals 244

Rice 248

Barley 248

Oats 249

Wheat 249

Maize „• 250

Potatoes 255

Milk , 256

Demonstration of vitamines in milk 256

Influence of food on the vitamine content and composition of milk 257

Vitamine content of milk 258

Influence of heat on the vitamine content of milk 259

Influence of drying and evaporation 260

Nutritive value of milk . 26]

CONTENTS 5

Meat 262

Table showing the vitamine content of the most frequently used
foodstuffs 263

PART III

THE HUMAN AVITAMINOSES — CONDITIONS IN WHICH THE VITAMINES

PLAT A R6LE

Chapter I. Beriberi 275

Mode of occurrence of and diets leading to beriberi 280

Symptomatology and types of beriberi 282

The sensory, motor form 283

The dry, atrophic form 287

The wet, atrophic form 287

Epidemic dropsy 288

The pernicious, acute form 289

Infantile beriberi 290

Pathology 291

General 291

Blood 293

Cerebrospinal fluid 293

Urine " 293

Therapy 293

General 293

Infantile 294

Ship beriberi 294

Relationship between beriberi and scurvy 295

Chapter II. Scurvy 297

Mode of development 300

Symptomatology and progress of scurvy 303

Infantile scurvy 306

Diagnosis 307

Hematology 308

Metabolism experiments 308

Pathology 308

Barlow's disease 309

Therapy 311

Chapter III. Rickets 312

Occurrence 313

Symptomatology and diagnosis 315

Pathological anatomy and chemical pathology of rickets 315

Metabolism 317

Therapy and therapeutic effect on metabolism 220

Etiology 323

Vitamine etiology of rickets 324

6 CONTENTS

Osteomalacia . 329

Symptoms 330

Anatomy 330

Metabolism 331

Therapy 331

Etiology 331

Chapter IV. Some nutritional disturbances in children — tetany, carbohy-
drate dystrophy (Mehlnahrschaden), atrophy 332

Tetany (Spasmophilia) 332

Etiology 332

Symptoms 333

Therapy 334

Carbohydrate dystrophy (Mehlnahrschaden) 335

Etiology 336

Atrophy 336

Other nutritive disturbances of children 337

Chapter V. Nutrition in man — An introduction to the study of pellagra

and hunger edema 340

Chapter VI. Pellagra 351

Geographical distribution 351

Progress of pellagra 352

Acute, malignant form 354

Light, sub-chronic form 354

Severe, cachetic sub-chronic form 354

Chronic form 354

Symptomatology and pathology 355

Gastro-intestinal tract 355

Skin 357

Nervous system 357

Circulatory system 359

Bones 360

Sexual organs 360

Other organs 361

Chemical pathology 361

Metabolism 362

Prognosis - 362

The relationship of pellagra to the accepted avitaminoses — beriberi

and scurvy 362

Mode of development of pellagra 363

Therapy 366

Etiology 366

Chapter VII. Sprue 369

Symptomatology 369

Pathological anatomy 370

Therapy 370

Pathogenesis . 371

CONTENTS 7

Chapter VIII. Hunger edema 372

Symptomatology and mode of development 373

Pathology 374

Metabolism 375

Therapy £75

Etiology 376

Chapter IX. Pathological conditions in which the lack of vitamines may

be suspected 378

Kallak 378

Trench Sickness 378

Intestinal stasis, etc 379

Sterility 379

Hemeralopia 379

Exophthalmic goitre (Graves' Disease), 380

Significance of vitamines in infections 380

Ophthalmia 381

Nature of the disease 382

Tuberculosis '. 384

Leprosy 385

Pneumonia 385

Chapter X. Influence of nutrition (vitamines) on the action of some

poisons and upon pathological conditions of non-infectious origin . . . 386

Anemia, etc •. . ." 386

Diabetes 387

Cancer 388

Development of teeth 391

Calculi : . . 393

General literature on the subject of the vitamines 395

Literature to the text 399

Index to Sectional Divisions of the Literature 476

Author index 477

Subject index 491

PREFACE TO THE FIRST EDITION

This initial attempt at a classification of our knowledge on vita-
mines and avitaminoses contains, besides a number of facts, the per-
sonal conceptions of the author, new questions, ideas and lines of
research. Much of the material is fragmentary while some of it is
purely hypothetical. This small book is to be regarded as the first
step in a new direction in the field of physiology and pathology.
Many facts are still lacking in this field of work so that hypotheses
must naturally fill the gaps. Even if the structure is skeleton-like,
at least the fundamentals are well established.

At this point, I want to thank Dr. Fraser, Dr. Stanton, Prof.
Hoist, Dr. Mott, Dr. Roberts, Dr. Standwith and Dr. Zeller, and the
London Society of Tropical Medicine, for the permission to reproduce
illustrations. I am thankful also to Dr. Macauley of Cape Town for
having called my attention to a South African cattle disease. In
particular, do I wish to express my deepest gratitude to my Father,
Dr. J. Funk, for his unvaried and tireless assistance.

I would appreciate it greatly if the investigators who are conducting
research on problems discussed in this book would be kind enough
to send me reprints of their work which, at times, is available only
with great difficulty.

THE AUTHOR.

Cancer Hospital Research Institute,
Brompton, London, S. W.

October, 1913.

PREFACE TO THE SECOND EDITION

This edition, totally revised and almost completely re-written,
appears seven years after the first edition, deferred because of the
World War. In the interim, we have received many communications
from our colleagues informing us that our effort had provided them
with a welcome stimulus to their work. We have watched the great
development of this field of research since the appearance of The
Vitamines and it is a source of great pleasure that our work has also
contributed to this progress. The views expressed at the time have
been tested, for the most part, and found correct. While the first
edition was published at a time when great differences of opinion
prevailed, our desire then being to take cognizance of all of them, we
feel justified now in reviewing only those investigations, the view-
point of which is not too far removed from ours. In this way, it
has been possible to make the book more comprehensive. The
subject of vitamines, already beyond the stage of hypothesis, is based
on a firm foundation and has received universal recognition. In
spite of this, however, we are well aware that great gaps exist in our
knowledge, so that we can not regard the chapter as closed.

These gaps may be explained by the fact that the field of vitamine
research gave many investigators the opportunity of making a name
for themselves with rather superficial work. Only seldom did they
seriously endeavor to get at the basis of the phenomena observed.
To permit of progress in the subject of vitamines, it would be very
desirable that at least some of the workers should abandon the beaten
path of exclusive animal experiments and break a fresh trail leading
towards the chemistry, physiology and pharmacology of the vitam-
ines, as well as the anatomical pathology of the avitaminoses.

For help in the preparation of this edition, we are indebted to
many of our colleagues; we wish also to express our thanks to the
Medical Research Committee (London) and to Dr. Alfred F. Hess
(New York) for permission to reproduce a number of illustrations.
In addition, we wish to thank those whose permission it was impos-
sible to obtain because of uncontrollable circumstances. We are
especially grateful to Dr. Richard Hamburger, First Assistant at the
Pediatric Clinic, Berlin, who took it upon himself to correct and

critically review the proofs.

11

12 PREFACE TO SECOND EDITION

In particular, however, we take pleasure in recording our apprecia-
tion of the assistance extended by the Hon. Herman A. Metz (New
York) who, although a layman in this field of work, understood its
significance and gave us the opportunity, after an enforced interrup-
tion of many years, of resuming the experimental work in the
"Research Laboratory of H. A. Metz."

This book will be of interest not only to nutrition investigators
and childrens' specialists, but to every physician who comes into
touch with questions of the physiology of nutrition. It treats many
problems which are closely allied to other lines of research and will
be of value to plant physiologists, bacteriologists and animal breeders.

CASIMIR FUNK.

College of Physicians and Surgeons,

Columbia University,

New York City.

December, 1921.

TRANSLATOR'S NOTE

The translator has been associated with the author for three years
in experimental vitamine research, and it is a pleasure to record at
this point the spirit which animates him in his work and his earnest
desire to see real progress made towards the solution of the many
problems of nutrition. It was this which prompted the preparation
of the present edition, intended rather as a stimulus to thought and
further research than as an elaboration of technicalities. Inasmuch
as the author has had the opportunity of reading the manuscript
before it went to press, the reader may be sure that the sense of the
book has been accurately preserved.

In conclusion, the undersigned wishes to extend his thanks to the
Hon. Herman A. Metz for the many facilities provided in the prepara-
tion of this book.

HARRY E. DUBIN.

Research Laboratory of H. A. Metz,
New York City.

December, 1921.

13

INTRODUCTION

The first edition of this book, drafted seven years ago, was intended
to serve the purpose of directing the attention of the medical and
chemical professions to this new and attractive field of investigation.
The findings upon which we based our views in 1913 appeared indeed
to be far from indisputable but time has shown the comparative
accuracy of our conception. Since then, we have made it a point
to note any work that might have some bearing upon the question
of the vitamines, and have now undertaken to point out what may
be regarded as erroneous and what may be looked upon as correct,
in the light of the present status of the subject.

Our original classification included beriberi, scurvy and Barlow's
disease as true avitaminoses, while pellagra, sprue, rickets, 'and some
metabolic diseases of animals, were held to be hypothetical avitam-
inoses. At present, beriberi, scurvy and Barlow's disease are uni-
versally accepted as true avitaminoses, while rickets is quite generally
acknowledged as such. The etiology of pellagra and sprue, on the
other hand, has not yet been definitely established. Regarding
pellagra, which we shall discuss later, the dietetic hypothesis is at
present in the foreground.

As for the chemistry of the vitamines, an examination of the
literature reveals little progress in this direction. To be sure, one
must bear in mind that the outbreak of the war was not particularly
conducive to the long and tedious research that is necessary, if this
phase of the subject is to be cleared up. From tune to time, word
comes from this or that laboratory to the effect that the puzzle has
been solved — that progress has been made even towards the synthesis
of the vitamines — but after much patient waiting, nothing more is
heard of the discovery. However, since a large part of the experi-
mental work is being carried on in many industrial research labora-
tories, it is quite possible that definite important progress has been
made, of which reports have not yet been published.

Incidentally, the war has given added impetus to vitamine research
in another direction. The vitamines have now become of great
importance from the point of view not only of pure science and medi-
cine, but also of political economy — much greater than could have

15

16 THE VITAMINES

been foreseen when the first edition of this book appeared. The
war, first of all, prevented the normal interchange of foodstuffs
among the various countries. Subsequently, this was followed by a
disturbance in the entire mode of living, due to the necessity of
making war, the utilization of farm labor for other purposes, and the
lack of means of transportation. Although, as this is being written,
more than two years have elapsed since the end of the war, conditions
have become worse instead of better, according to available reports.
Similarly, notwithstanding the national rationing of food both
during and after the war, the people have suffered because of the
mistakes made during the war. In the face of higher prices, general
unrest and unwillingness to settle down again to some productive
work, the abnormal conditions already mentioned will likely prevail
for several years to come. It is not our purpose to suggest that
knowledge of the vitamines will solve the present difficulties. Still,
we shall call attention to facts and principles which, at present, are
of universal interest — principles which may contribute to the allevia-
tion of the wretchedness of stricken Europe.

Considering more closely the dietaries of farmers and of the rural
populations in various parts of the world, it is easy to see that no
knowledge of vitamines is necessary to keep those people in good
health. From generation to generation, their nutrition has been
regulated according to the climate, the economic situation, and the
exigencies of the work performed. Of course, in some oriental
countries, where conditions are not so well known to us, instinct some-
times does not choose the correct food and hence the prevalence of
beriberi. All in all, we see that the white races have a wholesome
knowledge of their food requirements, which is only natural. In
certain provinces, before the war, it was possible to see examples of
particularly monotonous and simple dietaries accompanied, on the
whole, by no pathological conditions. We may be sure, however,
that since these same peoples apparently subsisted on a practically
unchanging diet for hundreds of years, they would most certainly not
have survived if their choice of diet had been anything but correct.

When the usual equilibrium is disturbed by extraneous conditions
such as war, a financial crisis or a catastrophe, then the practical
knowledge of centuries suddenly becomes useless to the people, and
they are obliged to seek some other basis of existence. Untold
hardships are endured till this is accomplished, the population func-

INTRODUCTION 17

tioning similar to experimental animals, used to establish the value
of new foodstuffs. Here, at least, there is the added advantage of
successfully applying the various principles of nutrition and sharply
terminating the unnecessarily protracted period. In this connection,
it is not impossible to make immediate use of the enormous amount
of information obtained on the subject, both before and during the
war, especially by the United States and England. During the past
few years, virtually all foodstuffs have been tested as to their nutritive
value and their vitamine content. Although most of the data secured
are the results of animal experiments, they are, with a few reserva-
tions, directly applicable to man — especially as it has been demon-
strated that there are no vital differences between the findings
obtained with man and those with animals.

In general, it may be said that during the war, and after, there
was a disarrangement of the nutritional elements resulting in a
decreased consumption of protein and fat and a greater intake of
carbohydrate, together with a diminution of vitamines. It should
be noted here that the more important dietary constituents, in com-
parison with the physiologically inferior ones, have increased greatly
in price, so that there is this danger to contend with in addition to
the element of scarcity. Now, when a well-planned animal experi-
ment is undertaken, all factors are controlled as far as possible except
that one whose influence upon the organism is being determined.
Quite another state of affairs confronts us in the case of nutrition
investigations on man, where conditions arise which are only rarely
met with in animal experiments. If there is a lack of one constituent,
then it is almost certain that the entire choice of diet is not correct.
For example, if the protein content of the diet is too low, it is at the
same tune apparent that the carbohydrates are present in excess,
that the vitamine content is diminished, and that the inorganic
elements are inadequately grouped. It is easily possible that right
at this point is where we must search for an explanation of the
pathologically occurring avitaminoses, the etiology of which it is so
difficult to establish. If in such a case as mentioned above, the
missing factor in the diet is supplied, good results do not follow, for the
reason that still other factors have not been taken into consideration.

Under certain circumstances, a seemingly well chosen diet may
prove to be inadequate, particularly when special demands are made
upon the individual, such as hard work, growth, birth, and nursing.

18 THE VITAMINES

All these factors must be taken into account when commenting upon
the pathological conditions which are of exceptional interest to us.
These are chiefly the conditions which give us an insight into the
causes, which are so difficult to determine in hunger edema and
pellagra. Even a well informed physician may easily be led astray.
He questions his patients about their diet, whereupon they enumerate
a long list of foodstuffs, from which apparently nothing has been
omitted. Immediately, his attention will be directed to obvious
things, such as mode of living, nature of work performed, and method
of preparing and cooking the food — everything which may be etio-
logically important and may help him solve his problem.

Despite the fact that a number of ideas originated by us are
credited to others, it is a source of pleasure to witness the great
progress that has been made in vitamine research. In our opinion,
the name "Vitamine", proposed by us in 1912, contributed in no
small measure to the dissemination of these ideas. The word,
"Vitamine", served as a catch word which meant something even to the
uninitiated, and it was not by mere accident that just at that time,
research developed so markedly in this direction.

Our view as to the fortunate choice of this name is strengthened, on
the one hand, because it has become popular (and a badly chosen
catchword, like a folksong without feeling, can never become pop-
ular), and on the other, because of the untiring efforts of other workers
to introduce a varied nomenclature, for example, "accessory food
factors, food hormones, water-soluble B and fat-soluble A, nutramine,
and auximone" (for plants). Some of these designations are cer-
tainly not better, while others are much worse than "Vitamine."

HISTORICAL SURVEY

In spite of the fact that the knowledge of the vitamines taken as
a whole is not older than ten years, and although until lately, the
idea was prevalent that for the complete nutrition of an animal
organism only proteins, fats, carbohydrates, salts and water were
necessary, there is nevertheless, in the older literature, no lack of
statements which of themselves should have given rise to an eager
search for additional dietary components essential to life. The
progress which has already been made by research in vitamines
removes all doubt as to the actual existence of such substances, and
every year brings forth new findings which enhance the importance
of the vitamines to life.

The scientific research leading to the conception of the vitamines
proceeded through many intermediate stages, which we shall shortly
describe. A great stimulus to the development of the modern science
of nutrition was furnished by the investigations into the chemistry
of the proteins which we owe, above all, to the classical work of
Emil Fischer and Kossel. These investigations not only contributed
to the knowledge of the composition of the proteins, but also gave rise
to the study of the relationship between the individual dietary con-
stituents. They demonstrated particularly, that the various proteins
exhibited, qualitatively as well as quantitatively, a varied composi-
tion, and that naturally occurring proteins have not the same physi-
ological value. Without going into details, which may be found in
any text-book, we shall take up only those facts bearing upon the
study of the vitamines. It has been shown by many investigations
that certain amino acids, such as tyrosine, tryptophane, arginine and
lysine, are more or less indispensable to the animal organism. This
question is not yet completely settled, but we know, however, that
some proteins, for example, zein (which is lacking in tryptophane),
or gelatine (which lacks several important amino acids), are not
sufficient for normal maintenance and growth. Latest developments
also indicate that an animal will utilize a diet containing animal
proteins, better than one made up of plant proteins. This conception
is based preeminently upon the supposition that animal proteins
have a composition more nearly related to body protein than do plant

19

20 THE VITAMINES

proteins. That is to imply that the animal organism needs a smaller
amount of animal protein to maintain its nitrogen balance. This
view was utilized by Thomas (1) to group the various proteins of
animal and plant origin according to their biological value. All
these questions have exerted their influence upon the development
of the study of the vitamines, and are even now closely related to
many of our problems to which we shall call attention later.

Another matter that is of interest, is the extent of the ability of
the animal body to synthesize many of the substances necessary for
its existence. The peculiar deficiency symptoms, of which we speak
in this book, have often been attributed to the apparent inability of
animals to synthesize some of the body constituents. The sub-
stances which, of all others, come to mind here are the nucleins and
the lipoids or phosphatides. If this were really the case, then these
substances would also have to be regarded as vitamines, a conception
which is upheld even now by some authors. The older animal
experiments, conducted with purified food substances, showed that
nucleins and lipoids actually could be synthesized. In spite of this,
we encounter in the work of the last ten years, the observation that
these substances have exhibited real therapeutic and nutritive
qualities, an observation that found its chief exponent in H.
Schaumann (2) who elaborated the theory of phosphorus insuffi-
ciency as an explanation of beriberi and similar diseases. This inter-
pretation was vigorously opposed by the writer and gave him the
chance to propound the theory of the vitamines. We shall speak of
the nucleins and lipoids in greater detail in another chapter, but we
shall also touch upon them lightly at this point.

Even before the vitamines were known, reports were published
dealing with the synthesis of lipoids in the body. McCollum,
Halpin and Drescher (3) working with chickens on a lipoid-free
diet, showed that the eggs contained a normal amount of lecithin.
Fingerling (4) and also Abderhalden (5) were able to show that
animals could build up the needed organic phosphorus compounds
from inorganic phosphates. Still, we find in the description of the
condition of the above animals, some observations that point to the
necessity of lipoids for life. It remained for vitamine research to
clear up this matter completely.

In this connection, we shall mention, first of all, the important
work of Stepp (6) in 1909. He made the significant observation that

HISTORICAL SURVEY 21

mice cannot maintain themselves on a diet composed of bread, and
milk which has been extracted with alcohol. If, however, the
extracted portion is recombined with the diet, then the animals
begin to grow once more. In another communication, Stepp (7)
was convinced that his findings were not due to the loss of salts
during extraction, and classified in the lipoid group, those substances
essential to life. In spite of the fact that this conclusion was not
quite justified, the work of Stepp really merited more attention than
it received at the tune of its publication. Unfortunately, this was
not the case, for the relegation of the substances necessary for life
to the class of lipoids, which already had been considered essential,
did not signify real progress.

Aside from the work of Stepp, attention could easily have been
focused upon the conception of the vitamines by research which had
for its purpose the study of the importance of salts in nutrition.
Relative to this, the school of Bunge is credited with a great deal of
merit. Another series of investigations dealt with the utilization of
purified foodstuffs, especially in the mouse, the rat and the dog.

In 1873, Forster (8) tried to determine whether or not dogs could
maintain themselves on an ash-free diet. For this purpose, he fed
the meat remaining after the preparation of Liebig's meat extract.
These residues were washed repeatedly with distilled water until
they contained only 0.8 per cent ash. They were then combined
with fat, sugar and starch, and fed to dogs, with the result that they
died sooner than starving animals. Experiments with pigeons, fed
with casein and starch (and occasionally a little fat), gave the same
results. The symptoms noted by Forster, in pigeons, were partic-
ularly interesting. They refused food, lost a great deal of weight,
showed weakness, opisthotonos and characteristic circular motions.
As we shall see later, these symptoms, which Forster attributed to a
lack of salts, were apparently identical with "poly neuritis galli-
narum," a disease of chickens described by Eijkman in 1897. This
view is further strengthened when we note the length of time that
the pigeons lived — 13, 26 and 31 days. In dogs, there was noted
tremor, peculiar gait and weakness similar to that of paresis.

The investigations from Bunge's laboratory, which are mentioned
in his book on physiological and pathological chemistry (9), are
worthy of attention. Lunin (10) reported experiments with mice,
fed on casein, fat and cane sugar. Out of five animals used, one

22 THE VITAMINES

lived 11 days; the others, 13, 14, 15 and 21 days, respectively, while
starving animals lived only from 3 to 4 days. The addition of soda,
which served the purpose of neutralizing the sulphuric acid arising
from the protein cleavage prolonged the life of the animals to a
certain extent; the ash of milk likewise had the same effect. Lunin
explained the results as being due to a lack of organic phosphorus
compounds (lecithin), and to a disturbance in the balance between
the inorganic and organic food components. Subsequently, Lunin
made the unusually important observation that mice could thrive
very well on milk powder even after two and a half months, and
therefore concluded that milk contained besides the known elements,
other unknown substances essential to life. That this conclusion
had already been arrived at in 1881 must appear to us as truly
remarkable.1 Socin (11), working in the same laboratory, also came
to a similar conclusion in 1891. His problem required him to find
out whether inorganic or organic iron is the better utilized in the
animal body. Mice were kept on a diet consisting of blood serum,
fat, sugar, starch, cellulose and ash (obtained from milk). The
missing iron was added in the form of hemoglobin, hematin, or iron
chloride, without the least effect being apparent on the duration of
life. All of the animals died after 32 days, while the controls, fed
on yolk of egg, starch and cellulose, were still alive after 99 days.
Bunge (12) himself obtained the same results. Hausermann (13),
working with rats, guinea pigs, rabbits, cats and dogs, kept on white
bread and rice, could see an improvement after the addition of iron,
although a normal state of nutrition could not be brought about.
The experiments of Socin were confirmed by Hall (14) and
Coppola (15).

Considering simple feeding experiments, we find the work of
Pasqualis (16) on chickens, receiving a diet composed of 14 per cent
protein, 65 per cent corn starch, 4 per cent dextrin, 4 per cent sugar,
9 per cent olive oil, 2 per cent wood chips, 1 per cent common salt

1 Lunin concluded that "mice can live well under these conditions when
receiving suitable foods (milk) but as the above experiments demonstrate
that they were unable to live on proteins, fats, carbohydrates, salts and water,
it follows that other substances indispensable for nutrition must be present
in milk besides casein, fat, lactose and salts." This conclusion was quoted
in Bunge's Physiological and Pathological Chemistry, but did not attract
the attention it merited.

HISTORICAL SURVEY 23

and 1 per cent salt mixture (in the form of corn ash). To his aston-
ishment be observed that the animals lost weight and died — in spite
of the large food intake. The first exact feeding experiments on
rats were carried out by Henriques and Hansen (17). The experi-
ments were, in the main, of short duration, lasting only from three
to four weeks. During this time, the rats gained weight and showed
a retention of nitrogen, but the experimental period was apparently
too short to discover any food deficiency. Different findings, how-
ever, were reported by Falta and Noeggerath (18) who repeated
these experiments. They fed various proteins of animal origin
together with fat, sugar, starch, salts, lecithin and cholesterol. It
was possible to determine the value of the various proteins, although
not one of the combinations was permanently sufficient, as was
apparent by the poor nourishment to be noted after varying intervals
of time. The deficiencies of the diet made their appearance only
after prolonged feeding. If the experiment had been terminated at
the end of four weeks, the false conclusion could have been made
that the diet had been adequate. The food intake was diminished,
and the interesting observation was made that the eyes of the rats
exhibited pathological changes, of which we shall have more to say
later. Working in the laboratory of O. Frank, Jacob (19) reported
some feeding experiments with pigeons and rats on a diet which at
that time was thought to be sufficient. The pigeons died after a
maximum of four weeks with symptoms of severe digestive dis-
turbance, although they lived somewhat longer when casein was
replaced by meat powder. Rats, on the contrary, lived longer on
casein and died after 43, 73 and 125 days. Undoubtedly, the cause
of death was the unsuitable composition of the diet, though Jacob
considered it to be due to the uniformity and lack of stimulating sub-
stances in the diet. The same trend of thought is often met with
in the older as well as the newer literature. For instance,
McCollum (20) explained the dietary deficiency of synthetic diets
as being due to lack of flavoring and stimulating substances,
although at present he is the most eager advocate of the new
ideas. Through the addition of these flavoring and stimulating
substances, he was unable to demonstrate an improvement in the
nutritive condition. A very interesting investigation was that made
by Watson (21). He fed rats, both young and full grown, on various
meat products. The young animals either died very quickly or

24 THE VITAMINES

showed inhibition of growth, while full grown animals suffered from
paresis and sterility. This last symptom was not to be attributed
to lack of tryptophane. The animals whose growth had been stunted
resumed normal growth on a mixed diet.

About 1906, there appeared the classical work of Hopkins and his
pupils, who displayed marked clearness in their course of reasoning.
Hopkins fed mice on a mixture which contained zein (one of the corn
proteins) which lacks tryptophane. The young animals were able to
live only 16 days, whereas upon the addition of this amino acid, life
was prolonged for 14 days more. Tyrosine exerted no influence. It
appeared from these results that the above diet was lacking in some-
thing else besides tryptophane. The appearance of animals so fed,
as described by Wilcock and Hopkins (22) was not very bright. The
animals were torpid, had cold extremities, half-closed eyes and slimy
fur. At that time, Hopkins thought that tryptophane could be
conceived of as the precursor of adrenaline. The chemical nature of
the missing components (even after the addition of tryptophane) was
not recognized at that time, although the experiments led Hopkins (23)
to the prophetic statement which is reproduced here verbatim:

But further, no animal can live upon a mixture of pure protein, fat and
carbohydrate, and even when the necessary inorganic material is carefully
supplied, the animal still cannot flourish. The animal body is adjusted to
live either upon plant tissue or other animals and these contain countless
substances other than the proteins, carbohydrates and fats. Physiological
evolution, I believe, has made some of these well nigh as essential as are the
basal constituents of diet; lecithin for instance, has been repeatedly shown
to have a marked influence upon nutrition, and this just happens to be some-
thing familiar, and a substance that happens to have been tried. The field
is almost unexplored, only it is certain that there are many minor factors in
all diets of which the body takes account. In diseases, such as rickets, and
particularly scurvy, we have had for long years knowledge of the dietetic
factor, but though we know how to benefit these conditions empirically, the
real errors in the diet are to this day quite obscure. They are, however,
certainly of the kind which comprises these minimal quantitative factors
that I am considering. Scurvy and rickets are conditions so severe that they
force themselves upon our attention, but many other nutritive errors affect
the health of individuals to a degree most important to themselves, and some
of them depend upon unsuspected dietetic factors.

All that Hopkins says in this short paragraph applies to this very
day, although from the point of view of that time it was held, for
example, that lecithin was essential for life. These assertions of

HISTORICAL SURVEY 25

Hopkins were unknown, because he offered no experimental evidence
in their support till 1912.2

Till a short time ago, there was no lack of opinions which denied
the existence of substances essential to life. We have only to recall
the work of Abderhalden and Lampe in 1913 (25), although lately
Abderhalden has changed his view upon the matter. He (26) con-
ducted many feeding experiments with various proteins, hydrolyzed
meat, etc., without once recognizing that these diets were lacking in
something that was essential.3

It is true that the experiments did not last -very long, and that the
various constituents were not carefully purified. Incidentally, it
must be noted that in these experiments short periods, in which a
more natural diet was given, were frequently interspersed in order
to abate for a time the distressing deficiency symptoms. Above all
things, we must recognize that in dogs, the symptoms manifested
because of a deficient diet, are not so pronounced as in pigeons and
rats. Some time after the vitamine theory had already become
familiar, Abderhalden (29) found indeed that digested casein had less
value for growing animals than digested meat. He immediately
assumed that glycine was the cause of the difference, whereupon he
started various experiments, adding this amino acid, but naturally
without any success. Similar criticism may be made of the work of

2 For instance, Rohmann (24) is mistaken when he asserts that the ideas
expressed by us originated with Hopkins. Inasmuch as we entered the field
in 1911, the statements of Hopkins were unknown to us.

3 While I was working in the city hospital at Wiesbaden, I carried out under
Abderhalden's direction, some metabolism experiments with dogs. The
animals were fed with edestin, gliadin, milk powder and meat, together with
other usual dietary constituents. Since the experiments were considered
unsuccessful, only a small part of the work was published by Abderhalden
and my self (27). At that time, most of my dogs died, so that the feeding
experiments had to be started anew a number of times. Even then, I made
the observation that animals fed on edestin or gliadin quickly recovered on
addition of milk powder or meat. I wrote to Abderhalden a number of times,
that the edestin and gliadin combination was not sufficient to make the animals
thrive. The symptoms which I observed then recall those noted by Chit-
tenden and Underbill (28) in dogs fed on peas. To my letters, I received the
na'ive reply from Abderhalden, that in such metabolism experiments, success
depends upon the manner in which the animals are taken care of. I conscien-
tiously attempted to follow this advice, but the dogs, on such a poor diet,
refused to manifest any friendship for me.

26 THE VITAMINES

Grafe (30) who studied the protein-sparing action of ammonium
salts. The same thing applies to the work of many others on animal
nutrition.

One of the last opponents of the vitamine theory was the late
Rohmann (31 and I.e. 24). He expressed the idea that the existence
of the vitamines was seized upon only because certain investigators
had had poor results with their experimental animals. The subject
was treated by Rohmann in a rather illogical manner, since in the
end he admitted the existence of the vitamines, under a different
name. His statements, in particular, have been discussed by so
many authors that we shall not consider them further here.

In spite of the great powers of observation possessed by such
investigators as Bunge and Hopkins, the conception of the vitamine
theory could never have attained its present importance, if a powerful
impulse had not been received from clinical sources. The findings
mentioned above have not been utilized in human pathology, since
most clinicians are not very much disposed to apply the results of
animal experiments to man. However, great interest in vitamines
was justifiably aroused when the applicability of the results to human
pathology and physiology was demonstrated. For a long time, long:
before the publication of the laboratory findings, the literature con-
tained views upon the etiology of scurvy, rickets and pellagra which
appeared to be very nearly correct. The conditions surrounding
these diseases were, however, too complicated to be suitable for a
direct experimental research. On the other hand, as regards beriberi,
the circumstances were entirely different. Here we had to deal with
a problem the etiology of which was comparatively simple, for the
disease could be brought logically into causal relationship with the
continued consumption of rice. Nevertheless, many years of effort
were necessary for these conceptions to gain a foothold in the litera-
ture. When the first edition of this book was being written in 1913,
it was still necessary for us to wage a hard fight in support of our
contention. Such is not the case at present, for the characterization
of beriberi as an avitaminosis has met with general recognition. At
the end of this chapter, we shall discuss the history of beriberi
research, since it may logically serve as an introduction to the study
of the vitamines.

We shall begin here with a discussion of scurvy, although this
disease really did not contribute any direct stimulus to vitamine

HISTORICAL SURVEY 27

research. Scurvy, however, is the first disease the etiology of which
was associated with a definite mode of nutrition. The reasons why
scurvy gave no immediate impulse to research were to be looked for
in the variety of the feeding, which might have been responsible for
the onset of the disease. It was difficult to conceive of the disease
as being due to a lack of one and the same substance in the diet.

It was Kramer (32), an Austrian army physician, who recognized
for the first time, the existence of scurvy. In 1720 with a field army
in Hungary, he was confronted with a severe epidemic of scurvy. He
wrote to the authorities and to his colleagues to secure help.
A shipment of dried antiscorbutic herbs was hurried to him in spite
of which thousands died of this disease. He then made the following
entry in his book:

Scurvy is a terrible disease for which there is no known cure. Medication
does not help, neither does surgery. Be careful of bleeding; shun mercury
as a poison. The gums may be massaged, the stiff joints may be rubbed
with fat — but all in vain. If one could only have available a supply of green
vegetables, or a sufficient amount of the vital antiscorbutic juices; or if one
could have at hand oranges, limes or lemons, or their preserved pulp or juice
so that a lemonade could be made out of them; or administered as such in
three or four ounce doses — then one could be in a position to cure this dreadful
disease, without other help.

As we see, Kramer selected a method of treatment which could
not be better chosen even today.

Bachstrom (33) recognized in 1734 that the incidence of scurvy
was not due to cold weather, sea air or salted meat, but to a lack of
fresh vegetables. The latter, he perceived, was the primary cause
of the disease. Lind (34), in his work on scurvy, has noted many
cases which were cured by administration of oranges or lemons.
Cider was next to oranges in its efficacy. At that time, he
made the important observation that very severe cases could be
cured in 6 days, and recognized also that hard work accentuated the
symptoms of scurvy. Coming now to the modern history of scurvy,
we must give prominence, above all others, to the name of Barlow (35)
who attributed the onset of infantile scurvy (also called, Moller-
Barlow's disease) to milk which had been heated for a long time. It
was only lately, in 1907, that scurvy in guinea pigs was discovered —
a discovery of the greatest significance to vitamine research. The
work of Hoist and Frolich (36) on this subject was repeated on all
sides, and their results were completely corroborated.

28 THE VITAMINES

Research on rickets was undertaken experimentally only in late
years, although Miller (37) had already associated rickets with a
particular dietary even before the vitamines were known. He
thought that many of his cases were due to a lack of butter, and

FIG. 1 FIG. 2 FIG. 3

FIG. 1. DECREASE IN THE NUMBER OP CASES OP BERIBERI IN THE JAPANESE
NAVT AFTER THE INTRODUCTION OP MEAT INTO THE DIETARY (TAKAKI)

FIG. 2. THE CALORIC VALUE OF THE FOOD DIMINISHED (TAKAKI) (CF. FIG. 1)
FIG. 3. THE BOOT WEIGHT INCREASED (TAKAKI) (CF. FIGS. 1-2)

consequently prescribed milk, cream, butter, egg-yolk and cod liver
oil. Schabad (38) saw a marked difference between olive oil, sesame
oil, and cod liver oil in their influence on rickets, and in a later
work (39) he discusses the question as to the nature of the curative
substance.

HISTORICAL SURVEY

29

He was convinced that it was not a ferment and that its efficacy
was not diminished after heating for one hour at 100°C. Since the
war, great strides have been made in our knowledge of scurvy and
rickets, and we shall discuss them in later chapters.

Beriberi is an avitaminosis about which we are best informed.
Although known for hundreds and perhaps thousands of years, it is
only in the last 25 years that actual progress has been made relative
to its occurrence and prevention. This is perhaps due to the increase
in the number of cases of beriberi, coincident with the introduction
of modern machinery for rice milling. It was quite properly pre-
sumed by Wernich (40) and van Leent (41) that there existed a
causative relationship between rice consumption and beriberi. In
1882, Takaki (42) proposed a change in the rice diet of the Japanese
navy, so that meat, bread, fruit and vegetables were added. Since
that time beriberi has almost completely disappeared.

Takaki's tables are exceptionally instructive. It is apparent from
Fig. 1, that upon the elimination of the rice diet in 1882, there was
an immediate drop in the number of cases of beriberi, which number
subsequently remained trifling. In Figs. 2 and 3 it may be seen
further, that although the caloric value of the new diet was smaller
in comparison to that of the previous one (causing beriberi), there
was a definite increase in the average weight of the men.4

Even greater strides were made in Java by Dutch investigators.
Based upon very rich statistical material collected at Eijkman's
suggestion by Vordermann (43) in a large number of Javanese prisons,
it was possible to demonstrate that the disease was associated with
the ingestion of white (polished) rice. The fluctuation in the number
of cases of beriberi due to various kinds of rice is well illustrated in
Vordermann's Table:

NUMBER OF

NUMBMR OF

BERIBERI CASES

PRISONERS

CASES

NUMBER OF PRISONERS

White rice

150,266

4,201

1:39

Rice with partial "silver skin". .
Unpolished rice

35,082
96,530

85
9

1:416
1:10725

The findings of Vordermann were corroborated by Braddon (44)
on a large number of cases in the Malay peninsula. He was able to

4 Figures 1, 2, and 3.

30 THE VITAMINES

show that some natives, like the races of Tamils, who live on "cured"
or "parboiled" rice, remain free from this disease. This rice was so
prepared that it was steamed before using; the husk came off easily,
thus obviating the necessity for polishing. To convince himself of
the accuracy of Braddon's observations, Fletcher (45) in 1905 under-
took, in the Kuala Lumpur insane asylum, some research on lunatics,
which lasted for one year. His purpose was to determine the differ-
ence between "cured" and ordinary decorticated rice. Throughout
the entire perod, the diet varied only qualitatively; out of 120
patients on polished rice, 36 contracted beriberi from which 18 died,
while from 123 patients on "parboiled" rice, only two developed
beriberi, and these two cases were admitted as such into the hospital.
In 1909, Ellis (46), working at the insane asylum in Singapore,
reported similar investigations, which extended back as far as 1901.
In his fruitful statistics, he showed definitely that the number of
beriberi cases decreased steadily from year to year, upon gradually
substituting steamed rice for polished rice. Analagous results were
also obtained on healthy laborers by Fraser (47) .

In 1897, Eijkman (48) (cf. Vordermann), found that beriberi is
brought about by a long continued consumption of white rice, since
polishing removes a substance which is protective against the out-
break of beriberi. The Dutch investigators were of the opinion
that the most important part of the rice grain, the so-called "silver-
vlissen" (silver skin), could neutralize the toxins of white rice. There-
upon, it fell to Fraser and Stanton (49) to make clear the point that
upon polishing rice, more than just the "silver skin" is lost.

Eijkman's (50) discovery of experimental beriberi marked a great
step forward. This finding was made accidentally since Eijkman
observed that chickens which fed upon the remains of the food used
in a hospital for beriberi died of a disease which he recognized oppor-
tunely to be similar to human beriberi This discovery, which was
made in 1896 (the disease was called "poly neuritis gallinarum"),
made it possible to get away from experiments upon man, and assured
the collection of more valuable experimental data in a shorter time.

After the discovery of experimental beriberi, Eijkman went a
step further. He found that the addition of the pericarp of the rice
kernel, or even the rice bran, to white rice made it possible to prevent
the occurrence of beriberi in animals. . For these observations,
however, he did not find the correct explanation; he believed that the

HISTORICAL SURVEY 31

starch of the grain gave rise to toxins which exerted a deleterious
action on the nervous system, and that this was prevented by the
addition of the pericarp. This conception should not surprise us
since in those days the nature of the disease was quite puzzling.
Eijkman (51) likewise made the important observation that the
watery extract of rice bran possessed therapeutic properties. Phytin
was found in the rice bran, but it was shown to be without effect on
beriberi. He noted also that the curative substance is dialysable
and not precipitated by the addition of alcohol. The observations
made in 1897 were, after all, those upon which the modern research
on beriberi is based. All credit is due Eijkman for having laid the
foundation for the conduct of future experiments.

Grijns (52), continuing the work, was able to confirm the experi-
ments of Eijkman completely. He was the first worker to express
clearly a conception of beriberi, which holds good to this day. Grijns
said that the disease developed when the diet was lacking certain
substances which were of importance in the metabolism of the
peripheral nervous system.5 Curative substances similar to those
occurring in rice bran were found by Grijns in a kind of bean called
"Katjang-idjoe" (Phaseolus radiatus), and in meat; he also demon-
strated that these foodstuffs lost their curative properties when they
were heated to 120°C. These experiments were of greatest signifi-
cance in the further development of the question, and they were
also confirmed by Eijkman (I.e. 51). Breaudat (54) used rice bran
in the treatment of human beriberi with good results. Fraser and
Stanton (55) sought to determine more definitely the nature of the
substance. They found that it was soluble in strong alcohol and
that it retained its activity after the removal of alcohol-soluble
proteins. They made analyses of various kinds of rice and believed
that a rice poor in phosphorus would cause beriberi. Thereupon,
they suggested the phosphorus content as a practical indicator
of the nutritive value of rice. For instance, rice which contained
a minimum of 0.46 per cent P2O5 was to be considered harmless.
However, we should not forget that Schuffner and Kuenen (56)
have shown that the method of preparation of the rice diet is
likewise of importance. That is, the rice should be partaken of
together with the broth, particularly in the case of whole rice.

5 Eijkman carried out a lengthy polemic on this point, but he admits now
(53) that Grijns was correct.

32 THE VITAMINES

If the broth is regularly discarded then beriberi can easily occur,
even with whole rice. Based upon the findings of Eraser and Stanton,
Schaumann (57) thought of beriberi as a disturbance in metab-
olism due to a lack of organic phosphorus compounds. This theory
was likewise applied to other avitaminoses, such as scurvy and ship
beriberi. It found disciples (Simpson and Edie (58)), and dominated
this field of pathology till the advent of the vitamine theory. At
that tune, it was quite plausible to regard the great difference in
phosphorus content between white rice and rice bran, and the
undeserved acclaim of organic phosphorus compounds, as thera-
peutic factors in medicine.

In the years that followed, there appeared in quick succession a
great number of experiments dealing with the chemical nature of the
curative substance. Various foodstuffs were used for this purpose.
Hulshoff Pol (59) showed that a watery extract of "Katjang-idjoe"
beans, clarified with lead acetate, contained the curative substance.
From this clear filtrate, Pol obtained a crystalline substance which
he called "X-acid;" there is a lack of further information as to
whether this substance possessed any characteristic physiological
properties.

Schaumann (I.e. 2), who extended the list of curative substances
to include yeast (which was used by Thompson and Simpson (60) in
the treatment of human beriberi), investigated the influence of the
already known yeast constituents. Among these he investigated
the influence of yeast nucleic acid and yeast lecithin, but without
obtaining definite results. Eijkman (61) showed that the active
substance of yeast could be extracted with 88 per cent alcohol. Funk
(62) had already noticed this before, but by this method only an
incomplete extraction could be effected. Teruuchi (63) extracted rice
bran with dilute hydrochloric acid, neutralized the solution, thereby
precipitating phytin, then evaporated the filtrate and extracted the
residue with alcohol. This extract was active and contained only a
small proportion of its original phosphorus content. Similar results
were also obtained by Chamberlain and Vedder (64). They found
that the curative substance was adsorbed with animal charcoal and
tried to develop a procedure based upon this finding, but failed.
Shiga and Kusama (65) found that the active principle of rice bran
was destroyed by heating to 130°C. with 0.5 per cent hydrochloric
acid or with 1 per cent soda solution, but not at 100°C.

HISTORICAL SURVEY 33

Research on the preparation of the active substance from rice
polishings were also conducted by Tsuzuki (66), but with little suc-
cess. Owing to the enormous content of phytin in rice bran, Aron
and Hocson (67) believed that it was curative; the good results they
obtained may be explained by the probability that the phytin was
contaminated by some of the active substance. Research on phytin
had already been carried out by Eijkman without results, and also
by Cooper and Funk (68).

To summarize our knowledge of the chemical nature of the active
principle prior to the introduction of the vitamine theory (till 1911),
the following may be set down with certainty:

1. The substance is soluble in water, alcohol and acidified alcohol.

2. The substance is dialysable.

3. The substance is destroyed at 130°C.

When we took up the question in 1911, it was not known whether
the active substance was organic or inorganic in nature, whether or
not it was a constituent of proteins, nucleins or phosphatides. It was
not certain that we were not dealing with a ferment, nor was it known
if the substance belonged to some chemical group already described,
or to some new unknown class of substances. We shall be in a
position to answer a good many of these questions during the course
of our discussions.

INTRODUCTORY

In the historical part, we have seen that the animal organism
cannot live very long upon an artificially prepared diet. Since
systematic investigations into the vitamine requirements of all classes
of plants and animals have not yet been made, it is not possible to
say with entire certainty that the above contention is generally true.
The firmly established importance of the vitamines for the existence
of certain animals and plants, organisms which are far removed from
each other genetically, makes it apparent that these substances are
of universal importance to life.

It is evident that there are differences in the qualitative and
quantitative requirements and it may later develop that various
organisms need various quantities as well as various kinds of vita-
mines. We shall briefly enumerate the facts that have led us to this
conclusion, and point out how many types of vitamines are known
up to the present. We choose to speak of vitamine types in this
case, and not of definite vitamines, for as long as these substances
remain unidentified, and till such time as they may be compared
with each other in the pure state, it is obviously impossible to talk
of their identity. Until now, we have differentiated three such
types. Whether with these three types we have finally reached
the possible limit, it cannot be stated definitely; however, it appears
unlikely that Nature, in the variety of its manifestations, essential
conditions and intensity of metabolism, should limit itself to only
three types. On the other hand, it is possible that after purification,
that which seemed to us a single substance might well prove to be
a mixture. We may be dealing with complex substances having
some chemical groups in common, and others, the significance of
which has not yet been demonstrated.

It would be quite premature to propose, with McCollum (69), that
only two or three vitamines exist in nature. He arrived at this
conclusion as a result of his numerous rat experiments, which showed
that for this species two types of vitamines were sufficient, and
thereupon drew conclusions that were to be applied to the entire
plant and animal kingdom. Although we are very well informed as
to the food requirements of rats, compared to those of other organ-

35

36 THE VITAMINES

isms, we cannot maintain that we know all that there is to be known
of the requisite food constituents, as long as we shall not have
obtained them (including vitamines) in the chemically pure form.
This applies naturally even more to other animals, about whose
metabolism we are still less informed. It is therefore of greatest
importance, at this stage of our knowledge, to disregard generaliza-
tions of this sort since they are detrimental to the development of
the questions that interest us.

We shall now consider the work leading to the present conception
of the three different vitamine types. When we pointed out in 1912
(I.e. 62) l that the animal organism needs vitamine for complete
nutrition, the term, "vitamine," was used only in a very general way.
At that time, we drew a distinction between the antiberiberi, anti-
scorbutic and antirachitic vitamines, although this classification was
based only upon the physical characteristics, origin and influence on
metabolism. New facts have been brought to light, but this classi-
fication has remained unchanged. We knew in 1912 that the pericarp
of grains contained something that was of importance in the metab-
olism of man and certain species of birds. Then Schaumann (I.e. 2)
found that yeast and some animal extracts contained a subtance of
similar nature. Furthermore, it was known that man as well as
the guinea pig was in need of a vitamine as a protection against
scurvy — a vitamine that was recognized by us to be quite different
from the antiberiberi vitamine. In spite of the above mentioned
facts and the work of Stepp (I.e. 6, 7) already referred to, and in
spite of the results of vitamine research in 1911, the new ideas were
not immediately applied to the general science of nutrition. The

1 I regarded it of paramount importance, that the then ruling conception
of the necessity of the lipoids or the nuclein substances was substituted by the
fundamentally different vitamine theory. At the same time, I must admit
that when I chose the name, "vitamine," I was well aware that these sub-
stances might later prove not to be of an amine nature. However, it was
necessary for me to choose a name that would sound well and serve as a catch-
word, since I had already at that time no doubt about the importance and
the future popularity of the new field. As we have noted in the historical
part, there was no lack of those who suspected the importance of still other
dietary constituents, besides those already known, for the nutrition of animals.
These views were unfortunately unknown to me in 1912, since no experimental
evidence had appeared in their support. I was, however, the first one to
recognize that we had to deal with a new class of chemical substances, a view
which I do not need to alter now after eight years.

INTRODUCTORY 37

only investigation on this subject was the classical work of Hopkins
(70) which appeared in 1912, demonstrating that the addition of a
small quantity of milk to an artificial diet induced growth in rats.
The amount of milk was so small as to be negligible, as far as its
energy factor was concerned. In these experiments, there was no
attempt to differentiate between the various vitamines, since milk
contains all that is necessary to life. While the vitamine conception
had at that tune attained a definite standing in England (not without
vigorous effort), the dissemination of these ideas in other countries
met with but poor success. For instance, Abderhalden and Lampe
denied the existence of vitamines in 1913, and Rohmann even in
1916. In the United States there appeared simultaneously with the
beginning of vitamine research the very important work of Osborne and
Mendel (71) on the artificial feeding of rats, which greatly advanced
our knowledge of the food value of the various kinds of proteins.
These investigations were indeed the first to be carried out with
such carefully purified proteins and over such a long period of time
(more than one year, one-third of the lifetime of a rat). These
experiments served two purposes, first, to determine the nutritional
value of various proteins, and secondly, to determine how long rats
can live on an artificial diet. At that tune, we emphasized the fact
that for such investigations it is particularly important to provide
the animals with vitamines, if clear results are to be obtained.
Besides this, we showed that in many nutrition experiments the diet
was in some unknown manner contaminated by vitamines which
were responsible for the length of time the animals survived. This
was later shown to be true; for example, we could demonstrate the
presence of some nitrogen -containing impurity in milk sugar; the
same is true of other products derived from milk. In the experi-
ments of Osborne and Mendel, the diets, taken as a whole, were
obviously lacking in vitamines, since many of the animals died
suddenly, or they would have died if the diet had not been changed
quickly. Young animals lived for a certain time, but mostly failed
to grow.

In another investigation, Osborne and Mendel (72) described
experiments in which rats were fed on a mixture that could have
been thought of as fat-free. It contained, among other things,
protein-free powdered milk extracted with ether. In this case also,
normal growth was obtained. The same authors then analyzed the

38 THE VITAMINES

protein-free milk for its inorganic constituents, and prepared an
artificial protein-free milk having the same composition; this also
permitted of good growth. In a later communication, these inves-
tigators (73) pointed out that the results were not so favorable with
the artificial preparation. Their animals lived from 114 to 277 days,
but after this they died without the post-mortem giving any plausible
reason for death. In one case particularly, the addition of natural
food was ineffective in preventing death.

Hopkins and Neville (74) reported that they attempted to replace
milk by a preparation made according to Osborne and Mendel.
They stated that when they used purified lactose, which had been
prepared from milk, the animals failed to grow. McCollum and
Davis (75) found that rats, weighing 40 to 50 grams, could grow
normally for about three months on the Osborne-Mendel diet. At
the end of this time the animals stopped growing, but were in good
general health. In the light of our work on vitamines, the authors
came to the conclusion that the failure to grow was not due to lack
of salt, fat and phosphatides, but to lack of vitamines. They found
the necessary substances in egg-yolk and butter, especially in the
ether-soluble portion. Besides this, they found that rats on the
above diet produced very little milk, and that the young were
stunted. Normal growth was restored upon the addition of the
ether extract of eggs or butter.

Almost simultaneously with the work of McCollum and Davis,
there appeared the paper of Osborne and Mendel (76) in which they
confirmed their earlier findings. As a result of these studies, they
showed that artificial milk, like the natural protein-free variety, is
not sufficient for growth. To obtain adequate growth, either milk
powder or butter had to be added.

In later publications, McCollum, as well as Osborne and Mendel,
disregarded the importance of the antiberiberi vitamine for growth
more and more, and reiterated the existence of a specific growth
substance in certain fats, like butter, egg-yolk and others. Little
by little, the entire vitamine structure became shaky, especially after
Osborne and Mendel (77) had started their studies on butter. By
centrifugation, they separated butter into three fractions. They
reported that the pure butter fat, supposedly nitrogen and phosphorus
free, still retained its activity. In particular, they were able to cure
ophthalmia (keratomalacia, a disease of which we shall speak

INTRODUCTORY 39

later) in rats kept on an artificial diet. These experiments were
intended to show that the growth substance could not be a vitamine.
We do not yet know the chemical nature of the substance in butter,
but there is nothing to disprove the conception that this substance
contains nitrogen and may be classified as a vitamine, especially
when it is remembered in what small amounts it may be active.2

Working with Macallum (81) on the fractionation of butter, we
demonstrated that when we followed the procedure of Osborne and
Mendel, and dissolved large amounts of butter fat (about 12 kilos)
in acetone and shook out this solution with dilute hydrochloric acid,
we found 23.4 mgm. of nitrogen in the extract; after hydrolysis of
the fatty residue with dilute hydrochloric acid, an additional 22 mgm.
was obtained. It is quite obvious that it is impossible to remove
all the nitrogen from butter with the above procedure. In spite of
these findings, it is true that butter is poor in nitrogen and therefore
it must contain a substance that is active in minimal amounts.
McCollum and Davis (82) endorsed this opinion, inasmuch as they
doubted the absence of nitrogen in butter. In order to answer the
question, Osborne and Wakeman (83) conducted another experiment
with butter and found that it contained traces of nitrogen and

2 Later on, McCollum and Kennedy (78) sought to introduce the classi-
fication, "water-soluble B" for the antiberiberi vitamine and "fat-soluble
A" for the antirachitic vitamine; lately, Drummond (79) used the term "water-
soluble C" for the antiscorbutic vitamine. Regarding this matter, I have
attempted to show (80) that these designations are incorrect, chemically
and logically. It would be totally inaccurate to differentiate substances
extracted from complicated mixtures by their solubility in certain solvents.
It would seem to be of importance to replace, in the English publications,
such designations as "growth-promoting, water-soluble, accessory B factor,"
by something more simple. For this purpose, I suggest, at least for the present,
the adoption of the following nomenclature:

Vitamine B for the antiberiberi vitamine
Vitamine A for the antirachitic vitamine
Vitamine C for the antiscorbutic vitamine

These letters are already used very often in the English literature, and the
nomenclature to be used in this book is suggested as being simple and time
saving, particularly in indexing the literature. Since this has been written,
a note by Drummond (80a) has appeared in which the same kind of nomen-
clature is proposed, except that he suggests "vitamin" instead of "vitamine"
for the English scientific literature. I cannot agree to this change since
I still believe in the nitrogenous nature of these substances.

40 THE VITAMINES

phosphorus. The question as to the nitrogen content of butter has
not been touched upon for some years, since the solution of the
problem could only be obtained through a purification of vitamine A.
In this connection, the amount of lecithin in butter, based upon the
phosphorus content, was calculated to be 0.017 per cent by Wrampel-
meyer (84) and from 0.04 to 0.07 per cent by Supplee (85) ; the latter
also demonstrated the presence of choline, trimethylamine, and
ammonia.

While Osborne and Mendel, as well as McCollum, regarded butter
and certain other fats as the only growth-promoting substances,
Funk and Macallum (86) went to the opposite extreme, stating that
vitamine B was the only growth-promoting substance. It appears
from the newer investigations, as we have always believed, that
vitamine B is far more important to life and is required in greater
amounts then vitamine A. Nevertheless, both substances are
necessary for growth as has been developed by the above controversy.
The Funk-Macallum experiments showed that butter, when added
to an artificial diet, does not cause growth, and it appeared that an
addition of yeast was imperative. To be sure, there was then no
apparent difference in the efficacy of butter and lard, but it should
be said here that in our investigations we found it necessary to add
a large amount of yeast in order to obtain good results; fresh yeast
worked better than dried yeast. It is not impossible that yeast, in
the fresh condition, contains some vitamine A; on the other hand, in
the light of the results obtained by Daniels and Loughlin (87), it is
possible that some fats, till now looked upon as vitamine-free, contain
enough of this vitamine to stimulate growth in rats.

In this manner, the existence of two vitamines, A and B, was
definitely established. In addition, it became apparent that the
substance playing the greatest part in the growth of rats is either
identical with the antiberiberi vitamine or belongs to the same type.
Coming now to vitamine C, its individuality and its differentiation
from the other two vitamines was determined in the following
manner. While we find vitamine C often associated with vitamine B
in Nature, there are products such as egg-yolk and cereals, which
are very poor in vitamine C but especially rich in vitamine B. The
evidence of their dissimilarity may be seen more clearly in the work
of Seidell (88), who showed that vitamine B could be adsorbed quan-
titatively with Lloyd's reagent (fuller's earth). Harden and Zilva

INTRODUCTORY 41

{89) then showed that from a mixture of autolyzed yeast and orange
juice, substances frequently used as a source of both vitamines,
vitamine B could be completely removed by adsorption with fuller's
earth, while the presence of vitamine C could be demonstrated in the
filtrate. These findings have been corroborated by Byfield, Daniels
and Loughlin (90) incidental to some other work. Vitamine C is the
most unstable of the vitamines, although it appears to be more
stable in juices that are slightly acid. In association with these
facts, there is the discovery of Fiirst (91), who was the first one to
show that if grains are allowed to germinate a formation of vitamine
C takes place, which disappears again upon drying and reappears
upon the addition of water. We have thought (1. c. 62) that in
this observation there was a possibility of finding a genetic relation-
ship between vitamine C and vitamine B. Through progress in the
methods of demonstrating the presence of vitamine B, it may be
possible to investigate the question, if with the formation of vitamine
C there is a corresponding diminution in vitamine B. The relation-
ship between these vitamines has become more interesting, since
Osborne and Mendel (92) showed that known antiscorbutics, such
as fruits, always contain a definite amount of vitamine B. Although
we know that this vitamine has no effect upon scurvy, the reverse of
this, the influence of vitamine C on beriberi, is not sufficiently estab-
lished. It might be possible for instance, that for the cure of scurvy
very small amounts of vitamine C are necessary, whereas to in-
fluence beriberi similarly much larger doses are required. This
matter must be cleared up by further research.

We must also put forth the question whether, with the enumeration
of the above three vitamines, we have exhausted all the possibilities.
There is no definitive answer to this at present, but it is frequently
touched upon in the latest investigations, and it came into prominence
through a review of the available data by Mitchell (93), pointing to
the idea that vitamine B is different from the growth-promoting
substance. In spite of this, we must note that the conclusions of
Mitchell are not indisputable. He compared earlier work done with
comparatively inefficient methods, with newer work that is more
liable to be accurate, and consequently it is not to be wondered at
that he found a difference; besides, he compared extracts made from
different materials. Somewhat later, there appeared the first experi-
ment on this matter of Emmett and Luros (94), which at least had

42 THE VITAMINES

the advantage that the source of material (unpolished rice) was the
same in the comparison of both characteristics. We shall comment
upon the questions referred to, particularly the vitamines, in greater
detail, but at this point we wish to emphasize that the functions of
growth and the curative action on beriberi were tested on two dif-
ferent animals, the pigeon and the rat. Heated unpolished rice still
retained its growth promoting property, although to a somewhat
lesser degree, while its influence on pigeon beriberi was completely
destroyed. We can find many explanations for this behavior without
necessarily accepting the existence of two vitamines of the same type.
Above all, we do not know just what are the vitamine requirements
of these types of animals. We shall see furthermore that all the
known facts point to the idea that, although it shall have been demon-
strated that both substances differ from each other, they would
still be related chemically. In this connection, it might be shown
that various types of animals can utilize the different stages of vita-
mine cleavage with varying degrees of success. This could proceed, on
the one hand, through a difference in synthetic abilities and, on the
other, through certain specific symbiotic intestinal flora.

A similar question is brought up in reference to vitamine A (anti-
rachitic vitamine). Mellanby (95) holds the view, that in the
etiology of rickets, that vitamine which according to our nomen-
clature must be called antirachitic vitamine (vitamine A) is of
importance. It is found in fats such as butter, egg-yolk, and cod
liver oil. Whereas Mellanby worked with young dogs, Hess (96),
working with rachitic children, found that milk fat had no effect,
while cod liver oil was found to be beneficial. From this, Hess drew
the conclusion that the growth-promoting vitamine found in butter
was different from the antirachitic vitamine. Here also, both sub-
stances belong to the same type, and we must bear in mind the main
question whether the vitamines, like other natural substances, do
not also deviate from each other somewhat chemically, in spite of the
possibility that they play the same or very nearly the same role in
physiology.

Much more important and interesting are the experiments which
deal with the part played by milk and milk products. We are
sometimes almost tempted to assume that milk, aside from the vita-
mines already mentioned, contains still other essential substances.
This applies not only to milk, but also to products made from milk,

INTRODUCTORY 43

such as casein, lactalbumin, and partly also to lactose. Obviously
another conclusion is possible here, namely, that milk contains the
most suitable combination of amino acids for the animal body. It
was pointed out by McCollum and Davis (97) that heating of casein
for one hour, at one atmosphere in an autoclave, materially decreases
its food value; Funk and Macallum (98), attempting to confirm this
observation, found that the facts were somewhat in accord, but that
the diminished nutritive value of casein could be remedied by the
administration of fresh orange juice.3

We believed, at that time, that heating of the casein destroyed the
adsorbed C-vitamine, and our view was strengthened by the later
work of Harden and Zilva (100) and Drummond (I.e. 79). We (101)
had already been able to show that beriberi in pigeons could be cured
by the addition of the vitamine fraction of lime juice, although we
were not certain if this effect was not due to the presence of vita-
mine C. Still later, Osborne and Mendel (I.e. 92) showed that the
favorable influence of orange juice on the growth of rats was due to
the presence of vitamine B. This observation was further confirmed
by the work of Byfield, Daniels and Loughlin (I.e. 90), in which they
showed that orange juice treated with fuller's earth exerted no
growth-promoting influence either on young or grown rats, in spite
of still containing vitamine C. Till now, there has been no explana-
tion of why, when the diet already contains apparently sufficient
vitamine B, upon the addition of more of this vitamine the organism
should respond with an acceleration of growth. There is a possi-
bility here that orange juice may contain a new vitamine which,
together with B vitamine, is adsorbed by fuller's earth.

Osborne, Wakeman and Ferry (102) showed that certain types
of proteins, for instance, edestin, manifest a greater capacity for
adsorbing vitamine B than do other proteins, and that the latter, by
means of thorough washings, could not be freed from it. It would be
plausible to assume that casein, as well as lactalbumin, might show a
selective adsorptive capacity for some new vitamine. As we shall
show towards the end of this book, the acceptance of a new and
important amino acid or of a new vitamine appears inviting for the
elucidation of the etiology of certain pathological conditions, and
also for the explanation of the difference in food value between
proteins of vegetable and animal origin.

3 Hogan (99) found that heating of proteins, casein in particular, does not
impair their effectiveness.

44 THE VITAMINES

In a great number of experiments, Osborne and Mendel (103)
showed that the nutritive value of lactalbumin is much greater than
that of casein, and they attributed it to the nature of the particularly
favorable amino acid composition of this protein.4 Edelstein and
Langstein (104) recently arrived at the conclusion that the superiority
of mother's milk over cow's milk for the child is due to the greater
content of lactalbumin in mother's milk.

In a series of papers, which are not altogether clear in their mean-
ing, Emmett and Luros (105) sought to show that while lactalbumin
of itself was a complete protein for growth, the specially favorable
results obtained must be attributed to the influence of the protein-
free milk added at the same time. In this, they coincide with
the view of McCollum, Simmonds and Parsons (106). In other
words, the favorable influence of lactalbumin on the growth of rats
was to be attributed to the presence of a vitamine in the protein-free
milk or the lactose. This vitamine was supposed to be different
from vitamines B and A.

Emmett and McKim (107) have also assumed the existence of
two vitamines in yeast — one that cures beriberi and one that pro-
motes growth in animals. They reached these conclusions from
adsorption experiments with fuller's earth. They showed that the
activated fuller's earth was protective against beriberi, but did not
influence the weight of the animal. Such experiments, unless well
controlled, can have no real significance, considering the possibility
of incomplete adsorption, the relative amounts used and other
factors.

Sugiura and Benedict (108) point out that certain diets with the
addition of yeast serve to increase the growth of young rats, although
the mothers appear to be lacking in milk. They conclude therefore
that for this purpose a special vitamine is desirable. As an illustra-
tion of this, the example is given that an addition of casein exerts a
much greater influence than a simple addition of extra protein. In
one particular case it was shown that purified casein could not be
replaced by purified meat. In a second experiment, they diminished
the amount of the added yeast to 0.5 per cent of the total diet and it
therefore appeared not impossible that the failure of milk production

4 Later, these investigators showed that this is true only when protein-free
milk is used as source of vitamine B. This addition seemed to have the effect
of completing the value of that particular protein.

INTRODUCTORY 45

was due to an insufficient amount of vitamine B. Unfortunately,
these investigators failed to determine whether an addition of B-
vitamine would not stimulate milk secretion. Invariably, all of the
above mentioned workers stressed the possibility of finding a still
unknown vitamine, especially in milk.

In conclusion we wish to mention that besides other possibilities
of clearing up the etiology of pellagra, we have already intimated
(I.e. 62) that there might be a specific antipellagra vitamine — a view
held as well by Goldberger (109)'. This vitamine could be a sub-
stance easily adsorbed from animal protein as we pointed out on
page 43. Still, for these or for other conceptions of pellagra, no
real experimental evidence is at present available.

After having discussed the definitely established types of vitamines,
we shall take up the vitamine requirements of various organisms.

PARTI

The Vitamine Requirements of Plants
and Animals

CHAPTER I
THE R6LE OF THE VITAMINES IN THE VEGETABLE KINGDOM

It was quite clear from the start that the importance of the
vitamines was not limited to animal life only, but was applicable
to all living matter. At present, the view is held that the animal
organism does not possess the ability of synthesizing these factors,
and in this respect it is dependent either directly or indirectly on
plant life. We must determine, however, which plants, and espe-
cially what parts of plants, possess the power of building up the
vitamines out of simpler combinations. Furthermore, we should
like to know what role these substances play in plant physiology. We
can immediately assert that although the work on this aspect of the
subject is still in its infancy, not all plants apparently are able to
build up the vitamines (this is perhaps not true of all vitamines).

Upon more careful consideration of the higher plants, we find the
vitamines localized in two divisions, in the seeds and in the green
leaves, parts of the plant which at certain tunes are of particular
importance in its metabolism. We find, with few exceptions, the
greatest amount of vitamine B in seeds, together with varying
amounts of vitamine A. It is quite apparent that in seeds, the
analogue of milk and eggs in the animal kingdom, all the substances
are present which are necessary for the sprouting of the new plant.
The vitamines play a great part in this process, yet we are dealing
here only with conjectures. As soon as seeds are planted under
conditions of proper moisture and temperature, the ferments, which
had been in a quiescent state, commence to function. These fer-
ments begin to marshal the stored up reserve substances. In this
instance, we may also consider the vitamines as reserve substances,
and we may conceive that they too undergo some chemical changes
through the influence of ferments. As an analogy to this example
we already know that vitamine C is generated upon the germination
of seeds. All other phenomena, like the division of the nuclei of
cells, remain unknown to us, although we are soon aware of the
appearance of the first green shoot. What part the vitamines play
in this case is not definitely known. Perhaps they are of importance

49

50 THE VITAMINES

in the first stages of metabolism; perhaps they are responsible for
the sudden onset of cell division. In organisms which multiply by
conjugation or copulation, the vitamines or similar substances exert
some influence perhaps in the subsequent nucleus and protoplasm
partition. When we compare the quiescent with the sprouting seed,
we see the difference between the living and the dead tissue, although
the gross chemical compositions in both conditions are not far
different from each other. The entire difference may be due to the
formation of an active vitamine modification. It is evident that no
great progress is made when an unknown matter is explained by
something equally unknown. It may nevertheless be of value, as
far as future investigations are concerned, to point out in which
class of chemical bodies the substances essential to life are to be
sought.

Clark (110) described the influence and characteristics of a sub-
stance which he called "Oocytin," and which was supposed to be
able to bring about the formation of the fertilization membrane of
sea urchin eggs, previously treated with strontium chloride. This
substance was prepared in powdered form from centrifuged and
defibrinated ox blood. The powder gave the reaction of protein,
purine, and pentose, and contained only a small amount of phos-
phorus. It was inactivated by heating to 73° to 80°C., or by long
contact with alcohol. Similar substances were demonstrated in the
sperm of the sea urchin by Robertson (111). These or other active
substances naturally play a similar part in the plant kingdom,
although they are for the moment unavailable for physiological-
chemical work.

The question as to the ability of certain types of plants and par-
ticular parts of plants to build up vitamines is still far from solution.
In higher plants, for example, there are three possibilities:

1. In seeds, stored up vitamines suffice to stimulate further
synthesis of vitamines in the growing parts of the plants.

2. It is possible that the small amount of vitamines in seeds suffices
to carry the plants along to a certain stage, from which there is a
symbiosis with certain microorganisms which provide the plants with
vitamines.

3. It is possible that some of the plant structures, for example,
the leaves, possess the ability to synthesize certain vitamines.

THE VITAMINES IN THE VEGETABLE KINGDOM 51

The subject of this chapter serves merely as a suggestion of the
important role of the vitamines in plant life, without which knowledge
of the vitamines can never be complete. The very few facts
mentioned here should serve to stimulate further work in this
direction.

YEAST

Since the discovery of its curative influence on beriberi by Schau-
mann (I.e. 2), yeast appears to be the best material available for the
chemical study of the vitamines. Of all natural products, it seems
to be the one most rich in vitamine B and therefore is worthy of
special attention. One could be tempted to assume that since yeast
cells are so rich in vitamines and possess such an energetic metab-
olism of their own, they should be able to prepare their own
vitamines. This is, however, only partially the case, as has been
already shown in some older neglected work which has recently
been repeated, and confirmed. Pasteur (112) made the obser-
vation in 1871 that the capacity of yeast-cell growth depended
very much upon the size of the inoculation. He found also that
addition of certain organic substances accelerates the fermenta-
tion of certain types of yeasts. It was Wildiers (113) who correctly
understood Pasteur's observation and developed it further. It may
well be said that Wildiers foresaw the existence of the vitamines
as far back as 1901. His important studies have demonstrated
that upon the addition of a small quantity of sterile yeast extract
to a nutritive solution, containing an ammonium salt as its sole
source of nitrogen, a small inoculation is sufficient; the growth
of the cells being measured in this case by the CO2 evolved. - He
recognized that this growth stimulus could not be attributed to
the presence of the hitherto known factors, and named this new
substance "bios." Its characteristics were not so far different
from that which we have come to know as B-vitamine. It was
soluble in water and also in 80 per cent alcohol, and insoluble in
ether. It was dialyzable and could not be precipitated by the well-
known precipitants, phosphotungstic acid included. The substance
was found to be stable in acid medium but on heating for a short
time with alkali it was destroyed. The investigation of the resistant
qualities of this substance gave no clear results. The presence of
bios was never shown in the decomposition products of egg albumin

52 THE VITAMINES

split by acids, and likewise not in yeast ash. The experiments by
Wildiers were then confirmed by Amand (114) who demonstrated
that the failure of the yeast to grow, with just a slight inoculation,
was not due to the toxicity of the nutritive solution. Devloo (115)
reported that he isolated pure bios from lecithin. From his own
data it is evident that his preparation was not very active, and at
present we can say with certainty that the activity noted was due
to the contamination of the lecithin with vitamine. In this connec-
tion, it has been also stated that lecithin is curative for beriberi ; we
shall have occasion to refer to this matter later.

We should not think that the important conclusions of Wildiers
found immediate acceptance in the literature. For example, H. H.
Pringsheim (116) took the stand that the bios of Wildiers was nothing
else than protein material, which was best used in the form that is
found in yeast cells themselves. This statement of Pringsheim's
was introduced into the literature, and therefore the conclusions of
Wildiers were given no further attention. For instance, Rubner
(117) said that the growth of yeast began only when there was a
certain relationship between the amount of food and the number of
yeast cells, an excess of food acting as a stimulus for yeast growth.
From time to time, however, reports appear in the literature dealing
with the observation of Wildiers. Vlahuta (118) prepared a peptone
from beer yeast, with cold suffuric acid, which could give rise to
fermentation. Kurono (119) foihid that a vitamine extract, prepared
from rice polishings, added to Hayduck or Nageli solution, accelerated
to a greater extent the growth of yeast cells and also the degree of
fermentation, than when peptone or asparagine were added. Brill
and Thurlow (120), on the contrary, could obtain no increase in the
growth of yeast cells with rice polishings. The ability of certain
brewery residues to accelerate fermentation can be attributed to the
presence of vitamines. Moufang (121) made the observation that
dead beer or baker's yeast exerted a marked catalytic action on
fermentation. Saito (122) was of the opinion that certain chemical
substances were important to allow of a complete development of the
reproductive capacity of yeast. Bokorny (123) observed an increase
in weight of yeast when it was grown on urine containing sugar
(we sliall see later that urine contains vitamine). Vansteenberge
(124) studied the influence of autolyzed yeast upon the growth of
yeast and lactic acid bacteria, and found that above a certain

THE VITAMINES IN THE VEGETABLE KINGDOM 53

optimum concentration, this autolysate is no longer active and even
inhibits somewhat the growth of the cells — a finding that is of greatest
importance for our arguments. He found later that the autolyzate
must be diluted to obtain the best results. Eeucine, tyrosine, and
asparagine have an inhibiting influence on the growth of yeast cells
but not on the lactic acid bacteria. Recently Lampitt (125) has
shown that the increase in the quantity of yeast cells depends on the
original number present. Evidently, this observation is to be
explained in this way : that due to excessive inoculation, a number of
dead cells are implanted at the same time and that the living cells
can utilize the vitamine set free.

Since the growth of yeast cells might be utilized as a means for
the determination of antiberiberi vitamine, systematic investigations
were undertaken during the last two years for the purpose of studying
the vitamine requirements of these cells. It was expected in this
way to measure vitamine B, but we shall presently show that this
is not the case. In spite of the possibility that the substance pro-
moting the growth of yeast belongs to the type of B-vitamine,
newer work has shown that the substance promoting the growth
of yeast is not identical with vitamine B. It is possible, however,
that for yeast, an organism characterized by greater synthetic
ability, the cleavage products of vitamines might be sufficient,
whereas in higher animals, on the contrary, the entire vitamine
complex must be administered.

In 1912, the author, using the fermentation method, endeavored
to show that vitamine could act as a co-ferment, but without success.
Abderhalden and Schaumann (126) were apparently more fortunate
and they described a phosphorus-free substance, obtained from
yeast, which could favorably influence the fermentation of certain
sugars. Abderhalden and Kohler (127) likewise described the
growth-promoting influence of some yeast fractions on the yeast
cells. The same authors also investigated the influence of yeast
extracts on growth of Flagellata (Colpoda). and Algae (Ulothrix)
with positive results. A whole series of experiments on this sub-
ject has lately been published in the United States. The method
of R. J. Williams (128) denoted progress in that it substituted for
fermentation a direct observation on the growth of yeast cells.
The method is as follows: A hanging drop is prepared so that it
contains the least number of yeast cells (one if possible) obtained by

54 THE VITAMINES

inoculating Nageli's solution with a needle-point of yeast . culture,
preparing at the same time the necessary controls. The cells are
counted after 5 to 6 hours and again after 20 to 24 hours. From
20 to several thousands of cells could develop from a single cell in
24 hours, depending upon the vitamine concentration. Williams
undertook some problems with the help of this method, for example,
the protective influence of fat upon the destruction of vitamines by
alkali. Vitamine extracts, prepared with acid or alkali, appeared
to be best utilized by yeast. The greatest influence was obtained by
a preparation made by shaking yeast with fuller's earth. The ac-
tive substance was also shown to be present in the alcoholic extract
of protein-free milk, wheat embryo, and pancreatin, and in the
decomposed phosphotungstic acid fraction from yeast. Casein
and lactose were likewise active, although in the former the ac-
tivity was not attributed to the amino acids but to the small amounts
of vitamines present. The active substance was partly destroyed
by heating in the autoclave for a half hour at one atmosphere. An
alcoholic extract of egg-yolk was active while the ether extract
containing vitamine A was inactive. The C-vitamine, as we shall
note later, appears to have no influence on the growth of yeast.
Bachmann (129) investigated the behavior of two kinds of yeast,
one apparently " Saccharomyces cerevisiae" and the other, isolated
from fermented pears. These two varieties showed a different
behavior towards the vitamine solutions. One of the yeasts grew on
the surface and was less dependent upon the vitamine addition (in
this she indicated the possible analogy of green leaves and influence
of oxygen). The Bachmann method is as follows: A yeast sus-
pension in a liquid nutritive medium is placed in a sterile fermenta-
tion tube, and the COg evolved is measured after a definite incuba-
tion period, being compared with controls in which no vitamine was
added. Orange juice, yeast extract, peptone, Liebig's meat ex-
tract, honey, certain vegetable extracts and milk were tried as a
source of vitamine, and the results were for the most part the same
as had been found in animal experiments. Certain discrepant re-
sults could be explained by the greater sensitivity of the method.
Pasteurization and sterilization decrease the activity of milk. It
was also interesting in this work to note the different behavior of the
two kinds of yeast as to their dependence upon the vitamine addi-
tion, one requiring more than the other. Abderhalden and Schau-

THE VITAMINES IN THE VEGETABLE KINGDOM 55

mann (I.e. 126) developed a somewhat similar method simultaneously
with the American investigators.

There seems to be no unanimity as to whether or not yeast
requires extraneous vitamine for growth. Thus, Fulmer, Nelson
and Sherwood (129a) believe that yeast does not require vitamine
B for growth. These authors were able to grow yeast on a
synthetic culture medium supposedly free from' vitamine. How-
ever, their findings are not as definite as their conclusions. Nelson,
Fulmer and Cessna (129b) have found that if the brewer's yeast
in a synthetic diet was replaced by yeast grown on a synthetic
medium in the absence of vitamines, the growth of rats proceeded
normally, showing that the above yeast synthetized its own
vitamine. Working with pigeons, Harden and Zilva (129c) also
found that yeast synthetizes vitamine on a synthetic medium
but to a lesser extent than in the presence of vitamine. Funk and
Dubin (129d) were unable to corroborate the findings of Fulmer,
Nelson and Sherwood, using the medium F described by the latter.
Poor yeast growth was obtained in the absence of vitamine while
a marked improvement was noted when vitamine was added. These
same conclusions were reached by Eddy, Heft, Stevenson and
Johnson (129e). MacDonald and McCollum (129f) maintain
also that they can grow yeast on a pure nutrient medium in the
absence of vitamine B. However, they obtained better growth
with this vitamine added. This subject became a matter of con-
troversy between Ide (129g) and MacDonald and McCollum (129h),
the former maintaining that yeast can grow in two ways — slow
growth without bios and rapid growth in the presence of bios,
while the latter hold that there is no necessity for assuming the
existence of a specific substance promoting the growth of yeast.
We personally are in accord with the view of Ide that at least cer-
tain yeast species do require a specific substance for growth. A
number of procedures such as improving the inorganic moiety of
the medium and the addition of glucose or amino acids may give
slight increase in growth but not to be compared with that ob-
tained on the addition of vitamine. Until we know how many new
cells can grow out of a few broken down cells, this question, as to
whether yeast can grow without vitamine, must be left open. From
the foregoing, it would seem that yeast cells can produce vita-
mines, if an original stimulus is provided.

56 THE VITAMINES

BACTERIA

While yeast extract was used in 1904 by Bertrand (130) for the
growth of B. xylinum, it did not occur to anyone that this nutritive
medium possessed such special characteristics. Most bacteria, how-
ever, thrive well on the commonly used media. With some organisms
on the contrary, difficulties arise and consequently the method of
Noguchi (131), recommending the addition of a small amount of
testicular substance for the cultivation of spirochetes, denotes impor-
•tant progress. What was responsible for the stimulating effect of
this tissue? The question remained unanswered till the war
came and the bacteriological laboratories were stripped of their
customary sources of supply of nutritive media. It became neces-
sary then, above all things, to replace Witte's peptone and nutrose.
Investigators found this difficult, till systematic work on this subject
was undertaken. On the whole, it may be said that most of the
nutritive media used before the war very likely owed their favorable
influence on the growth of bacteria to a small amount of vitamine
present therein. This conclusion was not apparent immediately,
since many investigators thought that bacteria could grow on a pure
synthetic medium. For instance, Vedder (132) cultivated the
meningococcus on starch prepared from cornmeal. Doryland (133)
reported success with pure synthetic media, which evidently con-
tained no vitamine. These experiments may be explained in that
the inoculation was perhaps so great, that, similar to yeast, growth
cook place. Pieper, Humphrey and Acree (134) reported great
success with synthetic media, while Lockeman (135) considered that
the factors necessary for the growth of the tubercle bacillus were
asparagine, as a source of nitrogen, glycerol and inorganic salts,
phosphorus, potassium and magnesium. Nevertheless, for other
microorganisms, other requirements appeared to be necessary, and
Bainbridge (136) believed that certain microorganisms could not
grow with protein as the only source of nitrogen. This view was
also maintained by Sperry and Rettger (137). Robinson and
Rettger (138) compared, in this connection, a protein obtained by
ferment action (which he called "Opsin"), with the products of
protein acid hydrolysis. Decolorization of the opsin by animal
charcoal decreased the value of that nutritive medium. Of the
proteins tested, the products of casein acid hydrolysis gave the best
results, but not so good as opsin. Extracts of beef worked very well.

THE VITAMINES IN THE VEGETABLE KINGDOM 57

It was of greatest importance to this entire subject that the
presence of vitamines in typhus bacilli was detected by Pacini and
Russel (139). They proceeded from the observation that in the
typhus convalescence period there is often noticeable quite an increase
in growth. The bacilli were grown on the usual vitamine-free
Uschinsky nutritive medium, and consequently they must have been
able to synthesize the necessary vitamine. Both the remaining
medium and an acid-alcoholic extract of the bacteria were admin-
istered to rats on an artificial diet, whereupon normal growth took
place. Since the presence of vitamines in bacteria was demonstrated
in this way, it was plausible to assume that for certain types of
bacteria, a vitamine-containing medium would be of value. Such a
preparation was introduced by Mackenzie Wallis (140) ; it consisted
of casein, peanut flour and soda, and served as a substitute for nutrose
in the composition of Conradi-Drygalski medium (for the typhus
coli group) . The bacteria grew well on the medium and the success
was attributed to the presence of vitamine in the globulin fraction
of the flour. A similar preparation, introduced by Huntoon (141)
was made from meat and eggs. In the description of this preparation
it was stipulated that all filtration, whether through filter paper or
cotton, must be avoided. A yeast extract as culture medium was
also described by Ayers and Rupp (142).

Meningococcus. The conditions for the growth of this micro-
organism were systematically studied by Miss Lloyd (143). She
found that growth was* impossible without vitamines. Although
amino acids were the chief components of the nutritive medium, it
could not be utilized except when a voluminous inoculation had
been made. In this way it was possible to note a difference in the
behavior of different laboratory stock toward vitamines — older
laboratory stock needed no special vitamine addition when a sufficient
amount of amino acids was available, while newly isolated bacteria
could not be cultivated without vitamine. Between the necessary
amounts of vitamine and amino acids, some relationship is apparent,
and therefore it is held that the role of the vitamines in the nutrition
of the meningococcus is traceable to the acceleration of the reaction
velocity of the proteolytic processes. A further function is attributed
to the vitamines in that the assimilation of iron, calcium, phosphorus
and iodine is facilitated. The vitamine in question was soluble in
water and alcohol and somewhat thermo-stable; it could stand heat-

58 THE VITAMINES

ing for 45 minutes at 120°C. This vitamine could be easily adsorbed
by filter paper but not by glass wool. It was shown to be present
in blood and in milk (and to a smaller extent in serum). On this
basis, a blood agar was prepared (144) which, in spite of the coagula-
tion of its protein by heat, still contained vitamine, but could not be
subjected to filtration. The precautions to be observed as regards
filtration pointed to the probability that they were dealing with two
vitamines, both of which were necessary and one of which was either
slightly soluble in water or not at all. The latter could be adsorbed
by protein, and of special interest, is the fact (145) that the meningo-
coccus, after from one to ten re-inoculations, does not need any
more vitamine additions, and this is apparently the reason why the
importance of vitamines for bacteria was not recognized sooner. At
the same time, somewhat similar experiments were carried out by
Gordon and Hine (146), and also by Flack (147). These investi-
gators were able to show that serum albumin, hemoglobin, fibrinogen,
legumin and human blood accelerated the growth of the meningo-
coccus on common agar, while the addition of glucose was without
any effect. Among others, they recommended trypagar, prepared
from pea flour and digested ox heart. The best results were obtained
with an agar containing bouillon and an extract of cooked pea flour,
or the fractions obtained after removal of starch and dextrin by
alcohol. Extracts of raw and cooked wheat embryo showed a
similar ability to promote growth and to prolong viability; these
authors believed that pea flour contained a vitamine acting on growth,
while that from wheat influenced the viability. Shearer (148) also
investigated the influence of some extracts. He found the active
substance in nasal mucous and believed that it acted not as a usual
dietary constituent, but as an activator. The substance was soluble
in water, less soluble in alcohol and not at all in ether; it was
unaffected by heating with strong hydrochloric acid for 12 hours.
Besides the meningococcus, it was active for pneumococcus, B.
typhosus, B. coli communis and other pathogenic bacteria. In the
continuation of his work, Shearer (149), noted that the cerebrospinal
fluid contained a substance which is still more active than blood or
nasal mucus. This peculiarity explained the rapid growth of the
meningococcus in the brain tissues and in the membranes and
cavities of the central nervous system. In this case too, the action
was observed not upon the meningococcus only, but also upon other

THE VITAMINES IN THE VEGETABLE KINGDOM 59

pathogenic bacteria, like pneumococcus,!?. typhosus,B. coli communis,
certain intestinal streptococci and throat bacteria. The proof that
the vitamine was actually necessary was not available, acceleration
in growth only being demonstrated. Eberson (150) likewise found
that a yeast extract could increase the viability of a meningococcus
culture for more than a month. The medium consisted of agar,
peptone and potassium phosphate. The yeast was first extracted for
two hours at not more than 100°C., and then sterilized for a half
hour at one atmosphere.

Gonococcus. Cole and Lloyd (151) carried out vitamine experi-
ments on gonococcus. Tryptamine blood (tryptic digestion product
of casein) was used for cultivation. The gonococci cultures required
a suitable ion concentration, a considerable amount of free amino
acids and two kinds of vitamines to permit of favorable development.
One of these, easily adsorbable, is found in blood corpuscles and
possesses the ability to promote growth; the other occurs in tissues,
is relatively little adsorbable, and -has the peculiarity of bringing
about a secondary plentiful growth. The method of demonstration
of both vitamines is given in great detail. They are precipitated by
protein, but not in the presence of agar. Filtration through paper
removes the first vitamine completely and we have a good basis for
the idea that the substance from the blood corpuscles belongs to the
type of A-vitamine (antirachitic vitamine) which is found naturally
associated with fats; or it may also belong to a vitamine type that
is easily adsorbed by protein. On the other hand, the vitamine of the
tissues belongs perhaps to the B-vitamine type. Blood seems to
contain both substances although, in sheep serum, the first one
appears to be lacking. Morini (152) grew the gonococcus on gelatin
and beer yeast.

The observations of Douglas, Fleming and Colebrook (153) suffice
to give us some idea of the conditions under which anaerobic bacteria
are best cultivated. Special measures for the exclusion of air are
shown not to be necessary, if the bacteria are given the opportunity
to provide themselves with local anaerobic conditions by the addition
of some porous material such as asbestos. It is very important,
however, to add to the bouillon culture, potatoes, carrots, cabbages,
and grapes, or better still, bran. An alcoholic extract of bran is
effective only when large amounts are used.

60 THE VITAMINES

Tubercle bacilli were cultivated by Sazerac (153a) and by Masucci
(153b) on digested casein (aminoids).

In regard to hemophilic bacteria, there is the work of Davis (154).
For this type, two factors in particular are of great importance. One
is hemoglobin, and the other is a substance which is found in other
strange bacteria, as well as in a variety of fresh plant and animal
tissues. This second substance is found in B. influenza when both
strains are cultivated together. The similarity of the factors neces-
sary for the growth of rats with those necessary for the successful
growth of hemophilic bacteria has been emphasized by Davis. He
explained it by the fact that hemoglobin serves as the source of iron,
while the second substance facilitates the assimilation of this element.
In a later publication, Davis (155) found that the growth-promoting
influence of hemoglobin, white and brown rice, pure wheat flour and
wheat bran, was shown by no other kinds of bacteria except by the
hemophylic. Germinated rice or wheat kernels exhibit a greater
influence than ungerminated. In this case also, the active substance
was adsorbable by filter paper and was quite thermo-stable, having
resisted a temperature of 100°C. for one to two hours.

The favorable influence of cooked meat on bacterial growth was
observed by Wolf and Harris (156), on the B. histolyticus,andby
Wolf (157) on the vibrion septicus, B. sporogenes, B. welchii and B.
proteus (158). Boyer (159) recommended a hydrochloric acid extract
of finely divided bones for the cultivation of streptococci, and Otabe
(160) suggested a wheat extract instead of meat for staphylococci.
We can see from this short list that the addition to the medium of a
natural product as a source of vitamine was necessary. For the
cultivation of B. coli, Dienert and Guillerd (161) prepared a medium
as follows: 500 grams of pressed yeast were heated to 50°C., the
optimum temperature for the action of endotryptase. In less than
24 hours the autolysis was finished, the yeast having become liquid,
yielding 397 cc. of filtrate which was then diluted to two liters. The
extract was sterilized for a half hour at 100°C., and finally diluted to
7500 cc. This preparation was used advantageously instead of
peptone.

Bunker (162) and particularly L. David and Ferry (163) have
made an exact study of the conditions that are necessary in order to
obtain a very active toxin from cultures fo B. diphtheriae. It was

THE VITAMINES IN THE VEGETABLE KINGDOM 61

shown by these investigators that a medium of amino acids and salts
did not suffice and that an addition of creatine, creatinine, xanthine
and hypoxanthine had no effect. Only when a 0.5 per cent of
bouillon was added, did it result in good growth. For the prepara-
tion of toxins, the amount of bouillon present must be increased to 10
per cent. Addition of peptone, on the contrary resulted in a slight
growth and toxin formation. Toxin is not supposed to be a synthetic
product of bacterial cells; but a product of metabolism which makes
its appearance only in the presence of certain amino acids and
vitamine.

Agulhon and Legroux (164) investigated the influence of vitamine
on B. influenzae (B. Pfeiffer). The growth-accelerating influence of
blood, serum and ascitic fluid, could not have been due to protein
addition but to the influence of vitamine. The vitamine extract of
defibrinated blood was obtained in two ways. By the first, blood
was precipitated by about four volumes of absolute alcohol; the
precipitate was extracted with the same volume of water, centrifuged
and filtered through a Chamberlain filter. By the second procedure,
blood was extracted with sodium chloride solution, and the extract
heated to 80°C. for 15 minutes. From 5 to 10 per cent of these
extracts, added to the usual medium, showed a great influence on
the growth of B. Pfeiffer. Even a one per cent solution showed a
definite effect. It might well be conceived that the vitamines in
the cells are present in combination and that alcohol or heating sets
them free. They seemed to be insoluble in alcohol, for a cold
alcoholic extract was inactive, while one prepared at 80°C. was only
slightly active. Acetone extracts, prepared either hot or cold, were
inactive. Warming to 80°C. in the presence of alcohol or acetone,
with subsequent drying at a lower temperature did not bring about
decomposition, since extraction of this powder with the water yielded
an active substance. The watery vitamine extract partially lost its
activity after heating to 90°C. for 15 minutes. It appears to be more
stable when the heating is done in the presence of gelose, although in
this case too, it is partially or completely inactivated, depending upon
whether the temperature is raised only to 100°or to 120°C. When
a totally defibrinated blood was used, it was observed that the vita-
mines were found in the corpuscles, since the watery extract of the
serum gave completely negative results, while the washed blood
corpuscles gave positive results. Hemoglobin and blood corpuscle

62 THE VITAMINES

stroma were negative. Cold extraction of blood corpuscles for 24
hours with a physiological salt solution gave only a weakly active
extract. If hemolysis is induced with distilled water before the
extraction, then the vitamines are found in solution. In this case,
contrary to what has been noted previously in this chapter, the
extracts may be filtered through a Chamberlain filter or through
paper without loss of activity; this characteristic permits of steriliza-
tion without the use of heat. Incidentally, the action of vitamines
has been erroneously ascribed to their peculiar physical condition.
Legroux and Mesnard (165) extended their experiments to include
extracts of kidney, liver and heart. Davis (165a) has pointed out
the necessity of two substances for the growth of B. influenzae, one
derived from the blood pigment and one of a vitamine nature. The
same conclusions were reached by Rivers and Poole (165b), by
Thjotta (165c) and by Thjotta and Avery (165d). Thjotta and
Avery differentiate between a thermo-stable substance contained
in blood, and called "A," and a thermo-labile substance "V" (vita-
mine) which is found in blood, yeast and vegetable extracts. Crys-
talline hemoglobin contains only the substance "A."

Finally, there are the vitamine studies of Kligler (166) on bacteria.
At first he used autolyzed yeast and found that pneumococci and
meningococci do not grow so well on it as on bouillon. In a later
work, he (167) investigated the influence of ox heart, goat blood,
rabbit and cat tissue and human excreta on gonococcus, meningo-
coccus, pneumococcus, Streptococcus hemolyticus, B. diphtheriae, B.
pertussis and B. influenzae. All of the investigated extracts exerted
a favorable action in exact proportion to the amount added. The
spleen, liver and kidneys were rich in vitamines, while muscle
tissue was only slightly active, and ether extracts were inactive.
Mueller (167a) found that streptococcus needed for growth a sub-
stance which he regarded as a new amino acid. The streptococcus
was cultivated on beef-heart extracts. If this medium was de-
colorized with "norit" it was unsuitable for growth, but permitted
of growth upon the addition of peptone or certain protein hydrol-
yzates. The growth substance was precipitated from these hydrol-
yzates with mercuric sulphate, but it was not identical with
tryptophane, tyrosine, cystine or histidine. In a subsequent
publication the same author (167b) accepted the necessity of three
substances for the growth of Streptococcus hemolyticus — one which

THE VITAMINES IN THE VEGETABLE KINGDOM 63

remains in the decolorized beef-heart infusion and two others which
were found in the hydrolyzate of the commercial casein preparation
"aminoids. " Of the latter two, the substance "x" was detected
in the silver sulphate precipitate and the substance "y" in the fil-
trate. In collaboration with Freedman (167c), we were able to
show that not all p\ire proteins (hydrolyzed) fulfill the nutritional
requirements of Streptococcus hemolyticus. As a rule, purification
of these proteins diminished their nutritive value for these bac-
teria. Other proteins obtained from sources rich in vitamines
are with great difficulty, or not all, freed from this growth substance.
On the other hand, this growth substance can be adsorbed in a way
similar to vitamines, since fuller's earth and norit remove it from
autolyzed yeast, hydrolyzed casein and a neutral solution of sodium
caseinate. This phenomenon will permit of further investigation.
The absence of the growth substance from a number of purified
proteins of various sources renders it unlikely that the above growth
factor is an integral part of the protein molecule. It is more than
likely that the growth substance is similar to that necessary for
yeast — and found in the same starting material — the only dis-
crepancy being that those hydrolyzed proteins which act on strepto-
coccus do not act upon yeast. However, this might easily be a
matter of inhibition due to the various protein cleavage products.
In this connection, Norris (167d) found that casein was a better
medium than brewer's yeast for culture.

The many points brought out in this small chapter, touching in
particular upon the physical characteristics of the substances which
act on bacteria, form, for the time being, the sole criterion for their
classification in the group of vitamines. By comparing the char-
acteristics here set forth with the description of the vitamines to
follow later, the reader will have the opportunity of making a decision
for himself as to which group these substances should be assigned.
Looking upon this chapter as a whole, we may conclude that at least
some types of bacteria respond to the addition of vitamine with
accelerated growth, showing possibly that a small initial quantity of
vitamine suffices to render the bacteria independent of any further
vitamine additions for a long time. As we shall see in the chapter
on the growth of higher plants, bacteria seem to exist entirely inde-
pendent of any vitamine supply. According to our present knowl-
edge, these are the bacteria which supply the whole organized world

64 THE VITAMINES

with the vitamines, or with the substances from which they are
formed.

All of the above mentioned observations conform preferably with
the conception of two or three different vitamines.

FUNGI «

Because of the lesser practical significance as compared with
bacteria, the question as to the vitamine requirements of fungi
received little attention. Lutz (168) made the observation that fungi
often show an interruption in growth when the nutritive medium is
exhausted. Still, this observation could be attributed to a general
exhaustion of nutritive constituents in the medium. According to a
paper by Currie (169), Aspergillus niger needs no extra addition in
order to grow, and yet he states that an extract of old mycellium, or
spore bearing portions of the same fungus, brings about quite a
definite acceleration in the growth of a new culture of the same
fungus.

The only work affording an insight into the vitamine requirements
of parasitic and saprophytic fungi is the recent investigation of
Willaman (170) . He worked with Sclerotinia cinerea [(Bon) Schroter],
which he sought to cultivate on a solution of purified salts, cane
sugar and nitrates. This experiment, and others in which there
was also an addition of asparagine and amino acids, turned out
negative. The results were different when fruit juices, yeast or
extracts of wheat germ were used. The method of procedure was so
chosen that the surface of the mycellium was measured in square
centimeters, and the number of spores noted. The juices from
peaches, plums and apples showed the greatest activity. Besides
these, a great number of natural products were tested and found
active, at least as regards the growth of this fungus. However, not
all of these extracts were equally active on the sporulation. For
this purpose, extracts had to be prepared from parts of plants which
displayed a very active metabolism. The following could be classed
as such: pollen, sporophores of the fungi and the sclerotinia itself,
terminal butts of the Phaseolus and sporulated mycellium of Asper-
gillus. This observation indicates, perhaps, the importance of the
two types of vitamine, although Willaman is inclined to assume, in
the meantime, the existence of a single vitamine, varying in its effect
according to its source of origin. Nevertheless, one must concede

THE VITAMINES IN THE VEGETABLE KINGDOM 65

that in the course of this work, the firmly established physical and
chemical characteristics of the active substances are best in accord
with the conception of a second vitamine.1 Working with Oidium
lactis, Linossier (170a) found that on addition of vitamine, the
activation took place only in the first few days.

HIGHER PLANTS

In the first edition of this book, in anticipation of a possible
analogy to animal growth, we wrote a small chapter on plant
growth without being able to find any supporting experimental
evidence; still, it was not necessary to wait for it a long time. In
1909, Clinton (171) made the observation that plant extracts have
the power to stimulate the reproduction of certain plants. Haber-
landt (172) found that parts of potatoes containing the morphological
structure, "Leptom," showed cell division when placed in a nutritive
solution. The embryonic structure only was able to undergo cell
division without the presence of leptom. This author suspected the
presence of a special hormone in leptom corresponding to the internal
secretions of animals.

Bottomley (173) was the first investigator to take up this problem
seriously. In 1912, he occupied himself with the problem of the
ability of certain bacteria to transform the humic acids of peat into
water-soluble combinations. He demonstrated at that time that
such peat contains all of the chemical combinations necessary for the
growth of young tomato plants, buckwheat, radishes, and barley.
In the further development of this question, which appeared at
about the same time as our earlier vitamine work, Bottomley (174)
extracted a substance from bacterized peat with water or alcohol.
The addition of phosphotungstic acid to this alcoholic extract yielded
a precipitate which, upon decomposition, acted favorably on plant
growth. The solution thus obtained was precipitated with silver
nitrate, and in the end the growth substance was found present in
the same fraction as the vitamine. All active fractions were tested
for their influence on the growth of young plants, and then, in order
to set forth the results more clearly, the cotyledons, which contain

1 Willaman defines the vitamines as a class of substances whose presence
is necessary for normal metabolism, but which do not contribute to the
requirements of the organism as regards inorganic constituents, nitrogenous
substances, and energy producing food constituents.

66

THE VITAMINES

reserve vitamine, were removed after germination without harm to
the plant. The results were very significant and showed that the
young plants, in a stage in which they could not live by their own
process of metabolism, are dependent upon the stock of vitamine
which is stored up in the seeds. In mature plants, the vitamines
are built up through symbiosis with bacteria, which prepare it
synthetically from the organic constituents of the soil and from
8

A

B

O 10 ZO JO 40 50

Time in days
FIG. 4. PLANT GROWTH
A, with vitamine addition; B, without vitamine addition (Bottomley)

fertilizers, and carry it to the plants through the roots. In the
investigations of Bottomley, the control plants (in the nutritive
solution above) underwent a decrease in weight of 10.9 per cent in 50
days, while with the addition of vitamine there was an increase in
weight of 59 per cent. The curves appended illustrate what has just
been stated. We see here the influence of the silver nitrate fraction

67

added to the nutritive solution in a dilution of 0.35 in 1,000,000.
Bottomley's results were soon confirmed by others (175). The sub-
stances influencing plant growth were called "auximones" by Bot-
tomley, who also described a method for their detection (176). This
method possesses certain advantages as against the use of higher
plants, and consists in an extraction of fermented peat, and precipita-
tion of the resulting extract with phosphotungstic acid. If this
extract is added to a culture of nitrifying bacteria and the whole
placed in an incubator at 26°C., a scum is formed on the surface
after 24 to 36 hours, and no nitrates are found in the solution. On
the contrary, without the addition of vitamine, the scum does not
appear and the nitrification proceeds rapidly instead. This method
seems to be specific for the vitamines, since no such effect has been
recorded after the addition of cane sugar, maltose, asparagine,
peptone, leucine, tyrosine and hordeine. The microorganism isolated
from this scum, similar to the nitrifying and sulphur- and iron-
assimilating bacteria, may grow without the addition of organic
carbon combinations. Originally, Bottomley was of the opinion that
the auximones differed from the vitamines in that the. former were
heat resistant. We believe, however, that a successful classification
cannot be built up on such differentiations. Chittenden (177)
repeated these flower-pot experiments in which the ratio of the
bacterized peat to the required amount of soil was very high, and
therefore was also able to produce an acceleration of growth. When
this method of experimentation was applied to greatly extended
soil experiments, the results were not so significant; the greatest
success was obtained when the rainfall was large. Bottomley (178)
himself repeated his first experiment more carefully. For this
purpose he used a type of lentil (Lemna minor) that could live in
water; this was cultivated on Detmer's nutritive solution, of which
the only source of nitrogen was potassium nitrate. This plant could
not live only upon inorganic constituents; it gradually became
weaker, and presented an abnormal appearance. Bottomley then
added to the nutritive solution various extracts prepared from
fermented peat. From the results recorded in the table below, only
one conclusion may be drawn — that the influence of these extracts
was due not only to the presence of certain necessary building stones
therein, but also to the presence of vitamines. These experiments
were then repeated by Bottomley (179) with other water-plants like
Selvinia natans, Azolla filiculoides and Limnobium stoloniferum,
with the same results.

68

THE VITAMINES

The extracts of the fermented peat were examined chemically by
Rosenheim (ISO).2 To give the reader some conception of the
quantities of the active substances used, we shall go into these experi-
ments more in detail: 15 grams of bacterized peat were extracted
with 2 liters of water and 500 cc. of this brown solution were diluted
to 1250 cc. This extract contained 0.045 per cent of dry substance,
0.034 per cent of organic and 0.011 per cent of inorganic material,
and 0.003 per cent of nitrogen. The plants investigated (Primula
malacoides) received in one dose 0.18 grams of peat, which contained
20 mgm. of organic material and 1.9 mgm. of nitrogen. The plants
so treated (figure 5) grew higher, showed leaves of a deep green color,
and were also generally larger than the controls, which is especially
apparent in the illustration. The substance which could bring about

VELOCITY OF WEIGHT
DOUBLING

UNITS OF TIME NECES-
SARY FOR DOUBLING
OF WEIGHT

Number
of plants

Original
weight

Number
of plants

Original
weight

Controls

1.0
2.05
1.91
1.63

1.24

1.0

3.09
2.71
2.01

1.48

100

48
52
61

SO

100
32

37
50

67

Watery extract of bacterized peat.. . .
The same without humus acid

Alcoholic extract of bacterized peat. . .
Phosphotungstic acid precipitate pre-
pared from the last fraction

such a great change must belong to the type of vitamines, according
to Rosenheim. From the active extract, it was possible to obtain
with phosphotungstic acid a precipitate which was only partly
soluble in acetone (like the vitamines, as we shall see later). Rosen-
heim hoped to extend his experiments to include yeast and milk.

Bottomley (181) investigated the nitrogenous constituents of the
peat, and although there was no nucleic acid present, he did find
their decomposition products, an uracil-adenine-dinucleotidc and the
components of guanine-cytosine-dinucleotide.

Appleman (182) believed that potato tubers contained substances
which were necessary for the growth of the shoots. It is therefore
of importance in planting potatoes, that not too small a piece of the

2 In this paper, Rosenheim claims priority as to the findings on the impor-
tance of vitamines for plants.

THE VITAMINES IN THE VEGETABLE KINGDOM

69

tuber be used. He thought that the disease of potatoes common in
the United States, and known as "spindling sprout disease," was due
to lack of these vitamine-like substances.

Bottomley (183) also investigated the influence of crude nucleic
acid derivatives upon the growth of Lemna minor. The action, in

FIG. 5. GROWTH OF PRIMULA AFTER VITAMINE ADDITION (ROSENHEIM)

this instance, was similar to the activity of a sterilized suspension of
Asotobacter chroococcum. The combined activity of the two products
the suspension and the nucleotides were complementary to each
other. Purified adenine-uracil fractions showed no activity.8 In

3 Papers published by Jones (184) and also by Stead (185) may be looked
upon as reviews.

70 THE VITAMINES

Bottomley's laboratory, Mockeridge (186) studied the effect of
extracts of bacterized peat upon nitrifying bacteria of the soil (Bacil-
lus radicicola and Azotobacter chroococcum) . The results are apparent
from the accompanying table, in which the milligrams of fixed
nitrogen are given.

1. Nutritive medium alone 2.6

2. Nutritive medium plus watery extract of ordinary peat 1.9

3. Nutritive medium plus watery extract of fermented peat 7.7

4. Alcoholic extract of fermented peat 6.5

5. Phosphotungstic acid precipitate of No. 4 5.6

6. Silver fraction of No. 5 6.5

We may see from these results that the above bacteria, having
synthetic ability to assimilate nitrogen from the air, are nevertheless
influenced by vitamines. While the vitamine action was favorable
to the fixation of nitrogen, the effect upon denitrification was reduced,
and the formation of ammonia was uninfluenced. The investigations
were then applied to other soil bacteria and it was found, as a rule,
that the growth of the nitrifying bacteria is activated by vitamines,
while that of the putrefaction, denitrification, and ammonia forming
bacteria is not affected. Those bacteria which have the power of
decomposing organic matter should in fact have no need for vitamines
or else should be able to synthesize them. These two great classes
of bacteria, one that begins the nitrogen cycle and the other that ends
it, differ from each other entirely in their metabolism if Mockeridge
is correct.

It must be especially emphasized that Bottomley and his school
always moved further and further away from the importance of the
vitamines for plant growth, and ascribed this remarkable action to
the nucleic acid derivatives. These conclusions followed in the paths
which Schreiner and Skinner (187) and their co-workers laid out
for the action of nucleic acid derivatives on plant growth. These
investigators isolated from the soil some purine and pyrimidine
derivatives and tested their activity. They showed that xanthine
could increase the weight of wheat sprouts kept in water about 21
per cent. Similar experiments were made by Macalister (188) with
allantoine and by Coppin (189) with other analogous substances.
Recently, Miss Mockeridge (190), experimenting with Lemna major,
showed that green plants need an addition of certain other sub-
stances to continue to live. In this series, the influence of natural

THE VITAMINES IN THE VEGETABLE KINGDOM 71

fertilizer was investigated. The results obtained in the fifth week
were as follows :

Average number
of plants

1. Control solution 27.0

2. Extract of rotted leaves 64.6

3. Extract of soil 64.4

4. Extract of fermented peat 132.6

It is evident that identical results were obtained with fertilizer
and with bacterized peat; indeed, fresh fertilizer was less active than
one in which marked bacterial decomposition had set in. Mocke-
ridge then found that nucleic acids and nucleotides predominated in
the less active extracts, while in the more active extracts, the amount
of free purine and pyrimidine bases was increased. Therefore, the
conclusion was made that the effect was brought about not by the
nucleic acids themselves, but by their decomposition products.
These conclusions are obviously not justified especially since the
investigations were not carried out with pure chemical products;
consequently the results could be only interpreted to indicate that
the substrate acted favorably on plant growth because of some
unknown changes that had taken place in the composition of
the substrate. Hydrolysis of the afore-mentioned nucleic acids
could well be thought of as a secondary occurrence without any
physiological significance. It is very likely that the growth-promot-
ing activity of peat and soil extracts is not due to the nucleic acid
decomposition products but to the presence of vitamines which are
carried down in the fractionation. Should it be established that in
the growth of plants, similar substances are involved as in the growth
of yeast, then it would be very improbable that they could be asso-
ciated with purine and pyrimidine derivatives.

Miss Mockeridge's idea of the vitamine cycle, was that the nitrify-
ing bacteria furnish the green plants with the growth-promoting
substances which are used in the metabolism of the plants, partly
transformed into the vitamines already known to us, and which can
then be utilized by the animals.

The investigations of Bottomley and Rosenheim have estab-
lished that higher plants also need a vitamine of the antiberiberi
type in their metabolism, and Bottomley expressed the idea that
this vitamine occurred in the plant through symbiosis with certain
bacteria. Whether this is the only vitamine that plants need,

72 THE VITAMINES

remains to be demonstrated. Still, it is apparent that the vitamine
of type A, which is found mostly in seeds and green leaves, is formed
in the leaves perhaps from primary products which must be furnished
to the plants from some outside source. The question, as to whether
old plants that already have leaves need extra vitamine in the same
degree, must eventually be answered and may perhaps be solved by
the use of methods for sterile growth, as proposed by Grafe (191).

Since the entire animal world obtains its vitamine requirements
either directly or indirectly from plant food, it is especially important
to know the vitamine cycle in its entirety.

CHAPTER II
THE ROLE OF VITAMINES IN THE ANIMAL KINGDOM

Coming to this important chapter, we must first of all satisfy our-
selves as to the relative importance of the vitamines in nutrition.
In fact, the role of the vitamines is placed in the foreground by some
investigators, so much so, that the impression prevails that these
substances can completely compensate for all dietary deficiencies.
Obviously, this is not the case. The requirements of the animal
organism for each of the food constituents have been determined
chiefly in dogs, and lately in rats; we need only refer the reader to
the modern text books on the science of nutrition if he wishes to study
the question more thoroughly. We must specifically emphasize, as
we did in the first edition, that most of the old and many of the new
experiments on this subject were not entirely convincing, since the
investigators were either unaware of the existence of the vitamines or
else ignored their significance. In the last six years, however, many
of the experiments were repeated, bearing in mind the progress that
had been made in vitamine research. The conclusions arrived at in
these experiments, for example, for rats, are as follows : For complete
nutrition, these animals need, above all, a biologically complete
protein — a protein of high biological value — one containing all of the
necessary amino acids; further, a carbohydrate of the nature of
starch, sugar or dextrin; a certain amount of fat, a salt mixture, so
chosen that it contains the necessary cations and anions in proper
proportion; and finally vitamines A and B.1 It must be held open
for further investigation to determine whether or not rats can get
along on a complete protein, a salt mixture and vitamines (without
carbohydrate and fat) ; for some animals, like the dog or cat, which
can get along on meat alone, this has already been definitely estab-
lished. The series of experiments by Spriggs (192) and Maignon

1 Parsons (191a) recently found that rats do not require a supply of
vitamine from outside sources. In spite of the lack of this vitamine in the
diet, the organs — spleen, kidney and muscles — of rats so fed contain appre-
ciable amounts of vitamine Cwhen tested on guinea pigs in the form of extracts.
Whether this vitamine C originates from the vitamine B of the diet has to
be left for future demonstration.

73

74 THE VITAMINES

(193), dealing with the harmful effect of an exclusive protein diet on
rats, can very likely be attributed to the insufficient amount of
vitamines supplied, although it might also have been due to a relative
lack of salts. In a special case, Maignon (194) noted that when he
fed rats on egg albumin, together with some salts, they developed
a disease which he held to be an acute intoxication of the central
nervous system, from which the rats died in a few days. It is not
improbable that it was some sort of an avitaminosis.

Regarding dietary factors such as nucleins, lipoids, etc., it is quite
apparent that they are synthesized in the animal body. It is often
specified that for certain animals, and also for man, the animal
proteins are preferable to the vegetable proteins. In practice this
is quite true, and it may be explained in many different ways. In
the first place, when we speak ordinarily of animal and vegetable
protein, we have in mind the natural products containing those
proteins. Now, there are great fundamental differences between the
two types of foods. When we feed an animal protein, like meat,
eggs, etc., we give the protein, which is very poor in other dietary
components, like fat, and especially carbohydrates, in very concen-
trated form. With plant products it is different; they contain
protein in a very much smaller concentration, accompanied, for the
most part, by large amounts of carbohydrates. That this may be of
importance, we shall see later. Furthermore, it is possible that the
amino acids of animal proteins, because of their close resemblance
in composition to those of the body proteins, are better utilized;
besides this, they may contain known and perhaps even unknown
vitamines carried along during the process of preparation. For
example, Osborne, Wakeman and Ferry (I.e. 102) found that it is
very difficult to free edestin from vitamine B. As to the relative
value of plant and animal protein, we shall go into it more fully in
discussing human nutrition.

When we elaborated the vitamine theory in 1912, we stated that
these new substances are important or even indispensable to all
plants and animals, predicated upon the then available data. The
number of facts which we can bring to our support in proof of the
above statement has been greatly augmented since the first edition
of this book appeared, although not all types of animals have been
investigated in this respect. It is quite true that some animals
placed upon a vitamine-free or vitamine-poor diet do not remain

THE VITAMINES IN THE ANIMAL KINGDOM 75

in good condition, even though no symptoms of any of the known
avitaminoses are apparent. Nevertheless, these exceptions cannot
militate against the general significance of the vitamines for the life
processes for many reasons. For one, so long as the experimental
animals are kept upon an artificial diet made up of purified con-
stituents, they do not live long if extracts of unpurified products
are not given. All experiments seeking to demonstrate the contrary
have been carried out by feeding a dietary containing either traces
of known or unknown vitamines. Then again, it has also been shown
that some species of animals can dispense with one or another of the
vitamines. This phenomenon may perhaps be explained by the
symbiosis of the host with certain bacteria. In the higher animals,
these microorganisms are found in the intestinal tract in large
amounts and are perhaps able to build up the vitamines from simple
substances and convey them to the animal. At the end of this
chapter we shall add a short paragraph dealing with the role of bac-
teria in animals, but at this juncture, we shall only remark that no
attempt has as yet been made to establish these new viewpoints
experimentally.

All these procedures bring us nearer to the point that interests us
mostly, namely, accurate knowledge of the subject of human nutri-
tion. At once, we are confronted with questions dealing with
economic and agricultural problems. Let us consider the breeding
of animals as an example. From everything that has already
been mentioned it is apparent that the condition of the soil, which
contains quantities of organic and nitrogenous matter, can be
brought into relationship with the study of vitamines. The indi-
cation for such problems is seen in the publication of W. A. Davis
(195) who thought that the poverty of the soil (in Bihar, India)
in phosphorus is the cause of an avitaminosis in cows, which is
responsible for a smaller yield of milk and is connected with the
under-nourishment of children. Similarly, a nervous disease of
horses was observed in these villages. In European countries, as
well as in the United States, where agriculture and cattle raising are
carried out rationally, these conditions are of course to be noted
very infrequently. It is different in countries like Australia, Argen-
tine and South Africa, where cattle raising is carried out on a large
scale without paying sufficient attention to the care of the soil, due
no doubt to the lack of laborers. There we often hear of diseases,

76 THE VITAMINES

which are not always considered as deficiency diseases, though they
might turn out to be such. Because of insufficient irrigation of the
fields and periodic dry spells, we see domestic animals, feeding on
wretched vegetation for months at a time, dying from diseases which
we shall describe more in detail further along in the chapter. Here,
we are perhaps dealing with conditions which follow each other
like the individual links of a chain. As a result of the unfavorable
condition of the soil and the paucity of soil bacteria, the vegetation
suffers. The animals which must feed on this vegetation either
become sick, or else are not in perfect health, though no unusual
symptoms may be apparent. Subsequently, their offspring and
the children drinking the milk of such animals are naturally poorly
nourished; the same is true of grown people who use the meat of
such animals. Indeed, these conditions could as well prevail in
European countries where, because of the war, the number of cattle
has decreased, leading to the exhaustion of natural fertilizer, which
could not be replaced by artificial fertilizer because of the difficulty
of obtaining it. The diagram, illustrative of the relationship be-
tween the above mentioned factors, is shown below.

^Unfavorable soil conditions
Lack of soil bacteria Insufficient fertilizer

I T

Wretched vegetation > Diseased animals

I

Meat and milk of incomplete
nutritive value.
Avitaminoses in man

LIFE WITHOUT BACTEEIA

As we have stated several times previously, the incomplete knowl-
edge of the role of the bacterial flora in animals is the cause of a
great gap in the conception of the general significance of the
vitamines. We know, or at least we suppose, that some bacteria
are independent in their metabolism of an outside addition of vita-
mine, and since we know that a great number of bacteria live in the
intestinal canal, either in symbiosis or else as a parasite, we can
easily imagine that their functions may have something to do with
supplying the host with vitamines.

THE VITAMINES IN THE ANIMAL KINGDOM 77

Long before the introduction of the vitamine theory in the study
of nutrition, Schottelius (196) sought to solve the problem as to
whether it was possible to raise animals in a sterile state, and to keep
them alive for a long time. After overcoming great experimental
difficulties, among which was the construction of specially designed
apparatus, he was finally able to raise and keep young chickens under
sterile conditions. The animals, however, soon became cachectic and
died. Using similar methods, Belonowsky (197) was no more fortu-
nate. The problem was then undertaken with lower and higher
animals by various workers. Of the lower animals, flies were chiefly
used. In this connection, there are the experiments of Bogdanow
(198) who attempted to raise larvae of meat flies (Calliphora vomi-
toria) in a sterile condition. He proceeded as follows : the eggs were
sterilized with sublimate and the larvse cultivated upon sterile meat.
They grew very slowly, and showed no metamorphosis. Since they
did not survive, Bogdanow concluded that bacteria are necessary for
this type of insect. In a number of experiments, Delcourt and
Guyenot (199), and also Guyenot himself, sought to reach the goal
by the use of a different method. They attempted to obtain aseptic
larvse through successive cultures on an acid medium. They noted
that flies could live on yeast and, after several generations, also on
sterile potato. Larvse grew well on peptone and salt, but showed
no metamorphosis. Wollmann (200), following the method of
Bogdanow, carried these investigations further, and came to the
same conclusions as the latter.

A number of animals, like the scorpion and winter lizard, are
supposed to have sterile intestines and hence are specially suitable
for such experiments. Mme. Metchnikoff (201) tried to raise frogs'
larvse in a sterile state but without success. Moro (202) had already
made similar experiments with larvse of the alliaceous toad. The
larvse could be kept alive for 35 days, but their growth was much
less favorable than that of the controls. Since all these experiments
appeared to be rather indefinite (at least for the pre-vitamine period),
Cohendy (203) undertook the experiments of Schottelius with
chickens. For this purpose, he built a very ingenious apparatus, in
which the food, together with the whole paraphernalia, was sterilized
at 115°C. for 25 minutes, or for 1J hours at 118°C. Cohendy
reported that he had obtained good results with this arrangement,
and came to the conclusion that life is quite possible in the absence

78 THE VITAMINES

of bacteria. Upon reading through his work, one receives the
impression that although the animals ate more food than the controls,
they weighed much less, and soon died. The longest duration of life
was 40 days (204). Schottelius (205), criticising the above work,
expressed the idea that Cohendy's experiments showed, on the con-
trary, that the intestinal bacteria were useful, or even indispensable.
Such experiments were likewise carried out with mammals.
Kiister (206), working with goats, was able to maintain young
animals in a sterile condition for 35 days, during which time no
difference from the controls was noted. It is not impossible that
these experiments did not last long enough. Charrin and Guille-
monat (207) carried out similar experiments on guinea pigs, while
Kianizin (208) used guinea pigs and rabbits. The latter made his
animals breathe sterilized air and eat sterilized food. The poor
results of this regime were noticeable in from 4 to 9 days; the animals
suffered from weakness and died soon thereafter. Nutall and Thier-
felder (209) conducted the well known experiments on guinea pigs
in which these animals were kept free from bacteria and symptoms
of sickness and even showed a gain in weight. In this case, too,the
experiments were of rather short duration.

These experiments required corroboration from the viewpoint of
progress in vitamine research. This view is held likewise by Weill
and Mouriquand (210) and Schaeffer (211). It must be pointed
out that while all the investigators working on the problem of bac-
teria-free existence thought that they were dealing with only a
single unknown factor (the role of bacteria), there were really two
to be considered. During the process of food sterilization, the
vitamines — according to the choice of food and the conditions of
temperature— were more or less affected by the heat, and probably
destroyed. It is obvious, therefore, that the problem of a germ-
free existence was much more complicated than was imagined by the
above mentioned investigators. From the facts known at present,
we may conclude that in some animals, in which it is not possible to
produce an avitaminosis by means of a diet either poor or lacking in
vitamines, the bacterial intestinal flora possibly assume the task of
preparing the necessary substances for the host. To solve the ques-
tion beyond all doubt, it is necessary to proceed so, that in the
first place we arrange for a bacteria-free growth; if this should fail,
the animals may perhaps be saved through the addition of vita-

THE VITAMINES IN THE ANIMAL KINGDOM 79

mine. It may then be possible to experiment with the pure cul-
tures of the intestinal bacteria indigenous to the particular species,
to produce a normal dietetic condition without the addition of vita-
mine. Only then shall we be in a position to state that vitamines
are necessary for the existence of the entire animal kingdom and
eventually for the whole organized world.2 Sometimes, however,
a poor idea has some truth in it, and this seems to be the case with
Portier 's hypothesis. It is possible, as we have already said, that
certain animals and plants obtain their vitamines through a sym-
biosis with bacteria. This idea was tested experimentally by
Portier and Randoin (217). They made the observation that
rabbits fed on sterilized cabbage and carrots developed an avitamin-
osis, while rabbits which received an addition of the excrement of
another rabbit on the same diet remained normal; still, it is not
quite clear to us why rabbits kept on the same diet should possess
a different intestinal flora than those receiving an addition of feces.
It appears that these experiments, serving to demonstrate the
importance of the symbiotic intestinal bacteria, could be much better
explained on the basis of the vitamine content of the feces.

PROTOZOA

Investigations on protozoa were carried out chiefly with paramecia.
Lund (218) has recently shown that he grew these single-cell organ-
isms on yeast extract, but the details of the experiment have not yet
been published. Calkins and Eddy (219) worked with Paramecium
aurelia kept on two different nutritive media — hay infusion and
cooked flour. As a source of vitamine, a preparation of vitamine B
of the pancreas was used. The number of daughter cells varied
within the limits of experimental error, and therefore it was con-
cluded that the vitamine exerted no influence on this type of animal.
Flather (220) obtained somewhat dissimilar results from experiments
in which pure cultures of species of paramecia were maintained
always under the same experimental conditions; in every experiment,

2 Portier (212) has recently formulated a hypothesis in which he assumes
that in certain plant and animal organs, there are heat resisting bacteria
(called symbiotes) which behave biochemically like vitamines (Bierry and
Portier (213). Just now, we shall not discuss this theory itself for it appears
to be lacking a real basis, and besides it has already been made the subject
of an earnest critique by Delage (214), Lumiere (215) and Ranc

80 THE VITAMINES

three cells were grown in two drops of water, with the addition of one
drop of a 1 per cent watery solution of polished or unpolished rice,
or malted milk. The division factor was taken as a measure of cell
activity and the first 24 hours were disregarded, to give the cells
the opportunity of adjusting themselves to the new conditions. The
following values were obtained: for white rice 0.34, for unpolished
rice 0.58, for malted milk 0.84. It was then possible, by the addition
of malted milk, to make the nutritive value of the white rice approxi-
mate that of the malted milk Other experiments were made with
various concentrations of malted milk and also with orange juice,
with the result that an addition of malted milk in higher concentra-
tions was of no further advantage, while orange juice showed a very
slight activity Addition of orange juice to white rice was followed
by better results than when added to unpolished rice, and with
orange juice alone, no paramecia cells could live after six days. In
these experiments, the influence of vitamines is easily recognized;
in fact, Flather was undoubtedly dealing in this case, not with one
vitamine but with the combined influence of the mixture of vitamines.

Chambers (221) carried out similar experiments which demon-
strated the slight effect of potato extract; this displayed some influ-
ence only if the nutritive solution did not contain sufficient dietary
constituents; the same was true of malted milk. Peters (222)
cultivated Golpidium colpoda on a nutritive solution which con-
tained, besides salts, a mixture of some amino acids and in some
cases only one amino acid, tryptophane. The mixture of amino
acids was more active than the tryptophane alone. The first
divisions took place very slowly, though it was possible to keep the
cultures alive for three months through sub-culture. Peters ex-
plicitly emphasized that he could find no symbiotic microorganism.
He stated that in certain stages of development, a vitamine-like
substance seems to play a definite role. In all these experiments,
having for the purpose the study of the influence of the vitamines,
it is important that the food mixture used should be proved to be
vitamine-free. Abderhalden and Kohler (I.e. 127) investigated
the action of yeast extracts on flagellates (colpoda) with positive
findings.

When we leave the protozoa, we find a dearth of available work
on other kinds of animals, although it should not be thought that
such work does not exist. This is specially true of the polycellular

THE VITAMINES IN THE ANIMAL KINGDOM 81

animals investigated at various biological experimental stations,
where observations which would be of interest to us have undoubtedly
been made on the modes of nutrition of many insects. Unfortunately,
these data were unavailable to us.

METAZOA

Coming to the polycellular animals, we shall first describe a series
of experiments undertaken for some other purposes. They deal with
the influence of lecithin upon growth and were carried out by Goldfarb
(223). In this respect, he investigated a whole series of animals, and
also tadpoles and eggs of the starfish. At first, he stated emphati-
cally that lecithin had a definite action, but in a later work (224),
he admitted that it was inactive. This is apparently associated
with a vitamine impurity, whose presence may vary with the mode
of preparation of lecithin.

It may not be amiss to say a few words here on Carrel's tissue
culture in vitro, which in the future may perhaps be of help in advanc-
ing the vitamine problem. These tissue cultures may be thought of
as polycellular organisms.

The growth of tissue in vitro

Most of the experiments of Carrel were made at a time when the
vitamines were either unknown or given very little consideration;
otherwise it might have been possible to choose still better conditions
for tissue culture. We must admit, however, that Carrel practically
foresaw the future trend of work, in that he chose the natural juices .
of the animal organism as the nutritive solutions. Inasmuch as the
author has little practical knowledge of this method, he is naturally
not in a position to judge whether or not Carrel would have obtained
even better results if he had tested his nutritive solutions for the
presence of all the necessary elements for nutrition. Because of the
great interest that experimental surgery of the future may have in
this method, it is very important to obtain the greatest success.
The tissue fragments used in these experiments, are removed from
their normal environment and must then lead an independent exist-
ence. For this purpose, they need all of the dietary constituents
(also vitamines) designated as essential by the modern science of

82 THE VITAMINES

nutrition. By means of frequent sub-cultures, Carrel prevented the
nutritive substances from becoming exhausted. That this had to be
resorted to frequently in order to keep a fragment of tissue alive for
a long while, shows perhaps that the conditions for growth could
have been better chosen.

In this respect, the method may be applicable to a new field of
work. We have in mind particularly Carrel's antiseptic method
of treating wounds, which came into use during the war. It is
apparent that the organism suffering from shock and other complica-
tions could not be in the best nutritive condition; and in the wounded
tissue itself, through the disturbance of the blood circulation, con-
ditions could not be the most favorable. For these reasons, it would
be of interest to apply to the treatment of the wound, those facts
that have been learned in the study of the composition of nutritive
fluids best suited for the tissues. Practically, this could be brought
about by the substitution of the antiseptic solution by a nutritive
solution from time to time, with the possibility that the wounded
tissue could be nourished and the process of healing hastened.

The method of Carrel (225) is as follows : Under appropriate con-
ditions, tissue fragments, and pieces of tumors too, may be grown for
more than four months, during which the tissue shows growth and
peripheral expansion. The tissue (226), antiseptically prepared, is
placed in a plasma diluted with J to \ of its volume of distilled
water. For this purpose, it is best to use the plasma of the animal
under investigation or that of a homologous animal. In from 3 to
4 days, the tissue culture (Carrel (227), Carrel and Burrows (228))
is placed for a few minutes in Ringer's solution and then transferred
to a fresh plasma. Some tissue cultures have been kept alive for
four and a half months, having been transferred as above 48 times.
A fragment of heart continued to beat after 140 days. Only embry-
onic tissues and tumors could grow in an artificial medium (Locke's
solution, agar and bouillon). In a later work, Carrel (229) investi-
gated in greater detail the influence of tissue extracts and body fluids
(as did Walton (230) too), and found that they increased the growth
of connective tissue in particular from 3 to 40 times. This was the
case especially with embryonic spleen extracts, and it was consider-
ably smaller with greater dilutions. It was true only of the same
type of animal, and the action of the extracts was weaker on heating
to 56°C. and entirely dissipated at 70°C. The active substance

THE VITAMINES IN THE ANIMAL KINGDOM 83

could be filtered through a Berkefeld filter, but not through a Cham-
berland filter. Regarding the foregoing, it appears likely that a new
field for vitamine research is opened up.

INSECTS

We have to deal here with the very interesting studies of Jacques
Loeb (231). He demonstrated that a type of fly (Drosophila) could
attain the larva stage upon a solution of cane sugar and salts, together
with the addition of filter paper. The larvae grew quickly, especially
on the addition of alanine or an ammonium salt. In this particular
instance, no attention was paid to the influence of the bacteria on the
substrate. Five successive generations of the banana fly (232) were
grown on a solution of glucose, cane sugar, ammonium tartrate, citric
acid, di-potassium phosphate and magnesium sulphate. As Loeb
himself observed, it was not quite certain that the solution was free
of bacteria; the presence of yeast in particular was not excluded.
In these experiments, the flies themselves were not sterile and there-
fore Loeb and Northrop (233) started experiments with flies hatched
from eggs sterilized by a sublimate solution. Twelve sterile genera-
tions were cultivated in this manner, carefully protected from every
possibility of contamination with yeast growth. The nutritive solu-
tion used was composed of 450 grams baker's yeast and 50 grams
citric acid (used for the purpose of decreasing the danger of infection
during the various manipulations) in a liter of water. In a great
number of experiments, the investigators showed that on another
medium, for instance, filter paper and cane sugar, with the addition
of salts, and likewise with the addition of casein, edestin, egg albumin,
milk, or a mixture of amino acids, the larvae attained normal size
but could not be brought to the stage of metamorphosis. Sterile
flies, grown on sterile bananas or potatoes, showed no sexual develop-
ment. The results with yeast as substrate were different; here, all
the substances are manifestly at hand, which are indispensable for
the growth and development of flies. Butter, nucleic acids, thymus
and thyroid extracts were without influence, and the authors believed
that these substances are different from those which are of importance
for pigeons, rats and other warm-blooded animals. This view must
naturally undergo a slight modification, since there is obviously no
reason to differentiate between flies and warm-blooded species. The

84 THE VITAMINES

yeast substance was heat-resistant but lost some of its activity on
treatment with cold or hot alcohol. Drosophila is a monophageous
animal and yeast seems to be the only adequate food for it. These
flies can live in any nutritive solution that is suitable for the growth
of yeast, and the medium is usually infected with this organism.
Loeb and Northrop rightly observed that in considering the synthetic
abilities of the higher animals, one must not lose sight of a possible
cooperation of the intestinal bacteria. Loeb and Northrop (234) and
also Northrop (235) showed by means of a specially planned series
of experiments that the developmental stages of flies (except imago)
can be prolonged from 8 to 17 days at will. They believed that the
three stages of metamorphosis could be regulated by the formation
and disappearance of three different substances.

In a later work, Northrop (236) investigated the role of yeast in
the nutrition of flies. He found that the number of flies that could
be developed on a certain quantity of yeast could be increased by
the addition of bananas, casein or sugar. The rate of growth on a
combination of yeast and bananas is greater than on yeast alone.
In mixtures containing a small amount of yeast, growth proceeds
more slowly, and in a dilution of 1 to 128, it becomes entirely abnormal.
Kidney, liver, dog pancreas, mouse liver, and bodies of flies them-
selves were suitable for the growth of the larvae, while sterilized
spleen, heart muscle, suprarenal glands, thyroid and blood of the
dog were unsuitable; the addition of muscle tissue, testicles and
sterilized thymus gland of the dog, rabbit and calf, resulted in the
development of some chrysalis, but growth proceeded slowly and
the flies appeared dwarfed. Robertson's tethelin (lipoid from the
hypophysis) was inactive.

Guye'not (237) attributed to the vitamines some part in the devel-
opment of fly larvae of the Drosophila ampelophila. Baumberger
(238) investigated some species of flies and came to the conclusion
that Drosphila melanogaster can live on fermenting fruits and also on
yeast protein. A combination of yeast, nucleo-proteids, sugar and
a salt mixture provides a suitable food for larvae. Other kinds of
flies, like Musca domestica, Desmotopa, Sciaria and Tyroglypha, can
all thrive on microorganisms.

From this chapter, we see, especially from the work of Loeb, that
flies need vitamines for their development, and that vitamine B
plays the most important part.

THE VITAMINES IN THE ANIMAL KINGDOM 85

FISH

As we progress in the animal series, we do not meet with any
vitamine experiments — at least to our knowledge — till we come to
fish, the single great group typical of the vertebrates, of which we
have no knowledge in this respect. There is only the work of Mor-
gulis (239) which might be considered to have some slight relationship
to our subject. He fed ox heart to fish and found that it is better
assimilated when cooked, while ox liver had no particular peculiari-
ties. Almy and Robinson (240), comparing dry and fresh fish food,
found that with the latter there was less mortality and a more definite
weight increase.

AMPHIBIA

Emmett and Allen (241) used frog larvae in their vitamine studies,
but the fact that the larvae consumed each other complicated the
experimental work somewhat. The larvae reacted poorly towards a
greater addition of fat, and developed better on a diet containing 5
per cent fat than on one containing 28 per cent. Both vitamines, A
and B, appeared to be necessary, although the latter seemed more
important. The variation of protein content from 10 to 30 per cent
was of no special significance, although the optimum growth was
obtained with higher concentrations. Lactalbumin or a mixture of
beef and oats was superior to corn gluten, indicating the importance
of the protein used. The growth of the hind legs depends upon
the addition of vitamine, according to the findings of Emmett, Allen
and Sturtevant (242) . For complete metamorphosis, the addition of
iodine also appears to be necessary.

Harden and Zilva (243) investigated both larvae and adult frogs.
With tadpoles kept in water to which was added fresh meat, a normal
development was noted, while an experimental diet made up of 20
per cent casein, 75 per cent starch, 5 per cent salts with the addition
of butter, yeast extract, and orange juice, did not permit of normal
development, since only two out of six animals attained full growth.
According to this a better experimental diet had to be prepared.
With adult frogs, another difficulty presented itself in the develop-
ment of a fungus-like disease. On a diet free from vitamine B, some
of the animals stretched out their legs, before death, and one suffered
from convulsions. Without A- and B-vitamines, seven frogs died in
about four months. Out of five animals on a diet containing the

86

THE VITAMINES

PCR/0D A4

n H f

Hi

PERIOD

ff H ft f} f

FIG. a

FIG. 6. GROWTH OF FROG LARVAE

Diet Hi contained A- and B-vitamines, while diet II2 was lacking in B-
vitamine (Emmett-Allen).

FIG. 7. GROWTH OF FROG LARVAE

Diet Ilia was complete while diet Viz was deficient in vitamines
(Emmett-Allen).

THE VITAMINES IN THE ANIMAL KINGDOM 87

above two vitamines, with or without vitamine C, only one died.
Out of six animals getting A-vitamine but not C-vitamine, all died
in about 155 days. Out of five animals receiving vitamine B, with
or without C, four survived for 250 days.

Accordingly, B-vitamine is necessary for the maintenance of
grown frogs, although the symptoms were noticeable only after six
months, while the C-vitamine appears to exert no influence on the
animals.

BIRDS

Chickens

As stated in the historical part, Eijkman (I.e. 50) and Grijns (I.e.
52) were the first to experiment with chickens. It was particularly
noted that Eijkman, by feeding chickens with the food discarded by
a beriberi hospital, observed that they developed a disease which he
called "Polyneuritis gallinarum." Eijkman soon found that other
foodstuffs, like sago and starch, could also bring on this disease.

Normal nutrition of chickens. We shall first consider the normal
dietary requirements of the chicken. One who has tried to raise
young chickens in the laboratory knows the great difficulties that
have to be dealt with. The first attempt in this direction was made
by the author (244) who investigated the influence of various kinds of
foods on the growth of experimental tumors. In spite of the great
eagerness with which the chickens ate the food, growth ceased and
they developed a characteristic disease of the legs; the toes lost their
vigor and were bent. Very often it was noticeable that the beak was
not closed properly, and the animals died because of their inability
to pick up food. The eyes were frequently closed, and the chickens
suffered from a disease which gradually led to complete blindness,
and which would have been regarded to-day as ophthalmia. A
change in the diet was not enough to better conditions since the
animals were already receiving quite a variety of food. An attempt
was made to feed the chickens with live grub-Worms, and for a time
the floors of the cages were covered with grass and earth, which was
frequently renewed. We believed then that the wooden floors had
some pathological effect on the legs of the animals. However, all
these measures were of no avail and the birds became sick, showing
symptoms which we then attributed to rickets. On this basis, we
used cod liver oil as a therapeutic measure. As the figures below
indicate, a young chicken could maintain itself for eight months on

88

THE VITAMINES

unpolished rice and cod liver oil (Funk and Macallum 245). The
animal was normally built but weighed only 160 grams while a chicken
of this age should weigh about two kilos. No symptoms that could
have been regarded as rachitic were noted.

The animal had a large beak and excessively long feathers for his
size; all secondary sexual indications were lacking and it chirped like
a four weeks old chick.

On white rice, the young chickens died (I.e. 244) after about two
weeks, while on unpolished rice it was possible to keep them alive for
more than 5 weeks. When the young chickens were fed Spratt's
chicken food, the weight increased more than twice, while on unpol-
ished rice for 4 weeks, there was no growth at all. These experi-

FIG. 8. SEVEN MONTHS OLD CHICKENS

Normal diet and a diet of unpolished rice and cod liver oil. (Funk -Mac-
allum). Left— 2500 grams; right— 160 grams.

ments were conducted with 15 days old chickens and were later
repeated, with the same results (Funk 246). In all exact feeding
experiments with chickens, it is necessary, particularly during the
summer, to cover the cages with muslin, otherwise they eat so many
flies that the results may be misleading. We have often observed
that a chicken which grew in spite of a deficient diet stopped growing
promptly as soon as the cage was covered.

Drummond (247)3 carried our experiments further and criticized
our results rather sharply. He found that the possibility of raising

3 After my departure from the London Cancer Hospital Research Institute
in 1915, my former assistant, J. C. Drummond, continued a great number of
experiments started by me, without communicating with me, and without
stating in his publications that the ideas had originated with me.

THE VITAMINES IN THE ANIMAL KINGDOM 89

chickens in the laboratory to full growth depended upon the age of
the chickens used. He observed that the animals often failed to grow
on a normal diet, and he accordingly believed that this suppression
of growth had nothing to do with the vitamines, although his results
with white and with red (unpolished) rice, as compared with a normal
diet, were identical with ours. Drummond then tried to improve
the condition of the animals by giving them earth, sand and wood
chips, but without success. We showed later (248) that it was
possible to decrease the mortality of the animals and to improve
their general condition, by adding animal charcoal to the diet and
substituting milk for the drinking water.

Buckner, Nollau and Kastle (249) demonstrated that young
chickens did not grow well on a diet poor in lysine, as compared to
one rich in lysine; the addition of butter was without effect. It is
not improbable that the diet rich in lysine possessed altogether better
nutritive value. Similar results with lysine were obtained by
Osborne and Mendel (250). In a later investigation, they (251)
occupied themselves with the problem of raising chickens in the
laboratory. They stated, in accord with our results, and in opposi-
tion to those of Drummond, that it is quite possible to raise young
chickens in cages. On a diet containing butter as a source of vita-
mine A, and an addition of filter paper, it was possible to raise some
several weeks old animals to normal size. Although most of the
animals suffered from weakness in the legs, the investigators believed
that as soon as all conditions for the growth of chickens were known,
it would be possible to conduct growth experiments similar to those
with rats. In these experiments, no green food was given and the
diet was thought to be completely carrotinoids-free, which was denied
by Palmer. Hart, Halpin and Steenbock (252) were of the opinion
that the disease of chickens mentioned above, designated "leg weak-
ness," could not be considered as an avitaminosis. Practical chicken
breeders attribute this disease to the close confinement, lack of green
food and over-eating. The above workers explained the condition
as being due to constipation and stated that the green food, because
of its vitamine content and cellulose, might have had a beneficial
influence on the constipation. They found indeed that green food
is not necessary, and that the same results could be obtained by giving
filter paper, dirt or animal charcoal instead; it was necessary to
mix these substances intimately with the diet. These additions

90 THE VITAMINES

served not only as roughage, preventing constipation, but also to
dilute the diet and make it more easily utilized. In most cases, the
diet consisted of 25 per cent casein, 38 per cent dextrin, 15 per cent
butter, 5 per cent salt mixture, 2 per cent agar and 15 per cent dried
yeast. With less casein, butter, yeast and salts, and more dextrin,
in making up a diet which is very good for rats, no growth was
observed in chickens. The experiments lasted from three to four
months and it still remains to be seen whether with this regime the
experiments can be duplicated, for only then can we regard the
problem of the synthetic nutrition of chickens as solved. Filter paper
seemed to be the best substance to use for roughage, although an
addition of 10 per cent animal charcoal was also effective.

Palmer (253) carried out a series of experiments in which he tried
to raise animals, and particularly chickens, on a diet completely free
from plant pigments. The experiments are of special interest because
of their relationship with the antirachitic vitamine, to be referred
to later. Palmer found that some of the animal pigments belonging
to the carrotinoid group, have their origin in the vegetable kingdom.
The species having a colored adipose tissue obtain this pigment from
the blood, in which it is easy to demonstrate its presence. In a series
of feeding experiments with chickens, Palmer and Kempster (254)
showed that in some foodstuffs a certain relationship exists between
the carrotinoid and the vitamine A content; they demonstrated,
however, that this relationship is only apparent and that the pig-
ments, per se, play no part in metabolism. They also experienced
considerable difficulty in raising the chickens in a closed room, and
further, in rinding a diet free from pigments. In the end, they chose
a diet composed of white corn, bran from the white corn, bleached
flour, centrifuged milk, and bone meal. The chickens used were
white Leghorns and weighed from 700 to 750 grams at the start;
the colored parts of the body, like the beak, and the feather quills,
were only weakly colored; the mortality was quite high. After six
months, there were only five out of eleven chickens still alive which
had doubled their weight and begun to lay eggs. The eggs were not
completely pigment-free, but were nevertheless only weakly colored.
Very young chickens could not be raised on this diet. Better results
were obtained when filter-paper was added, according to the method
of Osborne and Mendel.

THE VITAMINES IN THE ANIMAL KINGDOM 91

However, instead of using butter, which is not carrotin free, pig
liver was used as a source of antirachitic vitamine. Sixty chicks
were placed on this diet, upon which they grew well for about 6
weeks, but then began to lose ground. At this stage of the experi-
ment, young pigs' liver was given with the result that an immediate
improvement was seen. The animals, which were 3 months old, were
of normal size and weight and received during the summer an addition
of pigment-free vegetables, such as white onions. Palmer and
Kempster concluded that young chickens could live without carrotin-
oids, if the diet contained sufficient vifcamines. They believed also
that the possibility of discovering the nature of vitamine A by an
investigation of the carrotinoids was without foundation in fact.
The animals fed on a carrotinoid-free diet laid eggs after 6 months,
and yet the chickens in the second generation were few in number
and rather puny in appearance.

Hart, Halpin and McCollum (255) and Hart, Halpin and Steen-
bock (256) studied the nutritive requirements of chickens. For their
experiments, they used half -grown animals fed on a mixture of corn,
corn gluten and calcium carbonate, and also wheat, wheat gluten
and calcium carbonate. The birds grew well and were able to lay
fertile eggs. These findings stood out in marked contrast to the
results obtained with rats and pigs, and showed satisfactorily that the
nutritive needs of chickens and some mammals were totally dissimilar.
In chickens, an addition of salts, casein and butter were without the
slightest influence, while the protein concentrates acted favorably.
These results might be attributed either to the influence of pro-
tein concentration or to the addition of. unknown factors, which
are not present in cereals and casein, and which exert a favor-
able influence on egg production. The experiments with younger
animals (I.e. 256) resulted somewhat differently. In this case, corn
was a sufficient source of B- and A-vitamine, but not so with wheat,
upon which the animals lived only three months. It was only when
salts, casein and butter were added that the diet appeared to be
complete. In a similar manner, Harney (257) was able to show that
for the production of eggs, the use of plant foodstuffs, in spite of
the high protein content, could not be compared to that of animal
food, and besides this, Kaupp (258) held that an addition of skimmed
milk could influence the production of eggs favorably.

92 THE VITAMINES

In accordance with these experiments, we must recognize the
importance of two vitamines, A and B, for the growth of young
chickens. For the grown animal, however, the importance of
vitamine A had not been set forth so clearly. The recognition of
these related facts is important for the proper consideration of
chicken beriberi,4 a disease arising largely through feeding with
white rice. McCollum and Davis (259) were able to show that this
kind of rice lacks, besides vitamine B, protein of high biological value,
some salts, and vitamine A. Whether these results, obtained with
rats, are applicable to other animals without further work, is at best
questionable; yet we must consider the possibility that when we try
to cure animals, fed with white rice, by adding vitamine B, the
indefinite therapeutic results sometimes obtained might be due par-
tially to the lack of other dietary constituents.

In conclusion, we shall mention the work of Houlbert (260) who
kept chickens on white rice, wheat and barley, heated in an autoclave
to render them poor in vitamines; twice a week, cod liver oil was
added. After 40 days, it was found that the division of cells in the
sexual and hematopoetic organs had come to an end. However, if
the missing vitamine was replaced in the diet, the glands resumed
normal development.

Chicken beriberi. In relation to the experiments of Eijkman and
Grijns, chicken beriberi, besides being brought on by white rice, may
arise from feeding white bread (Hill and Flack 261), an observation
also made by Ohler (262) and by Weill and Mouriquand (263) . Ohler
also produced the disease by feeding hominy, but not that from whole
corn. Based upon our experiments, Wellman, Bass and Eustis (264)
used cane sugar and corn starch; Wellman and Bass (265) also used
macaroni. Naturally, these experiments are only of historical inter-
est since they showed that the etiology of beriberi is not related only
to the rice consumption. We have known for a long time that all
food poor in B-vitamine can give rise to beriberi.

In the work of Vedder and Clark (266), there is an excellent
description of chicken berberi. The first symptoms, on an exclusive

4 In agreement with some others, in the light of the facts established in
this field of work, I have dropped the designation "Polyneuritis gallinarum,"
since according to the newest developments, it has nothing to do with neuritis,
but perhaps with a general change in almost all of the tissues. For a long
time, I have used the term "experimental beriberi of animals" for this
condition.

THE VITAMINES IN THE ANIMAL KINGDOM

93

diet of white rice, appear in from 20 to 30 days. At first there is
paralysis of the extensors of the legs; the bird sits on a flexed tarso
raetatarsal joint. Paralysis soon extends to the wings, nape of the
neck and the entire musculature. The animal then lies motionless
on its side ; a deep prostration appears frequently on the second or
third day after the onset of paralysis — at the latest, in one week —
and is followed by death in all cases; the whole course of the disease
is run in a very short time. From the characteristic symptoms of
experimental beriberi, the following should be noted particularly:

The crest becomes bluish red very soon, and this appearance may
be considered as one of the initial symptoms.

FIG. 9. AVIAN BERIBERI
First day symptoms (Fraser-Stanton)

Paralysis of the wing musculature belongs to the later symptoms —
the wings then hang loose down to the ground.

Spasticity seldom appears in the initial stages of the disease, but
more often in the later stages. We notice here the retraction of the
head to the back; sometimes there is seen the spastic walk on tip-
toes with stiff knees. Dysphagia belongs to the early symptoms;
water given to the bird flows out again from the beak; on attempting
to feed forcibly, the pigeon chokes. According to Vedder and Clark,
the loss of weight is constant at about 20 per cent of the initial weight.

Progress of the disease. The above named investigators describe
the progress of the disease in the following manner: In acute cases,
with prostration and great loss of weight, all the symptoms are

94

THE VITAMINES

apparent in 24 hours. The bird lies on one side, shows dyspnoea and
cyanosis and dies in a few hours. The second type is more chronic;
in these cases we see paralysis of the legs, but good general condition.
The appetite remains good, the loss in weight is insignificant; the

FIG. 10. AVIAX BERIBERI
First day symptoms (Fraser-Stanton)

crest remains red; in this condition, the chicken may live for weeks.
According to our personal experience, the second type is not so suit-
able for curative experiments.

Segawa (267) also described two types of beriberi — one form which
he regarded as a pure polyneuritis, and another, possessing more of

THE VITAMINES IN THE ANIMAL KINGDOMS 95

the characteristics of starvation, and expressed by a distinct aversion
to rice. In 66 per cent of all cases, both types occurred together.
In Segawa's experiments, some animals remained in good condition
for about 219 days; this may perhaps have been due to the possibility
that the animals had eaten traces of other food, such as flies and
vermin. He erroneously considered the disease as a real polyneuritis.
Tasawa (268) observed that the marked emaciation and starvation
could be avoided by an addition of egg-yolk or cooked meat, where-
upon only clear beriberi symptoms are noted.

As we shall see later in pigeons, fatigue plays a big part in the
development of acute symptoms. In accord with this, Hulshoff Pol

FIG. 11. AVIAN BERIBERI
Second day symptoms (Fraser-Stanton)

(269) found that forced feeding accelerates the acute symptoms, and
Williams and Johnston (270) found that the same effect could be
hastened by increased temperature and exercise.

Pigeons

The problem as to the normal food requirements of pigeons appears
to be much simpler than that of chickens. Since pigeons may be
raised in the laboratory with little difficulty and since they do not
eat vermin and flies, the feeding experiments may be conducted with
greater exactness. Young pigeons, after they have been hatched, are
fed alternately by their parents, so that it is possible, through the

96 THE VITAMINES

administration of a special diet to the latter, to study its effect upon
the young from the very beginning.

We have personally kept grown pigeons on an exclusive corn diet
for more than a year, during whch time they were not only in splendid
health but reproduced normally.5 It cannot be emphasized suffi-
ciently, that whole corn provides sufficient nutrition for the maintenance
and growth of pigeons, which indicates that corn is quite sufficient for
at least one species. This is of particular importance for our later
conception of the etiology of pellagra, which has recently been attrib-
uted to the deficiency of corn proteins. That corn is a complete
food for pigeons has been shown by other workers. Voegtlin and
Myers (271), reported that they maintained pigeons in good health on
an exclusive diet of corn for at least four months. In another series
of experiments, the same workers (272) investigated the conditions
of growth in young pigeons which attained normal size in 40 days,
after having been fed by their parents who had been given a diet of
whole corn or whole wheat, with the addition of a calcium salt. The
same result was obtained when the diet was composed of white bread,
with the addition of vitamine B and calcium salts; and also wheat
starch, casein, A- and B-vitamine, and calcium salts. Because of the
possible interest in the study of the growth of young pigeons, there is
appended a diagram illustrating normal growth as compared with
growth on corn, taken from the work of Voegtlin and Myers.

It is shown that young pigeons require two kinds of vitamines,
namely, A- and B-vitamines, and can get along very well in the
entire absence of C-vitamine, which is in direct opposition to the
needs of most mammals. Regarding the requirements of grown pigeons
for vitamine A, this must be left for the future to determine. Should
it appear that adult pigeons also need vitamine A, it would be worth6
while to so change the usual diet used to demonstrate the onset of

6 Among these were also pigeons which after being cured of beriberi, still
showed normal fecundity.

6 Experiments were made by us (273), in which two pigeons lived for 49
to 54 days respectively on an artificial diet if B-vitamine was injected every
few days, and finally died of sepsis caused by contamination of the vitamine
solution. Later on, Stepp (274) carried out some similar experiments in
which he kept a pigeon in excellent health for 91 days on dog biscuits extracted
with alcohol, and rice polishings (orypan) ; when the orypan was discontinued,
the bird died in 37 days. In spite of the use of orypan, the bird could not
fly well and lost some weight.

THE VITAMINES IN THE ANIMAL KINGDOM

97

"beriberi, namely, white rice, that it would lack nothing but B-vita-
mine, although we were able to induce a typical beriberi with a syn-
thetic diet lacking vitamine A (275) . As we have already pointed out
in the case of the chicken, all these dietary deficiencies can complicate
the picture of experimental beriberi unnecessarily. Because of the

FIG. 12. GROWTH CURVES OF SQUABS

Parents were fed on a diet limited to the corn and wheat kernel, supple-
mented by the addition of calcium carbonate (oyster shells) (Voegtlin-Myers).

protein deficiency of white rice, Butcher (276), and others, used an
addition of 3 per cent casein, calculated on the dry weight of rice.

Pigeon beriberi. The method most commonly used to induce beri-
beri in pigeons is to feed white rice, although other foodstuffs and
mixtures of isolated substances are available for that purpose.

98 THE VITAMINES

Among others, Funk and Cooper (277) showed in 1911 that the dis-
ease could be brought about with pure sugar, inulin, dextrin and
starch. The pigeon is quite the best animal to use in the study of
beriberi. When pigeons are placed upon a diet of polished rice,
they eat it with great avidity during the first few days. Following
this, their appetite diminishes markedly, and they frequently attempt
to disgorge the rice out of their crops. This behavior is more often
noticed in animals that are fed forcibly, but sometimes this rice is
again eaten voluntarily. This condition makes it difficulty to carry
out exact rice feeding experiments with pigeons. One has the impres-
sion that the animals have an aversion for rice, which, however, dis-
appears if the necessary vitamine is administered at the same time
with the food. After a few days, it is best to put the pigeon in a box
with a wooden cover so that its head extends; in this way it is possible
for one person to manage when forcible feeding must be resorted to.
The beak is opened and through a small metallic funnel with smooth
•edges, the weighed rice is pushed into the crop with a glass rod.
Another method used by us was to prepare the food in the form of
pills, and feed them as such. For feeding purposes, it is best to use
healthy males weighing from 300 to 350 grams; 20 to 30 grams, of
rice per day may be given in three portions. If the crop should
happen to be full, it is necessary to wait till it is empty. We shall
speak of the effect of the food ingested when we come to the physi-
ology of vitamine B (p. 210). After several days of rice feeding, a
marked change is noted in the animals. Very little remains of the
usually predominating desire to quarrel when a number of pigeons
are kept in one cage. Likewise, the sexual instinct, such as the
strutting of the male around the female, seems to be held in abeyance,
and the animals sit on the perch in a sleepy, apathetic manner. They
experience increasing difficulty in flying on to the perch and a few
days later some of the pigeons sit on the bottom of the cage, regardless
of the fact that they are soiled by the feces of the animals on the
perch above. The fecal matter, normally of semi-solid consistency
and whitish color, becomes slimy and water-clear, or slight yellowish,
and the animals show no disposition to keep themselves clean. If
they are left to feed themselves, their appetite disappears gradually
and their condition is that of semi-starvation. About 30 per cent of
the animals which are permitted to feed themselves develop symp-
toms of beriberi, while the remainder die of general weakness. Of

THE VITAMINES IN THE ANIMAL KINGDOM 99

the animals forcibly fed, a greater proportion develop the typical
beriberi symptoms, after a period of time which is subject to great
deviations. We may say, however, that most of the animals develop
the disease in from 10 to 30 days; it is noticeable that the animals
mostly become sick within a few days of each other, so that there is
the false impression of the appearance of an epidemic. In the
individual animals, we note the development of various types of
diseases, as illustrated in the accompanying pictures, closely resem-
bling the conditions described in man. This analogy may well be
fallacious and the types described below may depend perhaps only
on the nutritive condition and the length of time the rice is fed.

In one type, which we shall designate as acute, we notice that the
ability to walk is decreased. If the animal is frightened, it runs a
few steps without difficulty, but when fatigue sets in, the animal
helps itself by flapping its wings, but sits motionless when left alone;
the heart beat is pronounced and behaves as though it had been over-
exerted. The next symptom is uncontrolled motions of the head.
If the animal is turned about several times in the air, then in many
cases the acute form is produced out of the latent. The phenomenon
appears quite suddenly — the head is pressed against the back by a
retraction of the neck muscle, the legs are drawn up to the belly;
the animal turns somersaults, and this may last a long time under
certain conditions. In this stage, the animal does not survive very
long. During the course of a day, a disturbance in breathing
develops; the animal opens its beak wide, apparently struggling
against suffocation, and finally death results in from 12 to 24 hours
after the appearance of the symptoms, with an easing up of the
spasms.

The second or chronic type (figs. 14-15) develops as follows:
After a few weeks the animal is found sitting in the cage; it moves
very reluctantly in spite of its ability to do so. After some days,
it loses even this capability, and sits motionless in one spot. Only
seldom does this form go over into the spastic. Usually, the animal
lives a few weeks longer and dies without moving from its place.
These cases are not suitable for curative experiments. For this
purpose, we must use only those animals in the first group.

The latter also show variations. In some, the spastic condition
persists till death while in others, it disappears for short intervals
only to reappeaT in a more severe form.

100

THE VITAMINES

FIG. 13. SPASTIC FOBM OF BERIBERI IN PIGEONS

FIG. 14. ATROPHIC FORM

THE VITAMINES IN THE ANIMAL KINGDOM

101

FIG. 15. PARALYTIC FORM

FIG. 16. SPASTIC-PARALYTIC FORM

102

THE VITAMINES

A further phenomenon, common to both forms of beriberi in
pigeons, is the loss in weight, which is apparent before the onset of
the symptoms. This loss, amounting to from 20 to 45 per cent of the -
original weight, occurs always in pigeons fed on rice, according to our
experience. The loss in weight is apparent whether the animal is
fed forcibly or not. Results to the contrary are due to the fact that
the pigeons, chiefly because of the paralysis of the muscles of the
crop, are unable to empty it. On post mortem, the crop is tightly
packed with rice, weighing about 100 grams, which ordinarily would
have been calculated as body weight. It cannot be emphasized
sufficiently that in experimental beriberi, following a strictly managed

400

350
300
250
ZOO

no

100

FIG. 17. DROP IN WEIGHT OF PIGEONS FED ON 30 GRAMS POLISHED RICE DAILY

diet, death ensues in 100 per cent of the cases. Theiler, Green and
Viljoen (278) have observed that starving pigeons, only on water,
may show spontaneous cures. This behavior was explained by the
mobilization of the vitamine in the tissues. The tissues are catabo-
lized more rapidly during starvation and therefore a greater supply
of vitamine may suddenly be made available for the animal. If rice
feeding is continued death must eventually ensue. The question of
malnutrition in beriberi was also taken up by Lumie're (279).

Besides chickens and pigeons, beriberi has also been described in
other birds, which might sometimes also be of practical importance.
For instance, Merklen (280) described a disease in 3 or 4 weeks old
ducks manifesting itself in symptoms of cramps, paralysis of the legs

THE VITAMINES IN THE ANIMAL KINGDOM 103

and inanition, which quickly disappeared on substituting the diet
by one more varied. Perhaps we are dealing with beriberi here
too. The appearance of beriberi in other birds is tabulated below:

Animal Observer

Ducks Eijkman (l.c.48) and Kulz (281?

Geese Eijkman (l.c.48)

Sparrows Fujitani (282)

Quail Toyama (283)

Jushimatsu Toyama (283)

Parrots Fink (284)

Rice birds Ottow (518)

Munia maja Jansen (284a)

Pathological anatomy and chemical pathology of beriberi in birds

Pathological anatomy. Till recently, everyone was of the opinion
that in avian beriberi the manifestations in the central nervous
system were in the foreground. The new investigations show, on
the other hand, that as a result of the lack of vitamine B, many, if
not all, of the organs suffer, and for this reason the disease cannot be
thought of as "polyneuritis." Despite many attempts, it has not yet
been possible to find out which organ or tissue is primarily responsible
for the manifestations of the disease. This is of course bound up with
our insufficient knowledge of the physiology and significance of the
vitamines.

Nervous system. We find a very good description of it in the
work of Vedder and Clark (I.e. 266), already mentioned, and
more recently in a report by Onari Kimura (285). We see from
these investigations, that not all nerves undergo pathological
changes in the same measure; for example, the vagus degen-
erates, but not in a high degree. However, in the opinion of
Kimura, beriberi is a general disease of the nervous system, and
this was our opinion too. We still believe that all of the hetero-
geneous pathological changes occurring in beriberi may best be
looked upon as of central origin, in so far as they may all be regarded
as atrophic changes. Naturally, we admit that still another inter-
pretation may be found.

The nerves of the lower extremities are selectively more affected
than those of the upper. Especially is this true of the sciatic and
peroneus nerves which show a fatty degeneration, though no definite

104 THE VITAMINES

paralysis is noted. These changes arise after 7 days of rice feeding.
The number of degenerated fibers bears no relation to the severity
of the paralysis. Cases of -light clinical symptoms often display
marked degeneration, while severe cases frequently show only from
4 to 10 per cent of degenerated fibers. As a rule, from 10 to 15 per
cent of the total fibers are found changed. Schnyder (286) on the
contrary, finds only slight degeneration in birds and' believes,
because of the therapeutic influence of the vitamine, that the paresis
could not be the result of degenerative processes in the brain. We
have frequently pointed out that if we kill an animal a few days
after it has been cured with vitamine, and make a histological study
of the nerves, the degenerated fibers are still to be seen and persist
for a long time. In the meantime, the normal fibers appear to
assume the functions of the diseased fibers.

The nerves, histologically examined, give the following picture:
According to Kimura, the first indication of degeneration is to be
sought for in the axis cylinder, while the earlier prevalent idea was
that the signs of degeneration are first noted in the medullary sheath;
the myelin sheath, according to Kimura, may remain intact. How-
ever, as soon as the latter degenerates, the axis cylinder can no longer
be differentiated. The myelin fragments thereupon are resorbed
in situ through the cells of Schwann's sheath. If a degenerated
myelin fiber still contains an axis cylinder, it is a regenerated axis
cylinder from the ribbon-like protoplasm. The new cylinder is
smooth and fragile and resembles that seen after a trauma.

The same changes are also to be noted in the dorsal and ventral
nerve roots, as in all the dorsal strands of the spinal cord. Further,
changes are apparent in the cells of the forward and rear horns of the
lumbo-sacral cord; here the tigroid bodies are invisible, and the
stainable substance (Nissl's method) accumulates over the axis
cylinder. In some cases, the nuclei are weakly colored. Similar
pathological nerve studies were made by Weill and Mouriquand (287) ,
Kato and Shizume (288) and Paguchi (289).

The muscles. The muscles exhibit atrophy and fatty degeneration,
but the changes disappear rapidly on returning to normal nutrition.

The heart. In most cases, this is unchanged; only seldom is there
any edema, pigmentation, and traces of parenchymatous degenera-
tion. Hypertrophy of the right heart, as in human beriberi, does not
take place in chickens. On the contrary, we have personally seen

THE VITAMINES IN THE ANIMAL KINGDOM 105

hydropericardium in pigeons. McCarrison (290) attributes the
edema to the enlargement of the suprarenals.

Endocrine glands. The first investigations in this direction were
made by Funk and Douglas (291). The findings in pigeons showed
that the thymus glands disappear; this was also noted by R. R.
Williams and Crowell (292). The behaviour of this gland in birds
was then investigated further by McCarrison (293). Funk and
Douglas made a histological investigation of pituitary, adrenals,

FIG. 18. FATTY DEGENERATION OF SCIATIC NERVE IN CHICKEN BERIBERI

ovaiy, testicle, kidney, liver, pancreas and spleen. In all these
organs, definite signs of degeneration were noted. The investigation
of thyroid in greater detail was made by Douglas (294) who observed
carefully the great variations and the tendency of the colloids to
disappear from the vesicles. McCarrison (295) studied the influence
on the thyroid of food poor in vitamine, and found that the size of the
gland, as well as the weight, decreased. The study of the endocrine
glands of pigeons was then undertaken by the same worker (296)
who largely corroborated the work of Funk and Douglas, and found

106 THE VITAMINES

that the organs undergo atrophy in the following order: thymus,
testicle, spleen, ovary, pancreas, heart, liver, kidney, stomach,
thyroid and brain. McCarrison made the very interesting and
important observation that the suprarenals are often hypertrophied,7
and that this hypertrophy bore a causative relationship to the onset
of edema (I.e. 290). He believed that in unpolished rice, butter
(particularly in butter, as will be noted later) and onions, there was a
substance which was protective against edema, and which might be
thought of as vitamine A.

McCarrison found also that the brain, suprarenals and the pitui-
tary, are very sensitive to a lack of vitamine. The sexual organs
in males showed a disappearance of spermatogenesis, with resulting
sterility; in females, there was a condition similar to amenorrhea.
This is in opposition to the fact that we mated pigeons, cured of beri-
beri, and secured a completely normal progeny. It is nevertheless
possible that in these cases, there may have been a regeneration.

During the course of his fruitful investigations, McCarrison also
examined the muscles and found them highly atrophied, while the
central nervous system was only slightly so; he attributed the result-
ant paralytic symptoms directly to the impaired functional ability
of the nerve cells. Because of the remarkable atrophy of the thymus,
testicles, ovaries, and spleen, much more apparent than in other
organs, resulting from the lack of vitamines, he believed that these
organs provided the distressed organism with the needed vita-
mine. When this reserve is exhausted then the bones are the
purveyors of the vitamine, in which case the marrow undergoes
marked changes. Red corpuscles are also diminished about 25 per
cent. Finally, McCarrison regarded the whole picture of beriberi as
a syndrome, arising from, (1) a chronic inanition; (2) a pathological
change of the organs of digestion and assimilation; (3) an abnormal
functioning of the internal secretory glands, especially of the supra-
renals; and (4) poor nutrition of the central nervous system. In
addition to all these factors, there is also a decreased resistance to
the bacterial invasion. McCarrison (297) at first erroneously
attributed to the latter condition the entire picture of beriberi, but
he has long since abandoned this view. The pathological histology
of beriberi in pigeons is described by McCarrison (298) in a special
article.

7 Unfortunately, in the pathological department, which took up this
phase of the work, this observation completely escaped notice.

THE VITAMIXES IN THE ANIMAL KINGDOM

107

Changes in the gastro-intestinal tract. McCarrison's (299) investiga-
tion on 153 pigeons yielded some very interesting results. The
upper part of the intestine, which is muscular in nature in the region
of the pancreas, is mostly involved. Atrophy had proceeded to
such an extent that the walls were almost transparent, especially
when white rice was fed for a long time. .The intestine showed a

FIG. 19. THE UITER PART OF THE INTESTINE OF A HEALTHY PIGEONT

(McCARRISON)

definite congestion; ecchymosis was not infrequently seen and
microscopic hemorrhages were almost always observed. Under the
microscope, the congestion led to a partial disappearance of the
epithelium. The myenteron was so severely atrophied, that its
motor functions suffered. Further, there was described degeneration
of the mesenteric plexus of Auerbach, which impaired the functions.

108

THE V1TAMINES

of the intestinal nerves. The villi disappeared and there was atrophy
and inflammation of the mucosa and atrophy of the lymphatic tissue.
The latter condition is primarily responsible for the decreased
resistance to infection. According to McCarrison (300), the severe
intestinal infection is the chief reason why some animals cannot be

•

FIG. 20. SECTION OF UPPER PART OF INTESTINE FROM A PIGEON DYING OF

BERIBERI

Thinning of muscular coat, intense atrophy and necrosis of the mucous
membrane.

saved by an addition of vitamine. Pigeons fed on autoclaved rice,
butter and onions develop the same pathological picture, with the
difference that the changes are not so far advanced. After the
elimination of butter, it may be shown that onions have the power
to protect the intestinal canal against degeneration. By the
addition of peas, containing vitamine, the pigeons may be cured,

THE VITAMINES IN THE ANIMAL KINGDOM 109

while the pathological picture of the intestine remains the same. In
his opinion, the cure is so rapid that it cannot be due to the vitamine
absorption; the therapeutic effect may be compared, in its velocity,
only to the sudden manifestation of the growth impulse in germinat-
ing seeds.

Chemical pathology. The chemical investigations are not yet so
numerous. Schaumann (I.e. 2) investigated the brains of a beriberi-
pigeon and found that the phosphorus content was not decreased.
In contrast to this, we (301) found that the nitrogen and phosphorus
content of these brains is below the normal content; indeed in
normal pigeons, we found 9.77 per cent nitrogen and 1.84 per cent
phosphorus, while in beriberi pigeons, there was 9.31 per cent nitrogen
and 1.53 per cent phosphorus. Our results were confirmed byWle-
land (302) and byMathilde Koch and Voegtlin (303), although it is
not certain just how ordinary starvation would affect these figures.

Funk and v. Schonborn (304) also showed that in pigeon-beriberi,
there is an increase in blood sugar and a decrease in the glycogen
content of the liver almost to the point of complete disappearance.
Because of this, we were very much inclined to bring these facts
into causative relationship with the observation of McCarrison on
hypertrophy of the adrenals.

It must also be recorded that Fuji (305) found the diastase content
of the blood decreased in chickens suffering from beriberi ; on admin-
istration of vitamine, this value returned to normal. Analogous
results were obtained with glyoxalase of the liver by Findlay (305a).

MAMMALS

In 1913, we stated that certain animals do not develop beriberi
when they are put on a diet of white rice; we attempted to explain
this by its dependence on the specific purine metabolism of the
particular species. With this in view, we classified mammals into
two broad divisions, one in which the end product of purine metabo-
lism is uric acid, and the other in which it is allantoine. The first
division included man and birds, manifesting real beriberi, while the
second included the rat, the dog,8 the monkey — which can contract

8 Bearing upon this, it would be interesting to determine if a different
reaction to the lack of B-vitamine is manifested by Dalmatian dogs, which,
according to S. R. Benedict (307), excrete uric acid chiefly instead of allaijtr
toine, and by anthropoid apes, found by Hunter and Ward (308) to have :a
purine metabolism similar to that of man.

110 . THE VITAMINES

scurvy — while guinea pigs with their characteristic purine metabo-
lism occupy a middle position. Whether these conceptions may
still be regarded as correct is not certain, for in the first place, a
type of beriberi in dogs and rats has been described and secondly,
there is a possibility of another explanation of the above mentioned
observations. At all events, we wish to leave the question open for
further investigation, particularly since beriberi has not been defi-
nitely demonstrated in certain animals, and may even be confused
with other pathological conditions. The criteria for the recognition
of beriberi, such as the fatty degeneration of the nerves, are not
specific symptoms of beriberi. We have fed white rice for along
time to rats, rabbits and guinea pigs, without noticing any kind of a
symptom to suggest beriberi. The other explanation, of the differ-
ent behavior of certain animals on a diet of white rice, we find in
the unknown role of the bacterial intestinal flora, the significance of
which has already been mentioned (p. 76) in the chapter, "Life
Without Bacteria." We come now to the consideration of the impor-
tance of the vitamines for various mammals.

Rats

Rats have been extensively used as experimental animals in the
last 10 years in the attempt to solve the problems of nutrition, and
for vitamine research. The reason for this is that rats breed easily,
do not consume much food and are not particular in their taste. The
fact that a greater number of these animals may be used at one time
for an investigation makes the results much more certain. Since in
most problems of nutrition, the food must be specially prepared, as
a rule, the small amount ingested is of practical significance. Rats
live about 3 years and reach sexual maturity very early. The
pregnancy lasts 3 weeks and the offspring is numerous; the number
of young varies from 6 to 12 according to the age of the parents and
their nutritive condition. The young rats begin to eat of their own
accord after 3 to 4 weeks, and may be used for feeding experiments
when they weigh 30 grams, though it is better to wait till they
weigh from 40 to 60 grams. It has been shown by many investiga-
tors, among them McCollum, Simmonds and Pitz (309), that inade-
quacies of the diet are manifested in a diminished milk secretion of
the mother, or else in a poor state of health of the young, although
the parents show no such signs. It is therefore desirable, if one

THE VITAMINES IN THE ANIMAL KINGDOM

111

wishes to demonstrate the nutritive value of a certain diet to carry
out feeding experiments with the same material on the next genera-
tion. For such a purpose, the rat is naturally specially suited.
Thus, it is possible to study, in a short time, not only the effect of
the diet upon the parents, but also to extend the observations on the
procreative power, the number of living young, and the lactation of
the mother. Judging by our experience, we deem it inadvisable to

FIG. 21. RAT CAGE (MACALLUM)

6, water cup; 7, flask for urine collection; 8, bulb by means of which urine,
is separated from feces (urine flows on the outside of the bulb into the flask
and feces falls into beaker) ; 9, food cups of various sizes; 10, funnels of various
sizes adapted to the age of the animal, so that the food may not be scattered.

buy rats for experimental purposes; it is far better to raise them in
the laboratory. In this way we have the advantage of knowing the
parentage of the animals to be used. When one obtains a litter, it is
sometimes difficult to differentiate between the male and female.
To obviate this, we can utilize a method described by Jackson (310).
It is likewise of importance to use a cage so arranged as to be able to
control accurately the amount of food given, especially since rats

112 THE VITAMINES

have a tendency to scatter their food. This difficulty may be over-
come if we use such a cage as has been described explicitly by our
former co-worker, A. B. Macallum (311). The cage is equipped
with funnels of various sizes through which the rat may reach its
food without scattering it. The cage is arranged for two rats and
permits of separating the urine from the feces, so that an exact metabo-
lism experiment may be carried out.9 Great care is necessary in the
preparation of the diet; above all. it is important that the basal
diet is vitamine-free. This precaution was not sufficiently observed
in the older investigations, thus giving rise to differences of opinion.
We need only quote one example, lactose, widely used as a constituent
of an artificial diet. Regarding this, we showed10 that lactose, which
does not undergo any particular purification process, contains traces
of nitrogen. These traces may come from the milk and permit of
the growth of rats, because of its vitamine content. This was
demonstrated experimentally by McCollum and Davis (I.e. 97) and
also by Drummond (313). This is likewise true of the protein of
protein-free milk, as we have already seen. It is necessary to subject
the protein to be used to a purification process before it is fed. In
most feeding experiments casein is used, and it is rendered vitamine-
free by extracting with alcohol a number of times, according to the
procedure of Funk and Macallum (I.e. 81); we found that this
procedure did not decrease the nutritve value of casein, in spite of
statements by McCollum and Davis (I.e. 97) to the contrary. It is,
of course, possible to purify the casein by dissolving it in alkali and
precipitating with acid, and also by continued washing with water.
In using meat, according to Osborne, Wakeman and Ferry (I.e. 102),
it suffices to extract it repeatedly with water. From edestin, as we
have seen, this procedure removes the vitamine only with great
difficulty. Of the other commonly used dietary constituents, cane
sugar is vitamine-free, as are also most of the varieties of starch.
Regarding fat, we have used ordinary lard and under certain con-
ditions it can be entirely dispensed with. The purification of lard is
made in our laboratory by autoclaving at 30 pounds for 3 hours; we
shall have occasion to show later that this lard is then vitamine-free.
For the entire dietary mixture, about 3 per cent of agar is added to
diminish the possibility of constipation. If it is desired to use a sub-

' Ferry (312) describes the technique of rat feeding experiments.
10 The Vitamines, 1st ed., p. 159.

THE VITAMINES IN THE ANIMAL KINGDOM 113

stance hitherto not utilized, it is necessary, by means of a specific
control, to test for the presence of vitamines. For an inorganic salt
mixture, that described by Osborne and Mendel, somewhat like the
inorganic salts of milk, or the one used by McCollum and Simmonds
is available. The mixtures have the following composition :

Osborne and Mendel (314) McCollum and Simmonds (315)

CaCO3 134.8 NaCl 0.173

Na2CO3 34.2 NaH2PO4.H2O 0.347

H3PO4 103.2 K2HPO<. 0.954

H2SO4 9.2 CaH4(PO4)2.H2O 0.540

Citric acid • H2O 111.1 Iron citrate 0. 118

MgCO3 24.2 Calcium lactate 1.390-1.300

K2C03 141.3 MgSO4 (anhydrous) 0. 266

HC1 53.4

Iron citrate • 1.5H2O 6.34

KI 0.02

NaF 0.248

K2A12(SO4)2 0.0245

The food mixture is best prepared so that it has more or less of a
pasty consistency, in which form it is least scattered by the rats.
As an example of a diet now thought to be complete, one having the
following composition is given :

Substance Per cent Substance Percent

Casein 22 Lard 20

Cane sugar 10 Agar 3

Starch 27 Salt mixture 3

Butter 10 Yeast extract 6

In older investigations, for example, in the work of Hopkins in
1912 (I.e. 70), or even in that of Funk and Macallum (I.e. 8.6), not
only was the amount of food consumed taken note of, but it was
estimated how much of the food intake was assimilated, by means
of calorimetric determinations of the food and of the excreta. In
the newer investigations, the tendency has been to get away from the
above time-consuming procedure, and to dispense with records of the
quantity of food ingested. The rat has become, so to speak, a
reagent for the testing of the value of the diet, and since many
thousands of experiments have been made in the last 10 years, the
investigators sought to simplify the experimental conditions as much
as possible. Nevertheless, the weighing of the food seems to us to

114 THE VITAMINES

be of the greatest significance, particularly in those experiments
yielding negative results. It does not suffice merely to feed the
animal; one must also be certain that it eats enough. If no food
control is provided for, it is easy to make false conclusions; it would
be possible to designate a certain diet as insufficient, whereas the
true reason for the misleading experiments lies in the unsuitability
of the animal. This applies in general to rats that are bought; it
may be worth while mentioning that we have had particularly good
results with black and white rats, raised in the laboratory; this strain
seems to be unusually resistant.

It must be emphasized once more that not all problems of nutrition
can be solved by the use of rats as experimental animals. To illus-
trate, Osborne and Mendel (316) have shown that when the growth
of rats is prevented by a specially chosen diet, growth may be resumed
after they have reached two-thirds of their size, upon the addition
of the missing factor. While Jackson and Stewart (317), in con-
firmation of these findings, concluded that this ability to resume
growth, after a period of repression, depends upon the age of the
animals and the duration of the growth inhibition (in agreement with
the work of Briining (318) and Aron (319) ), and under certain cir-
cumstances may be appreciably affected, we must nevertheless admit
that the rat does possess this ability for the most part. Now, it
would be futile to try to achieve the same results in man or other
mammals of a corresponding age. As a matter of fact, various kinds
of animals behave quite differently. In the same way, it is evidently
impossible to study the nature of scurvy and pellagra in these
animals as some investigators have done. The individual animals do
not compare in regard to nutrition and nutritional diseases.

An interesting question is involved in experiments with inadequate
diets to show how far rats have the power to choose out of two diets,
the one that is adequate. This was investigated by Slonacker (320)
with a primitive method, and repeated later by Osborne and Mendel
(321). In the latter experiments, it could be shown that the rats
chose correctly almost invariably.

The vitamine requirements of rats. Although the individual phases
of this question have been briefly considered in the historical part and
other sections of the book, it appears necessary to treat systematically
this matter which has assumed such practical as well as theoretical
importance. We have already said (even though the question should

THE VITAMINES IN THE ANIMAL KINGDOM 115

not belong to the subject of vitamines) that for the correct feeding
of rats, it is necessary, above all, that the food should have the proper
composition. We know that a protein is necessary whose content
of amino acids is above reproach. We may say that casein, lactal-
bumin,11 egg-albumin, muscle protein, as well as some plant proteins
have been shown to be adequate for this type of animal. Suzuki
and his co-workers (323) demonstrated that an artificial mixture of
amino acids does not permit of growth in the rat, while digested
meat (erepton) possesses this ability because of its vitamine content.
Aside from this, it may be connected with a more favorable relation-
ship of the amino acids to each other. In addition, we refer to the
excellent work of Osborne and Mendel, who have carefully studied
the value of various proteins on rats.

Furthermore, it is necessary that the rats receive a sufficient and
properly composed salt mixture, and that the individual constituents
of the diet are present in proper proportion. This point, to be
mentioned again shortly, is of great significance. It cannot be said
with certainty that these basic conditions are always accurately
maintained; it is only in the work of the last six years that they have
been given their true value. All of the above mentioned factors must
be taken into account before undertaking a vitamine problem.

We have already called attention to the work of Osborne and
Mendel in 1911-1912, in which they still clung to the idea that they
had solved the question of artificial nutrition. At first they did not
see that the addition of "protein-free milk" to the diet was an
unknown factor whose significance for nutrition could not be accu-
rately gauged at the time. This happened in spite of the work of
Stepp, who had already affirmed the presence of essential lipoids in
milk. Through the classical work of Hopkins and through our (324)
simultaneous demonstration of vitamine B in milk, it became clear
that something could adhere to the products made from milk; this
might explain the striking results obtained by Osborne and Mendel.
Hopkins showed that a small quantity of milk (a few cc.),

11 Sure (322) has recently found that lactalbumin is not a complete protein
since it does not contain enough cystine and tyrosine. According to this,
it is obvious that the results of Emmett and Luros (I.e. 105), who suggested
the probability of a new vitamine, and who found that lactalbumin was
rendered complete when supplemented with protein-free-milk, could be
explained by the presence of cystine and tyrosine in the protein-free milk.

116

THE VITAMINES

added to a carefully purified food mixture, resulted in normal growth
of rats; at the same time the food intake increased markedly. The
favorable effect of these small amounts of milk is best shown in the
accompanying curve. Since Hopkins fed milk in the natural state,
its effect could be due to the cumulative influence of several unknown
factors — which was found later to be the case. Subsequently, Funk
(325), and also Osborne and Mendel (326), showed that milk does

0 2.5- 50

FIG. 22. GROWTH CURVES (HOPKINS)

Lower curve (up to eighteenth day), vitamine-free diet; then 3 cc. milk
daily; upper curve, the reverse.

not exert such a great influence as Hopkins first found; however, we
must not forget that the vitamine content of milk and other food
stuffs does not represent a constant mathematical unit and can under-
go great variations.

McCollum and Davis (I.e. 75), and shortly thereafter also Osborne
and Mendel (I.e. 76), showed that protein-free milk was not sufficient
for the growth of rats and that the diet needed certain fatty con-
stituents, found in- egg-yolk and in butter. The lack of these factors

THE VITAMINES IN THE ANIMAL KINGDOM 117

was evidenced by a general undernutrition, a rough coat and a
characteristic eye disease, now called ophthalmia (keratomalacia).
Since this disease does not occur only in rats, we shall have
occasion to speak of it again. It was difficult to bring these new
findings into accord with the then prevailing knowledge of vitamines,
the more so since Osborne and Mendel (I.e. 72) obtained good growth
with fat-free mixtures. Later on, Stepp (327) showed that his
"life-sustaining" substances did not belong to the lipoids, and Lander
(328) demonstrated that lecithin and cholesterol played no particular
part in nutrition. Hence we could not understand why the role of
vitamine B was relegated to the background, and even denied.
Subsequently, Aron (329) also reported on the importance of butter.12
Although we had already at that time assumed the existence of a
special vitamine, associated with fats (in cod liver oil, we assumed
the existence of a specific antirachitic vitamine because of the partial
lack of vitamine B and the therapeutic influence on rickets) we were
somewhat skeptical of the existence of a vitamine in butter. In this
we were strengthened by our experiments together with Macallum,
particularly since butter was shown to be free from B-vitamine and
since it was possible (330), by substituting fresh yeast for dry yeast,
adding orange juice, and treating the eyes with zinc sulfate and boric
acid solution, to maintain animals in good condition for 150 days.
The substitution of butter for lard was without influence in our
experiments. All these results are now easily explainable, and yet
we must assert that we have observed instances of ophthalmia on a
diet containing butter.13 Our findings showed that the substance
present in butter was not the only growth factor, asMcCollum would
have it, but that vitamine B is at least quite as important as vita-
mine A. This is well illustrated in the curve taken from the work of
Funk and Macallum.

12 Aron (I.e. 319) introduced the method of giving the vitamines as a medicine,
so to speak, independent of the diet. This procedure has the advantage
that the vitamine addition has nothing to do with the food consumption,
although it is not always easy to give the necessary dose to the animals, as
we ourselves can verify.

13 Lately, in association with Dubin (331), we have noted, among 30 rats,
2 that developed ophthalmia on a diet containing the usual amount of
vitamine A. One of them was improved by the addition of yeast to the diet.
The other improved without medication.

118

THE VITAMINES

Effect on rats of the lack of vitamine B. If a diet is chosen, complete
in everything else but lacking in vitamines A and B, a nitrogen
balance can be maintained for 24 days, according to the findings of
Desgrez and Bierry (332). If the experiments are extended for a
longer time, we obtain the following curve, reproduced in the Report

CMS

100

80

60

40-

20

I

10 20 <30 40 DAYS

FIG. 23. SIGNIFICANCE OP VITAMINE B FOR GROWTH OF RATS

Vitamine A was present as butter; addition of yeast indicated by * (Funk-
Macallum).

of the English Medical Research Committee (333) on vitamines. It
may be seen that growth ceases completely if both vitamines are
lacking, and the animals would have died if these factors had not
been administered in time. The addition of vitamine A only had no
influence on the growth, while the addition of vitamine B resulted in
slight growth, which, however, stopped again after a short time.

THE VITAMINES IN THE ANIMAL KINGDOM

119

The difference in the mode of action of both vitamines was explained
by the supposition that there is only a small reserve supply of the
antiberiberi vitamine in the body, and hence a lack of it is almost
immediately noticeable; on the other hand, the reserve of antirachitic
vitamine was thought to be greater. This observation may naturally
be explained in another way, namely, that vitamine B is of greater
significance in metabolic processes and growth, and hence larger
amounts are necessary. In accordance with this, is the fact that
grown rats require less vitamine A than young rats, whereas vitamine
B is necessary throughout the course of life. When there is a lack
of vitamine B, symptoms arise which have been likened to those of
beriberi by Funk and Macallum. Be that as it may, these symptoms

<cuu
150
IOO

50
FIG. I

/

™

>

/

<?/

' .

••

>

r

£

'•' -

-7

x

f

t

/

i

"*^

7

M. GROWTH OF RATS SHOWING THE DEPENDENCE ON VITAMINES A ANI
B. (REP. MED. RES. COM.)

Diet without vitamine B
Complete diet

Diet without vitamines A and B

x x x x Diet without vitamine B

may be obviated just as in avian beriberi. Whether it really is
beriberi remains to be demonstrated. A paresis of the legs is no
rare occurrence in these rats. Voegtlin and Lake (334) observed
that rats are much more resistant to beriberi than are cats and dogs.
Since beriberi is not an infectious disease, we cannot speak of a
resistance ; rather must we say that the lack of vitamine B in rats is
not followed by characteristic symptoms. It is interesting to follow
the influence of an addition of vitamine B upon the food intake, as is
shown in an experiment by Osborne and Mendel (335). Osborne is
of the opinion that a daily addition of 0.2 gram yeast is quite suffi-
cient to provide for the vitamine B requirement of rats. Osborne
and Mendel (336) rightly maintain that the effect of vitamine B

120

THE VITAMINES

^oo

150
100
50

cT

Z

Fic.^25. SHOWING FAILURE TO GROW ON A DIET DEFICIENT IN VITAMINE
B, AND RECOVERY ON ADDING THAT FACTOR (REP. MED. RES. COM.)

Diet without vitamine B

Complete diet

140

FIG. 26. INFLUENCE OF VITAMINE B ON THE FOOD-INTAKE (OSBORNE)

THE VITAMINES IN THE ANIMAL KINGDOM

121

•extends to the entire metabolism, and that the specific influence does
not depend upon the food intake.

The effect of the lack of vitamine B on rats was studied by Drum-
mond (337). Among other observations, creatinuria was noted; this
was connected with the cleavage of muscle tissue. Grown animals
often exhibited subnormal temperatures, while paralytic symptoms
were rarely noted. The right heart in some cases was enlarged and
the testicles showed a suppression of spermatogenesis.14 Emmett and
Allen (339) investigated the organs and glands of rats kept on a diet
poor in vitamine B. They noted atrophy of the thymus and hyper-

,2. 50

200

150

1

JO
0

(VDRMAL
itoWTtf "^

MONTH

50

T?IQ. 27. SHOWING PRELIMINARY GROWTH AND EVENTUAL DECLINE ON DIET
DEFICIENT IN VITAMINE A

Also recovery on addition of a source of vitamine A. In the small diagram,
the lower curve shows growth of breast-fed young when the mother is fed on
a diet deficient in A-vitamine; the upper curve shows the same when the
mother is fed on a complete artificial diet. (Rep. Med. Res. Com.)

trophy of the adrenals, similar to the findings in birds. Liver, heart
and other organs, particularly certain intestinal segments, were found
inflamed and infiltrated. Frequently, a fatty degeneration of some
organs was noted. Lack of vitamine A, however, led to no patholog-
ical changes, at any rate not of such a nature as were easily demon-
strated. In this case, the symptoms were least apparent in older

14 Osborne and Mendel (338) held that yeast as the sole source of proteins
causes sterility in rats, but it still remains to corroborate this finding. These
investigators confirmed our statement (I.e. 325) that for rats, yeast protein
can completely replace the protein of the diet.

122 THE VITAMINES

animals. The animals showed a coarse and sparse coat, and a-
decreased resistance toward infections of all kinds, such as ophthal-
mia, and pulmonary affections, like pneumonia. Tsuli (339a)
investigated the action of deficient diets on thyroid, gonads, parotid,
pancreas and other organs. Aron (340) observed these conditions,
including neuro-paralytic changes in the skin, to which he also
attributed ophthalmia. Morgulis and Gies (340a) found the calcium
content of rachitic bones and teeth lower than in the normal. Mattill
(340b) compared the creatine, creatinine, and urea content of the
blood in fasting rats and those deprived of B-vitamine. The non-
protein N was increased in both cases — in fasting animals, mostly in
the form of urea, while in the others, it was in the form of creatine.
Briining (340c) saw in rats fed on a carbohydrate-rich diet a patho-
logical condition arise resembling "Mehlnarhrschaden."

A question that has greatly interested the author for a long time,
was to determine if with both of the above mentioned vitamines, all
of the nutritive requirements of the rat have been fulfilled. Looking
through the many reports dealing with this problem, one is tempted
to answer this question in the affirmative. First of all, we must see
whether the growth that has been obtained till now on artificial
diets, represents the optimum growth of rats. It is obvious from
most of the growth curves that growth on the experimental diet
compares favorably with that on a normal diet. To satisfy ourselves
on this point, we (I.e. 325) followed the growth of a number of rats
on a normal diet, which contained, among other things, yeast and
condensed milk. The experiments showed that the growth, regarded
by Osborne and Mendel and also McCollum as normal, was not the
optimum for this type of animal. For comparison, we append the
normal weight curves obtained by us, Osborne and Mendel, and
McCollum. It should also be noted that in a large percentage of
these investigations, particularly those of McCollum and his co-
workers, still more complicated food complexes were fed, which, at
least theoretically, might still contain one or more unknown factors.
On the other hand, we must admit that in laboratories greatly experi-
enced in rat nutrition, for example, those of Osborne and Mendel,
the condition of the artificially fed rats, in relation to the state of
health and the ability to rear their young, leaves nothing to be
desired. The explanation here may be that through considerable
experience a dietary composition has been chosen, the components

THE VITAMINES IN THE ANIMAL KINGDOM

123

of which by chance may supplement each other. In our earlier work,
we believed at first that we could conclude from the symptoms (dis-
position to bleeding, petechiae and rosary) and from the favorable
influence of orange juice, that we were dealing with an antiscorbutic
vitamine deficiency. Macallum (341) later adopted the same view.
Recently, there appeared the papers of Harden and Zilva (I.e. 100) and
Drummond (I.e. 79) completely corroborating the above observa-
tions. Still, some doubt has lately arisen, since Osborne and Mendel

210

180

150

err 120

o

90

60

FUNK

/

OSE

MENDEL

)RNE

DAYS

MCCOLLUM

30

FIG. 28. COMPARISON OF NORMAL RAT GROWTH CURVES OF OSBORNE AND-
MENDEL, McCoLLUM, AND FUNK

(I.e. 92) as well as Byfield, Daniels and Loughlin (I.e. 90), showed that
vitamine B is found in appreciable quantities together with vitamine
C in fruit juices. This is in agreement with our latest observations,
noted in our work with Dubin, that rats on a diet containing sufficient
vitamine nevertheless show improvement upon the addition of extra
vitamine. Besides, in our earlier and later work we have often found
that an addition of freshly prepared dietary mixture is followed by
marked stimulation of growth.

124

THE VITAMINES

Reviewing the many publications dealing with the growth of rats,
it is evident that only strictly positive or strictly negative results are
recorded. On the other hand, we have often seen rats that were
not in the best of health after 90 to 100 days on a diet that could be
regarded as complete in every respect. To give a concrete example-

I9Q

170

150

130

C5

90

70

NOfl

MAL

*!DAYS:

NUTf ITIOM

face

so

TIG. 29. RETARDED GROWTH SOMETIMES NOTED IN RATS ON A SUPPOSEDLY

ADEQUATE DIET, COMPARED WITH THE NORMAL

CURVE (FUNK-DUBIN).

we have only to record a diet used in the work of Funk and Dubin
(I.e. 331):

• Substance Percent Substance Percent

Extracted meat ... 22 Lard 20

Cane sugar 15 Starch 22

Butter ....10 Agar 3

Substance Percent

Autolyzed yeast ... 4

Orange juice 3

Salts.. ..3

Despite the fact that this composition is above reproach, in the
light of present-day knowledge of nutrition, our animals did not
grow very well. Thereupon, we tried to bring about normal growth

THE VITAMINES IN THE ANIMAL KINGDOM 125

by the addition of vitamines A and B, tomato juice, small quantities
of milk, and by the substitution of casein for meat. Although some
of these modifications were effective for a short time, the growth
curve was nevertheless flat. That the animals were still able to
grow, was demonstrated by placing them on the usual normal diet.
Hopkins (341a) recently also reported that rats failed to grow on ade-
quate food mixtures, and he ascribes this to the seasonal variation
of the growth impulse. Osborne and Mendel (I.e. 336) also showed
that it is not so easy to raise rats with an addition of yeast as with
the addition of protein-free milk; it was particularly noticeable that
there was a difference in the ability of the rats to raise their young.
Sugiura and Benedict (I.e. 108) obtained similar results, as did also
Freise (342), who ascribed a particular r61e to milk.

In collaboration with Dubin we (342a) have recently demonstrated
that what has up to now been known as vitamine B does not appear
to be sufficient for the growth of rats. Thus, it seems that autolyzed
yeast contains besides vitamine B something else of a vitamine nature
without which the rat does not grow. This will be discussed more in
detail in the chapter on the "Chemistry of Vitamine B." Judging
from the data recorded here, it appears desirable to study the nutri-
tive requirements of rats still further.

Mice

As we have already seen, the pioneer work of Stepp, Rohmann,
Lunin, Socin and others on artificial feeding was done with mice.
Getting away from the older work, which has more of a historical
interest, and coming to the modern investigations, we see in the work
of Brailsford Robertson (343), statistical data in relation to growth
of mice, which could be useful in judging the effect of experimental
diets. Thompson and Mendel (344) studied the food requirements
necessary for the growth and maintenance of the mouse, the suppres-
sion of growth, shown by undernourishment, and the amount of food
necessary to overcome this suppression. Mendel and Judson (345)
working with mice analyzed the total dry substance, fat and ash of
various dietary constituents. M. B. Schmidt (346) made the inter-
esting observation that if a deficient diet has no effect on the first
generation in these animals; its results are apparent in subsequent
generations. Morpurgo and Satta (347) worked with mice in
parabiosis in which one animal received only cane sugar while the

126 THE VITAMINES

other was given a complete diet. Both mice were kept alive, showing
that the one animal received with blood, not only the usual dietary
constituents, but also the vitamines.

Of the important investigations on mice, we shall first mention
those of Stepp, upon whose work of 1909-1911 we have already
touched lightly. He extracted the whole diet, thereby partly remov-
ing and partly destroying the vitamines. In this way, he obtained
mixed avitaminoses that could be attributed to the lack of at least
two vitamines. He proved (348) that lecithin, cholesterol, kephalin
cerebron and phytin could not replace the substances extracted by
alcohol-ether mixture. He found also that while the important sub-
stances could not be removed by means of the ether extraction alone ;
they could be removed with alcohol alone. When egg-yolk is
extracted with alcohol, these substances go into solution, but not
when acetone is used. Following this, Rohl (349) found that ordinary
commercial lecithin contained these substances, while pure lecithin
did not. In a publication appearing in 1914, Stepp (350) drew close
to the prevailing view of the importance of two vitamines in the life
of mice although he still thought of the fat vitamine as a lipoid.
In 1916, Stepp (351) advanced a step further in that he used two
different extracts, one of egg-yolk and the other of rice polishings
(orypan). However, the experiments in this case were not very
clear, since at first Stepp worked with a vitamine mixture (egg-yolk),
believing he had to deal with a lipoid. Furthermore, he showed (352)
that a mixture of lecithin, kephalin, cerebron, cholesterol and orypan,
added to dog biscuit which had been extracted with alcohol, could
at least maintain mice at a certain level. It is, however, not impos-
sible that this was due to a cumulative action of vitamine A con-
tamination. Dezani (353) conducted experiments for the purpose
of determining the source of cholesterol. He fed mice on flour and
casein, extracted with alcohol and ether. The animals died after
18 to 19 days, showing a 41 per cent loss in weight. Cholesterol
was found in the feces, and Dezani (354) was of the opinion that mice
could be kept alive on a lipoid-free diet. We may mention here one
of the typical experiments of Rohmann (355), in which mice were
raised on a diet of casein, chicken protein, nucleoprotein from liver,
potato starch, margarine and a salt mixture. This food proved to
be inadequate and the animals grew less than on milk and rolls. The
substitution of vitellin for nucleoprotein did not yield any better

THE VITAMINES IN THE ANIMAL KINGDOM 127

results, and Rohmann regarded this diet as adequate only for grown
animals. Meat extract and dried meat powder had no particularly
favorable effect; egg-yolk on the contrary, was favorable but not
sufficient for growth, while egg-fat was not well tolerated, even in
older animals. In a second series of experiments, the food was
supplemented with yeast and baked, with the addition of egg-yolk.
The results were veiy good and were attributed to the yeast ; lecithin
was shown to be unnecessary. It may be seen that Rohmann's
findings coincide with those on rats. Wheeler (356) carried out some
experiments with mice, planned in accord with the first experiments
of Osborne and Mendel on rats. Here, too, protein-free milk was
used, and found insufficient as the sole source of vitamine; artificial
protein-free milk proved to be entirely inadequate. It seems that
mice need more protein than do rats in order to grow; yet the former
grow more rapidly after the addition of a milk preparation than do
the latter. MacArthur and Luckett (357) investigated the influence
of lecithin, kephalin, cerebrosides and cholesterol as additions to an
artificially prepared diet, and came to the conclusion that these
substances are superfluous. Furthermore, they studied the influence
of egg-yolk and found that the necessary substances (apparently
vitamines, as they remarked at the time) are insoluble in ether,
soluble in cold alcohol and thermo-stable. Butter and olive oil were
inactive, without any other additions. Mitchell and Nelson (358)
found that mice do not grow on an artificial diet till protein-free milk
was added. From the foregoing, it is evident that mice behave like
rats in their vitamine requirements.

Guinea pigs

Next to the pigeon and the rat, the guinea pig is one of the most
widely used animals for the purpose of vitamine research, especially
in the investigation of scurvy. It has been shown unsuited for other
avitaminoses, though there are no lack of investigators who propose
the use of the guinea pig for the study of pellagra and rickets, despite
its demonstrated unsuitability. This animal is likewise little used
for beriberi research although Schaumann (I.e. 2) reported, out of
65 guinea pigs fed on dry grain, two typical cases of beriberi, with
subsequent histological demonstrations of multiple neuritis. These
results have never been confirmed. On the whole, it is not easy
to conduct planned feeding experiments with these animals, and

128 THE VITAMINES

hence the difficulty of studying problems of nutrition. We know
comparatively little of their vitamine requirements, particularly as
to vitamines A and B, since in the study of experimental scurvy,
dietary mixtures are used containing both of these vitamines. Hume
(358a) has recently found that guinea pigs require vitamine A and
hence these animals can be used for the detection of vitamine A in
green foods but not in fats. Tozer (358b) , in a pathological investi-
gation of these animals could not find any difference between experi-
mental scurvy and the changes produced by lack of vitamine A.

FIG. 30. LONGITUDINAL SECTION OF A RIB OF A SCORBUTIC GUINEA PIG^

WITH FISSURES, HEMORRHAGES IN THE PERIOSTEUM AND PALE

MARROW (HOLST-FROLICH)

We owe the discovery of experimental scurvy to Hoist and Frolich
(359) in 1907, although in 1895 a similar condition was observed in
the same species by Theobald Smith (360). Hoist and Frolich
investigated scurvy in various animals and chose the guinea pig as
most suitable for this purpose. It appeared to be unnecessary to
choose a special diet, for these investigators noted that scurvy could
be produced in guinea pigs with any diet that would do the same
in man; this was also the case with any diet that was autoclavedat
a high temperature. They recognized, with great perspicacity, that
the symptoms were closely analogous to those in man. Guinea pigs

THE VITAMINES IN THE ANIMAL KINGDOM

129

fed with rye, or wheat bread (and water), or with oats, rye, wheat,
barley, and rice flour, died within a few weeks. Hoist and Frolich
used animals weighing about 350 grams since they could better with-
stand the great loss in weight (about 40 per cent) . In a later publica-
tion, the same investigators (359) came to the conclusion that human,
as well as guinea pig, scurvy may be attributed to the same causes;
they sought also to differentiate between experimental scurvy and
simple inanition. Starvation, in which there is a loss of from 30 to
40 per cent in weight, yields no scorbutic symptoms; on the contrary,
scorbutic symptoms ordinarily appear before the
animals show any appreciable emaciation.

Not all workers accept the simple etiological ex-
planation of scurvy in guinea pigs. Jackson and
Moore (361) and Jackson and Moody (362) be-
lieved that there was an infectious factor in the
etiology. Moore and Jackson (363) strengthened
their belief after they had seen that raw milk
could not protect these animals from scurvy.
The explanation for this is to be sought in the
probability that the milk used did not contain FIG. 31. SCORBUTIC^E
enough vitamine C for this type animal. In con-
trast to the above investigators, Givens and Hoff-
man (364) were able to demonstrate the absolute
sterility of the blood of the diseased animals.
Further, McCollum and Pitz (I.e. 69) believed that
scurvy in guinea pigs was due to constipation, and made the
sweeping statement that scurvy, as an avitaminosis, was non-existent,
and that the known antiscorbutic, properties of fruit juices could be
explained by their laxative action. They stated that other laxatives,
like phenolphthalein and mineral oil, could be used with the same
success. These statements were further developed by Pitz (365) a
co-worker of McCollum's, who found that lactose acted favorably,
and that its function was to modify the bacterial intestinal flora.
The observations of McCollum and his co-workers immediately
aroused a storm of protest.15 Shortly thereafter, Harden and Zilva
(367) showed beyond a doubt that the results of McCollum were
due to the fact that he gave the guinea pigs milk, ad libitum, and

18 Torrey and Hess (366) have investigated the bacterial intestinal flora
in guinea pigs and in infants, but failed to find any abnormalities.

MUROF A GUINEA PIG,

WITH PALE MARROW

AND. ATROPHIC

TRABECULAE
(HOLST-FROLICH)

130 THE VITAMINES

in this manner prevented the onset of scurvy. When milk was
omitted, lactose had not the least influence upon the progress of
the disease. Later, these observations were confirmed by many
others, and McCollum himself abandoned his view.

When we (368) spoke with doubt as to the identity of scurvy in
guinea pigs and in man, it was based upon failure to obtain ther-
apeutic results with milk, lime juice, potato and lemon juice. All of
the difficulties mentioned by us at the time have been overcome by
countless investigators, and at present there is no reason to doubt
that in guinea pigs, we are actually dealing with scurvy.

As we have already said, the vitamine requirements of the guinea
pig, aside from vitamine C, have been insufficiently investigated.
Hausermann (I.e. 13) extended his experiments on the influence of
iron salts on guinea pigs, without noting any results. Heim (369)
found that guinea pigs could not live on raw or cooked cow's milk.
Of eight animals, one died after a month, two after 2 months, and the
other five remained alive for 3 months (the whole experimental
period), but showed loss of weight. In another series of experiments,
the animals died in from 4 to 14 days; the addition of cellulose was
without effect, while malt extract and alcoholic extract of germinated
barley exerted a definite favorable influence. Rondoni andMontag-
nani (370) conducted similar experiments with corn and oats, and
with complete starvation. Guinea pigs on corn showed milder
scurvy symptoms than those on oats. Of particular interest, is the
work of Ingier (371) who investigated the influence of diet on
pregnant guinea pigs. Here, too, a diet of oats and water showed
scorbutic influence on the embryo. When this diet was used in the
later stages of pregnancy, no pathological changes were found in the
bones. A short feeding with the milk of the scorbutic mother, con-
verted the latent scurvy into the active form. In pregnant animals,
the disease developed more rapidly, with more marked symptoms,
ihan in those that were not pregnant, and in the first stages of preg-
nancy lead to death. Ingier described a fatty degeneration of the
nerves in these guinea pigs.

Conditions for the demonstration of experimental scurvy. — The
foodstuff mostly used for the development of this disease is oats.
According to the investigations of McCollum, Simmonds and Pitz
(372), oats are poor in adequate protein, salts and vitamine A. It is
undoubtedly true that under the working conditions maintained by

THE VITAMINES IN THE ANIMAL KINGDOM 131

Hoist, the guinea pigs soon lost their appetite and died, presenting
mixed symptoms of scurvy and starvation. Therefore, it signified a
step forward when, according to the procedure of Chick and Hume
(373), the animals were given an addition of sterilized milk. This
addition did not delay the development of the disease — the general
condition of the animals being markedly improved — and corre-
sponded with the view advanced by McCollum and his school as to
the nutritive value of oats. The demonstration of experimental
scurvy was no longer difficult, especially after the introduction of the
milk addition, and investigations on this subject were easily carried
out. For this purpose, Cohen and Mendel (374) used animals
weighing from 110 to 250 grams. They believed that cod liver oil
had no influence on scurvy, so. that a complication of rickets was
no cause for concern. Scurvy can be produced by feeding soya bean
flour, which contains enough of vitamines A and B, supplemented
with cellulose and salts. Hess and Unger (375) used oats, hay and
water (to this may also be added cod liver oil) as a basal diet and
found that guinea pigs weighing from 200 to 300 grams developed
scurvy in two to three weeks.

In the Lister Institute in London, where many investigations on
guinea pigs are being made, the original method of Hoist and Frolich
was modified so that the diet consisted of oats, bran and 60 cc. jof
milk for guinea pigs weighing 350 grams. The milk is autoclaved
for one hour at 120°C.; on this diet the animals grow normally for
about 15 to 20 days, after which the first symptoms of scurvy are
noticed. The animals lose weight, and after 30 to 40 days they die
of acute scurvy. In the presence of sufficient vitamine C, growth is
uninterrupted.

The symptoms, already largely described by Hoist and Frolich,
are as follows: the molars become loose, with bleeding of the gums.
Sometimes a bluish hyperemia is noticed and in rare cases, ulceration.
Apart from this, hemorrhages are common in the soft parts of the
knee joints and under the periosteum of the sternal ends of the ribs.
Very often the connection between the ribs and the cartilage is
loosened; there is a severing between the epiphysis and the shaft of
the bones, especially the tibia. Later there occur duodenal ulcers,
brittleness of the bones, hematuria and edema. In addition to these
symptoms Chick and her co-workers (376) observed extreme pain and
swellings of the joints, even in the first stages. The animal assumes

132

THE VITAMINES

a characteristic posture ("scurvy position"), lying on one side, the
painful limb stretched out into the air. In other cases, the animal
lies with its head on the floor of the cage, a position caused by the
pain in the jaws and gums ("scurvy face-ache position"). As soon
as the teeth become loose, the animals refuse their food and death
usually follows within the next few days. If the animals receive not
quite enough vitamine C, they recover and begin to grow, but the
joints do not become normal. An abundant addition of vitamine
C before the beginning of the experiment has no effect on the
time of development of scurvy, according to Hess (377); hence it
would seem that the body has no reserve supply of this vitamine.

The post-mortem re-
veals rarification of the
long bones, with fractures
between the epiphysesand
the bones themselves; frac-
tures are also evident in
the ribs. The bones show
arrested ossification, and
atrophy of the existing
bone tissue and of the
ossification areas; the bone
marrow loses its lymphoid
character at the diaphyseal
ends, becomes poor in cells,
and sometimes presents a
homogeneous appearance ("Helles Mark"). Hess and Unger (378)
called attention to their observation of rosary in scurvy, which
might be mistaken for rickets. Rondoni and Montagnani (l.c.370)
described pathological investigations of guinea pigs fed on corn.
The symptoms seemed to resemble those of experimental scurvy
although they were more mild. The organs mostly found altered
were the spleen, thyroid gland and adrenals; the central nervous
system, liver, digestive tract, kidney and bone marrow were slightly
changed, while the myocardium and the lungs were still less
affected. The changes in the thyroid and adrenals go hand
in hand with a hypertrophy of the islands of Langerhans. Thy-
roid and spleen gave evidence of sclerosis; the adrenals, a diminu-
tion of lipoids; and in the cortex, atrophy and degeneration.

FIG. 32. "FACE-ACHE" POSITION OF SCORBUTIC
GUINEA PIG (REP. MED. RES. COM.)

THE VITAMINES IN THE ANIMAL KINGDOM 133

Rondoni (379) had already observed that the adrenals were somewhat
enlarged, though containing less adrenaline. This signifies a great
difference between scurvy and beriberi, for in the latter, the amount
of adrenaline is increased. Rondoni believed that the picture of
"maidismus" (corn-feeding) is different in scurvy and pellagra.
McCarrison (380) studied the influence of a diet of oats and auto-
claved milk. While the weight of the normal adrenals in guinea pigs
is about 0.467 gram, that in scurvy was about 0.955 gram. Histo-
logical examination revealed hemorrhagic infiltration (which perhaps
causes an enlargement of the gland) and atrophy of the cellular
elements of the cortex and medulla. In spite of the size of the
gland, the amount of adrenaline is diminished and has about half of
its normal value. The quantity of adrenaline per gram of body
weight dropped to about one-quarter of the normal. These symp-
toms arise before the other symptoms are apparent. LaMer and
Campbell (381), working in Sherman's laboratory, were able to con-
firm the results of McCarrison completely. The thyroid gland,
according to McCarrison (I.e. 295), may also be found increased in
size, due to infiltration. Aside from this, McCarrison (382) found
that the bladder had undergone pathological changes. Inflammation
of the mucosa and the musculosa of the bladder and signs of degenera-
tion of the epithelium were noted, associated with hemorrhagic
infiltration.

As regards the metabolism in scurvy, we have the data of Bauman
and Howard (383), differentiating it from simple starvation. The
animal were fed on oats, and the inorganic metabolism was
controlled. Nitrogen, sulphur and phosphorus elimination were
decreased, sodium and chlorine were unaffected, potassium was
retained, while the calcium and magnesium output was increased.
These investigators observed the typical "white line" of infantile
scurvy, noted by Frankel. In comparison, there are the normal
figures obtained by Smith and Lewis (384) with guinea pigs.

Through a determination of the alkali reserve of the blood of these
animals, McClendon, Cole, Engstrand and Middlekauff (385) showed
that scurvy of guinea pigs has nothing in common with acidosis.
Funk came to the same conclusion somewhat earlier (I.e. 368), on
the addition of sodium bicarbonate.

Lewis and Karr (386) found the blood and organs of scorbutic
animals much richer in urea, as compared with those of the controls;

134 THE VITAMINES

this could not be explained by hunger or loss of water. Karr and
Lewis (387) studied the amount of conjugated phenols excreted. The
figures obtained were normal and hence their conclusion, that in
scorbutic guinea pigs there is an abnormal bacterial decomposition
in the intestine, seems unjustified. According to McCollum and
Parsons (387a), the prairie dog behaves not like the guinea pig but
like the rat, in its vitamine C requirements.

Rabbits

Rabbits have been infrequently used in the solution of some of the
vitamine problems. The reason for this is that these animals are
much less affected by a lack of vitamines, which we have personally
demonstrated (I.e. 368). We found that a rabbit may live on oats
alone for more than three months, exhibiting individual variations in
their behavior. The explanation for this may be found in a paper
by Portier and Random (I.e. 217). In this investigation, rabbits
were fed on cabbage and carrots, autoclaved for one hour at 125°C.
The animals usually died after 11 to 17 days, with symptoms of an
avitaminosis. When the experiments were repeated with a larger
animal, these results could no longer be obtained; the animal was
observed for three months during which time it gained weight. This
observation found no explanation, till it was noted one day, that the
animal was eating the feces. Two series of experiments were then
arranged, in one of which the animals received, in addition to the
diet, the feces of the other, kept also on the de-vitaminized food. The
animals receiving feces were in good health after 100 days and gained
weight. The investigators explained this result by saying that with
the feces, bacteria were ingested which prepared vitamine for the
animal by symbiosis. It must nevertheless not be forgotten, that
feces may contain vitamine, as we shall see later on. Nevertheless,
the observation of Portier and Randoin was of greatest interest.

Schaumann (I.e. 2) thought that he had produced beriberi in
rabbits, fed on corn, which could be cured by yeast or Katjang-idjoe
beans. Since corn is very rich in vitamine B, this condition was
evidently confused with scurvy. Abderhalden and Lampe" (I.e. 25)
have observed paresis in a rabbit fed with rice, but beriberi has never
been actually demonstrated in rabbits. On the other hand, scurvy
in rabbits has been described by Hoist and Frolich (I.e. 36), identical
with scurvy in guinea pigs. In opposition to this, Morgen and

THE VITAMINES IN THE ANIMAL KINGDOM 135

Beger (388) regarded the disease of rabbits fed on oats as an acidosis
which could be obviated by the addition of calcium carbonate and
sodium bicarbonate. With these additions, some of the animals
lived up to 190 days, and often showed a gain in weight. We (I.e.
368) have repeated this work and have satisfied ourselves that the
addition of alkali actually does excite the appetite and prolong life,
but does not protect from death. McClendon, Cole, Engstrand and
Middlekauff (I.e. 385) obtained the same results. Kurijama (389)
found that the alkali reserve of the blood depends very much upon
whether the food gives rise to bases or acids, although McClendon,
v. Meysenbug, Engstrand and King (390) rightly observed that
his results were complicated by scurvy, since he fed his rabbits on
oats.

From what has just been said, we see that rabbits are hardly the
proper animals to use for the study of scurvy. According to the work
of Nelson and Lamb (391) rabbits seem to be adapted for the study
of ophthalmia. Two rabbits, kept on commercial casein, dextrin,
lactose, wheat germ, salts and an alcoholic extract of alfalfa grass,
developed, after 60 days, an eye disease which they thought to be
similar to ophthalmia; one of the animals was cured on the addi-
tion of butter.

Cats

Schaumann (I.e. 2) fed two cats on denatured meat heated in an
autoclave with sodium carbonate at 120°C. One of the animals
died after 42 days, the other, after 58 days with complete paralysis.
It seems, according to the newer investigations, that what Schaumann
saw was really beriberi. The experiments were repeated by Weill,
Mouriquand and Michel (392) and they showed that the same
symptoms could be obtained with cooked as well as with sterilized
meat. While the animals died after 45 days on small amounts of
fresh meat, nervous symptoms appeared after 35 to 39 days with
sterilized meat. Voegtlin and Lake (393) produced typical beriberi
in cats on fat-free meat, digested with 10 per cent sodium carbonate
solution for three hours at 120°C. The beriberi symptoms were
completely removed with vitamine B, sometimes after only 12 hours.
The disease could be prevented by daily additions of 2 cc. of autolyzed
yeast per kilo body weight, but not by the addition of 5 per cent
butter or 10 per cent purified casein. On beef, heated without soda,

136 THE VITAMINES

the animals remained healthy, except during pregnancy and lactation,
when the vitamine requirements are greater. Osborne, Wakeman and
Ferry (I.e. 102) found very little vitamine B in lean beef, and noted
that cooking several times in water removes this substance, leaving
only traces. Mackay (393a) and Tozer (393b) did not succeed in
producing experimental rickets in cats in the absence of vitamine A.

Lions

Bland-Sutton (394) observed rickets in young lions in captivity
when they are weaned too soon and fed on raw meat. Apparently,
there is a lack of vitamine A, since the animals were cured with milk
and cod liver oil.

Dogs

Dogs are now frequently used for vitamine studies and therefore
we shall devote a little more space to this subject. The dog has
become particularly important for experiments on rickets. In the
historical section, we have already discussed the metabolism of dogs
to some extent. It frequently happened, in these experiments, that
the various investigators used a diet poor in vitamine although the
purpose of the experiments was aside from the question under con-
sideration. For instance, there is the work of Cahn (395) in 1886,
who studied the results of chlorine hunger. Cooked meat was
used as a food, and Cahn described a number of symptoms of chlorine
hunger, among which was blood in the stomach contents, which might
perhaps have been due to decreased capillary resistance. Similar
experiments were also carried out by Trappe (396) with extracted
meat, to study the significance of hydrochloric acid in the stomach.
Rosemann (397) too, experimented with the same end in view, as did
also Bonniger (398), using a diet of extracted meat. Bonniger
observed that the animals refused the food and manifested vomiting,
apathy and distress. It was indeed remarkable that these symptoms
should have disappeared on the addition of sodium bromide. Pfliiger
(399) observed enteritis in dogs fed on sterilized meat, poor in fat.

A whole series of experiments, which have only historical interest
at present, were made by Wolfgang Heubner (400) and his school
(Lipschiitz, Durlach) on the significance of phosphorus in dogs.
The diet usually used for this purpose consisted of tapioca or white
rice, egg-albumin, palmin, cane sugar and salts. Heubner thought

THE VIT AMINES IN THE ANIMAL KINGDOM 137

to obtain a phosphorus-free diet in this way, whereas in reality, he
was dealing with a vitamine-free diet. Influenced by our work,
Heubner later recognized the importance of the vitamines. From the
work of Schmorl (401) who undertook the pathological examination
of these dogs, it may perhaps be noted that Heubner was dealing
with a mixed avitaminosis. In young dogs, rachitic symptoms pre-
vailed, although it is not clear whether the bone changes were of
scorbutic or rachitic origin, or due to a combination of both.

Still less satisfactory were the investigations of Masslow (402),
who studied the same problem as Heubner, but on dogs, and on
practically the same diet. Although the work was done in 1913,
Masslow evidently knew nothing of the significance of the vitamines.
His only observation of interest to us was the general emaciation of
the animal body on a diet actually free of vitamines.

In discussing vitamine problems, we must note, first of all, that
dogs seem to be sensitive to a lack of all three vitamine types. Never-
theless, although Hoist and Frolich (I.e. 36) produced experimental
scurvy in dogs, they seemed to be least sensitive to a lack of vitamine
C. Schaumann (I.e. 2) fed dogs on denatured meat and described
a condition similar to beriberi, with paresis and changes in the
pharynx and tongue; the latter appears to be more likely of a scorbutic
nature. Despite this, Schaumann (403) held that all these symptoms
could be removed by administering yeast, and since yeast contains
no vitamine C, he must have been dealing with beriberi. It should
be pointed out here that the findings on beriberi in dogs are not
concordant. Theiler, Green and Viljoen (I.e. 278) stated that they
could not produce beriberi or scurvy in dogs by feeding with white
rice; the animals merely suffered from malnutrition. Voegtlin and
Lake (I.e. 393), on the contrary, described true beriberi in dogs fed
on meat, extracted with soda.

Karr (404) fed dogs on lard, cane sugar, salts and protein, in the
form of casein or wheat gluten. After 3 to 9 weeks, the animals
refused the food. It was noted that grown dogs needed very
little vitamines A and C, since yeast alone, in amounts of 0.1
gram daily, restored the appetite to normal, while an addition of 0.2
gram cured beriberi in 8 to 12 hours. The utilization of protein
seemed uninfluenced by the presence of vitamine B.

Proceeding to the rachitic-like disease in dogs, we come first to
the work of Stilling and v. Mering (405) in 1889, who fed dogs on

138

THE VITAMINES

extracted meat and fat in order to study osteomalacia and the effect
of a calcium-poor diet. The experiment lasted 126 days. Hebrant
and Antoine (406) described a typical osteomalacia in dogs. Gue'rin
(407) observed rickets in young dogs fed on meat, while animals from
the same litter, which were suckled, remained unaffected, Some of
these experiments were made not only with a vitamine-poor but also
with a calcium-poor diet and the pathological condition described
was nothing but osteoporosis. To this category, belongs the work
of Roloff (407a) in 1879, and also the communication of Reimers
and Boye (407b). Bull (408) described a severe occurrence of rickets
in young dogs fed on cooked meat and pollard, in Adelaide, Australia.
However, the systematic investigations of Mellanby (I.e. 95) are
of chief interest in this chapter. This investigator fed young dogs
5 to 8 weeks old on diets of various composition, poor in vitamine A.
These diets were more or less suitable for the production of rickets
and in the end, the following were chosen for this purpose.

DIET A

DIET B

Skimmed milk

175 cc.

250-350 cc.

Wheat bread (white) . .

ad libitum

ad libitum

Linseed oil

10 cc.

10 cc.

Yeast

10 grams

5-10 grams

Orange juice

3 cc.

NaCl

1-2 grams

1-2 grams

Some 200 dogs were used in this work; at first, they were kept on
the diet for 6 months but later, a shorter time, for it was demon-
strated that dogs on diet B developed rachitic symptoms after 6
weeks. Mellanby investigated the behavior of various foodstuffs of
animal and vegetable origin, and found that those that are protective
against rickets are rich in vitamine A. Still, meat poor in fat had
some effect in delaying the onset of rickets. Quickly growing dogs
develop rickets soonest, which apparently shows that the more
active the growth the more vitamine A is necessary.

Symptomatology. The ossification of the bones proceeds in an
abnormal manner; hence the bones bend and the ligaments become
loose, making the deformities even more apparent. Swelling of the
epiphyses of the bones was easily noticeable and according to the
severity of the condition, deformities of the thorax developed with a

FIG. 33. RICKETS IN A DOG (MELLANBY; REP. MED. RES. COM.)

FIG. 34. X-RAY OF BONE OF RACHITIC PUP (MELLANBY; REP. MED. RES. COM.)

139

140 THE VITAMINES

typical rosary. The animals were very little resistant to infections,
were apathetic, lethargic and extremely unwilling to exert them-
selves, at a time when the existing bone changes could not yet have
been responsible. This was undoubtedly due to the impairment of
the muscular tonus.

After death, the bones were found to be very poor in calcium and
showed,, in the X-ray, an appearance characteristic of rickets, as
may be seen in the illustration. Of two dogs of the same litter,
one animal received only the "rachitic" diet, the other, vitamine A
in addition. The malformation of the cartilage epiphyses and the
more frequently occurring osteoid tissue are the two chief symptoms.
According to Henderson (408c) the muscle of the rachitic dog con-
tains less creatine than the normal. Findlay, Paton and Sharpe
(408b) conducted metabolism experiments on rachitic dogs. A cal-
cium-poor diet caused osteoporosis, but not rickets, while in rachitic
dogs the blood and the tissues showed the normal calcium content.

The significance of Mellanby's experiments for the study of rickets
will be discussed further in the description of this disease.

In conclusion, we shall mention an investigation of Chittenden and
Underhill (I.e. 28). They fed dogs on cooked peas, zwieback and
cottonseed oil, and noted a disease which they believed to be anal-
ogous to pellagra in man. The disease developed after 2 to 8 months,
showing that a larger amount of peas was less harmful. The dis-
ease, which could be cured by adding meat to the diet, produced
ulcerations of the mouth; in particular, the entire enteron was
hemorrhagic. Besides, there was observed a great loss of weight,
peculiar gait and diarrhoea. If a part of the peas was substituted
by casein, the symptoms appeared sooner. This shows, perhaps,
that the condition is associated with a lack of vitamines, since peas
contain some vitamine but not enough. To our knowledge, this work
has not yet been repeated by any other investigator.

Domestic animals

We come now to the consideration of domestic animals, among
which we shall discuss goats, sheep, horses and pigs. In this field,
there are questions which are not yet quite clear, but which possess a
definite significance in their social and economic aspect. We have
very little accurate knowledge of the vitamine requirements of these
animals. In various parts of the world, we encounter diseases,

THE VITAMINE IN THE ANIMAL KINGDOMS 141

which, though not yet explained etiologically, nevertheless always
give some indications showing that they may be avitaminoses. In
most cases, they occur on a large scale in Nature, and consequently
are specially deserving of our interest.

Sheep and goats. Experimental investigations with these animals
were first made by Schaumann (I.e. 2). A goat, fed on corn and
white rice, lost weight rapidly and showed, according to Schaumann,
paresis, although corn certainly contains sufficient vitamine B. The
animal was then given yeast, Katjang-idjoe beans, and green fodder
(the latter for one month), with a resultant gain in weight. This
effect was attributed to the yeast and the beans, although the green
fodder very likely played a greater role. This experiment was not
clear and consequently is of little significance. Henriques and
Andersen (409) administered pa^enterally hydrolyzed meat, glucose,
sodium acetate and salts to goats. For 20 days, a marked nitrogen
retention was noted. They then heated the food mixture for 20
minutes at 110°C., and were able to show that it was no longer
possible to maintain the nitrogen balance, except when the heating
was not carried beyond 100°C. These results were attributed to the
susceptibility of tryptophane to heat. Fingerling (410) fed lambs on
inorganic and organic phosphorus combinations, without observing
any particular differences. He made the correct observation that
the frequently noted improper nutrition with inorganic phosphorus
compounds was not due to the character of the phosphorus, but to
the composition of the food itself.

Theiler, Green and Viljoen (I.e. 278) were unable to demonstrate
an avitaminosis in sheep and goats. These animals tolerated the
rice diet better when an addition of autoclaved hay or straw was
made. Very soon, enteritis made its appearance, though in the
opinion of these investigators, this had nothing to do with the diet.
At all events, the requirements of vitamine B appear to be very
modest.

The naturally occurring nutritional disturbances were described by
Hoare (411) as a form of edema, which develops because of an insuffi-
cient fodder and poor climatic conditions. A disease of sheep, known
as "staggers," was described by Jones and Arnold (412) in South
America. It is a nervous disturbance of sheep, as well as horses and
cattle, caused by the consumption of coarse grass (Poa Argentina).
They were not clear as to the etiology, and were more inclined

142 THE VITAMINES

towards the conception of a toxic phenomenon. The observed
symptoms were general weakness, muscle tremors, peculiar movement
of the head, stiffness of the limbs, intermittent paralysis with spastic
appearances, particularly after irritation, impairment of the visual
capacity and conjunctivitis. The disease could be produced experi-
mentally by feeding pampas grass. The time necessary for this
varied between 2 and 21 days, averaging 10 days, during which
spontaneous cures often occurred. By changing the diet, it was
possible to effect a cure. Young animals were far more susceptible
than grown ones, and the disease was attributed to a poison in the
grass, present at all seasons and in all parts of plants. The post-
mortem revealed nothing positive and afforded no definite indications
of the nature of the toxin. Reid and Aston (413) observed a bone
cachexia in sheep, which they referred back to a lack of inorganic
factors of the diet.

Reindeer. Ibele (414) noted a disease called "Lecksucht" supposed
to be due to the lack of alkaline earths.

It is difficult, without having personally seen these animals, to
draw a picture of the nature of the disease. The observation that
in one case the disease could be produced after two days feeding,
would speak against its being an avitaminosis, if the investigators
had not explicitly emphasized that this is possible only in certain
susceptible animals. This lack of resistance can be explained, at
least theoretically, by a previously occurring deficient dietary.

Pigs. The nutrition of these animals was studied from two view-
points— first, from that of the animal breeders in connection with the
food requirements, and secondly, from that of laboratory men in
relation to the vitamines. It is true, as Eward (415) pointed out,
that if these animals are permitted to select their own food, their
natural instinct will lead them to choose a correct diet. Conditions
are different in winter, however, when a prepared diet must be
provided. Hart and McCollum, as well as their co-workers, studied
the exact food requirements of pigs in relation to growth, maintenance
and reproduction. Such studies were published by Hart, McCollum
and Fuller (416) in 1908. They showed that pigs fed on rice and
washed bran suffered from osteoporosis, while those fed on unwashed
bran did not manifest this disease. At that time, the conclusion
was made that the phosphorus compounds were washed out with
water — which of course does not hold today. Hart and McCollum

THE VITAMINES IN THE ANIMAL KINGDOM 143

(417) observed that between wheat and corn, there are great differ-
ences as regards the growth-promoting properties. While the
animals attained a weight of 100 pounds on wheat, on corn they
weighed from 200 to 300 pounds. McCollum (418) fed young pigs
on wheat, oats and corn and found that casein, as well as skimmed
milk, was a better source of nitrogen. After three months, the
animals declined and McCollum left the question open as to whether
this condition was due to lack of salts or vitamine, or to the presence
of a toxic substance. Hart and McCollum (419) could not keep
young pigs alive for a long time on corn and gluten, while the addition
of casein or milk appreciably improved growth. Hart and Steenbock
(420) investigated a series of plant products, with the addition of
the same animal foods as above, and came to the conclusion that
rice, wheat, corn, potatoes, and cabbage possessed little nutritive
value, but it was not further investigated whether the vitamines
played a role. Later, Hart and Steenbock (421) sought to settle
the question as to whether the presence or absence of vitamines has
any effect. For this purpose, grown pigs were kept for a long time
on corn and oats. Eventually, symptoms of the disease appeared,
such as stiffness of the legs and difficulty of getting about, these
conditions being specially manifested in pregnancy and lactation.
A great number of young were born dead. If these symptoms were
not noticed in the first litter, they were present in the second. In
most cases, an improvement was seen when the cereal diet was sub-
stituted by a corn-alfalfa mixture, which is much richer in vitamines.
The dietary deficiency was however attributed largely to the unfavor-
able composition of the inorganic constituents. These exact experi-
ments are particularly important for the recognition of the etiology
of some very frequently occurring diseases of cattle which are of
practical importance, and which have lately been quite often attrib-
uted to the effect of a poison. We see, however, that feeding in
the stable may lead to dietary deficiencies, without it being necessary
to assume a toxic cause.

The superiority of skimmed milk over all other food mixtures was
also set forth in a number of publications of Klein (422). This
favorable influence may be explained either by a better utilization of
the protein, by the vitamine content, or by both factors.

Despite all these investigations, and still others to be mentioned,
we are not yet accurately informed as to the vitamine requirements of

144 THE VITAMINES

pigs. This is so because few investigations in which only the vita-
mine factor is lacking, while all other factors are controlled, have been
carried out. All of the above-mentioned investigations are com-
plicated by mixed symptoms, and it is therefore impossible to form
an unbiased opinion as to the nutritive requirements of these animals.
This is all the more regrettable, since the pig, which eats anything,
is peculiarly adapted for the study of human nutrition.

In 1907, Hoist and Frolich (I.e. 36) described, in rice-fed pigs,
symptoms which were thought to be mixed forms of beriberi and
scurvy. Since white rice is lacking in all three vitamines, it is impos-
sible to say which one is most important for these animals. Theiler,
Green and Viljoen (I.e. 278) on the contrary, could observe no
specific symptoms on this food. In one case, gall stones were found;
in others, enteritis was noted, which may perhaps be regarded as a
sign of an avitaminosis. Plimmer (422a) observed scurvy in pigs
fed on cooked food, the symptoms of which were relieved by the
administration of uncooked food. Green (423) carried out investiga-
tions with a diet poor in calcium, and in this way found two kinds
of undernourishment; one kind depends upon protein which was
qualitatively shown to be inadequate, while the second could be
attributed to insufficient protein and salts addition. The animals
were in poor nutritive condition and showed defective bone growth
with insufficient calcification. Substitution of lard by butter exerted
no favorable influence. It is one of the few investigations on the
nutrition of domestic animals, in which vitamine A was given special
attention. Another investigation, in which the presence of vitamine
A in the diet was provided for, was that of Hart, Miller and McCollum
(424) A typical composition of the diet used by them was 95.5
parts wheat meal, 2.5 parts wheat gluten, 2 parts butter, di-potassium
phosphate and calcium lactate. On this diet, the animals developed
symptoms quite similar to those of beriberi. Among other symp-
toms, pathological changes were noted in the spinal cord which
showed fatty degeneration. The symptoms made their appearance
after about 9 months, when a decrease in weight, difficulty of forward
motion, rough coat, labored breathing and muscle tremors were
observed. There was also stiffness of the extremities and dragging
of the legs; if the animals were helped to their legs they again sank
at the knees. Since McCollum, at this time, denied the existence
of vitamine C, the diet used was considered as complete, and the

THE VITAMINES IN THE ANIMAL KINGDOM

145

disease was attributed to toxic action of wheat. Addition of whole
milk had only a slight effect upon the general well-being of the
animals, but the addition of alfalfa grass soon permitted of the
development of a normal condition. In all these investigations,
growth was interrupted when the general condition of the animal
became worse. It seems to us that without further indisputable
evidence, it is difficult to accept the view as to the toxicity of wheat
kernel. It may be possible that food mixtures were used, the vita-

FIG. 35. BERIBERI-LIKE DISEASE IN A PIG ON A DIET CONSISTING OP 45 PARTS

CORN, 45 PARTS OATS, 10 PARTS OIL SEED PRESS-CAKE AND 5 PARTS

BUTTER (HART, MILLER AND McCoLLUM)

mine A content of which was barely sufficient, while vitamine C was
almost completely lacking. Experiments with the addition of these
vitamines were not made.

With products derived from cotton seed, the behavior was naturally
different. These contain a toxic substance called "gossypol." Such
products, fed to pigs, cause death in 50 to 80 days, according to
Roberts (425) . But even in these cases, there seems to be a vitamine
deficiency, according to Rommel and Vedder (426). In 8 to 15 days,
this food produces in these animals a picture of disease resembling

146 THE VITAMINES

perhaps the wet form of beriberi in man, and quite analogous to the
symptoms which pigs develop when fed white rice. Zilva and col-
laborators (426a) found that the pig was susceptible to the develop-
ment of experimental rickets, but in this animal, the lack of vitamine
A (of milk) was not the sole causative factor.

In spite of this series of results we are not yet clear as to the vita-
mine requirements of pigs, so that further work is necessary.

Horses. In 1908, Friedberger and Froehner (427) described edema
in horses and oxen which performed heavy work in the sugar refiner-
ies, and were fed on sugar beet residues. These residues contained
only a slight amount of protein, about 0.5 per cent, very little dry
substance (5 per cent) and high water content (95 per cent). This
diet which, aside from the protein, appeared to be lacking in other
constituents, together with the hard work and high water content of
the food, was supposed to be responsible for the development of this
disease. Similar pathological conditions were likewise noted in
horses by Hutyra and Marek (428).

Scheunert, Schattke and Lotsch (429) observed a disease in horses,
resembling osteomalacia, in the poor mountainous districts in Saxony,
Kawakami (430) observed in horses, a disease called "sukumi" or
"gokusukumi," which developed when rice, barley and oats were fed.
The disease exhibits a certain similarity to experimental beriberi and
starts with gastro-intestinal disturbances. Theiler, Green and
Viljoen (I.e. 278) fed horses on white rice for six months without
noticing any signs of an avitaminosis. This is the more remarkable
since white rice is supposed to be lacking in protein, salts and vitamine
A, besides vitamine B.

Cows and oxen. In 1913, the late Dr. Donald Macauley called our
attention to a South African cattle disease, known there as "Stijf-
ziekte" and "Lamziekte." On perusing the available literature, it i&
apparent that similar conditions are known in all parts of the world.
Stewart (431), for instance, reported a disease known in Australia
mostly as "rickets."16 In the poor mountainous districts of Sax-
ony, according to the report of Lotsch (432), there is not so infre-
quently a pathological condition called "Stallm angel," supposed to
be caused by a diet poor in salts. This severe disturbance of metabo-
lism reminds one strongly of human rickets and osteomalacia.

18 Stewart was disposed to consider the disease as an avitaminosis.

THE VITAMINES IN THE ANIMAL KINGDOM 147

From New Zealand, there was a report by Aston and Reakes (433)
on progressive anemia, known locally as "bush-sickness." It was
supposed to be caused by lack of salts, and differed from lamziekte.
In addition to these naturally occurring pathological conditions, we
have the experience gained by the experimental feeding of cattle.
Russel and Morrison (434) found that cattle fed on oats, straw and
butter, or casein, butter and oat-straw, gave birth to very miserable
looking offspring. This condition was not regarded as an avitam-
inosis but as a lack of calcium, since an addition of these salts seemed
to improve the condition of the animals. Fleischmann (435) noted
that calves fed on hay often became sick, and he proceeded to study
the chemical changes attending the drying of the grass. He showed
that lecithin and phosphoproteid, as well as protein, underwent
some decomposition during the process of drying. However, since
the total nitrogen showed no change, we may assume that there was
no loss of inorganic constituents. The real difference observed must
be looked for in the vitamine content.

We find a somewhat different conception in the work of Henry
(436). In the description of an Australian cattle disease known as
"impaction paralysis," resembling lamziekte, he attributes the cause
of the disease to the poverty of the soil in calcium and phosphorus.
The disease appears at the end of the dry season, in places where the
vegetation is injured by the Australian rabbit-plague. The disease
was found where there was an abnormal desire to eat bones and
where osteomalacia had also prevailed for some time. A favorable
effect was noted on adding a nourishing diet, such as skimmed milk.
We have already mentioned the interesting assumption of Davis
(I.e. 195), who held that in the last analysis the poverty of the soil
was the cause of diseases of domestic animals. When the vegetation
is so impoverished through extraneous conditions that it no longer
provides sufficient calcium and phosphorus for the animals, it is
conceivable that other important substances, for example, vitamines,
might also be lacking. It must likewise not be forgotten that in the
avitaminoses there is frequently a loss of inorganic constituents,
and this condition might manifest itself in an abnormal desire for
bones or other materials rich in calcium and phosphorus. For
example, Forbes (437) stated that domestic animals often display an
abnormal craving for mineral substances. Pigs and cattle sometimes
eat large quantities of bone meal, with resultant improvement in

148

THE VITAMINES

health. Cattle growers know that the condition of the legs in horses
depends upon the fodder and largely on the nature of the soil. Place
(438) believed he saw among cattle not only diseases similar to beri-
beri but also scurvy-like conditions in cattle, horses and sheep,
infantile scurvy in calves and sheep, and diseases of the type of
pellagra, with skin and gastro-intestinal symptoms.

Stijfziekte and lamziekte. These diseases of cattle and goats occur
widely in South Africa, and are described by Theiler (439). These
conditions, particularly lamziekte, have become of such economic
importance that a great number of well-known investigators have

occupied themselves with this problem.
In some severely affected districts, the
lamziekte problem has become so acute
that it no longer pays to breed cattle
because of the great mortality. Of
the two diseases, stijfziekte has been
least investigated, apparently because
of its lesser practical importance. It
is a bone disease of young animals,
accompanied primarily by swelling of
the joints, the metatarsals and meta-
carpals, and secondarily, the epiphyses
of the long bones. These swellings are
painful and since the fore-legs are most-
affected, the animal tries, as far as
possible, not to use them, and runs
around with forward-bent hind legs
and hunched back. Muscular weak-
ness and abnormal appetite (pica) are
among some of the symptoms of the disease. The temperature
remains normal. The diaphyses of the diseased bones, on sectioning,
appear deep red and blotched with bloody-serous liquid.

Some forms of stijfziekte resemble lamziekte very much, without
necessarily indicating a relationship between the two, in the opinion
of Theiler, Green and Viljoen (I.e. 278). Theiler admitted that
stijfziekte might be an avitaminosis. If this were so, it would help
clear up the problem of lamziekte, since it could serve as a proof that
the nourishment of cattle in some sections of South Africa could be
improved. As it happens, stijfziekte may be cured by a change of

FIG. 36. STIJFZIEKTE (THEILER)

THE VITAMINES IN THE ANIMAL KINGDOM 149

diet, according to available data. Feeding of good grass, green
barley and green millet is supposed to be accompanied by beneficial
results.

Lamziekte or gal-lamziekte. The history of lamziekte research is
particularly instructive since it shows us how easy it is to go from one
extreme to another. In times of great popularity of the vitamine
theory, one is tempted to classify every condition that bears some
analogy to the already known avitaminoses in the same group.
Lamziekte, according to the present stage of investigation at least,
is not an avitaminosis and would not have found a place in our
treatise if it were not for the possibility of explaining one of the
etiological factors through a lack of vitamine.

This disease is prevalent in all breeds of cattle without regard to
species or sex. Young, pregnant or milking animals appear to be
specially susceptible. Lamziekte occurs particularly after long
periods of draught, and is manifested by paralysis and contractions
associated with the degeneration of the peripheral and central
nervous system, roughly resembling avian beriberi. We note here
paralysis of the legs, opisthotonos, dysphagia and paresis of the
tongue. Lack of appetite and pica (abnormal craving), excessive
salivation, loss of milk, constipation and also diarrhoea — all these
constitute the earlier symptoms. The temperature appears to be
subnormal. The usual progress is sub-acute and lasts some weeks;
the animals sometimes show an apparent improvement, but fatal
results ensue when a second attack occurs. In spite of a cure, the
contractions may still persist for a long time. Acute forms last
from two to seven days, and yet sudden developments are noted, in
which apparently healthy animals fall into a comatose condition in
10 to 20 hours, which, accompanied by sub-normal temperature,
results in death. This condition has been frequently confused by
South African farmers with another called "poverty," and resembling
malnutrition. This condition, "poverty," may eventually prove to
be a concealed avitaminosis.

Pathological anatomy of lamziekte. The findings have been spe-
cially described by Hedinger (440). They consist of hydrothorax,
and ecchymoses of the pleura, epicardium, endocardium and thymus;
hyperemia was observed in all the abdominal organs, in the mucosa
of the fourth stomach, intestines and lungs; a moderate hydroperi-
cardium was frequently noted; enteritis of the small intestine,
coupled with hemorrhages and ascites, with a dilatation of the heart,

150 THE VITAMINES

occurred not infrequently. In the bones, on the contrary, at least
microscopically, no changes were found.

Etiology. We may well imagine that because of the great practical
significance of this disease no effort has been spared to discover its
cause. One of the older theories of Theiler (I.e. 439) assumed the
presence of toxins in the grass. This theory was experimentally
tested by Viljoen (441) in that he fed a series of animals on
various kinds of grass, found in regions affected by lamziekte, with
negative results. The theory of infection was tested by Spruell
(442); cows and goats received intravenous injections of the blood,
lymph glands and ascites fluid of sick animals; here too, the experi-
ments proved to be negative. Mitchell (443) and Walker (444)
sought to bring about the disease in healthy animals by using carcass
material of lamziekte animals, but in vain. We formerly held that
lamziekte was similar to beriberi, but this view was modified by us
in 1915 (445), in that we considered it to be similar to rickets.
This view is also held by Hart, Steenbock and Hoppert (445a), who
arrived at this conclusion from experiments with goats. Here cod
liver oil had a distinctly favorable action on the calcium metabolism.
In spite of this, our first opinion was carefully tested, and Stead (446)
attempted to show that lamziekte was analogous to beriberi. He fed
pigs on meat obtained from animals which had died of lamziekte, in
the belief that it was poorer in vitamine B than normal meat.
Cattle were also fed on white rice, samp (a product made from
decorticated corn), white flour and oats; they showed pica, stiffness
of the legs and pains on walking. Theiler, Green and Viljoen (I.e.
278) have investigated a series of animal species, already mentioned
previously, as to their vitamine B requirements; horses, dogs, goats,
sheep and pigs were used. They were kept on a diet consisting chiefly
of white rice, and in no case was there any symptom characteristic
of beriberi. Cattle were kept on white rice for a year without the
appearance of any beriberi symptoms. In the worst case, symptoms
were noted, in accord with the findings of Stead, which were thought
of as laminitis and which could not be attributed to a lack of vitamine
B. Latterly, Stead seems to have abandoned his view in favor of
Theiler's.17 One cow fed on white rice and autoclaved straw or

17 Theiler said that no grass could be so poor in vitamine as was the diet
which he used and he believed, therefore, that it had nothing to do with an
avitaminosis. The survival of the animal, he attributed to a bacterial
symbiosis (p. 75).

THE VITAMINES IN THE ANIMAL KINGDOM 151

hay, gave birth to a calf that was blind. The validity of the vitamine
hypothesis was tested in two ways. First, through a repetition of
Stead's experiments, by feeding to healthy animals the meat of
animals which had died of lamziekte, definite indication as regards
impoverishment of vitamine B could not be found. Secondly, the
influence of substances rich in vitamine B was studied. An addition
of orypan (extract of rice polishings) and yeast had no effect, and
we may therefore assume, with certainty, that lamziekte cannot be
caused by a lack of vitamine B. Experiments with other vitamines
were not tried.

In the meantime, Theiler and his co-workers (447) have apparently
completely cleared up the etiology of lamziekte. They maintain that
in the etiology of this disease, six phases prevail, which are responsible
for its occurrence, and are related to each other, like the links of a
chain.

1. A toxin or poison which causes the disease.

2. Toxin-forming saprophytes.

3. The availability of carcasses from which the toxins are formed.

4. Pica, or abnormal craving, which leads animals to eat carcasses,
not present under normal conditions.

5. The character of the vegetation and the soil (and climate) upon
which pica depends.

6. Susceptibility of the animals to the toxin.

The toxin was systematically investigated and seemed to be
similar to the botulinus toxin. It is very active, inasmuch as 0.00001
cc. per kilo body weight is a fatal dose. Lamziekte may also be
produced experimentally by administration of this toxin. The pro-
phylaxis and therapy consist of the elimination of one of the six
above mentioned links of the chain, at least theoretically. Practi-
cally, however, it seems that the best starting point is the pica and
since this condition is of greatest interest to us, we shall speak of
it in greater detail.

Pica. It has been demonstrated in South Africa, that not only
cattle may exhibit this condition, but also ostriches, goats and
poultry. Pica is supposed to be a nervous ailment, which arises
only where the condition of the soil is such that the water quickly
passes through the upper layers. Because of this, the soil becomes
poor in phosphorus and hence produces a vegetation poor in this
element. Pica itself is not dangerous, but becomes so when the

152 THE VITAMINES

animals find infected carcasses to eat. This condition is tested by
giving the animals rotten bones; if they chew these bones, then the
diagnosis of pica is verified.

Cure of pica. To overcome pica, a mixed diet may be used which
will cure the disease in one month. Two pounds of wheat bran cures
it in three weeks, and 112 grams of bone meal is supposed to have
the same effect. Besides this, the addition of calcium phosphate
and sodium phosphate, and even phosphoric acid in drinking water,
may be used.18

Etiology of pica. As we have seen above, Theiler thought that
the chief cause of pica was the poverty of the diet in phosphorus,
accompanied perhaps by too much calcium, although it is admitted
that other dietetic factors also may play a role. Theiler also stated
that on analysis of the diet, the phosphorus was found within the
limits of the accepted standards for such animals. He concluded
therefore, that the accepted figures had to be revised. We know
from the study of deficiency diseases, that under such conditions
there is often a negative calcium and phosphorus balance, and it has
been stated from time to time, that the addition of phosphorus (we
refer only to phospho-cod liver oil, hypophosphites, etc.) may result
in a definite improvement. This may be the case with pica. The
animal perhaps has sufficient phosphorus in the food but is unable to
retain it because, we may say, the diet lacks vitamine A. An addition
of phosphorus, in whatever form, may perhaps lead to a temporary
improvement and, in the light of the modern conceptions of lamziekte,
may also protect against this disease, since the animals would not
eat the carcasses. But the cause of pica and the metabolic disease
may endure till the arrival of a better season and a recovery of the
vegetation. For it can be shown definitely that pica prevails only
at certain times of the year, when the vegetation thrives but poorly.
If this is so, then vitamines, as well as salts, may be lacking; a normal
vegetation should provide everything for the animal that is necessary
to life. The simultaneous development of undernutrition, in the
form of "poverty," and perhaps also stijfziekte, in lamziekte regions,
is illustrative of the profound dietetic deficiency of the local vegeta-
tion. As to the etiology of pica, this can be cleared up only by exact
investigation, especially by metabolism experiments. Such accurate

18 In this connection it is interesting to compare the influence of phos-
phorus on pica with that on experimental rickets in rats (p. 327).

THE VITAMINES IN THE ANIMAL KINGDOM 153

investigations appear still to be lacking. Above all, it must be
determined, in the event that the disturbance of metabolism may be
cured by the addition of phosphorus, how vitamine A and C, which
may well be present in poor vegetation in sub-optimal amounts,
would act. The influence of these vitamines on pica has been very
little investigated.

Monkeys

Only the lower types of monkey have been used of late for vitamine
studies; the first work was that of Schaumann (I.e. 2) who fed
monkeys on washed white rice, with the result that the animals died
in 74 days. Before death, there was paresis of the hind legs, the
appetite had disappeared and the animal showed a 27 per cent loss
in weight. The histological nerve findings were not so typical for
experimental beriberi, and controls with the addition of vitamine B
were lacking, to prove that this condition really was caused by lack
of this substance. Shiga and Kusama (448) have described a real
beriberi in monkeys, with anasarca, hydropericardium and edema of
the lungs, while Noe (449) was unable to induce beriberi in these
animals by feeding rice. McCarrison (450) tried then to produce
beriberi in Macacus sinicus, by feeding autoclaved, ground white
rice, while in another experiment extra butter was added. The first
group died in about 23.4 days and the second (with butter), after 15
days. None of the animals lived longer than 100 days. Although
typical beriberi or edema was not noted, the clinical symptoms
consisted of progressive anemia, gastro-intestinal disturbances and
progressive asthenia. Stomach, intestinal walls and omentum were
very thin and had lost their fat content. Congested, necrotic and
inflammatory changes were apparent in the entire gastro-intestinal
tract. Aside from this, there were degenerative changes in the
neuro-muscular apparatus of the intestines, leading to the distention
of the stomach and to other parts of the intestine. Signs of necrosis
were seen in the secretory layers, and attenuation of the elements
protective against infection was noted. These findings need not of
course be attributed only to a lack of vitamine B, since in the diet
utilized, vitamine C was also lacking.

The monkey is of greater value in the study of scurvy, as was
found by Hart (451), and later by Hart and Lessing (452). Young
monkeys kept on condensed cooked milk, rice and white bread,

154 THE VITAMINES

developed a condition which resembled infantile scurvy in its symp-
toms, while in grown monkeys, the symptoms were more like those in
grown people. In their book, they gave a detailed enumeration of
the symptoms and the pathological findings (with many illustrations),
so that we must refer the reader to this book for greater details.
Particularly were the changes in the bones accurately described. The
experiments of Hart were confirmed by Talbot, Todd and Peterson
(453), and a few years later by Harden and Zilva (454). The latter
produced scurvy on a diet containing only traces of vitamine B, but
concluded that in future investigations, it would seem desirable that
the diet should contain sufficient vitamine A and B. In our opinion,
too, this is very important, since in vitamine studies, it must be so
arranged that only a single factor is lacking. Harden and Zilva
were able to cure the disease by means of antiscorbutics. In a later
publication, they (455) showed that it took two months to develop
scurvy in monkeys. The diet used by them consisted of 300 grams
of rice, 50 grams of wheat germ, 2 grams of salts and 5 grams of butter.
A daily administration of 2 cc. orange juice per os sufficed to protect
the animals against scurvy. If smaller doses were given, the animals
developed scurvy, with paralysis of the hind legs, which appeared to
be a pseudo-paralysis. It is interesting to note that the monkey
requires less vitamine C than does the guinea pig. A monkey
weighing from two to three kilos needs as much as a guinea pig
weighing 300 to 400 grams.

Among other symptoms, Hart and Lessing (I.e. 452) noticed an
exophthalmus on the left side, with severe hemorrhagic edema of the
upper eye-lids. The same observation was also made by Zilva and
Still (456).

Chemical studies of the mineral metabolism of the scorbutic
monkey were made by Howard and Ingvaldsen (457). In this
case, the disease was produced with condensed milk in 4 months.
Loss of salts, usually so marked in man and guinea pigs, was not
noted here.

Harden and Zilva (458) reported an experiment on three monkeys,,
two years of age, fed on 250 to 300 grams of rice, 10 grams of yeast
extract (marmite), 2 grams of salts and 4 cc. orange juice. One of
these animals received no fat and no vitamine A, and developed an
edema with diarrhoea in 289 days. The second animal received
butter and lived 299 days, whereupon the experiment was interrupted.

THE VITAMINES IN THE ANIMAL KINGDOM 155

The third animal was given olive oil, and died after 262 days without
characteristic symptoms. Butter had no growth-promoting influ-
ence. Since the diet was very poor in fat, and since the significance
of fat for monkeys is not known, the investigators wished to await
further results before saying what role vitamine A plays in the
development of this edema. However, Hewlett and de Korte (459)
stated that in 1907 he had described edema in monkeys which had
received sufficient milk to provide for their vitamine A requirements;
nevertheless, he might have been dealing with a lack of vitamine B.
No albumin was noted in the urine of these animals.

In conclusion, we wish to point out that Chick and Hume (460)
have described, in a monkey, a condition which might be considered
to be identical with human pellagra. The diet used consisted of
sugar, corn starch, salts and corn gluten, in varying amounts, so
that in this way more or less protein was given. All of the three
known vitamines were present in the diet. We shall speak again of
this experiment in discussing the etiology of pellagra.

For the investigators who are interested in nutrition experiments
with these animals, it may be of interest to point out how monkeys
are raised by dealers and fed in our zoological gardens. Schaumann
(I.e. 2) states that in the Institute for Ship and Tropical Hygiene in
Hamburg, monkeys are kept on a porridge made of cooked white
rice; in addition they receive carob seed, peanuts and fruits. On
personal inquiry at the New York Zoological Garden, we learned that
the monkeys were given cooked rice, a cup of milk, bananas, apples,
sweet potatoes, onions and carrots; once a week they were given
some powdered sulphur. The chimpanzee received two raw eggs in
milk, bananas, two oranges and a dozen cooked prunes every two
days. The monkeys lived for years on these diets in very good
health.

Vitamine requirements of man

After having discussed the vitamine and food requirements of
various animals, we are prepared to study the above relationship in
man. It is not infrequently seen that clinicians are unwilling to
apply the results of animal experimentation to man, and they are
justified in being sceptical. Yet we must emphasize that all we have
discussed in the preceding chapters is based upon a solid foundation.
We are dealing here with the fundamental laws of cell nutrition

156 THE VITAMINES

applicable to all living organisms. As little as the modern clinician
can afford to ignore the physiological significance of the proteins or
of certain amino acids, just so little need he doubt the importance of
the vitamines for human nutrition. It is true that the conditions
leading to deficiency diseases in man are perhaps not so clear cut as
in animal experiments, since in the latter, we may choose such con-
ditions as will permit of the study of the significance of a single
constituent exactly. In spite of this, we do not yet know all there
is to be known of the laws of nutrition. It has been frequently
necessary to proceed empirically; for example, we fed white rice to
chickens and pigeons without knowing at the time tjiat this food-
stuff lacks other important constituents besides vitamine B. We
were still in the empirical stage when we wrote our first edition, but
since then, vitamine research has made great strides; nevertheless,
many of the findings obtained at that time, with few exceptions,
still endure to this day. Should it appear later that the human
avitaminoses are associated not only with a deficiency of the corre-
sponding vitamines, but with other correlated factors, then the
significance of vitamines for life would still retain its complete
justification. That the etiology in human avitaminoses may be more
complicated than appears at first glance, we have already pointed
out. Should it happen that there is a deficiency of one vitamine,
we may be almost certain that it is not the only fault in the diet
leading to the particular disease. Along with the lack of one vita-
mine, there may be a sub-optimal quantity of other vitamines; there
may also be a protein and salt deficiency. In addition to this, it is
possible that the quantitative relationship of the individual con-
stituents may not be correct; it may likewise be (and this is very often
the case in practice) that the vitamine is indeed present in the diet
but not in quite sufficient quantities. Related as it is to all these
factors, we must not wonder that the picture of an avitaminosis often
seems to us to be clouded. Taking rickets as a concrete example,
we see that although this disease is not of rare occurrence among
well-to-do people, it nevertheless occurs most frequently in the poorer
stratas of society, where poor living conditions and hereditary factors,
together with dietary causes, may play a part. It is also not likely
that the lack of vitamine A only is the cause of rickets, as is now
thought to be the case; it may be associated with a partial
deficiency of other factors, which chiefly consist in that the balance

THE VITAMINES IN THE ANIMAL KINGDOM

157

between the proteins and the carbohydrates is disturbed in favor of
the latter. All these conditions may affect the clinical picture of
rickets as we understand it.

Most of the pathological conditions which we designate as human
deficiency diseases find their replica in animal affections already
described. The diseases have been produced experimentally and are
regarded by our greatest specialists as identical with human avitam-
inoses. However, this does not exhaust our data on the subject.

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OF 60 DAYS GAVE UNIFORM WEIGHT INCREASES

During period I, an increase in the food produced no corresponding increase
in weight. When the usual quantity of orange juice (15 cc.) was given at
B, a second increase in food was made. The effect of the two food increases
(dextri-maltose D.M.) was distinctly less than that produced by the larger
quantity of orange juice. Orange juice from which the B-vitamine was
removed by kaolin showed no such effect (Byfield, Daniels and Loughlin).

Exact experiments have also been conducted on man, having the
convincing value of a well-planned animal experiment. Thus, we are
familiar with clinical cases in which the food intake was controlled,
so that the course of the particular disease could be followed quite
accurately. From what has been said, it is clear that we are no longer
concerned with conjectures in the classification of human avitami-
noses. We are in possession of proof, which makes it certain that
man needs at least three vitamines — antiberiberi (B), antirachitic
(A), and antiscorbutic (C).

158

THE VITAMINES

Especially from children's clinics, do we have observations that
are in accord with the results of animal experiments. In this regard,
we wish particularly to mention the work of Hess (461), who studied
the influence of vitamine C on the growth and weight of infants. To
be sure, Byfield, Daniels and Loughlin (I.e. 90) believed that his
results must have been due to the simultaneous presence of vitamine

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WEEKS

FIG. 38. BREAST-FED CHILD SHOWING STATIONARY WEIGHT

Mother received an addition of 50 grams butter and 30 grams turnip juice
(vitamines A and B), at the point indicated (Dalyell; courtesy of Brit.
Med. J.)

B, but the value of the experiment of Hess is not lost for our purpose.
Byfield, Daniels and Loughlin were able to show that when children
are given orange juice shaken with fuller's earth, thereby removing
vitamine B, there is no influence on growth, whereas the untreated
orange juice has a far greater effect than an addition of extra
calories, in the form of an increased basal diet. This is evident
from the illustration (Fig. 37). It is interesting to note that this

THE VITAMINES IN THE ANIMAL KINGDOM

159

•experiment with children was accompanied by a similar experiment
with rats, with identical results. A second example of this kind is
seen in the work of Chick and Dalyell (462), who studied the effect
of vitamines particularly on under-nourished children in Vienna.
Here, too, the growth-promoting influence of vitamine C may

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FIG. 39. TWINS, BOY AND GIRL, WITH SCUKVY

The girl received at the point C an antiscorbutic in the form of raw milk;
later at AC, lemon or turnip juice, with butter or cod liver oil were given.
The boy was suffering from subacute scurvy and did not get any antiscorbutic
except at the place marked C, when his growth markedly improved; he suffered
also from rickets. (Dalyell; courtesy of Brit. Med. J.)

perhaps be explained by the apparent presence therein of vitamine
B. The administration of vitamine to the nursing mother, with
effect on the growth of the child, can find its parallel only in the
most exact animal experiment.

Logically, we should begin here with the description of human
avitaminoses, like beriberi, scurvy, rickets, osteomalacia, and some

160 THE VITAMINES

which are not yet classified as such, like pellagra and war edema.
Other conditions also, like ophthalmia, dental caries, and some
infectious and constitutional conditions, the progress of which seems
to be influenced by vitamine additions, should be mentioned here.
The application to human pathology is obviously the objective point
of the entire vitamine problem. We believe, therefore, that after the
demonstration of the chemistry and physiology of the vitamines, as
well as the vitamine content of the various foodstuffs, and the influ-
ence of the treatment these foodstuffs undergo before consumption,
the reader will be better able to perceive the real significance of the
vitamines in human pathology.

PART II

Chemistry, Physiology and Pharmacology
of the Vitamines

CHAPTER I
THE ANTIBERIBERI VITAMINE — VITAMINE B

In the foregoing chapters, we have discussed facts which permitted
of the assumption of the existence of at least three different vitamine
types. A great number of investigations of a chemical nature have
already made their appearance, dealing with the characteristics of
these important substances; it may indeed be said that no single
division of physiological chemistry has latterly been productive of
more publications than that of the vitamines. The reason is that
new facts and observations are obtained with an ease that is truly
astounding. Because of this, many investigators have failed to
penetrate more deeply into this subject. They have satisfied them-
selves, as a rule, with results obtained by indirect conclusions. This
condition of marking time and useless quibbling provides an excellent
opportunity for the representatives of destructive criticism which,
here more than in any other field of research, is effective in hindering
the solution of our problem. In spite of these difficulties, the knowl-
edge of the nature of the vitamines progresses gradually, and we may
hope that with the resumption of scientific research, after the war,
the desired goal — the chemical isolation of the vitamines — is not far
distant.

A question which we must touch upon is the comparison often
made between the vitamines and the ferments, a relationship
specially emphasized by Seidell (462a). Since the nature of the
ferments, in spite of the many years of study, still remains to be
cleared up, this comparison raises a poor prognosis as to the outcome
of our vitamine investigations. However, we find many far reaching
differences between these two classes of substances. Whereas we
may observe in ferments that in a temperature interval between 50°
and 70° C., there is a sudden snapping, like the over-wound main
spring in a clock, we note in vitamines that the inactivation proceeds
gradually with an increase in temperature. In general, the complete
destruction of vitamines is seen after a much higher temperature
than in ferments. The vitamines appear to us, as compared with
ferments, to have a relatively simple structure, and it would be highly
desirable to have more pure organic chemists take up this problem.

163

164 THE VITAMINES

We come now to the discussion of the difficulties involved in the
field of vitamine fractionation. Starting from any crude material
and subjecting it to a variety of chemical and physical manipulations,
we usually find that the vitamines are quite resistant. If now a
chemical fractionation is undertaken, we note that some procedures
cause either a partial or total disappearance of vitamine activity.
What then is there in the natural combination which protects the
vitamines against destruction? It has long been thought that it
was an acid which served to protect these substances, but this is
apparently only partially the case. A second possibility is that the
natural combination is protective against oxidation; this may be
tested experimentally and will be spoken of later.

The main difficulty of isolation, in our opinion, is that in most of
the starting material used till now, the vitamines are associated with
large amounts of inactive substances. When we mix a small quantity
of a known substance with a large quantity of another known sub-
tance, we find it hard to effect a separation. Here, too, we are
dealing with the law of mass action, and only those substances which
occur in large amounts in the mixture may be easily separated. Now,
if we recall that we are dealing with cellular constituents, the chemical
nature of which is still insufficiently known, then it is clear that our
problem becomes much more complicated. Just how little the con-
stituents of the cells are known becomes more evident when we
attempt a fractionation of the vitamines; we immediately encounter
cell components totally unknown to us. It is sought to avoid these
difficulties by the choice of starting material which, in addition to a
definite vitamine content, has less inactive substances. Even this
procedure is uncertain, since upon concentration of the vitamines,
there is a corresponding concentration of the impurities. There is
no method whereby we may, in one operation, remove the vitamines
from the bulk of its associated impurities, and which would be
applicable to a large quantity of crude material. Besides, it is not
impossible that the delay attending the usul chemical procedures
may have some destructive influence on the vitamines. For example,
if it should be necessary to exclude air, it is not very difficult to
provide for this for a few days, but it is another matter when the
procedures employed involve weeks instead of days. It is necessary,
therefore, that we should try to develop some color reaction or
pharmacological test for the demonstration of the vitamines, which

THE ANTIBEKIBEEI VITAMINE 165

may provide an answer for the question as to which fraction contains
the vitamine. It sometimes takes weeks before we are certain of
this, and during this time the fractions obtained must await further
manipulations. The means used till now for the study of the nature
of the vitamines (almost exclusively the more stable vitamine B)
are as follows :

1. Fractionation of the active material according to the known
methods.

2. Attempts to separate the active material by solubility and
selective adsorption.

3. Testing of vitamine-like action of combinations of known or
unknown composition.

4. Synthetic experiments with substances obtained in vitamine
work.

Up to the present, none of these four procedures has yielded very
substantial results, although the first of these methods, which is
most direct, has been productive of the most instructive findings.
By this means, some precipitation reactions have been found which
lead to a vitamine fraction which appears to be quite simple in com-
parison with the complexity of the starting material. From these
fractions, there have been isolated well defined substances which
possess a very marked curative action while in the impure state, but
which, upon further purification, lose much if not all of their activity.
Nevertheless, it has been possible to isolate a substance that has been
crystallized to constant melting point, at the same time retaining a good
deal of its activity; this was the case with our work on yeast. How-
ever, the objection is justified that the vitamine is present in
such a small quantity that it has no effect on the melting point. We
must determine now whether we are dealing with inactive material
that has adsorbed the vitamine, or with one or more cleavage prod-
ucts serving as the inactive basic structure for the vitamines. It is
obviously very unlikely that the vitamines should disappear without
leaving a trace. When radium, for example, undergoes complete
decomposition, the end product is inactive lead. It is not impossible
that in vitamines we encounter a similar problem. Our view is
further strengthened in this respect when amongst the substances
usually accompanying vitamine B, we constantly meet with products,
having the pyridine ring, which has been recognized as a cell con-
stituent for the first time in vitamine work. If we were certain

166 THE VITAMINES

that these pyridine derivatives are indicative of decomposition
products of vitamines, then it would be possible to demonstrate their
formula and chemical constitution with comparative ease; subse-
quently the synthesis could be made. Although the amount of these
pyridine derivatives is small enough to permit of a possible relation-
ship with the vitamines, our facts are not sufficiently certain to
proceed to such work, since in the end we may be dealing with
incidental impurities.

The second method of isolation, by extraction with various solvents
or treatment with adsorptive agents, has given some interesting
results, though it could finally be shown that such fractions still
contained complexes. Hence a further fractionation would again be
necessary, whereby the above mentioned difficulties would again have
to be dealt with. .

The third way, naturally only adapted to exclude from our con-
sideration substances of known composition, did not prove of value
in the studies of the nature of vitamines. Obviously, a positive
result might have been due to a contamination with vitamine, or to
a secondary effect that might be able to simulate a vitamine action.

The fourth, or synthetic, procedure has yielded no results till
now, since the investigations have been based on weak foundations.
What then is the best method for the chemical study of the vita-
mines? First of all, we must stabilize the vitamines, steps towards
which have already been taken. If this should prove to be impracti-
cable, we must look for another vitamine in Nature which is more
stable. We shall see directly that in general, vitamine B is more
stable than the two other vitamines. It would suffice if, after the
chemical purification of this substance, there remained a single
function which would characterize the substance as a vitamine. We
have already learned that all three vitamine types are present in
seeds, or that they may eventually arise through chemical re-arrange-
ment. Although we have no positive proof that the three vitamine
types are related chemically, it is not impossible that the identifica-
tion of a single vitamine may be of immeasurable help in the study
of the other vitamine types.

The vitamines have this in common, that they are all necessary
to complete a synthetic diet. Further, none of the vitamines can be
identified with any of the dietary constituents already known; they
are active in very small quantities and none of them possess any too

THE ANTIBERIBERI VITAMINE 167

great stability. It is possible, on the one hand, that we may need
new chemical methods in order to discover their chemical nature;
on the other hand, it is not unlikely that our older methods, which
have been of such use to us in the identification and isolation of so
many naturally occurring products, may lead us to the desired goal.
We have already pointed out the status of this question, up to
1911, in the historical part. We stated that till then, the investiga-
tors in this field were in doubt as to whether the factor curing beriberi
was really a chemical substance. We have shown, together with
Cooper (I.e. 68) that when pressed yeast is hydrolyzed by boiling
vigorously with 20 per cent sulphuric acid for 24 hours, a id then the
sulphuric acid completely removed with baryta, the evaporated
filtrate still exhibited a very marked curative action. This stability
in the presence of acid makes it appear that the substance in question
is an organic base, which would distinguish itself by the known
chemical characteristics of this class of substances. Based upon this
assumption, we (463) undertook a systematic investigation of rice
polishings.

CHEMICAL INVESTIGATION OF RICE POLISHINGS

Since no chemical reaction was known for the demonstration of
this vitamine, every one of the investigated fractions was tested on
beriberi pigeons. The stage at which vitamine was administered
(mostly given per os in the beginning) was the appearance of con-
tractions of the neck, wings and legs. Left to themselves, the
pigeons died in about 12 hours from the onset of the above symptoms.

A series of preliminary experiments were made in order to secure
an active solution of the simplest possible composition. This was
obtained by shaking rice polishings, which consist chiefly of cellulose,
phytin and fat, with alcohol saturated up to a certain point with
gaseous hydrochloric acid. This procedure possessed the advantage
that the solution was completely freed of alcohol insoluble material.
The alcoholic solution was then concentrated in vacuum, yielding a
fatty residue. This residue was melted on the water bath, extracted
with hot water, and the layers separated in a separatory funnel
while still hot.. The watery extract, which was very active, was
treated with sulphuric acid till a 5 per cent solution was obtained,
and then completely precipitated with a 50 per cent phosphotungstic
acid solution. The precipitate was decomposed with baryta and the

168 THE VITAMINES

resulting filtrate, after removal of the excess baryta, was tested for
its curative power on beriberi pigeons. The solution was very active
and was free of phosphorus, protein, and carbohydrates (I.e. 463).
The first difficulty encountered was that of determining the exact
dose for a sick pigeon. As will be shown later, the solution con-
tained much free choline which is very toxic for pigeons. This
difficulty could be overcome by calculating the dosage on the basis of
the quantity of rice polishings used. With every additional fraction,
the naturally occurring loss must be made up by an increased dosage.

The fact that the solution was phosphorus-free served as a final
blow to Schaumann's hypothesis regarding the lack of phosphorus,
since it was demonstrated for the first time that a solution completely
free from phosphorus, was active. Since the phosphotungstic acid
filtrate was absolutely inactive, all of the active substance must have
been contained in the precipitate.

The usual methods were used in the working up of such precipitates.
The resultant solution, which contained a large amount of potassium
and hence reacted strongly alkaline, was neutralized with hydro-
chloric acid and then concentrated to a syrup in vacuum. The
residue was taken up in alcohol, leaving the greatest part of the
potassium chloride behind. The alcoholic solution was precipitated
by sublimate, the precipitate being recrystallized from water, with
the addition of some sublimate. The crystals were then suspended
in water and decomposed with hydrogen sulfide. This solution, as
well as that obtained after the elimination of excessive sublimate, was
active (the filtrate, however, being more so). Hence it was not
possible to effect a separation with sublimate. The solution obtained
from the precipitate was concentrated in vacuum, the residue dis-
solved in alcohol and precipitated with an alcoholic solution of
platinic chloride. A series of fractions were obtained which, on
analysis, proved to be pure choline. The alcoholic solution was still
active after complete elimination of choline. It could not be doubted
therefore that the curative substance did not belong to the choline
group. In the mother liquor remaining after precipitation with
sublimate, various precipitants were tried in order to precipitate the
active substance.

For this purpose, silver nitrate in alkaline solution was used by the
author (corresponding to the precipitant for the histidine group).
The solution was freed from chlorine by silver sulfate. After the

THE ANTIBEKIBERI VITAMINE 169

removal of silver and sulphuric acid, the solution was acidified with
nitric acid, and then treated with silver nitrate till it gave a brown
precipitate with baryta.

A saturated baryta solution was then added so long as a drop of
the clear solution gave even a slight white precipitate on addition of
ammoniacal silver nitrate solution. The precipitate was then
washed free of nitric acid, decomposed with hydrogen sulphide, the
last traces of baryta removed with very dilute sulphuric acid, and
concentrated in vacuum. The solution thus obtained was very
active. By means of the gradual evaporation of the solution in the
desiccator, with the addition of alcohol, a colorless crystalline sub-
stance was obtained. This weighed 0.4 gram out of 50 kilos of rice
polishings (secured from Malay). The very small yield prevented
further work, it being possible only to make a series of reactions.
The needle-like crystals, melted sharply at 233°C. (uncorrected) and
were rather insoluble in cold water and cold alcohol. At that time,
no other substance could be found in this fraction.

As to the therapeutic action of the product, we expressed ourselves
only with great caution, but beyond all doubt there was a curative
action present. The amount used for a curative dose contained only
four milligrams of nitrogen. Smaller doses were not tried at the
time.

In a later publication (I.e. 62), we called this active substance
"beriberi vitamine," to make it evident that it contained nitrogen
and was indispensible for life.

To illustrate our procedure, we append a schematic diagram.

Rice polishings > Alcohol extract > Watery extract

S
Phosphotungstic acid precipitate > Filtrate

. / \ \

KC1 Sublimate Silver nitrate and baryta.

4 i.

Choline Vitamine.

Still later, we attempted to simplify our procedure (I.e. 324). In
the first place, the rice polishings were extracted with ordinary alcohol
instead of acid alcohol. The watery extract was then immediately

170 THE VTTAMINES

precipitated with silver and baryta, instead of phosphotungstic acid.
We observed that this extract behaved quite differently from the
previous one; it appeared to contain less potassium and no free
choline. On decomposing the silver precipitate, a substance was
obtained (not the vitamine fraction), which melted at 231°C. and
was identified as allantoine. Schaumann (464) described the isola-
tion of a substance containing nitrogen, which was perhaps allantoine
but was not investigated either chemically or therapeutically.

We have therefore isolated from rice polishings two known
nitrogenous substances, allantoine and choline, and the vitamine
fraction. The method described in our first paper on rice polishings
seems to be peculiarly adapted for the isolation of the vitamine
fraction, although it has shown the problem to be more complicated
than was apparent at first sight. We met with particular difficulty
when the rice polishings were not extracted in the laboratory but in
a chemical factory, and in large quantities. The investigation was
begun with 380 kilos of rice polishings (465) using the method
described in our earlier work on the same material. For technical
reasons, however, it seemed necessary to remove the fat first. Ordi-
nary instead of hydrochloric acid alcohol was used, and the evaporated
extract was hydrolyzed for two hours with 5 per cent sulphuric acid.
In other respects the procedure was the same as above; the individual
fractions were not investigated but careful experiments were made
with the vitamine fraction. The other substances, of no direct
concern in our problem, were kept for later work. We thought at
first that we had isolated two different substances by means of frac-
tional crystallization of the vitamine fraction, although they seemed
to crystallize out together, especially when the concentration was too
great. When the first fraction was recrystallized, needle-like crystals
were obtained which melted at 233°C. (uncorrected) and gave the
following figures on combustion according to the method of Pregl :

4.796 mgm. substance yielded 10.34 mgm. CO2 and 1.685 mgm. H2O corre-
sponding to 58.80 per cent C and 3.93 per cent H.

4.212 mgm. substance yielded 9.095 mgm. COz and 1.465 mgm. H2O, corre-
sponding to 58.89 per cent C and 3.89 per cent H.

4.367 mgm. yielded (at 713 mm. 18°C.) 0.420 cc. N corresponding to 10.58 per
cent N.

Only a small part of the nitrogen could be determined by Van
Slyke's method. Nitrogen could not be estimated by Kjeldahl's

THE ANTIBEBIBEKI VITAMINE 171

method. From the above figures, the following formula was calcu-
lated: C26H2oN409. Still, as we shall see later, these figures are
almost equally applicable for the formula of nicofcinic acid, C6H5O2N,
containing 58.3 per cent C, 4.08 per cent H, and 11.34 per cent N.
The substance seems to have been identical with that found in this
fraction earlier (I.e. 463). While the weight of the original crude
product was 2.5 grams, that of the recrystallized substance was 1.8
grams. On additional purification, the melting point was no higher.
The mother liquor was further fractionated, yielding a substance
which appeared to be more soluble in water than the first. After
many recrystallizations, it melted at 234°C. (uncorrected). When
treated with a watery solution of picric acid, this substance yielded
a somewhat insoluble picrate, which could be recrystallized from
water, melted at 218°C. (uncorrected), was light yellow in color and
crystallized in prisms and plates. The following results were
obtained on analysis:

4.217 mgm. yielded 9.625 mgm. CO2 and 1.48 mgm. H2O; 58.37 per cent C, 3.93

per cent H.
4.276 mgm. yielded 9.16 mgm. CO2 and 1.55 mgm. H2O; 58.45 per cent C,

4.06 per cent H.

3.11 mgm. yielded 0.315 cc. N (714 mm. 19.5°C.); 11.11 per cent N.
3.608 mgm. yielded 0.362 cc. N (712 mm. 19.5°C.); 10.97 per cent N.

For the formula C6H502N (123.05) there is 58.3 per cent C, 4.08
per cent H, and 11.34 per cent N. The picrate had the following
composition:

4.471 mgm. yielded 6.74 mgm. CO2 and 0.895 mgm. H2O, 41.12 per cent C;

2.24 per cent H.
2.404 mgm. yielded 0.349 cc. N (707 mm. 19°C.); 15.79 per cent N.

These figures correspond to the formula Ci2H809N4 (352.12),
which contains 40.9 per cent C, 2.29 per cent H, and 15.91 per cent N.
The composition of the substance, as well as its picrate, characterizes
it as nicotinic acid (m-pyridine carbonic acid). This substance was
first found in Nature by Suzuki, Shimamura and Odake (466), and
was later described in detail by Suzuki and Matsunaga (467). We
shall see later that the same substance was found by us in the vita-
mine fraction of yeast.

From the mother liquor of the nicotinic acid, another substance
was isolated which gave the Millon reaction in the crude state.

172 THE VITAMINES

Because of the small yield, it could not be investigated further. After
the publication of our findings, Schaumann (468) undertook some
experiments with rice polishings, using our method. He obtained
a small amount of a crystalline substance from the sublimate fraction,
and demonstrated that it was active. He was able to confirm our
work, but he could not effect a complete isolation. Later, a great
number of investigators tried to verify our findings and succeeded,
at least as far as the vitamine fraction was concerned. Among these,
we wish to mention Wellman, Eustis and Scott (469), as well as
Eustis and Scott (470). Vedder and R. R. Williams (471) obtained,
with our method, a crystalline base that was therapeutically active.
They found that unhydrolyzed extracts acted more slowly than the
hydrolyzed. Besides this, they observed that the vitamine was not
completely soluble in 95 per cent alcohol, and was destroyed by strong
alkali; it appeared also that it could not be precipitated by basic
lead acetate. The same results were obtained by R. R. Williams and
Saleeby (472), substantiating the silver nitrate-baryta method.
Later, Williams (473) described various modifications of our method.
Issoglio (474) also found that phosphotungstic acid precipitates the
vitamine. Subsequently, the isolation of vitamine from rice polish-
ings was attempted by Kondo and Gomi (475) and by Murai (476) .*
Brill (479) tried to concentrate the vitamine by adsorption with
infusorial earth, but this procedure was not very suitable. Fraser
and Stanton (480) also conducted some experiments, attempting to
extract and isolate the vitamine.

A much more interesting paper was published in 1912 by Suzuki,
Shimamura and Odake (I.e. 466). They extracted fat-free rice

1 Some patents have been taken out on the isolation of vitamines from rice
polishings. That of Tsuzuki (477) was nothing more than a concentrated
alcoholic extract of rice polishings. In addition, there is the patent of Gams
and Schreiber (478) and the identical patent of the Society for Industrial
Chemistry at Basle, which briefly is as follows : the purification of the extract
of rice polishings, which is supposed to contain very little inactive material,
consisted of a precipitation with lead acetate, first in acid and then in neutral
solution. Then the filtrate was precipitated by alkaloidal reagents, like
phosphotungstic acid or oxalic acid, and the resulting precipitate decom-
posed in a watery solution with calcium carbonate. By this means, a syrup
was obtained which was soluble in methyl but not in absolute ethyl alcohol,
ether and acetone; it gave a strong diazo reaction. The product is sold under
the name of "Orypan."

THE ANTIBERIBERI VITAMINE 173

polishings with alcohol in a reflux condenser for three hours, using
fresh solvent, till the extraction was complete. The alcoholic
extracts were combined and concentrated. The residue was diluted
with water, sulphuric acid added (till a 3 per cent solution resulted)
and precipitated with 30 per cent phosphotungstic acid. The solution
obtained by decomposing the phosphotungstic acid precipitate was
very active and was called "crude oryzanin I." The yield was 1.2
grams from 300 grams of fat-free rice polishings. This fraction cured
pigeons promptly in amounts of 3 to 4 centigrams, while the filtrate
of the phosphotungstic acid precipitate was entirely inactive. The
active fraction was soluble in water and alcohol, was strongly acid
and gave a marked Millon and diazo reaction. On the addition of
lead acetate, a slight precipitate was obtained, increasing on the
addition of ammonia. The substance was partially precipitated
by sublimate, mercuric acetate, and mercuric nitrate. An attempt
to separate this active fraction resulted in a complete loss of
activity. This was never the case in our experiments; the vitamine
was very resistant to acids.

The hydrolysis was accomplished by heating for two hours with
3 per cent hydrochloric acid. On cooling, yellowish brown crystals
separated out, which were recrystallized from hot alcohol. In
this way two substances were obtained, the first of which was less
soluble than the other. Both were difficultly soluble in water and
gave an acid reaction; they were soluble in alkali, and could be
reprecipitated on the addition of acid. On analysis, the first product
gave the formula Ci8H16N209 and was called a-acid; the second prod-
uct gave the formula Ci0H8NO4 and was called /3-acid; both showed
the Millon and diazo reactions. Unfortunately, the melting point and
the investigation of the derivatives was not recorded in the report
of the Japanese investigators. From the hydrolyzate, choline and
nicotinic acid (m.p.214°C.) could be isolated as picrates; glucose
was also found. One hundred parts of crude oryzanin gave 10 parts
of a- and /3-acids, 30 parts of choline and nicotinic acid, and 23
parts of glucose. One gram of crude oryzanin yielded 0.044 gram
nitrogen, of which 0.035 gram could be precipitated by phospho-
tungstic acid, of 0.009 gram was in the form of a- and /3-acids.

As this composition shows, the investigation of rice polishings by
the Japanese workers yielded fundamentally different results from
ours. The crude oryzanin I. was then subjected to further

174 THE VITAMINES

purification; it was dissolved in water and precipitated with a
20 per cent tannin solution till only a slight cloudiness was seen.
The precipitate was filtered off and quickly washed with a 1
per cent tannin solution. It was then rubbed up in a mortar
with 3 per cent sulphuric acid till a complete solution resulted.
This was treated with an excess of baryta, the precipitate filtered
off, and the excess of baryta removed from the filtrate with
dilute sulphuric acid. The solution was shaken out with ether and
concentrated. In this way, a preparation was obtained, called crude
oryzanin II., which was three times as active as the first preparation.
When a concentrated watery solution of this preparation was
rubbed up with a slight excess of dilute picric acid, a flocculent
precipitate settled out which became crystalline on standing in the
cold. These crystals still occluded some nicotinic acid picrate, but
with careful technique the latter remained in solution. The oryzanin
picrate was recrystallized by dissolving in cold acetone and allowing
this to evaporate in the dessicator; yellowish brown microscopic
needles, grouped in star formation, were obtained. An amount
corresponding to two centigrams of picrate was very active for
pigeons; the substance was given only to two pigeons. The amount
of picrate obtained was so slight that there was not enough for a
melting point. The question as to whether the pure oryzanin would
give the same decomposition products as oryzanin I. was therefore
left open by the authors. Since the publication of this work in
1912, nothing of a corroborative nature has been printed by the
Japanese investigators (at least to our knowledge). In the mean-
time, Drummond and Funk (481) tried to confirm the above findings,
but all attempts to isolate the substance as a picrate failed. Above
all, it was evident that an extract of rice polishings is still quite a
complicated mixture. Of the many substances it contains, we were
able to isolate, besides the previously mentioned choline, allantoine
and nicotinic acid, also betaine, adenine, guanine and apparently
guanidine. An observation, made accidently, showed us how careful
one must be in drawing conclusions from such fractionations. In this
instance, an apparently pure substance was isolated, which had a
constant melting point on recrystallization. The substance consisted
of betaine and nicotinic acid, which could not be separated from each
other by recrystallization. Only when the nicotinic acid was
separated as a copper salt, was the betaine apparent. It was also

THE ANTIBERIBERI VITAMINE 175

clear from this work, that Barger (482) was correct in his statement
that the substance described by us in 1913, to which we then gave the
formula C26H2oN4O9, was in reality nicotinic acid. However, the
view expressed in the Report of the Medical Research Committee
(I.e. 333), that the substance isolated was nicotinic acid contami-
nated with vitamine is erroneous, since the analysis indicated, at the
time, pure nicotinic acid for which no curative action was claimed.
Summarizing our work with rice polishings, we were able to differ-
entiate the curative substance of 1911, but only when we undertook
the extraction of the rice polishings ourselves. When this was done in
the factory no curative substance was obtained, and hence no pub-
lication was made, thus explaining the non-appearance of the
protocols of the animal experiments. Therefore, it is not justifiable
to apply the conclusions drawn from our negative results with rice
polishings to our positive yeast findings, which we shall describe in
the next chapter.

In conclusion we wish to call attention to an investigation by
Hofmeister and Tanaka (483). The impression was given that the
active vitamine had been isolated in the pure state from rice polish-
ings. It will be well to describe Hofmeister's method, different
from any previously used. He began his work with the notion that
our vitamine is, in reality, nicotinic acid, but this is not the case,
since in every case where nicotinic acid was isolated there was no
curative action.

Hofmeister shook rice meal three times with double the volume of
80 per cent alcohol on the shaking machine. The filtered solution
was evaporated in the presence of a stream of air, in vacuum. Then
the residue was acidified up to 3 per cent with hydrochloric acid, the
fatty acids extracted with ether, the ether removed, the solution
concentrated to a syrup in vacuum at a low temperature, and again
taken up with 80 per cent alcohol to free it from colloidal impurities.
The clear filtrate was made faintly alkaline with sodium carbonate,
taking care to prevent an excess of alkali, and precipitated by bismuth
potassium iodide (prepared according to Kraut) with constant
stirring. It is necessary here to avoid a strongly acid reaction, else
the active substance precipitates too. The dirty grayish-yellow
precipitate of the choline fraction was filtered after standing for 5
hours; to the filtrate, was added one-tenth of its volume of 20 per
cent hydrochloric acid, and the active substance precipitated out

176 THE VITAMINES

with the same reagent. The powdery, brick to scarlet red precipitate
is quickly filtered off by suction, thoroughly drained, carefully rubbed
up in a mortar with silver carbonate, and immediately filtered.
The weakly alkaline solution, containing silver, is acidified at once
with hydrochloric acid, filtered free from silver chloride and evapo-
rated in vacuum almost to dryness. The residue crystallizes in weakly
colored, radiating deliquescent crystals. From the crude hydro-
chloride, of which 5 to 10 mgm. was sufficient to cure a pigeon
in 24 hours, and which retained its activity for 8 to 10 days, a yellow
precipitate was obtained with gold chloride, which was amorphous
at first and then crystalline, and which crystallized from water in
plates and flat prisms. The quantity of aurate obtained corresponded
evidently to the amount of hydrochloride used (m. p. 273.5°C.,
which on further recry^tallization rose to 277° C., uncorrected).
This aurate was not yet entirely pure and was, therefore, converted
back to the hydrochloride and recrystallized from water; from the
purer crystals, a preparation was obtained, poor in ash, which melted
at 240°C. and gave the formula C5HuNO2. HC1. From this prepara-
tion, the pure chlor-aurate was then prepared and analyzed. The
free base, called "Oridin," is a white, powdery, hygroscopic substance,
ea ily soluble in water with a slight acid reaction; it is slightly soluble
in cold absolute alcohol but quite so in hot. From the latter, the
base may be obtained in crystals with ether. When these are heated,
pyridine-like vapors are given off. In water, precipitates were
obtained with phosphotungstic and phosphomolybdic acid, bismuth
potassium iodide and gold chloride, but not with bromine water,
platinic chloride or sublimate. The phosphotungstic acid precipitate
is soluble in acetone, but not very soluble in water. The watery
solution does not dissolve any copper carbonate and gives no color
reaction with iron salts. Isonitrile and mustard oil reactions were
negative. On boiling with hydriodic acid and phosphorus, alkaline
vapors are evolved, but not with sodium hydroxide. With calcium
hydrate a pyridine odor is obtained. The purest preparation gave
only a very faint pine splinter reaction. No color was obtained with
formaldehyde and sulphuric acid, or on heating with nitric acid and
on addition of alkali. The formula is isomeric with betaine, and the
substance is perhaps a dioxypiperidine. The pure substance was
inactive, and Hofmeister seemed uncertain as to whether the active
substance had been destroyed or had passed into the filtrate. We

THE ANTIBERIBERI VITAMINE 177

have often tried to precipitate the vitamine with gold chloride but
without succeeding. Besides, as we have lately again noted, a num-
ber of substances including nicotinic acid, are precipitated by bismuth
potassium iodide. According to Efofmeister (I.e. 4,83) colamine,
proline, hematine, papaverine, trigonelline and stachydrine have no
influence" on beriberi in pigeons. We have tried the above pro-
cedure with tomatoes but found it impractical.

CHEMICAL INVESTIGATION OF YEAST

The chemical aspect of vitamine B has been neglected in most of
the books and reviews on the subject. We propose to describe only
the indisputable facts in order to give the reader the opportunity of
forming his own opinion of the status of the question. It is too
soon to say whether the isolation of this vitamine has been successful
or not. In any case, the investigation on yeast extract made by us
is one of the few researches upon which we may build further. We
have investigated yeast in various ways, and yet we do not know if
the vitamine occurs free or combined. The facts uncovered up to
the present point to both possibilities.

In our first attempt to isolate the vitamine from yeast (I.e. 324),
75 kilos of dried yeast were extracted with alcohol, the extract con-
centrated in vacuum and the residue hydrolyzed with 5 per cent
sulphuric acid for five hours; after cooling, it was filtered from
separated fatty acids, the filtrate diluted with an equal volume of
water and precipitated with phosphotungstic acid. The precipitate
was decomposed in the usual manner, and the resulting filtrate con-
centrated. A concentrated watery solution of silver nitrate was then
added till a drop of the clear solution gave, with a saturated baryta
solution, instead of a white precipitate, a brown precipitate of silver
oxide. The solution, freed from the heavy precipitate of purine
bases, was treated with saturated baryta so long as it gave a white
precipitate with ammoniacal silver nitrate solution. The silver
precipitate was decomposed with hydrogen sulphide and the filtrate,
after the removal of the last traces of baryta, concentrated to small
volume in vaccum; the residue was transferred to a dish with a little
alcohol and allowed to stand in the dessicator, whereupon crystals
separated out weighing 0.6 gram. They were recrystallized from
dilute alcohol yielding 0.45 gram of colorless needles (see fig. 40)

178

THE VITAMINES

melting at 233° C. This substance from yeast was therapeutically
active; 2 to 4 centigrams were sufficient to cure a sick pigeon in a
short time; smaller doses were not tried. The substance could be
precipitated by mercuric acetate but not by the nitrate or sulphate.
It gave no precipitate with a nitron solution — showing the absence
of nitric acid — and had a neutral reaction. No blue cop*per salt
was obtained with copper oxide, which would indicate that the
substance is not an amino-acid. The filtrate from the silver pre-
cipitate was inactive.

FlG. 40. MlCROPHOTOGRAPH OP CRYSTALS OBTAINED FROM VlTAMINE FRACTION
OF YEAST, POSSESSING CURATIVE ACTION

Another batch of yeast was treated in a somewhat different manner.
The alcoholic extract, after concentration in vacuum, was simply
taken up with water, and not hydrolyzed as above. Thus the
vitamine was obtained in watery solution. This was then pre-
cipitated with phosphotungstic acid and treated as above; in this
case however, the silver fraction did not yield an active product, a
substance being isolated which could be identified as uracil. All the
vitamine, in this case, was found in the filtrate from the silver nitrate
precipitate, which might mean that the vitamine of yeast occurs

THE ANTIBERIBERI VITAMINE

179

chiefly in combination. This filtrate was then hydrolyzed with
sulphuric acid, and the subsequent procedure again repeated. Here
too we met with a mixture of pyrimidine bases, among which thymine
could be demonstrated, so that finally after a long procedure, only a

FIG. 41. PIGEOX BEFORE TREATMENT

FIG. 42. THE SAME PIGEON (CF. FIG. 41) CURED WITH 8 MGM. OP YEAST
VITAMINE AFTER Two HOURS

trace of active substance was found. This observation shows that
in working with vitamines, the number of manipulations had best
be kept down to a minimum, if results are to be obtained.

Later on (485, and I.e. 465), we again investigated the vitamine
fraction, using 2.5 grams of the crude fraction, obtained from 100

180

THE VITAMINES

kilos of dried yeast, according to the method described. The sub-
stance was obtained from the decomposition of the silver baryta
fraction, and possessed a crystalline structure, melting at 210° C.
(uncorrected). This crude vitamine fraction was a splendid thera-

FIG. 43. PIGEON BEFORE TREATMENT

FIG. 44. SAME PIGEON

(CF. FIG. 43) AFTER THREE HOURS, WITH 4 MGM.
OF YEAST VITAMINE

peutic agent in a great number of beriberi pigeons. As the accom-
panying photograph shows (fig. 41-46), the preparation cured pigeons
completely in a very short time (2 to 3 hours). Further data on this
subject will be found in the protocol appearing at the end of this chap-
ter, showing, in the form of a table (p. 185), the action of substances

THE ANTIBERIBERI VITAMINE

181

obtained from yeast. The dose of crude fraction used was 4 to 8
mgm., and the preparation was administered intramuscularly. The
product gave a strong uric acid and phenol reaction with the Folin-
Macallum reagents (486).

FIG. 45. PIGEON BEFORE TREATMENT

FIG. 46. SAME PIGEON (CF. FIG. 45) AFTER THREE HOURS, WITH 8 MGM.

YEAST VITAMINE

This 2.5 grams of crude fraction was then recrystallized from
dilute alcohol, and gave, at first, 1.6 grams of a substance crystallizing
in microscopic needles. Even after several recrystallizations, the
melting point could not be raised above 229° C. (corrected). The
first crystallization still gave the reactions mentioned above; only

182 THE VITAMINES

when the substance was recrystallized twice did these reactions dis-
appear. The purified substance was compared with that obtained
before, and mixed melting points were taken, whereby the new
product melted four degrees lower. Mixed in equal quantities, both
substances melted at 229°C. (corrected). They were absolutely
identical as regards solubility, reactions and crystalline form. The
substance gave a white precipitate with Millon's reagent, but the
color reactions were always negative. In a 0.74 per cent solution,
a definite rotation in the polarimeter could not be shown. On
drying in vacuum at 100° C. and analyzing, the following figures
were obtained:

3.267 mgm. yielded 6.63 mgm. CO2 and 1.05 mgm. H2O; 55.35 per cent C, 3.60

per cent H.
4.224 mgm. yielded 8.425 mgm. CO2 and 1.36 mgm. H2O; 55.72 per cent C, 3.69

per cent H.
4.256 mgm. yielded 8.66 mgm. CO2 and 1.36 mgm. H2O; 55.50 per cent C, 3.58

per cent H.

3.048 mgm. yielded 0.373 cc. N. (710 mm. 15.5°C.) 13.53 per cent N.
3.627 mgm. yielded 0.430 cc. N (712 mm.; 14°C.) 13.21 per cent N.
3.286 mgm. yielded 0.401 cc. N (705 mm.; 18°C.) 13.28 per cent N.

These figures correspond to the formula C24Hi9O9NB (521.24),
which gives 55.25 per cent C, 3.68 per cent H and 13.44 per cent N.
Since a definite crystalline substance is noted on the tube when drying
at 100° C. in vacuum, analyses were made after the substance was
dried in vacuum over sulphuric acid; the following figures were
obtained :

3.733 mgm. gave 7.775 mgm. CO2 and 1.185 mgm. H2O; 56.80 per cent C, 3.55

per cent H.
3.538 mgm. gave 7.35 mgm. C02 and 1.195 mgm. H2O; 56.66 per cent C, 3.78

per cent H.
3.174 mgm. gave 0.372 cc. N (702 mm.; 17°C.) 12.74 per cent N.

These figures correspond to the formula C26H2i09N5, which gives
57.01 per cent C, 31.87 per cent H and 12.81 per cent N. From this,
it appears that on drying at higher temperatures a substance rich in
carbon is lost. This characteristic and the great difference in
elementary composition excludes the possibility that this substance
was nicotmic acid. We have recently attempted to secure further
evidence in this connection by the use of the yeast method. If the
nitrogen in this substance is estimated by the method of

THE AJSTTIBERIBERI VITAMINE 183

Kjeldahl, only a third of it is determined. 0.1328 gram needed
4.0 cc. -rk H2S04; found, 4.21 per cent N. If the substance is
titrated with sodium hydroxide against phenolphthalein, then
0.0373 gram requires 2.8 cc. TTT NaOH. For a four basic acid
of the formula C24Hi909N5, the calculated amount is 2.8 cc. That
this substance is of an acid nature, is very important and we shall
consider this phase of the matter again. As regards the therapeutic
action of substance I, examples are given in the table (p. 185).

The filtrate and mother liquor combined, of substance I, which
gave a strong phenol and uric acid reaction, were precipitated by
picric acid; a crystalline precipitate was obtained, amounting to 0.9
gram; which was first recrystallized from a mixture of acetone and
alcohol and then from water. The once recrystallized substance still
gave a positive phosphotungstic acid reaction. On purification (light
yellow needles), the melting point was raised from 217° to 219° C.
(unconnected) and gave the following figures on combustion :

4.70 mgm. gave 7.115 mgm. CO2 and 0.98 nag. H2O; 41.29 per cent C; 2.33 per

cent H.

2.966 mgm. gave 0.436 cc. N (708 mm., 19.5°C.), 15.99 per cent N.
4.212 mg. gave 0.61 cc. N (711 mm., 18°C.), 15.89 per cent N.
0.1061 grain gave 0.1652 Nitron picrate, 65.91 per cent picric acid.
0.1261 grain gave 0.1951 Nitron picrate, 65.49 per cent picric acid.

Calculated for Ci2H809N4 (352. 12) -40.9 per cent C, 2.29 per cent
H, 15.91 per cent N and 65.05 per cent picric acid. This picrate was
quantitatively decomposed with nitron, and on concentrating gave
0.45 gram of silky, lustrous, colorless needles, the melting point of
which, after recrystallization, rose to 235° C. (uncorrected) . The
crystals were very easily soluble in water and gave no color reactions.
When dried at 100°C. in vacuum, the analysis gave the following
figures :

3.432 mgm. yielded 7.345 mgm. CO2and 1.10 mgm. H2O; 58.37 per cent C;3.62

per cent H.
3.784 mgm. yielded 8.11 mgm. CO2 and 1.325 mgm. H2O; 58.45 per cent C.

3.92 per cent H.

3.415 mgm. yielded 0.343 cc. N. (702 mm., 18.5°C), 10.87 per cent N.
2.562 mgm. yielded 0.252 cc. N (15.5°C.), 10.92 per cent N.

Calculated for C6H5O2N (123.05) -58.3 per cent C, 41.08 per cent
H and 11.34 per cent N. The substance was evidently nicotinic acid.

184

THE VITAMINES

Mixed with the nicotinic acid from rice polishings, it showed the same
melting point, and the picrates behaved similarly.

The mother liquor of the picrate precipitate, which gave a strong
uric acid and phenol reaction, was shaken out in acid solution with
ether in order to remove the picric acid. After elimination of the
hydrochloric acid, the solution was slowly evaporated, yielding
0.4 gram of a substance melting at 210° C. and when recrystallized
from dilute alcohol to constant melting point, melted at 222 to 223° C.
(microscopic needles, somewhat more soluble in water than substance
I). The pure substance no longer gave the uric acid nor the Millon
reaction, but the phenol reaction was still positive. For analysis,
one sample was dried in the vacuum dessicator over sulphuric acid at
ordinary temperature; another was dried in vacuum at 100° C. and
another, at 115° C. The substance dried over sulphuric acid gave
the following figures:

3.559 mgm. gave 7.745 mgm. CO2 and 1.21 mgm. H2O; 59.35 per cent C; 3.80

per cent H.
3.445 mgm. gave 0.375 cc. N (706 mm. 17°C.), 11.90 per cent N.

Dried in vacuum at 100°C.
4.172 mgm. gave 9.065 mgm. CO2 and 1.535 mgm. H2O; 59.26 per cent C; 4.12

per cent H.
2.716 mgm. gave 0.295 cc. N and (711 mm. 15°C.) 12.04 per cent N.

Dried in vacuum at 115°C.

3.694 mgm. gave 7.99 mgm. CO2 and 1.22 mgm. H2O; 58.99 per cent C; 3.70 per
cent H.

These figures correspond equally well to two formulas, though
somewhat better to the second.

CALCULATED FOB

C2«Hi8O7N4 (462.22)

CjoHwO.Ns (585.28)

C

per cent

59.71
3.84
12.12

per cent

59.46
3.96
11.97

H

N :

To recapitulate briefly the yeast fractionation, we have been able
to isolate three different substances from the vitamine fraction,
which gives a strong uric acid and phenol reaction.

THE ANTIBERIBERI VITAMINE

185

SUBSTANCE

MELTING
POINT

URIC ACID
REACTION

PHENOL
REACTION

REMARK

C.24Hi9O9N5 (dried at 100°C)
C-26H2iO9N6 (dried at ordi-
nary temperature)

°C.

229
223

-

-

Gives no picrate
Gives no picrate

C29H23Oi,N5

222-223

+

Gives no picrate

C6H6O2N

235

_

Gives picrate

Nicotinic acid

Aside from this, the first substance, on heating in vacuum, loses
a small amount of substance rich in carbon, giving lower analytical
results. Although the statement is often made that we had obtained
only impure nicotinic acid, it is not justified, since the analytical
figures are too far apart from the composition and the characteristics
of this acid. Regarding the therapeutic action of these substances,
many experiments were performed, some of which naturally turned
out negative. In the table, the product C26H2iO9N5 is called
substance I, and Cagl^sOgNs, substance II.

NUM-
BER OF
ANI-
MALS

SUBSTANCE

DOSE

ACTION

DATS
SUR-
VIVED

mgm.

1

Crude crystallization

4

Cured in 3 hours

4

2

Crude (fig. 41-42)

8

Cured in 2 hours

6

3

Crude (fig. 43-44)

4

Cured in 3 hours

4

4

Crude (fig. 45-46)

8

Cured in 3 hours

4

5

Substance I

8

Partially cured in 7

hours.

3

6

Substance I

4

Improvement

4

7

Substance I

2

Cured in a few hours

3

8

Substance I + Nicotinic acid

5+2

Cured in 2J^ hours

4

9

Substance I + Nicotinic acid

4+2

Cured in 3 hours

5

10

Substance I + Nicotinic acid

3+2

Cured in 2 hours

6

11

Substance I + Nicotinic acid

4+2

Cured in 2J/2 hours

7

12

Substance I + Nicotinic acid

3+2

Cured in 4 hours

4

13

Nicotinic acid

10

Negative

2

14

Nicotinic acid

5

Slight

3

15

Nicotinic acid

4

Negative

2

16

Nicotinic acid

4

Negative

1

17

Substance II

5

Negative

—

18

Substance II

5

Negative

1

186 THE VITAMINES

All these substances were administered to the pigeons intra-
muscularly, and from the results, it is evident that substance I had
the greatest therapeutic action.

Others also investigated this question. Barsickow (487) concluded
that the therapeutic action was inherent only in the living or dead
yeast cells, and for the most part, he found the extracts inactive;
this has, of course, been shown to be incorrect. Edie, Evans, Moore,
Simpson and Webster (488) did not hydrolyze the yeast, but treated
the alcoholic extract, after the concentration of the solvent, directly
with silver nitrate and baryta. They obtained in this way a hygro-
scopic syrup which was very active for pigeons, 6 mg. being suffi-
cient, as a rule, for a cure. On further purification, a crystalline
substance was obtained which, because of the small yield, could not
be purified further. This crystalline substance was analyzed but the
data on its therapeutic action were lacking.

Another method, useful for the purpose of removing a large mass of
inactive material, was described by us (489) in 1916. This method is
based upon the separation of the phosphotungstates by means of
the varying solubility in acetone. The residue, obtained from the
alcoholic extract of 100 kilos of dried yeast, was extracted with 10
per cent sulphuric acid, this extract-was then diluted half again with
water and precipitated with phosphotungstic acid. The resulting
precipitate, weighing 2237 grams was triturated several times with
dry acetone, leaving in the end 144 grams of insoluble material. This
portion, which represents 5.1 per cent of the total precipitate, con-
tained all the vitamine, according to our animal experiments. The
precipitate was decomposed with neutral lead acetate2 and the
resulting solution precipitated with picric acid. The voluminous pre-
cipitate consisting of adenine picrate was filtered off. It was impos-
sible to crystallize the vitamine from the mother liquor, making it
necessary to continue the fractionation with sublimate, platinic
chloride, and picrolonic acid. The course of the fractionation, which
was controlled by animal experiments, is shown in the diagram
below, in which the double lines indicate the fraction containing the
vitamine. According to a patent by Bohringer and Sons (491) the

2 We also attempted to decompose the precipitate by shaking it out with
amyl alcohol in acid solution according to Vah Slyke (490), but the method
was not quantitative because the last traces of phosphotungstic acid are
removable only with great difficulty.

THE ANTIBERIBERI VITAMINE 187

Phosphotungstic acid precipitate from yeast.

Acetone insoluble part Acetone soluble part

Picrate Picric acid filtrate

l|

II \

Sublimate precipitate Sublimate filtrate

Platinic chloride precipitate

Filtrate

Sublimate precipitate Sublimate filtrate

phosphotungstic acid in the above method was substituted by phos-
photitanic acid. Cooper (492) then showed that when pressed yeast
is allowed to stand for 35 hours at 35° C., the mass becomes
semi-liquid and on filtration from the cell residue, a. liquid called
"autolyzed yeast" is obtained, which contains almost all of the
vitamine of the yeast. It was shown later that the yeast con-
stituents are not completely autolyzed, since if the liquid is heated
to 60 to 70° C., an appreciable amount of protein separates out.
When the acetone method was applied to this liquid, the amount of
the acetone insoluble portion was much larger — about 1125 grams
from 5 kilos of pressed yeast. We then showed that when the
phosphotungstates of choline, betaine, stachydrine, guanine, adenine,
guanidine, creatinine and nicotinic acid were tested for their
solubility in acetone-water solutions of various dilutions, the sol-
ubility was as follows :

25 per cent acetone — > 100 per cent — > 50 per cent — > 75 per cent

Choosing these four concentrations, the solubility was least in 25
per cent acetone and greatest in 75 per cent acetone. We were then
able to show, in our work with Dubin (493), that autolyzed yeast
contains much more vitamine than vitamine extracts obtained from
yeast by various solvents. For this reason, autolyzed yeast would
be superior to all other extracts for the purpose of vitamine fraction-

188 THE VITAMINES

ation, if a large quantity of inactive extractives did not go into
solution at the same time. If the phosphotungstic acid precipitate
from autolyzed yeast is treated directly with dry acetone, almost a
third of it remains insoluble, including a large amount of purine
derivatives. If the acetone-water method is used, as outlined above,
we obtain from the above-mentioned precipitate, weighing 1125
grams, the following fractions:

ACETONE CONCENTRATION

AMOUNT SOLUBLE

per cent

grams

per cent

25

522

46.4

100

190

16.8

50

155

13.7

75

108

9.6

Total precipitate — 1125 grams.

Insoluble residue — 150 grams = 13.3 per cent.

This method might be of importance but in this case, no animal
experiments were carried out.3

A method indicative of great progress at the time was described
by Seidell (495). If, for example, 1 liter of autolyzed yeast is shaken
with 50 grams of Lloyd's reagent (fuller's earth), allowed to stand for
several hours and the residue filtered off and washed with dilute
hydrochloric acid, vitamine B is quantitatively removed from the
solution, and the filtrate is completely inactive. The great hopes
held out for this method, however, have not materialized. In the
first place, it is very difficult to remove the vitamine from the fuller's
earth, and secondly, a number of other substances, difficult to remove,
are adsorbed together with the vitamine. In the end, R. R. Williams
and Seidell (496) were able to extract the vitamine from the fuller's
earth with 5 per cent sodium hydroxide, at the same time showing
that the vitamine was contaminated with considerable adenine.
Osborne and Leavenworth (496a) showed that dilute alkali has
no destructive action of vitamines, especially if the contact is of
short duration. For liberation of vitamine from activated fuller's
earth all those methods are suitable which are used in freeing alkaloids

3 In the meantime, this method was elaborated by Drummond (494) in
that he investigated the solubility of various phosphotungstates, and dis-
cussed the practical applicability of these data.

THE ANTIBERIBERI VITAMINE 189

from fuller's earth preparations by Rhodehamel and Stuart (496b).
Recently, Seidell (496c) has combined the fuller's earth method
with our silver method and has obtained some promising results.
He liberates the vitamine with a baryta solution, the procedure
being hastened as much as possible. From the filtrate, the purine
bases are eliminated with silver nitrate while the vitamines are
precipitated by ammoniacal silver nitrate. This fraction can be
reprecipitated in the same way and can be analyzed for its silver
content. A crystalline substance was isolated from this fraction,
but it proved inactive, showing it to be a fairly complicated mixture.
We found that nicotinic acid is also precipitated by ammoniacal
silver nitrate. For the liberation of vitamine from activated fuller's
earth, we have likewise used ammonia and pyridine; we were also
able to confirm the findings of Eddy, Heft, Stevenson and Johnson
(I.e. 129e) that vitamine can be adsorbed with a specially pre-
pared charcoal from which it can be liberated with glacial acetic
acid. Voegtlin and White (497) adsorbed the vitamine with mastic,
and with colloidal arsenic sulfide, in which case the impurity was
guanine instead of adenine.

Drummond (498) attempted to confirm our findings on vitamine
B, and although he was able to do so for the most part, he concluded
that our chemical work was of no particular significance, because he
held that the vitamines were not precipitated by the various pre-
cipitation methods, but were adsorbed by the precipitates. The
characteristic, adsorbability, is peculiar to many pure chemical sub-
stances. We need only recall Lloyd's reagent which, as we have
personally proved by tests on rats, quantitatively adsorbs strychnine
and other alkaloids. Pure glucose is also adsorbed in very appre-
ciable amounts by various kinds of blood charcoals. Besides, Drum-
mond carried out the silver nitrate-baryta precipitation in a manner
different from ours, and used rats as experimental animals, which,
as we shall see, behave differently than do pigeons and chickens.

Abderhalden and Schaumann (499) have recently published con-
siderable work on yeast, which unfortunately is described so
unclearly that it is hard to state whether or not progress has been
made. These authors declare, in accord with the phosphorus theory
of Schaumann, that vitamine B of yeast occurs as a nucleoproteid.
They were able to isolate an active nucleoproteid from yeast.
Pigeons receiving a half gram of this substance daily lived for 70

190 THE VITAMINES

days. We have already seen that such findings possess no special
significance, since the activity of the nucleoproteid might have
been due to adsorption, as with fullers earth. The authors also
assume that the mother substance of the vitamine varies according
to the source of material. The vitamine was obtained from yeast
extracts by precipitation with acetone, and purified by several
reprecipitations. This crude fraction was investigated physiologi-
cally, and found curative for beriberi, but it could not keep pigeons
alive for any prolonged period. Another preparation was obtained
from yeast by treatment with alkalis. This substance did not cure
beriberi, but when added together with the acetone preparation, its
effect was to prolong life. A number of substances were isolated from
the acetone preparation, by reprecipitating with sublimate, among
which there was apparently only a single new substance obtained in
the pure state. This base, called "Aschamine" had the structure of
dimethylpropenylamine, the formula being

CH = CH • CH2

/
C5Hi3O N = OH— N— CH3

\

CH3
H

it was inactive for beriberi. In the other fractions, betaine was
found as an impurity. All of these substances were prepared from
hydrolyzed yeast, and the authors believed that the active substance
belonged to the betaine group, having its characteristic structure.
In a later experiment, Abderhalden (500) states that the active
substance cannot be quantitatively removed from yeast by absolute
alcohol or absolute alcohol and acetone.

Sugiura (501) placed yeast and also carrot extracts in collodion
sacs, and permitted the contents to dialyze through the membrane,
whereupon the product crystallized on the outside of the membrane.
It was evidently hoped to prepare the active substance in the pure
state by this simple procedure, but naturally the entire yeast mixture
dialyzed out.

Osborne and Wakeman (502) described a method which promised
greater success. The procedure consisted chiefly in eliminating
the autolysis of the yeast cells by washing them and boiling with
water, to which some acetic acid has been added, thus coagulating

THE ANTIBERIBERI VITAMINE 191

the protein and obtaining an extract free from the products of auto-
lysis. Concretely, 4.5 kilos of fresh yeast, corresponding to 830
grams of dry yeast, was crumbled into boiling water, the mass centri-
fuged, the residue washed with water and again centrifuged. Alto-
gether, 15 liters of water were used, the combined extracts were
evaporated to two liters and poured into three liters of 93 per cent
alcohol, giving a 53 per cent alcoholic content by weight. The pre-
cipitate, which was inactive, was filtered off and the filtrate, together
with the washings, evaporated to 300 cc. This liquid was poured
into 1960 cc. of 93 per cent alcohol, giving a 79 per cent alcoholic
content. The second precipitate was dissolved twice in water and
precipitated with alcohol so that the alcoholic content should be
90 per cent. This fraction was supposed to contain almost the
entire quantity of vitamine B present in the yeast, and amounted to
6.2 per cent of the dry yeast. The preparation had an acid reaction
and gave a heavy precipitate with lead acetate and sublimate; 25
per cent of this fraction could be precipitated with silver nitrate-
baryta. This work shows that the vitamine of non-hydrolyzed yeast
is insoluble in strong alcohol. In our experiments this method did
riot prove successful, as in each fresh sample of yeast the limits of
precipitation have to be established anew.

There is also the report on the chemistry of yeast by Myers and
Voegtlin (503). They showed that vitamine B could be extracted
from yeast by shaking the latter with olive oil; the procedure obvi-
ously possesses no particular advantage over the usual extraction, so
that later they used methyl alcohol acidified with hydrochloric acid
for extraction. The resulting extract was concentrated in vacuum
and the residue extracted several tunes with dilute hydrochloric acid.
In this extract, the purine bases were precipitated with silver acetate.
The precipitate was filtered off and more silver acetate added to the
filtrate, followed by a saturated solution of baryta. This precipitate
was decomposed in the usual manner, and the histidine removed from
the filtrate with mercuric sulphate. The vitamine was then pre-
cipitated from the filtrate with absolute alcohol. The precipitate
was dissolved in water and freed from mercury, whereupon a very
active solution was obtained. This solution gave a brown diazo
reaction and contained a product similar to histamine. When the
solution was concentrated, crystals were obtained which were active
only so long as they contained some mother liquor. On washing the

192 THE VITAMINES

crystals with alcohol, the crystalline form changed from spindles to
prisms. If these prisms were recrystallized from water, spindles
were again obtained. In this experiment, the vitamine was inac-
tivated by drying. The adsorption methods were shown to be
unspecific.

OTHEE SOURCES OF SUPPLY

Hulshoff Pol (I.e. 59 and 504) reported further experiments with
theX-acid of Katjang idjoe beans (Phaseolus radiatus) without giving
more details on the chemistry of the substance. In 1912, we inves-
tigated milk (I.e. 324) with this point in view. As a source of supply,
we used a dried milk preparation, sold under the name "Trumilk."
This was extracted with alcohol and ether. The combined extracts
were concentrated and the residue hydrolyzed with 10 per cent
sulphuric acid for 5 hours. In this way, 51 grams of phosphotungstic
acid precipitate were obtained from 1398 grams of dried milk. The
silver-baryta fraction was prepared from this precipitate in the usual
manner. The decomposed precipitate yielded a small amount of
crystals melting at 230° C., and was curative for pigeon beriberi. In
the same way, it was possible to prepare from 2180 grams dried ox
brain (I.e. 324) 220 grams of phosphotungstic acid precipitate, and
to obtain a trace of a crystalline substance from the silver-baryta
fraction, melting at 203° C. and having curative properties. Voegtlin
and Towles (505) investigated extracts of spinal cord and found that
autolyzed extracts were more active than natural extracts. .

From 42 liters of commercial lime juice (I.e. 324), we obtained 1200
grams phosphotungstates, from which there resulted 5.9 grams of a
vitamine fraction. Although this fraction did not crystallize out, the
solution was very curative for pigeon beriberi. From our informa-
tion of the vitamines in the above mentioned publication, the
presence of vitamine B could indeed be demonstrated, and yet the
crystalline substance itself may perhaps have been impure nicotonic
acid contaminated with vitamine.

Sullivan and Voegtlin (506) fractionated wheat chaff and extracts
of peas and ox liver. The residues from the respective alcoholic
extracts were hydrolyzed with 5 to 10 per cent sulphuric acid for five
hours in a stream of CO2. The precipitates obtained with phospho-
tungstic acid were then decomposed, either directly or after dissolving
in 50 per cent alcohol, with neutral lead acetate. After the elimina-

THE ANTIBERIBERI VITAMINE 193

tion of lead, the filtrate was precipitated by an excess of silver acetate
and treated with baryta, after removal of purine precipitates.

Steenbock (507) prepared from egg-yolk a substance soluble in
water-acetone, active and not quantitatively precipitated by phospho-
tungstic acid. McCollum and Simmonds (I.e. 315) sought to con-
centrate vitamine B by extraction with various organic solvents.
They found that beans could be extracted with ether, benzol, or
acetone, without dissolving the vitamine. Subsequently it can be
extracted with alcohol and the solution, when concentrated, may be
taken up by benzol. The same method was applied to wheat germ
and pigs' liver.

SYNTHETIC EXPERIMENTS

Although we undertook some synthetic experiments with nicotinic
acid, based on its occurrence in the vitamine fractions obtained from
various sources, we perceived that theoretical reasons for this trend of
thought were hardly justifiable. Nevertheless, Williams (I.e. 473),
instituted such experiments. He condensed p-oxy nicotinic acid with
itself or with nicotinic acid, and believed that the first condensation
product was much more active for avian beriberi than all other
similarly synthesized condensation products. Thereupon, Williams
(508) proposed the very attractive theory that oxy-pyridine and
pyridine carbonic acid occur in two isomers, which possess different
crystalline forms, and which may be transformed into each other
under certain conditions. The labile form remained intact for some
days and in this case was therapeutically active; then it changed into
the inactive stable form. Later, Williams (509) extended his theory
to include such pyrimidine and purine derivatives as were theoreti-
cally capable of forming the betaine ring. Williams and Seidell (I.e.
496) tried to convert adenine obtained from autolyzed yeast by ad-
sorption with fuller's earth, by boiling with acetic acid and acetic anhy-
dride, into an isomeric form. This labile form was supposed to have
a therapeutic action; after recrystallization, this peculiarity was sup-
posed to disappear, the substance being changed back again to the
usual adenine. The theory of Williams explained the slight stability
of vitamine B very beautifully, but unfortunately it could not be con-
firmed. Voegtlin and White (I.e. 497) sought in vain to produce an
active product from adenine, while Harden and Zilva (510) tried to
do the same with o-oxypyridine and adenine. Since the course of

194 THE VITAMINES

reasoning pursued by Williams could not be confirmed, and since
he has published no further related work since 1916, the subject may
apparently be considered as closed.

STABILITY OF THE ANTIBERIBERI VITAMINE AGAINST HEAT AND
CHEMICAL AND PHYSICAL AGENTS

As far back as the work of Grijns (I.e. 52) we knew that the
vitamine B from Katjang-idjoe beans lost its curative properties on
heating to 120° C. Ever since, a number of investigators have
occupied themselves with this problem. It was soon evident that
the stability was dependent upon a number of factors. Among these
was the vitamine content of the starting material used, the chemical
reaction of the substrate and the duration of the heating period.4
Although these points are of great practical importance as regards
the etiologic and dietetic significance, they are only of moderate
theoretical interest, so long as the vitamine itself has not been pre-
pared in the pure state and its characteristics determined. Only
then shall we be in a position to conduct such experiments exactly.
From the description of the isolation experiments, it is clear that this
vitamine is quite stable and can withstand a number of chemical and
physical manipulations. Therefore, it is hard to determine at which
stage of the work this substance loses its activity. In the fraction-
ation, there is very little loss of activity till we come to the last
stages, where it decreases rapidly. One almost has the impression
that the substances accompanying the vitamine serve to stabilize it.
We must admit that we are still in the dark as to the cause, and
although this may eventually find quite a simple explanation, it
seems to us for the present remarkable and mysterious.

As regards the solubility of vitamine B, this has already been
spoken of in the preceding pages. All investigators apparently agree
that this substance is insoluble in strong alcohol. There are also
some experiments which indicate that prolonged boiling in alcoholic
solution may inactivate this vitamine. As to the effect of heat, the
findings vary ; similarly, with the effect of alkalis. On the other hand,
the investigators are one in stating that this vitamine is very resistant
to acid. Among other chemical reagents, diazotization, according to

4 Emmett and Luros (I.e. 94) give a good bibliography relative to this
question.

THE ANTIBERIBERI VITAMINE 195

McCollum and Simmonds (I.e. 315), does not affect the activity of
vitamine B, and this has recently been confirmed by Funk and Dubin
(I.e. 493) who also showed that the substance stimulating the growth
of yeast was not destroyed by reduction with palladium and hydrogen,
as well as with zinc and hydrochloric acid ; neither was it affected by
oxidation with finely divided platinum and oxygen for 7 to 8 hours.
Besides this, we (512) showed in 1916 that the action of radium in
doses used in radium therapy is without destructive influence on vita-
mine B originating from autolyzed yeast. In accord with this, there
is the work of Zilva (513) who investigated the effect of ultra-violet
light on this substance. Contrary to the above findings, Sugiura
and Benedict (514) report that large doses of X-rays of radium almost
obliterates the vitamine of autolyzed as well as dried yeast. Weill
and Mouriquand (515) carried out analogous experiments with
Rontgen rays on barley kernels.

DEMONSTRATION AND ESTIMATION OF ANTIBERIBERI VITAMINE

At the outset of vitamine investigations, only one method was
available for the demonstration of vitamine B. Beriberi was pro-
duced in chickens and pigeons (the latter being more commonly
used) by feeding white rice, and then the preparation to be tested
was given either per os or intramuscularly.5 The latter method had
the advantage of giving a more speedy effect and of making it certain
that the solution was utilized by the animal, and had not trickled
out of the beak. A negative finding of this therapeutic measure
did not entirely indicate that the solution to be tested was inactive.
Negative results may be due to the fact that the animal had pro-
gressed too far to be affected by the above measures or they might
be due to the presence of toxic products. It was chiefly through the
efforts of Osborne and Mendel, as well as McCollum and his co-
workers, that the second method was then developed, of which we
have already spoken in the chapter on the vitamine requirements of
rats. This method consists in adding the preparation to be tested
to a diet complete in everything but vitamine B. If growth was
obtained with young rats, then the presence of this vitamine was

6 We were one of the first to make use of the parenteral administration of
this vitamine. Unfortunately, we have been unable to discover in the liter-
ature by whom this method was first introduced.

196 THE VITAMINES

demonstrated. This method is more certain, though it has the draw-
back that much more time is required for its application. Both of
these methods have the great disadvantage that in isolation experi-
ments, every new fraction must await the findings of the animal
experiments, the vitamine preparation in the meantime being subject
to more and more decomposition. For these reasons, efforts have
long been made to develop a method that would permit of conducting
a test in the shortest possible time. For practical reasons too, such
a method would be of advantage in determining the vitamine- content
of various foodstuffs.

The first step in this direction was made by Fraser and Stanton
(I.e. 55) . Based upon Schaumann's theory of the lack of phosphorus,
they believed that the vitamine content of rice could be estimated
by the amount of phosphorus present. Voegtlin and Myers (516)
also suggested the phosphorus content of wheat and corn as a some-
what reliable indication of the amount of vitamine contained therein.
On the other hand, Green (517) showed that if the phosphorus
content of American corn is taken as a unit, then all South African
varieties of corn, which are very poor in phosphorus, might be
regarded also as very poor in vitamines, which is, however, not the
case. Ottow (518) is likewise of the opinion that the estimation of
the phosphorus content alone may lead to false conclusions. He
believed that the determination of the quantity of alcohol-soluble
fraction of rice was more reliable, at the same time regarding the use
of animal experiments as the only method to be relied upon.

We (519) sought, at first, to form some conception as to the
amount of vitamine present, by determining the nitrogen in foodstuffs
having a slight non-protein nitrogen, such as milk. For this purpose,
we dried some milk, made an alcoholic extract of the portion precipit-
able by phosphotungstic acid, and analyzed it for its nitrogen content.
The only result apparent was that on centrifuging the milk, a
large part of the residual nitrogen is lost and evidently goes over
into the cream. Whereas in a non-centrifuged milk this fraction
amounts to about 2.2 mgm. per liter, in the centrifuged milk it is
only 1.4 mgm. This shows perhaps that milk fat (butter) may
contain nitrogenous vitamine. Furthermore, since we noted that
vitamine fractions contained substances whose nitrogen could be only
partially estimated by means of a Kjeldahl determination, we wanted
to make use of the difference between the analyses, according to

THE ANTIBERIBERI VITAMINE 197

Dumas and to Kjeldahl, as an indication of the vitamine fraction.
However, since this fraction contained a relatively large amount of
pyridine derivatives, the method is valueless so long as we do not
know the relationship between these substances and the vitamines.
Brill and Alincastre (520) tried, in this way, to average the maximum
vitamine content from the phosphotungstic acid precipitates of
various vegetables. Seidell (521) sought to determine the vitamine
content of yeast by estimating the nitrogen in activated fuller's
earth. Eddy (522) tried to do the same with an activated fuller's
earth out of pancreas. Since fuller's earth adsorbs also a large
amount of inactive material, this method is of no special consequence.
In discussing the chemistry of vitamine B, it will be recalled that
we have often made use of a reaction introduced by Folin and
Macallum (I.e. 486) for the determination of uric acid and phenol.
It was shown by us, and later by others, that all vitamine-containing
extracts give this reaction. It depends upon the development of a
blue color with phosphotungstic and phosphomolybdic acids on the
addition of sodium carbonate. The chemical nature of the sub-
stances giving this reaction was investigated by Funk and Macallum
(523), as well as by Lewis and Nicolet (524). With Macallum, we
also showed that if an incision is made in the maize kernel and the
test applied in situ the blue color develops in the part known to be
rich in vitamine. Naturally, these parts are even richer in other
extractives. As this method came into use for the chemical analysis
of various corn meal products, we (525) showed that in extracts,
prepared either cold or warm, the developed color increased with the
vitamine content. In the warm extracts, the color was weaker,
showing that the substances responsible for the color reaction are
destroyed by heat. If it should appear later, with greater certainty,
that this reaction has nothing in common with the vitamine, it might
still be useful in vitamine fractionation as an index of the purity of
the isolated substances. These reactions could conceivably also
serve as an index of the purity of the isolated lipoids.

Green (526) tried to estimate the vitamine content of various food-
stuffs by means of a biological method (animal experiments) . The
method depends on the length of time pigeons survive on various
diets, rich or poor in vitamines. As a result of these studies, Green
proposed the following formula :

C 1

S =

V - X K

198 THE VITAMINES

in which S is the duration of life; C, the amount of vitamine that
the animal may lose without developing beriberi; V, the amount of
vitamine necessary to provide for the metabolism of a given quantity
of food; X, the quantity of vitamine in the given diet; K, the con-
stant depending upon the quality and quantity of the diet fed. By
means of this formula, the vitamine content of various foodstuffs,
especially corn, was determined. The figures obtained correspond in
general with experience gained by other investigators with the same
products. The weak point of the formula is that it shows the pres-
ence of vitamine in white rice despite all evidence to the contrary.
Aside from this, it is obvious that the survival of pigeons depends on
many unknown factors.

We come now to the consideration of a method, briefly discussed
in the chapter on the vitamine requirements of yeast, which was
indicative of real progress. This method is based upon the fact
that certain yeasts require an addition of substances containing
vitamines in order to grow. We have already spoken of the work of
Williams (I.e. 128), Bachmann (I.e. 129) and Abderhalden and
Schaumann (I.e. 499) — all of which led to the development of this
method. Soon after, papers were published on the practical applica-
tion of the method. Eddy and Stevenson (527) tested the method
of Bachmann and found that it was not so very reliable. Thereupon
they turned to the method of Williams, by introducing exactly
calibrated micropipettes (similar to those used in opsonic index
determinations) for drawing up the yeast cell suspension and the
vitamine solution, which made the method somewhat more exact.
The yeast and vitamine units, with the necessary controls, were
deposited on a counting slide and kept in the incubator. After a
certain time, the slides were removed from the incubator, stained
and the cells counted. The objection to this method is the fact
that clumps of yeast cells are often introduced with the cell suspen-
sion. In spite of shaking the yeast cell suspension and partial
centrifugation to remove the clumps, this objection was not quite
overcome and the results were therefore uncertain. Among other
substances, Eddy and Stevenson also tested the products isolated by
us from yeast in 1912 and 1913 (see chapter on the Chemistry of
Yeast) and found them active, while nicotinic acid was inactive. The
active substance was not destroyed by heating to 100°C., but partially
so at 120°C. Using this method, more vitamine was found in the
mammary vein than in the jugular.

THE ANTIBERIBERI VITAMINE

199

The methods of R. J. Williams (528) and of Funk and Dubin (I.e.
511) then appeared almost simultaneously, both signifying a step
forward in the demonstration of the part played by the vitamines
in yeast growth. The method of Williams is as follows : 0.3 gram of
ordinary fresh bakers' yeast was removed from the center of a yeast
cake and suspended in a liter of sterile water. Of this suspension,
1 ec. was introduced into the nutritive solution and the cultures kept
in the incubator at 30°C. for eighteen hours, after which growth was
stopped by the addition of formalin. The yeast cells were then

12
E '°

O)

f 6

CD

•I 6

-(-»

° 4

CO

<D „

1 2.. 3 4 „ -5

Vitamine solution — cc.

FIG. 47. INFLUENCE OF VITAMINE B CONCENTRATION ON YEAST GROWTH

(WILLIAMS)

filtered on a Gooch crucible, washed with water and alcohol and
dried at 103°C. for 2 hours. With a vitamine solution of a source
not mentioned, a growth curve was obtained, reproduced herewith.
He showed that with higher vitamine concentrations, the quantity
of cells rose proportionally and that under certain conditions, the
curve approaches a straight line. Williams also observed that certain
extracts such as autolyzed yeast, or extracts heated to a high tem-
perature in the presence of acids, could exert an inhibiting influence
on the growth of yeast. In such cases, an abundant dilution is of
great help and yields better results.

200

THE VITAMIXES

The method of Funk and Dubin is still more simple and is briefly
as follows: A yeast suspension is prepared by shaking 100 cc. of
Nageli solution inoculated with a platinum loop-full of a 48 hours
pure yeast culture, preferably bottom yeast. Two test tubes are

FIG. 48. CENTRIFUGE TUBES USED IN THE DETERMINATION OF THE EFFECT
OF VlTAMINE ON YEAST GROWTH

The tube at the left shows the reading of the blank, while the other two
tubes show the readings obtained with different amounts of autolyzed yeast
(Funk-Dubin).

prepared, containing the unknown vitamine solution to be tested;
in addition, there are two controls — one with yeast suspension alone,
and one with the vitamine solution alone. The last control is not
very important and serves only to detect the formation of a pre-

THE ANTIBERIBERI VITAMINE

201

cipitate or cloudiness at the beginning of the test. All of the test
tubes are placed in the incubator for 20 hours at 30°C., after which
the cells are inactivated by immersing the tubes in a water bath at
70 to 80°C. for a few minutes. The contents of the tubes are then
quantitatively transferred to centrifuge tubes, the lower part of
which is in the form of a capillary, graduated in mm. The tubes
are centrifuged for 15 minutes at about 2600 r.p.m., and the volume
of cells read on the graduated capillary of the centrifuge tube. The
reading should be taken at once, since there is a tendency for the
yeast cells to swell up, after a time. The control, without the
addition of vitamine, must be subtracted from the reading.

We found that the controls with yeast alone were quite constant
and almost independent of the number of yeast cells in the suspension

Mm.
5

<
o

^

L_JL_, I i I ,

j

X

0.5 1.0

1.5

2.0 2.5 3.0 3.5 4.0 45 5.0
Cc. of yeast suspension

FIG. 49. EFFECT OF VARYING AMOUNTS OF YEAST SUSPENSION (FUNK-DUBIN)

used, as may be seen from the curve. Experiments with increasing
amounts of autolyzed yeast gave a curve markedly different from
that of Williams, but which is perhaps better applicable to the study
of natural extracts. We feel that our results have been considerably
influenced by the presence of inhibiting substances. For this and
other reasons, it appears somewhat premature to estimate the
vitamine content of various foodstuffs by the use of these methods.
A number of problems were undertaken with the help of the above
method, and we shall record some of the results. We showed that
analyzed substances, isolated from yeast seven years ago by the
author, still retained a definite activity, although they had been
recrystallized to constant melting point. On the other hand, nico-
tinic acid obtained from yeast was inactive. White polished rice

202

THE VITAMINES

showed no influence on yeast growth; this was also true of vitamines
A and C (after vitamine B had been completely removed by adsorp-
tion with fuller's earth). The active substance was found also out-
side of the wheat and corn germ, though in smaller amounts. The
substance influencing the growth of yeast was very resistant towards
heat; it could be diazotized, oxidized and reduced without loss of
activity. With this method as a control, a fractionation of yeast
was undertaken; partial results have already been published (Funk
and Dubin (I.e. 511) ) but the work is not yet completed.

Mm.
35

30
25
20
15
10

7

t

0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0
Cc. of autolyzed yeast

FIG. 50. VITAMINE ACTIVITY SHOWN BY INCREASING AMOUNTS OP ATJTOLYZED
YEAST (FUNK-DUBIN)

The use of these methods was criticized by de Souza and McCollum
(529). They employed the first method of Williams, slightly
modified, for their investigations and found, as a rule, that with
increasing amounts of vitamine B, the growth of yeast is favorably
affected. They believed, however, that the method could hardly be
applied to the determination of vitamine B, because glucose and
amino-acids also show a growth-promoting action. Despite this, if
we examine their findings more closely we see that the addition of a
10 per cent glucose solution had no more influence during the first
24 hours, as compared with the controls; it was only after 48 hours —

THE ANTIBERIBERI VITAMINE 203

and not always so — that some effect was noticeable. This protrac-
tion of the influence showed that the accelerative action of glucose
was of a secondary nature, and might perhaps be explained by the
plasmolysis of some yeast cells. When de Souza and McCollum speak
of the favorable action of amino acids, they mean by that a product
resulting from the sulphuric acid hydrolysis of meat. This prepara-
tion, as well as an alcoholic extract of wheat germ, was shown to be
free from vitamine B in rat experiments, while they were still active
for yeast. Hydrolyzed meat gave only double the amount of cells,
so that the action was not so pronounced. These investigators
admitted that their experiment did not demonstrate that vitamine B
played no part in the growth of yeast, they likewise showed that the
growth of rats and yeast does not go parallel with the same extracts.
The question as to the identity of vitamine B with the substance,
promoting the growth of yeast will be discussed in the next section.
Newer investigations have shown that the yeast test is of value
in determining the relative richness of extracts in water-soluble
vitamines, but cannot be used for the quantitative determination
of vitamine B.

THE POSSIBLE IDENTITY OF VITAMINE B WITH THE SUBSTANCE
STIMULATING THE GROWTH OF ANIMALS AND OF YEAST

We have already (p. 41) touched upon the question raised by
Mitchell (I.e. 93) as well as by Emmett and Luros (I.e. 94), as to
the identity of these substances. As regards the comparison of
vitamine B with the substance promoting the growth of rats, Funk
and Macallum (530) showed that the latter substance could be
precipitated with phosphotungstic acid, whereupon further fraction-
ation was tried. Since the diet we used at the time did not contain
sufficient vitamine A, the results were not particularly fruitful, and
seemed to indicate that pigeons require less vitamine B for protection
against beriberi than do rats for growth, or that vitamine B is more
stable than the growth vitamine. Drummond (I.e. 498) corrob-
orated our findings and came to the conclusion that, in all prob-
ability, both substances are identical. Emmett and Luros (I.e. 94)
seem to have a different opinion on this matter. They investigated
the effect of autoclaving at various temperatures for various periods
of time. On the one hand, these products were given, as such, to

204 THE VITAMINES

pigeons and on the other, together with lactalbumin, butter, salts and
lard, to rats. Vitamine B, which is necessary for pigeons, appeared to
be unstable and was totally destroyed by heating for 2 hours at
1 20°C . , while rats could still grow on the above food mixture . Emmett
and Luros came to the conclusion that these vitamines are different
from each other. Nevertheless, we saw in our investigations on
yeast, contrary to the above experiment, that vitamine B was resis-
tant to heat; furthermore, in the experiments of these workers one kind
of animal was fed rice alone while the other received various addi-
tions. As we shall see later, protein may easily take the place of a
vitamine addition, besides which, these animals may differ with
respect to their vitamine requirements.

As regards the identity of the growth substance for yeast with
vitamine B, the question is of particular importance since we must
know, when using the yeast method described in the previous chapter,
which substance we are determining. There is no doubt, however,
that the substance important for the growth of yeast is much more
stable to heat, and yet we saw in the chapter on the chemistiy of
vitamines, that vitamine B resists many manipulations, and still
retains its activity after many weeks of work. Emmett and Stock-
holm (531), on the contrary, saw that strongly heated unpolished
rice, although inactive for pigeons, is just as active as the natural
product for yeast cells. The same findings were obtained with fil-
trates resulting from the treatment of extracts with fuller's earth.
In these cases, the filtrates were active for yeast but not for rats.6
Emmett and Stockholm believed that further research must establish
either the identity or dissimilarity of these substances, and it would
not be correct in the meanwhile, to regard the yeast method as
suitable for the determination of vitamine B.

Recently, Funk and Dubin (I.e. 129d) have corroborated the
statement of Emmett and Luros that the yeast substance is not
identical with vitamine B. If autolyzed yeast is shaken with a
small amount of fuller's earth vitamine B is usually removed almost
quantitatively. Now if the filtrate from the above is repeatedly
shaken with larger amounts of fuller's earth, the substance acting

'Karr (I.e. 404), working on dogs, noted that vitamine B of yeast is very
stable. Autoclaving for 3 to 4 hours with subsequent heating at 108°C. for
72 hours destroyed only a part of this substance.

THE ANTIBERIBERI VITAMINE 205

on yeast — and which we call vitamine D — is likewise almost com-
pletely removed. We have not succeeded thus far in separating
D-vitamine from B-vitamine quantitatively. The animal experi-
ments performed thus far in which vitamine B preparations were
used have to be repeated, since such preparations contain at least
two vitamines — B and D. While pigeons get along very well on
vitamine B alone, rats require vitamine D besides vitamine B.
Other animals are being investigated in this respect.

Vitamine D is one of the substances necessary for the growth of
certain bacterias, as we have already pointed out. The isolation
has been attempted by Goy (53 la) who thought it was a non-
nitrogenous substance but the pure substance he isolated proved
to be inactive. The isolation also was attempted by Frankel and
Schwartz (53 Ib).

We also feel that this subject needs further investigation. For
in every one of the cases investigated, we tested the activity after
the naturally occurring substances had been altered by the manipu-
lation undergone; we do not know what the action of the substances
in the natural state would have been. It may, for example, be
established later that vitamine B is a uniform substance, but that
parts of it may be utilized differently by yeast, pigeons and rats.
The analogy between the three modes of action, in relation to its
occurrence in Nature, is so far extended that we are perhaps justi-
fied in attributing to it a genetic relationship. The findings of
Funk and Dubin (I.e. 493), and others, as to specificity, also point
to this conception.

SPECIFICITY OF ANTIBERIBERI VITAMINE

A short time after the appearance of our chemical vitamine studies,
a number of publications appeared describing the curative action on
beriberi of various quite heterogeneous substances. These papers
had the regrettable effect that doubt arose as to the specificity of
vitamine influence. It almost seemed as though practically all
chemical groups, under certain conditions, could act as vitamine.
Although most of these results were either disproven or else aban-
doned by their respective authors, we see unfortunately that these
results gained acceptance in the literature. We must not forget
that the cure of acute beriberi symptoms may be accomplished
theoretically in two ways, first, by the addition of the specific vita-

206 THE VITAMINES

mine, second, by measures which will suddenly mobilize the reserve
vitamirie present in organs and tissues. As regards spontaneous
cures, which we personally have not seen (but which seem to be
plausible), occurring ostensibly in starvation, we may explain them
by the catabolism of the tissues and the consequent liberation of the
vitamine. Every substance, having pharmacological action, which
stimulates metabolism may also contribute to the above-mentioned
action.

Eijkman (532) stated that pigeons, used by us for the demonstra-
tion of vitamine B, were not suitable for this purpose, since they
show a different behavior from chickens. He maintained that a
mixture of potassium chloride and sodium chloride (3:1) in doses of
20 to 40 mgm. can cure beriberi in pigeons, but not in chickens. We
(533) tested this out on pigeons, but could not corroborate Eijk-
man. In spite of this, we note that his findings are frequently
quoted in the literature. Later, we showed (534) that certain purine
and pyrimidine derivatives exerted a life-prolonging action on pigeons
fed on rice. This observation was confirmed by some, Williams and
Saleeby (I.e. 472), and refuted by others. Since these products are
frequently not indifferent pharmacologically, their behavior, as
described above, may perhaps be explained in that light. Cooper
(535 and 536) found that quinine and strychnine exhibited a definite
action on experimental beriberi. Later, he showed that quinine
contained vitamine as an impurity; as for the action of strychnine
it may be explained by its effect on the metabolism. Abderhalden
and Lampe (I.e. 25) stated that castor oil had a curative effect, but
later on Abderhalden and Ewald (537) reported that they could not
confirm the earlier finding. They believed, however, that vitamine
B was analogous to B-imidazolethylamine. Dutcher (I.e. 276)
investigated the behavior of thyroid, thyroxin (chemically pure
active substance of thyroid), tethelin (lipoid from the hypophysis)
and pilocarpine and found that these substances have a therapeutic
effect on beriberi. Pilocarpine was found inactive by Abderhalden
and Ewald; thyroid was regarded as inactive by Vedder and Clark
(I.e. 266). Seaman (537a), on the contrary, could demonstrate the
presence of vitamine B in thyroid. Inasmuch as thyroid substance
acts as a powerful stimulus to the metabolism, the mobilization of
the vitamine from the tissues may, under certain circumstances,
simulate a curative action, although we convinced ourselves that

THE ANTIBERIBERI VITAMINE 207

death is hastened because of it. Regarding the action of tethelin,
when one considers its method of preparation, it may contain vita-
mine as an impurity. Dutcher, Holm and Bierman (511) tested
various indol derivatives of thyroxin and found them active, but
their findings require further corroboration. To complete our data
on this question, we may mention that it was impossible for Chamber-
lain, Vedder and Williams (I.e. 64) and Abderhalden and Schaumann
(I.e. 499) to influence beriberi by the administration of known sub-
stances, and especially a mixture of amino acids. Viewing all state-
ments to the contrary in a critical light, we must recognize that
vitamine B has a well defined and specific action, and it is not to be
substituted by other products.

PHYSIOLOGY AND PHARMACOLOGY OF ANTIBERIBERI VITAMINE

To draw a general picture of the nature of beriberi, we see, first of
all, a very definite retardation of the life processes,7 similar to that
observed in inanition. In this respect, Ramoino (538) showed that
in rice-fed pigeon, the respiratory quotient is very low but may be
raised again by an addition of vitamine. Similar observations were
made by Jansen and Mangkoewinoto (539). This is evident also
in McCarrison's (I.e. 300) investigations of avian beriberi, in which
he described pathological changes in the intestines and some glandular
organs. In hunger, however, the whole organism is attuned to
endure the least possible loss of energy and substance. On the
other hand, in experimental beriberi only a diet free from or poor in
vitamines is fed. In this way, energy is expended, digestive juices are
secreted and the assimilative processes are continued. At present, we
are not quite in a position to set forth the primary causes of the whole
disturbance. It is not yet possible to say, of the pathological con-
ditions observed in this disease, which may be regarded as primary
and which secondary. We only know that the animal requires a
minimal amount of vitamine in order to regain its normal condition.
With one stroke, the entire condition is altered, and it can hardly
bo, believed that this is brought about by such a small quantity of

7 In accord with this, there are the observations of Drummond (I.e. 33
and Abderhalden (I.e. 500) on the subnormal temperatures of beriberi
pigeons.

208

THE VITAMIXES

substance.8 In order to find an analogy, we may compare an animal
on a vitamine free diet with a machine, running without oil. At
first the machine operates faultlessly, but gradually trouble arises
and then, in due time, greater defects occur which can nevertheless
still be corrected by proper oiling. In the end, a condition results
which can no longer be be remedied and the machine falls to pieces.
In animals, we know only the primary cause, lack of the specific
vitamine, similar to the oil in the above mentioned analogy. It
still seems tempting to attribute all of the observed phenomena to
changes in the central nervous system, but another explanation is
that the vitamine might be thought of as the mother substance of an
important product of the endocrine glands — a conception which we,
among others, thought to be plausible and which has again been
put forward by Massalongo (540). Another relationship may be
seen in the origin of the digestive juices. Lumiere (541) believed
that the primary cause of beriberi is to be noted in the lack of secre-
tions. The same view was held by Damianowich and Pilado (541a).
Other investigators sought for a relationship with the oxidation
processes in the body. Dutcher (542) and Dutcher and Collafcz
(543) demonstrated a decrease in the catalase in the organs of beriberi
pigeons, but almost the same thing was described in starving animals
by Burge and Neill (544). Particularly in the observations as to
the nature of beriberi, is the picture of this disease somewhat obscured
by complication with symptoms of inanition. Jansen (545) studied

8 The influence of small quantities of vitamines on the metabolism of
pigeons is well illustrated in the metabolism studies of Schaumann (I.e. 403).

6S.

B .

o

PERCENTAGE RELATION OF OUTGO

•

TO INTAKE

02 \f, K

p

•

o

g 2 g

1

h

I

o

00 '""'

1

E

&.

D a «

o

I

N

PsO.

Ash

CaO

MgO

P

0

3

grams

•percent

per cen<

Without

vitamine

204.3

658.7

20.7

11

47

69.3

45.26

40.13

36.99

238.89

With vitamine . . .

295.1

951.6

9.41

8

.53

101.4

63.56

62.12

118.9

347

The nitrogen balance was positive and the balance of the inorganic con-
stituents was likewise improved. The weight of the feces, in comparison
with the food intake, showed that the food was utilized much better.

THE ANTIBERIBERI VITAMINE 209

the question as to the identity of secretin with vitamine and came
to the conclusion that they are not identical. Voegtlin and Myers
(546) also took up this question and showed that both substances
occur in the same fraction; they believed that both substances at
least resemble each other, if they are not entirely identical. The
results of their experiments do not seem to lead to such conclusions,
since they noted in the same fraction a strong secretin and a weak
vitamine action, whereas a vitamine preparation showed a consider-
able vitamine and only a slight secretin content. In this connection,
they showed later that vitamine fractions were contaminated by a
histamine-like substance. That the substances are not identical was
shown by the work of Cowgill (546a) and of Anrep and Drummond
(546b) . Bickel, Eisenhardt and Djenab (547) have demonstrated the
presence of a secretin in spinach, which stimulates the stomach juice
and pancreas secretion. The substance occurred in the arginine-
histidine fraction and was inactivated by heating to 140°C. It
occurred in combination and could be liberated by hydrochloric
acid hydrolysis. The relationship of this substance to vitamines
was discussed by the authors, and their results were confirmed by
van Eweyk (548).

Boruttau (549) believed that yeast, or the aleurone layer of oat
kernels, contains a substance which acts as a specific antidiabetic,
since their extracts greatly decreased the output of sugar. Uhlmann
(550) investigated the 'pharmacological action of orypan (extract of
rice polishings), as well as the extracts of various vegetables and food-
stuffs. The action noted was similar to, but not identical with,
that of choline, and pilocarpine. Uhlmann ascribed this to the
presence of vitamine. It is nevertheless apparent that in his cases,
he was dealing with histamine-like substances. A vitamine prepara-
tion made by us, was shown by Prof. Cushny to have no action on
blood pressure, respiration and the heart. No relationship to the
endocrine glands, aside from that already described in avian beriberi,
was noted.

When an animal is fed a vitamine-free food, what happens to the
vitamine reserves stored up in the organs? It would be plausible to
assume that the animal would hold on to this valuable substance,
but this is not the case. Cooper (I.e. 536) showed that when feces
of chickens, fed on unpolished rice, or of rabbits, fed on bread and
cabbage, are extracted with alcohol and the concentrated extracts

210 THE VITAMINES

given to beriberi pigeons, a cure could be obtained. This shows
that all the vitamine of the food is not assimilated, but that a portion
of it appears in the feces. Another explanation might be found in
the vitamine content of the bacterial intestinal flora, although Brad-
don and Cooper (551) noted only a small quantity of vitamine in
these bacteria. Portier and Random (I.e. 217) believed on the con-
trary that the vitamine of the intestinal canal arises from the bacteria.
Muckenfuss (552) found vitamine B in bile and urine, and was con-
firmed, as regards the latter, by Gaglio (553) and by Funk and Dubin.
Funk showed that when pigeons, which had died of beriberi, were
minced, extracted with alcohol, the extract concentrated, the residue
taken up with water and given to other beriberi pigeons, the latter
were promptly cured. This experiment, corroborated by Theiler,
Green and Viljoen (I.e. 278) showed definitely that animals develop
beriberi in spite of the fact that they still have vitamine in their
tissues. It seems that a portion of the vitamines in the tissues (per-
haps as a constituent of the cell protoplasm) is so combined that the
animal itself cannot utilize it. Another explanation of this is to
be found, perhaps, in the work of Green on pigeons (I.e. 526); he
noted that the animal organism requires a rather definite amount of
vitamine for normal health. This really seems to be the case, and
as the quantity drops below the minimal need, it leads to an
avitaminosis. The quantity of vitamines that may be liberated from
the organs normally is very small. If the animal is given a diet poor
in vitamine, it might be thought that the animal would consume
more of the food in order to obtain the necessary vitamine; but this
is not the case. It was supposed by Osborne and Mendel (554) that
some foodstuffs contain sufficient vitamine for rats, if these animals
would but ingest more of this food mixture. But it is right here
that we meet with a difficulty that is also of practical importance,
namely, the question of vitamine dilution in the natural foodstuffs.
With every diet prepared, the needed amount of vitamine must be
supplied if the diet is to prove satisfactory.

THE INFLUENCE OF DIETARY COMPOSITION ON THE VITAMINE
REQUIREMENT

Regarding the influence of a large or small addition of a vitamine-
free diet on the occurrence of beriberi, the literature is full of conflict-
ing opinions. While Maurer (555) and Cooper (I.e. 536) stated that

THE ANTIBERIBERI VITAMINE

211

the addition of large quantities of rice hastens the onset of the
disease, Chamberlain, Bloombergh and Kilbourne (556) noted that
the disease developed sooner on a smaller rice feeding. They found,
in fact, that beriberi arises in hunger. These observations were later
corroborated by Eijkman and Hoogenhuyze (557) . They found that
the outbreak of beriberi could be obtained by starvation, especially
when the precaution was taken to wash out the organism with water.
The starvation beriberi resulting thereby could be cured on the
addition of vitamine. At all events, Theiler, Green and Viljoen (I.e.
278) noted spontaneous cures in animals receiving only water. These
therapeutic results were attributed to the mobilization of the vitamine
present in the tissues. Walshe (558) found no beriberi in chickens
that had been starving a long time ; these observations have not yet
been made on pigeons, and it is an open question as to whether
starvation beriberi may develop in other animals, and also in man.
The influence of the composition of the diet on the occurrence of
beriberi was investigated by the author (559) in 1913. At that time,
there appeared the paper of Abderhalden and Lampe (I.e. 25), in
which they formulated the "toxic" theory of beriberi. The symp-
toms of the disease were apparent later on cooked rice than on raw
rice. This observation, which in itself was correct, but which was
thought to be due to the elimination of toxins, may be explained in a
very simple way. Cooked rice takes up so much water that it is
impossible to feed an amount corresponding to uncooked rice, and
the results appear as follows:

COOKED RICE

RAW RICE

Appearance of beriberi

Dead

Appearance of beriberi

Dead

days

44

days

44

days

25

daft

26

However, if pigeons were fed cooked rice corresponding to 10 grams
of raw rice, then the results were somewhat different :

COOKED RICE

RAW RICE

Appearance of beriberi

Dead

Appearance of beriberi

Dead

days

27

days

29

days

28

days

30

212

THE VITAMIXES

Here, we see that the disease develops almost simultaneously when
comparable amounts of rice are fed. Later, we (560) showed that
when pigeons are fed 5, 10 and 20 grams of rice daily that the disease
develops the sooner, the more rice is fed :

AMOUNT OF RICE

5 grams

10 grams

20 grams

Beriberi

days

39

days

36

days
22

Dead

42

38

22

We also tested this observation on a synthetic diet, the pigeons
receiving equal quantities of food (12.5 grams) in every case:

DIET

SALTS

CASEIN

SUGAR

LARD

STARCH

BERIBERI

A

4

60

12

12

12

30 days

B

4

12

12

60

12

40 davs

C

4

12

12

12

60

24 days

D

4

12

60

12

12

28 days

From these findings we concluded, at that time, that vitamine B
plays a particular part in the carbohydrate metabolism, since on
feeding large amounts of carbohydrates, the pigeons developed the
disease sooner. Our observations on the influence of rice feeding
were confirmed by Braddon and Cooper (I.e. 551 and 561) in 1914,
by Weill and Mouriquand (562) and also by Green (I.e. 526). Green,
however, assumed that not only carbohydrates but the entire metabo-
lism plays a role in the vitamine requirements. However, other
investigators were not in accord with our views. For example,
Eijkman and Hoogenhuyze (I.e. 557) found that an addition of
meat did not prevent the appearance of beriberi symptoms. Inas-
much as this disease may occur likewise on an exclusive meat diet,
they used this in combating our views. Since about 40 per cent of
the protein in the body is transformed into glycogen, and since the
destruction of the tissues proceeds similarly in starvation, these
facts, in reality, cannot be used to discredit our findings. Vedder's
(563) conclusions were similar to those of the Dutch investigators.
Altogether he carried out five experiments, which are perhaps less
convincing than appears at first sight.

THE ANTIBERIBERI VITAMINE 213

In the first experiment he fed 11 chickens, varying in weight between 1800
and 3800 grams on rice, which they ate voluntarily; the uneaten rice was
weighed the next day. No relationship was evident between the appearance
of beriberi and the amount of rice utilized. Since the birds varied consid-
erably in age and vitality, so that the energy expended and required were
different, and since this experiment was carried out on two series of experi-
mental animals, in two different months, with a possibility of the dissimi-
larity of the external temperature, therefore this experiment is not quite
significant. In a second experiment, Vedder fed forcibly 100, 50, and 25
grams of white rice daily to chickens averaging 1200 grams in weight. Vedder
then stated that the pigeons could die as well from over-feeding as from under-
feeding, because rice, collecting in the crop, mechanically caused disturbances
not related to the stimulation of metabolism. If we assume, with Vedder,
that chickens fed 100 gm. of rice really die as a result of over-feeding, and
only consider the birds receiving less rice, then chickens on 50 grams of rice
developed beriberi in 20 days and those on 25 grams of rice, much later, which
naturally completely corroborates the results of other investigators. In a
third experiment, unpolished rice was used, but in this case, the animals did
not die as a result of over-feeding, although they were given 50 and 75 grams
of rice daily. In a fourth experiment, chickens were fed on sterilized meat
and eggs, and the results show definitely, that eggs contain some vitamine
while meat contains a greater quantity. Vedder believed that if beriberi
could occur in the absence of carbohydrates, then it was certain that vitamine
B played no part in the metabolism of this dietary component. We have
already seen that this conclusion is, in all probability, incorrect. If these
foods were vitamine-free, the retardation of the beriberi symptoms would be
the best demonstration for the correctness of our observations. In the fifth
and last experiment, chickens were fed on sterilized eggs and meat, with the
addition of white rice. The eggs, apparently egg-white and egg-yolk, were
evidently not entirely vitamine-free, and the residual vitamine might perhaps
suffice for 25 grams of rice. The eggs and meat used here, according to Vedder's
statements, were supposed to be identical with the preparations used in the
fourth experiment. This is very unlikely, since the chickens, in the fourth
experiment, developed beriberi in 61 days, when fed on 50 grams of eggs,
while in the fifth experiment, the animals developed no beriberi symptoms
on an addition of 25 grams of eggs. As we see from the above, Vedder's objec-
tions do not carry any weight. Aside from this, upon examining our pigeon
experiments more closely and disregarding those fed on larger quantities of
rice, the results on lesser quantities of rice still prove our contention. The
amount of food was kept constant (12.5 grams daily) in another experiment,
where a synthetic diet was used which confirmed our rice results, so that the
food only varied qualitatively but not quantitatively.

The chief difficulty encountered in this experiment was that the
amount of vitamine needed for metabolism depends not on the
ingested quantity of food, but on the portion assimilated. This con-

214 THE VITAMINES

dition is controlled with difficulty in pigeons and chickens, because
of the scattering of the food and the ease with which they vomit.

It would, therefore, be desirable to repeat these experiments on
rats, in which it is easy to control the food. Such experiments,
which were carried out by Funk and Dubin (I.e. 331), and which con-
firmed in general the results with pigeons, will be described in another
chapter. Whether our view, that vitamine B plays an important
role in carbohydrate metabolism, may be regarded as correct must
remain for the future to show, since simultaneously with an enrich-
ment of the carbohydrates in the form of starch or sugar, there is a
decrease in protein. That protein plays a particular role in nutrition,
is well known. Maignon (564) fed rats on diets which varied con-
siderably in their protein, starch and fat content, but unfortunately
the vitamines were not provided for. While each of the protein-fat
combinations could maintain the animal at a constant weight for 50
days, this was true of the protein-starch mixture only when both
components were present in equal quantities. From this, it was con-
cluded that protein is better utilized in the presence of fat than in
that of starch. The best nutrition was observed when a mixture
consisting one one part protein, one part fat and 1.33 parts starch
was fed. Bierry and Portier (565) obtained similar results, and
expressed the view that there must be a very definite relationship
between the protein and fat of any diet. Bierry (566) noted that
disturbances in metabolism could be prevented only when the individ-
ual dietary constituents were present in proper proportions.

While we believed, at first, that the delaying effect of a protein
addition on the development of beriberi was due to the presence
therein of vitamine B, this view is perhaps no longer tenable. We
shall see later that the "vitamine sparing" action of protein, or the
presence of a special substance essential to life may play a part in
this phenomenon. For this reason, we must be conservative with
our conclusions when extra protein is added to the diet. To illustrate,
let us consider the work of Johns and Finks (567), who investigated
the nutritive value and B-vitamine content of war bread, which
was milled to the extent of 74 per cent. Working with rats, they
found that the above food contains sufficient B-vitamine but not
enough protein, since the animals resumed normal growth on the
addition of casein. This conclusion is perhaps not quite correct,
for the growth-inhibiting influence may possibly still have been due

THE ANTIBERIBERI VITAMINE 215

to a lack of vitamine B. Similar conditions are frequently met
with in the study of pellagra and hunger edema, to which we shall
refer again in the chapter dealing with the question. We have
already seen in the investigation of Funk and Dubin (I.e. 331) that
a larger addition of carbohydrates increased the vitamine B require-
ments. In this respect, rats are similar to pigeons except that in
rats, an excess of fat causes much greater disturbances than in
pigeons.

Besides the above mentioned results, Funk and v. Schonborn (I.e.
304) found that whereas in normal pigeons, the glycogen and blood
sugar content of the liver is 1.17 per cent and 0.1 per cent, respec-
tively, on an artificial vitamine-free diet, the glycogen content drops
to 0.48 per cent and the blood sugar rises to 0.15 per cent.9 Pigeons
fed on an excess of sugar showed 4.5 per cent glycogen and 0.15 per
cent blood sugar. On an excess of starch, there is no glycogen and
0.26 per cent blood sugar. In the last case, if vitamine B is added,
the values for glycogen and blood sugar approach normal. These
results were later repeated several times and corroborated by the
author (568). 10

Besides this, we also studied the influence of substances having a
known action on carbohydrate metabolism. Among them, the
effect of glucose, phlorizin, adrenaline, pituitrin, thyroid and para-
thyroids on pigeon beriberi was tested, normal and rice-fed pigeons
being used. Of special interest, was adrenaline, which shortened the
life of the animal; this was also true of thyroid. The influence of
parathyroids was quite different. While thyroid raised the blood
sugar content and the amino nitrogen in the serum, and lowered the
glycogen content of the liver, the addition of parathyroids increased
the glycogen and amino acid content, but had little effect on blood
sugar. We must admit, however, that Elias and Kolb (569) also
demonstrated glucosuria in starving dogs, which was attributed
to acidosis, and which could be remedied by the administration of
alkali. We have already emphasized that it is not known, at present
how many of the starvation symptoms may be regarded as results of

9 Scott (567a) has found the normal blood sugar content in pigeons higher,
but the difference may be due to a different method of bleeding the animal,
a fact which has been frequently observed by us.

10 The finding of glucosuria in pigeon beriberi has not yet been confirmed
in human beriberi.

FIG. 51. COMPARISON OF THE ADRENALINE CONTENT— TESTED ON SHEEP — OF

THE SUPRARENALS OF PlGEONS IN BERIBERI, NORMAL

NUTRITION AND INANITION

I, normal pigeons; II, injection of 0.01 mgm. adrenaline into sheep as
control. Ill, inanition; IV beriberi (McCarrison) .

216

THE ANTIBEBIBERI VITAMINE 217

an avitaminosis, and how many of the beriberi symptoms must be
attributed to a condition of starvation.

The discovery of glucosuria in pigeon beriberi may evidently be
-connected with some observations made by McCarrison (I.e. 296).
He found that in avian beriberi, the suprarenals undergo considerable
enlargement and are characterized by a high adrenaline content.
Here too, an increased adrenaline content was found in starving
pigeons, but not so much as in beriberi pigeons. The adrenals were
also somewhat enlarged. In the same publication we find numerous
data on temperature, respiration and weight of the individual organs
in avian beriberi, which are also of physiological interest.

In concluding this chapter, we wish to point out that in considering
the vitamine B requirements, we must take note of the composition
of the diet as regards protein, carbohydrate and fat. These require-
ments are possibly dependent also on the simultaneous presence or
absence of other vitamines. If, for example, the quantity of vitamine
A is too little, the requirements for vitamine B are perhaps increased.
However, there are as yet no available data on this point.

CHAPTER II
THE ANTIRACHITIC VITAMINE — VITAMIN E A

The history of the discovery of vitamine A has already been dis-
cussed, so that we may take up the chemistry of this substance at
once. We have noted that for a long time, vitamine A was supposed
to be free from nitrogen, but we feel that in the interest of future
investigation this question should be left open. We must also leave
for the future, the question as to whether the designation, vitamine
A, includes several substances of the same type, or only a single
substance. The chemistry of vitamine A has advanced but little
so that we shall only discuss its resistance to physical agents, solu-
bility and occurrence.

CHEMISTRY OF COD LIVER OIL

Cod liver oil is the only starting material from which attempts
have been made to isolate the vitamine. Although Qsborne and
Mendel (570) showed in 1914 that this oil contains vitamine A, and
although, because of its relationship to the therapy of rickets, it has
commanded attention for a long time, we are still in the dark as to
the nature of the vitamine contained therein. However, when we
consider the known facts about the resistance of vitamine A from
various sources, and the technical preparation of cod liver oil (571),
it is obvious that because of the stability of the vitamine, cod liver
oil is the best material to use for the chemical isolation of vitamine A.
Iscovesco (572) believed that almost all of the phosphorus and nitro-
gen of cod liver oil occurs as a lecithide or as a lecithalbumen. If
acetone is added to cod liver oil, with stirring, a precipitate is formed,
which, on reaching a maximum, is redissolved. The oil, freed from
some of its lecithin by acetone, was shaken with an equal volume of
95 per cent alcohol. To the alcoholic solution, which contains all
of the phosphorus of the oil, absolute alcohol was added, giving two
separate fractions. The soluble part was concentrated at a low
temperature, the residue dissolved in ether and precipitated with
acetone. In this way, from 1 kilo of oil, 0.02 gram of the pure
lecithide was obtained, which was soluble in benzol and chloroform,

218

THE ANTIBACHITIC VITAMIXE 219

melted at 70°C., and contained 60 per cent C, 7.3 per cent H, 2.1
per cent N and 4 per cent P. A part of this semi-solid mass was
used in an experiment on rabbits kept on a normal diet. The leci-
thide in olive oil was injected subcutaneously every second day for
130 days The result were as follows:

Gain in weight
per cent

Controls 33

Cod liver oil 55

Cod liver oil without lecithide 37

Olive oil 33

Olive oil with 0.5 per cent lecithide 56

It would appear from these results that the total active substance
was present in the lecithide, but it is still necessary to corroborate
these findings in rats fed on a diet free from vitamine A.

Other investigators turned their attention to the nitrogenous por-
tion of the oil. In 1888, Gautier and Mourgues (573) showed that
even a purified oil contains a small amount of organic bases which
at that time were thought to be ptomaines or alkaloids.

One hundred kilos of oil were treated with an equal volume of 33 per cent
alcohol, to which 4 grams oxalic acid per liter were added. The watery-
alcoholic solution was saturated with calcium hydroxide, filtered and the
filtrate evaporated in vacuum at 45°C. At the end of the distillation, calcium
carbonate and hydrate were added, the mixture evaporated to dryness aud
the residue extracted with 90 per cent alcohol. To this extract, after the
removal of the alcohol in vacuum, concentrated potassium hydroxide was
added and the alkaline solution extracted with ether. An ether solution of
oxalic acid was then added, to precipitate the bases present as oxalates. The
yield was 52 to 65 grams oxalate from 100 kilos of oil. The oxalates were dis-
solved in dilute potassium hydroxide, and the bases set free in the form of an
oil, which was dried and fractionated, giving the following fractions:

Fraction I, 87-90°C. butylamine; fraction II, 96-98°C. amylamine; fraction
III, under 100°C. hexylamine; fraction IV, between 198-200°C. hydrotoluidine.
The distillation was then continued to 215°C. whereupon the brownish distil-
lation residue was extracted with ether. The ether solution was evaporated,
the residue treated with HC1 and precipitated with a solution of platinic
chloride, giving a base as a double salt, called "aselin," having the compo-
sition C25H32N4. From the mother liquor, a second base was isolated, called
"morrhuin," having the formula CjgH^Na. It is not impossible that both of
these bases were rather condensation or decomposition products, arising
secondarily during the distillation.

This work, which is of little more than historical interest, shows
nevertheless that cod liver oil contains nitrogenous substances which
are of interest to us because of the vitamine A.

220 THE VITAMINES

The author (574) has investigated a crude oil and found that the
quantity of extractives therein was much greater than in the purified
oil. [Crude oil was found more active than refined oil in animal
experiments by Zilva and Miura (574a)]. 23.5 kilos of the oil were
extracted three times with about 660 cc. alcohol, 2 liters of water and
50 cc. concentrated HC1; the extracts were concentrated in vacuum,
the oily residue hydrolyzed with sulphuric acid and precipitated
with phosphotungstic acid. The practically dry precipitate weighed
969 grams. The oil, hydrolyzed with sulphuric acid (2 hours) gave
an additional 186 grams of this precipitate, while the oil remaining
after the concentration of the water-alcohol solution gave only 19.5
grams of the precipitate. A second portion of oil, weighing 25
kilos and treated with sulphuric instead of hydrochloric acid, gave
1117 grams phosphotungstate, of which 61.2 grams was insoluble in
acetone. This fraction, after the decomposition with lead acetate,
gave a precipitate with sublimate in alcoholic solution. A great
number of fractions were prepared by us, but could not be tested for
various reasons.

One of the common conceptions regarding cod liver oil was that
the mysterious action of unsaturated oils was ascribed to it. How-
ever, this view has not yet been experimentally tested. Paal and
Roth (575) reduced cod liver oil with colloidal palladium and hydro-
gen to a solid, whereby the reduction of all the unsaturated bonds
was almost completely quantitative. The general nutritive value
of such an oil was investigated by Suzuki and his co-workers (I.e. 323)
and found favorable. Chapman (576) and a number of Japanese
workers including Tsujimoto (577) and Kubota (578) isolated from
cod liver oil a very unsaturated hydrocarbon called, "spinacen" or
"squalen," which, however, was found valueless as a substitute for
vitamine A by Suzuki. Bull (579) obtained a mixture of C2oH3oBrio02
and C2oH28Bri202 on brominating cod liver oil. For use in the
therapy of leprosy, such products as sodium morrhuate were prepared
by Rogers (580).

CHEMISTRY OF VITAMINE A IN BUTTER AND OTHER SOURCES

The data on the chemistry of this vitamine in butter are very
uncertain and not infrequently contradictory. The first experiments
on the isolation of vitamine A from butter were made by McCollum
and Davis (I.e. 82). They reported that when a solution of butter

THE ANTIRACHITIC VITAMINE 221

in petroleum ether is hydrolyzed with alcoholic potassium hydroxide
at ordinary temperature, and the soap solution neutralized, the active
substance may be concentrated in olive oil by extracting with the
latter. Osborne and Mendel (581) dissolved ox fat and butter in
alcohol at 40°C. and allowed it to crystallize out on freezing at
— 20°C. The A-vitamine could not be demonstrated in the crystal-
lized portion, but was found in the oily, uncrystallizable portion.
Drummond (582) attempted to repeat the work of McCollum and
Davis on the stability after saponification. After butter had been
saponified at the usual temperature, the solvent was removed in
vacuum, the soaps dissolved in water and the unsaponified portion
extracted with ether. All of the fractions thus obtained were tested
on rats and found inactive. Recently, Steenbock, Sell and Buell
(582a) repeated the saponification experiments and have found
that cod liver oil can be treated with 20 per cent alcoholic potassium
hydroxide solution for 4 hours at 37°C. without losing its potency.
Since the active substance after dilution with water can be extracted
with ether, it was concluded that vitamine A is neither a fat nor
an ester.

Numerous experiments were carried out for the purpose of con-
centrating this vitamine from various vegetables and grasses by
extraction. McCollum, Simmonds and Pitz (583) thought that the
vitamine could not be removed by extracting plant tissue with ether,
because it is in all probability bound to the protein. Still, Osborne
and Mendel (584) were able to do this. They dried spinach and
clover in a stream of air at 60°C., and then extracted it with ether,
obtaining a very active substance. These results were confirmed by
Zilva (585) who obtained an active alcoholic and ether extract from
carrots. Steenbock and Boutwell (586) were unable to obtain an
active substance by extraction of carrots with ether alone, but could
do so with carbon disulphide, chloroform and benzol. In spite of
this, the residues showed almost the original activity. Using alfalfa
grass as a source, a simple ether extraction gave no result, while a
mixture of alcohol and benzol gave almost quantitative extraction.
The same workers then tried to make a fractionation by means of
solubility. Alfalfa was treated with cold alcohol, the latter distilled
off on the water bath and the residue allowed to stand over night
with 20 per cent alcoholic potassium hydroxide. When the saponifi-
cation had been treated with ether and filtered from the ether-

222 THE VITAMINES

insoluble portion, the ether solution, after washing with water, was
shown to be active. From this residue, the carrotin was separated
from xanthophyll by extraction with petroleum ether, showing the
carrotin fraction to be active and the xanthophyll fraction, inactive.

THE NATURE OF VITAMINE A

Drummond (I.e. 582) in one of his papers proposed the conception
that vitamine A was of the nature of a ferment and was a more or
less unobtainable body. He carried out experiments with various
known substances and found that fatty acids, glycerol, cholesterol,
lecithin, sphingosine, phrenosine, kephalin, lipochrome and carrotin
could not simulate the action of vitamine A. Steenbock (587) made
the interesting observation that the vitamine A content of maize
goes parallel with the content of yellow pigment; white maize was
found free from this vitamine, and the same was also true of most
kinds of vegetables. According to Steenbock and Boutwell (588)
when white corn is supplemented by butter, clover or spinach, the
rat resumes growth. Palmer (589), on the contrary, was opposed
to this finding. In the first place, he showed that chickens could
live on a diet free from carrotinoids, at least through one generation,
although it is of course quite possible that chickens may live without
vitamine A, or at best with very little. Second, it was possible for
him to raise mammals (rats; Palmer and Kennedy, 589a), the blood
and subcutaneous tissue of which were pigment free. Despite the
fact that the milk of these animals was practically pigment-free,
it was possible for them to suckle and rear their young. Palmer
observed also that there are oils, such as cottonseed oil, which,
although containing pigment, show a lack of vitamine A. Rosen-
heim and Drummond (590) took up this question and in general
were able to confirm the fact that, as a rule, foodstuffs containing
lipochrome also contain vitamine A. Still, it was found that a
pure carrotin solution is free from this vitamine, and the authors
believed that although the latter must be different from carrotin
and xanthophyll, it is nevertheless associated with these pigments.
As to what action chemically pure carrotin has on rats, opinions
differ. While Stephenson (590a) finds that pure carrotin (m.p.
172-173°C.) is entirely inactive and while butter decolorized with
charcoal and free from pigment is still active, Steenbock, Sell,
Nelson and Buell (590b) have obtained normal growth with pure

THE ANTIRACHITIC VITAMINE 223

carrotin. They prepared vitamine A from alfalfa and found that
it withstood saponification with heat, oxidation with hydrogen
peroxide and reduction with nascent hydrogen. From the unsaponi-
fiable fraction, active crystalline acetylated products have been
prepared. Liver tissue contains, besides the two known yellow
pigments, another substance which gives the lipochrome reaction
and, in addition, the color reaction with sulphuric acid which is
regarded in the literature as being specific for cod liver oil. This
substance is not a casual impurity as it may always be demon-
strated in cod liver oil and in liver fats. Hijmans van den Bergh
and Muller (591) have described* methods which may be used for
the separation of the lipochromes in blood. In order to liberate
them, it was necessary to precipitate the serum proteins with alcohol.
It was assumed that other chemical substances were associated with
the pigments. Drummond and Coward C592) have investigated
the pigment and vitamine A content of a number of oils and fats,
and found that these two factors do not always occur together. In
butter, the pigment content was found to run somewhat parallel to
that of vitamine A, whereas certain pork fats and dog fat contain
no pigment, but a marked amount of vitamine. They came to the
conclusion that a relationship between the pigment and vitamine A
could not be maintained unless we assume, with Steenbock, that in
some cases the pigments occur in the leuco-form. As evidenced by
the above, the whole question still rests on an uncertain foundation
and can be solved only when a purification of this substance is made.
Properties of vitamine A. The characteristics of this vitamine are
as yet very little known. As for the question whether the substance
is soluble in water, it must remain open for the present. McCollum,
Simmonds and Steenbock (593) have stated that fat-free milk still
contains appreciable amounts of vitamine A, a finding which seems
to be in accord with the latest results. According to their work, the
vitamine may be extracted with small quantities of water from melted
butter by repeated extraction, the remaining butter being inactive.
On the other hand, Steenbock, Boutwell and Kent (594) showed that
both the residual butter and the watery extract were inactive. They
believed that heat and not oxidation is the reason for the destruction,
although we shall see that there is still another complication to be
considered. The finding of McCollum, Simmonds and Steenbock
(I.e. 593) that fat-free skimmed milk still contains vitamine A was
again confirmed by Hopkins (595).

224 THE VITAMINES

The first statement on the stability of the active substance from
butter was made by Osborne and Mendel (I.e. 581). They found
that when steam was passed through butter for 2| hours, the
vitamine was unaffected. That this substance in egg-yolk is resistant
to heat, was shown by McCollum and Davis (596) . In this connec-
tion, Drummond (597) found that vitamine A from butter and other
sources is veiy unstable to heat, thus corroborating the statements of
Steenbock. Drummond also reported that oxidation does not seem
to play any part in the destruction of this substance. Steenbock and
Boutwell (598) found yellow corn and alfalfa active after heating in
the autoclave for three hours. Osborne and Mendel (599) found
butter, as such, much more stable against storage and the influence of
light, than the uncrystallizable part — butter oil.

All of these apparently conflicting statements may be well under-
stood when the new observation of Hopkins (I.e. 595 and 599a)
is taken note of. He found that in the destruction of vitamine A
oxidation, not temperature, plays the chief part. This has been
confirmed by Drummond and Coward (599b) and is in agreement
with the observation of Osborne and Mendel (600), who found butter
stable when steam was passed through it, but not when the butter
was heated in the absence of water. The stability in this case may
be explained by the replacement of air by steam, although even dry
butter could be heated at 96°C. for 15 hours without noting any loss
of activity.

As regards the action of physical factors, Zilva (I.e. 513) showed
that butter lost its activity on exposure to ultra-violet rays for 8
hours. The possibility of the formation of ozone was considered in
a later paper, and it was actually found that ozone is destructive
to vitamine A (600a).

Although the destruction of vitamine A by oxidation has been well
explained by various statements, it is more difficult to understand
the destruction of fat on reducing at a temperature of 75° to 120°C.,
as carried out by Fahrion (601). For, in the first place, vitamine A,
according to the newer findings, is quite resistant to heat, and
second, the substances sensitive to oxidation are, for the most part,
resistant to reduction. Nevertheless, according to the findings of
Drummond and Coward (I.e. 592), as well as of Fahrion, the reduced
fats are totally inactive. Fahrion believed that vitamine A is almost
insoluble in fat solvents.

THE ANTIRACHITIC VITAMINE 225

Considering the manner of preparing commercial cod liver oil,
we must marvel, in the light of the above mentioned observations,
that the oil still has therapeutic properties. In this connection, it
would be interesting to observe the action of oxy-cod liver oil — a
preparation which, according to Freudenberg and Klocman (602),
is made by oxidation of cod liver oil with hydrogen peroxide— as well
as cod liver oil emulsions prepared by shaking in alkaline medium.
The influence of such a procedure on the therapeutic action is dis-
cussed by Hess and linger (603). If the vitamine A of cod liver oil
really withstands the above mentioned treatment and long storage,
then we must assume that the vitamine from this source is exception-
ally stable, a characteristic which should bs significant in further
studies. According to Zilva and Miura (I.e. 574a), cod liver oil
contains 250 times as much vitamine A as butter, and 3 milligrams
of this oil per day is sufficient for rats. Drummond, Coward and
Watson (603a), found that certain samples of butter contained
no more vitamine A than refined vegetable oils. The above data
undoubtedly explain the differences observed in the value between
cod liver oil and butter in the treatment of rickets.

DEMONSTRATION OF VITAMINE A

Till now, the only method used, as we have already seen, is the
behavior of the rat. To make the demonstration definite, it is neces-
sary, according to Drummond and Coward (604) to provide, above all,
that the basal diet is completely free from this substance. The basal
diet used for this purpose is composed as follows:

Purified casein 18 Yeast extract 5

Purified rice starch 52 Orange juice 5

Purified reduced vegetable oil. .15 Salt mixture 5

The casein is heated in open dishes for 24 hours at 105°C. Rice
starch has been shown to be pure for this purpose and may be used,
as such. Completely reduced vegetable oil, usually cotton seed
oil, or steam distilled palm-kernel oil (Stammers, 604a) is used as
a fat. If one is uncertain as to the freedom of this oil from vitamine
A, then the addition of fat should be omitted entirely. Orange
juice is free from vitamine Aj while yeast extract has not yet been
investigated sufficiently in this respect. Rats weighing 50 to 70
grams, and 4 to 5 weeks old should show no growth at all, on this

226 THE V1TAMINES

diet. If growth ensues, it may be concluded that the basal diet
was insufficiently purified. Older animals, weighing over 100 grams
should not be used since the vitamine A requirement diminishes
with age. If the growth remains stationary during 10 to 14 days,
the substance to be tested may be added, preferably apart from the
diet. If growth is seen, it may be concluded that vitamine A was
present in the substance added.

Significance of vitamine A in physiology and pathology. From a
study of this vitamine in various nuts, Coward and Drummond (605)
came to the conclusion that it occurs only in small amounts in seeds,
but that during the process of germination it appears in greater
quantities. We have already mentioned the possibility that green
leaves may synthetize vitamine A under the influence of sunlight,
and in this connection Delf (605a) has found that the inner white
leaves of cabbage are free from this vitamine, while the outer green
leaves contain the substance. Recently, Coward and Drummond
(605b) have experimentally substantiated these findings, showing
that green leaves form vitamine A only in the presence of sunlight,
while the lower organisms devoid of chlorophyll are unable to syn-
thetize this vitamine.

Hughes (605c) fed a number of animals on diets rich and poor in
vitamine A, but on examination of the organs, no relation could be
shown between the vitamine A content of the food and that of the
adipose tissue. Drummond, Golding, Zilva and Coward (605d)
explain the poverty of pig's fat in vitamine A in two ways (1) inade-
quacy of the diet in this substance, and (2) losses by oxidation during
the melting out process of the fat.

We have already stated that the lack of vitamine A in rats is fol-
lowed by no characteristic pathological changes, and hence it is
difficult to point out just what is the importance of this substance.
Drummond (606) tried to solve this question. Since this substance
is associated with fats, as a rule, it was easiest to assume that it
played a part in the fat metabolism, but it was not possible to find
such a relationship. Rats lacking both this vitamine and fat showed
no different behavior from those animals lacking only one of these
constituents. The temperature of these rats was normal, so that
this substance has nothing to do with the regulation of body tem-
perature. In another experiment, Drummond tried to see whether
or not vitamine A plays a role in the synthesis of neutral fats from

THE ANTIRACHITIC VITAMINS 227

fatty acids. For this purpose, one series of animals were fed with
fat, another with fatty acids. When vitamine A was absent from
the diet, the fatty acid mixture was well assimilated, and the animals
showed a normal fat content. In our experiments, together with
Dubin (I.e. 331), we fed rats on a diet containing a large amount of
neutral fat. These rats, in spite of the administration of vitamine
B, ceased growing and lost markedly in body weight. We tried to
improve the condition of the animals by adding greater amounts of
vitamine A in the form of cod liver oil or butter, but without success.
Unlike vitamine B, vitamine A, given parenterally, is without any
action.

The role of vitamine A in pathology was treated by Mendel (607) .
It was particularly noted that this substance was of less importance
for adults than for young growing animals. It was also shown to
be of significance in the manifestation of ophthalmia, as well as
in the occurrence of urinary calculi, conditions which arise chiefly
when vitamine A is absent from the diet. McCarrison (608) holds
that vitamine A plays a part in the prophylaxis of edema. All of
these statements will be discussed in relation to each other in another
chapter.

CHAPTER III

THE RELATIONSHIP OF THE ANTIBERIBERI AND THE ANTIRACHITIC

VlTAMINES TO LlPOIDS

The object of this chapter is to show that vitamines B and C have
presumably not the slightest relation to the lipoids. It is, of course,
more difficult to demonstrate this as regards vitamine A. The asso-
ciation of these two classes of substances, the lipoids and the vitam-
ines, came about through the conception that lipoids, and especially
lecithin, are not pure products but may contain vitamine as an
impurity. It has often been stated by Cronheim (609), and by
others, that lecithin is of significance in metabolism, while Robertson
(610) noted only an inhibition of growth on feeding this substance
to mice.

In biochemistry, the idea has long been prevalent that when
animal or plant tissue is extracted with alcohol, or better still, with
ether, the fraction in the solvent can be regarded as a lipoid. In our
first investigation, carried out in the Lister Institute in 1911, which
had for its purpose the isolation of vitamine, we convinced ourselves
that an extraction with alcohol removes a great deal of nitrogenous
matter (purines, pyrimidines, etc'.), in which the vitamines are also
to be found. This observation was made use of by McLean (611),
working in the same Institute at that time, to remove nitrogenous
impurities from lecithin. For this purpose, a crude lipoid from horse
meat was finely divided and repeatedly extracted with water. In the
first alcoholic extract, as well as in the watery extracts similarly
obtained, the presence of carnosine, hypoxanthine and vitamine was
demonstrated with certainty. For the purpose of vitamine isolation,
these mother liquors were given by McLean to Cooper (I.e. 535) who
readily fractioned out the vitamine. For the above mentioned
reasons, it is likewise evident that the growth-promoting influence
of the lecithides of the liver, isolated by Iscovesco (612), was due to
the presence of vitamine. The same is perhaps true for tethelin,
isolated from the hypophysis by Robertson (613). This lipoid,
regarded as pure, gave reactions for purine bases, as well as a diazo
reaction, so that it is certain that we are not dealing with a pure
lipoid in this instance.

228

RELATIONSHIP OF VITAMINES TO LIPOIDS 229

As regards vitamine B, Cooper (614), and later Sullivan and
Voegtlin (615), showed that there is no relationship between the
lipoids and vitamine B. As for vitamine A, the behavior is quite
different, but still there is no reason for saying with Aron (I.e. 329,
616) and Stepp (617), that lipoids are essential for life. In the first
place, with a mixture consisting of the known lipods, lecithin,
kephalin, cerebron and cholesterol (Stepp, I.e. 352), only a slight
effect could be discerned, due perhaps to the cumulative action of
all the impurities. Secondly, Stepp (I.e. 351) showed that at lease
one type of animal, the pigeon, could live in the presence of vita-
mine B, without lipoids. Recently, Stepp (618) stated that dogs
and rats cannot live on a lipoid-free diet, upon which the animal
develops paralysis of the hind extremities, but not beriberi. An
addition of vitamine B prolongs the life of the animal but cannot
save it from death. This experiment was supposed to show that
animals die because of a lack of lipoids. Regarding the matter
from the viewpoint of the relationship of vitamine A to the lipoids,
we see that while at first the view was held that this vitamine was
connected exclusively with the animal fats, it was found later that
green leaves likewise contain large amounts of the same substance.
Vitamine A is supposed to be present in skimmed milk, too, and
since the latter is believed to be free from lipoids, the relationship
between vitamine A and lipoids must necessarily be non-existent.
In particular the question as to the solubility of vitamine A in water
has not yet been cleared up; besides, in the extraction of a fat with
water, as we have already observed, the possible oxidation is to be
taken into account. Myers and Voegtlin (I.e. 503) shook out the
vitamine of yeast with olive oil, and showed that besides the vitamine,
other nitrogenous substances are taken up in the oil. In this case,
we must regard the total oil soluble portion as lipoid or, at any rate,
as fat-soluble. Since it has never been shown that vitamine A is a
constituent of a pure lipoid, and since up to the present all purely
isolated lipoids have given negative results when tested for their
activity, there is no reason for considering vitamine A as being
associated with lipoids; it may ultimately be shown that this associa-
tion is only incidental. Cramer (619) described a new type of
glandular fatty tissue, which he called "lipoid gland" or "cholesterol
gland." This tissue, which is very rich in cholesterol and other,
lipoids, loses these constituents when the animal is placed on a

230 THE VITAMINES

vitamine-free diet. Cramer believed that this tissue plays a part
in the development of avitaminoses, in that it functions as a vitamine
reserve of the body. The exact functions of this new fatty tissue
are not yet known, but Cramer thinks that a close relationship
exists between this tissue, the thyroid and the suprarenals.

CHAPTER IV
THE ANTISCOEBUTIC VITAMINE — VITAMINE C

THE CHEMISTRY AND THE NATURE OF VITAMINE C

We owe to Hoist and Frolich (I.e. 359) our first knowledge on this
subject. They showed that fresh potatoes, cabbage, dandelion,
carrots, raspberry juice, lemon and sour dock juice contain this
vitamine. They observed the interesting fact that the various anti-
scorbutics differ markedly in the stability of the vitamine present,
evident on heating, storage and drying. As to the reason for this,
we are still in the dark. We do not know if in these cases we are
dealing with different or with the same substances, and yet the varied
behavior towards external factors is not necessarily due to the pres-
ence of distinct substances. The ' different combinations of the
vitamine and the characteristics of the juice may be responsible for
this behavior.

It may be said in general that antiscorbutics exhibit less activity
when boiled than when in the natural condition; furthermore, heating
to 110° or 120°C. is harmful, even more so than boiling at 100°C.
It is not only the heating that exerts a harmful effect, but also the
incidental drying, which is in accord with the experience gained in
the study of human scurvy. According to Hoist and Frolich, the
relative moisture of the drying chamber plays a part, since the anti-
scorbutic activity of potatoes, carrots, dandelion and cabbage is
more quickly lost at room temperature than in an incubator at 37°C.
The dried dandelion, unlike cabbage, is completely inactivated.
Vegetable juices behave differently; for example, cabbage juice loses
its activity when heated for 10 minutes at 60°, 70° or 100°C.;
similarly, when it is stored at ordinary temperatures (in the presence
of preservatives) or in the ice-box. In the next chapter we shall
speak of the effect of the cooking on the antiscorbutic properties of
various foodstuffs.

Lemon juice behaves quite differently from the above mentioned
substances, as does also milk. The former may be heated to 110°C.
for 1 hour without any noticeable loss of activity. Now this juice
contains 7 per cent citric acid, and Hoist and Frolich held that

231

232 THE VITAMINES

vitamine C was more stable because of the acid present. This
idea was confirmed by the fact that other acid juices, like raspberry
and sour dock juice, show the same stability towards heating. The
heat stability of cabbage and dandelion juice was markedly increased
on the addition of acid, but permanent stability could not be obtained.

The last finding was made use of in extractions. If freshly dried
cabbage is extracted with alcohol to which a little citric acid is added,
instead of with pure alcohol, the extracts thus obtained are much
more active on guinea pigs. Other isolation experiments, such as
dialysis or extraction with petroleum ether, were valueless. Hoist
and Frolich were unable to favorably influence scurvy in guinea pigs
by intraperitoneal injections.

In a later communication, Hoist and Frolich (620) stated that
cabbage dried at 37°C. and stored in a desiccator still contained
vitamine C after 13 months. It was possible also to obtain an
active substance by extracting cabbage with a mixture of alcohol
and glycerine. This was also the case with extractions made with
dilute alcohol, to which 0.5 per cent citric acid had been added. Hot
80 per cent alcohol with 0.5 per cent citric acid likewise extracts
vitamine C from freshly dried cabbage. Since the experiments of
Hoist and Frolich in 1913, no real progress has been made, in spite
of the numerous publications that have appeared.

Shortly thereafter, the author undertook a chemical fractionation
of vitamine C, using the same methods as in the yeast fractionation,
but the results obtained were not very satisfactory. The investiga-
tion failed chiefly because of the lack of knowledge of scurvy in
guinea pigs, as well as of the behavior of vitamine C which at that
time had hardly been recognized. Although more firmly established
data are available on this subject at present, the chemistry of this
vitamine has thus far not progressed. In 1912 we investigated, in
this respect, milk, potatoes and lime juice, using guinea pigs and
rabbits kept on water and oats. The guinea pigs died in about 20
to 30 days with marked scorbutic symptoms. After the addition of
fresh potato juice, the animals died after 36 days, while after the
purification of this juice with lead acetate, the animals died after 27
days. A daily addition of 25 cc. milk had a definite inhibitory effect
on the scorbutic symptoms, while on addition of 50 cc. milk, the
symptoms of scurvy were absent even after 50 days, the animals
sometimes gaining weight; we shall discuss the dissimilar results of

THE ANTISCORBUTIC VITAMINE 233

Chick and Hume in the chapter on milk. Protein-free milk, prepared
by precipitation of casein with acetic acid at 50°C., lactalbumin
removed by colloidal iron solution, and concentrated by freezing
(this may perhaps be a practical method of concentrating vitamine
C), exhibited, in amounts corresponding to 200 cc. of fresh milk,
only a very slight activity. An alcoholic extract of milk dried in
vacuum by us, possessed no activity in large doses, while the residue,
given in doses of 30 grams showed definite activity in some cases.
We then turned our attention to lime juice (I.e. 101). which was
universally known as a splendid antiscorbutic. For this reason we
were unable at that time, to explain why it was inactive in guinea
pigs. The same result was also obtained with the decomposed
phosphotungstate, prepared from lime juice. We were beginning to
doubt the identity of scurvy in guinea pigs with that in man, though
recently the situation was completely explained by Alice Henderson
Smith (621). In a historical sketch, she showed that the statements
as to the protective action of lime juice on scurvy were not true for
the sour limes of the West Indies used to-day, but were so for sweet
limes and lemons growing in the Mediterranean regions.

The lime juice used to-day was shown to be inactive, but we
demonstrated the presence therein of vitamine B. Aside from this,
we isolated a terpene having the formula ds^Os, a purine base,
CcHrC^Ns (m.p. 282°C.) and a substance from the histidine fraction,
melting at 188° to 189°C. and having the formula C9H18O6N2. From
the choline fraction, a base was isolated as the double platinum salt,
which melted at 220°C. and which was given the formula, C8H1502N,
though a later investigation showed that the substance was identical
with stachydrine C7Hi302N. These data are quoted here because
it may later be shown that one of these substances is a cleavage
product of vitamine C.

The experiments of Hoist and Frolich on the extractability of
vitamine C with alcohol, were later confirmed by Freise (622) on
turnips, as weU as by Freudenberg (623) . Subsequently, Harden and
Zilva (624) showed that when orange juice is freed from citric acid
by precipitation with calcium carbonate, and the mixture treated
with an equal volume of alcohol, filtered, and the filtrate concentrated
in vacuum at 37°C., an active solution is obtained which, however,
loses its activity in time. If lemon juice is treated similarly, and
concentrated in acid reaction, an active residue is also obtained.

234 THE VITAMINES

Later, Harden and Robison (625) showed that vitamine C is not
volatile on distillation, that the dry residue has almost the same
activity as the original juice, and that it may be kept in a dry con-
tainer for six months. Apple juices were found to be much poorer
in vitamine C. This applies to juices concentrated in the labo-
ratory; when prepared in large amounts, these preparations lose 63
per cent of their activity, according to Harden and Robison (626).
Vedder (626a) prepared active extracts by successive extractions
with absolute alcohol acetone and ethyl acetate. Vitamine C was
destroyed by phosphotungstic .acid and the last extract exhibited
about one-seventh of the initial activity and contained only traces
of nitrogen. This led Vedder to the somewhat premature conclusion
that vitamine C is nitrogen-free. Without going further, we see
that active preparations of vitamine C may be obtained also in the
absence of citric acid, and we are therefore not yet certain whether
these acids possess protective qualities.

Givens and McClugage (627) dried orange juice in two different
ways. First, in shallow dishes at 55° to 60°C. for 50 hours; second,
according to a method used in drying milk (Just-Hatmaker process),
in which a quick drying at 75° to 80°C. is obtained. Whereas the
second preparation was almost as active as the original juice, the
first lost a great part of its effectiveness. The active products were
kept for 3^ months without loss of activity. Stable preparations
of vitamine C were made by Dubin and Lewi (628) and also by
Bassett-Smith (629). As regards the stability of vitamine C to
alkalis, a number of papers have been published. Harden and Zilva
(630) made orange juice alkaline with a /<r sodium hydroxide and
kept this for 24 hours, resulting in a loss of activity. Hess and
linger (631) carried out similar experiments and believed that, for
the most part, storage in alkaline medium exerted an unfavorable
influence on the activity. Accordingly, if the juice is boiled up in a
slightly alkaline solution and immediately neutralized, the activity
is only slightly diminished. McClendon and Sharp (632) investi-
gated the reaction of natural juices to determine if they are not some-
what alkaline, which would explain the destruction of vitamine C
on boiling; however, most of the juices were found slightly acid.

Sommer and Hart (633) stated that the destructive action of boiling
on vitamine C was not due to the precipitation of citrates, while
Faber (634) found that an addition of sodium citrate (even in a con-
centration of 0.25 per cent) has a destructive action on this vitamine.

THE ANTISCORBUTIC VITAMINE 235

As regards the oxidizability of vitamine C, Fowler (635) showed
that milk preserved with hydrogen peroxide is scurvy producing.
Hess (636) believed that milk or canned tomatoes shaken with air
lose considerably in their antiscorbutic activity.

According to Zilva (I.e. 513) ultra-violet light has no influence on
the activity of vitamine C. Adsorption likewise does not affect this
vitamine. It may be treated with fullers earth or colloidal iron,
according to Harden and Zilva (I.e. 89), and it may also be filtered
through a Berkefeld filter without loss of activity. Zilva and
Miura (636a) were unable to separate vitamines B and C by means
of differential dialysis.

As for the significance of vitamine C in the organism, very little is
known, and what little knowledge there is available will be described
under human scurvy. The question as to whether the parenteral
administration of vitamine C is effective has been answered differently
by different investigators. Hoist and Frolich (I.e. 359) gave this
vitamine intraperitoneally, and Harden and Zilva (I.e. 624) found
it inactive on subcutaneous injection. Hess and Unger (637), on
the contrary, working with children, showed that intravenous
therapy with weakly alkaline orange juice is possible, and they
recommend this procedure.

Here, too, as with vitamine B, Hess (638) accepted first that a
large addition of carbohydrates hastens the onset of scurvy, but
he (639) has changed his view recently. Hess has tried by addition
of 3 per cent flour to the milk to hasten the onset of scurvy in chil-
dren, with negative results. We do not believe, however, that this
amount of carbohydrate is sufficient to influence the effect one way
or the other.

CHAPTER V

VITAMINE CONTENT or VARIOUS FOODSTUFFS, IN THE NATURAL
AND PREPARED CONDITIONS

We have already shown that during the last few years most food-
stuffs have been investigated for their nutritive value and vitamine
content. The results obtained give us an insight into the compara-
tive values of the above, and yet we must be careful in interpreting
the data. First of all, it should not be forgotten, as frequently
happens nowadays, that the results obtained with one animal species
cannot, ipso facto, be applied to others.

For instance, it seems to us unjustifiable to determine the nutritive
value of an animal diet by using animals that normally live on plant
diet; this may be the cause of the wide differences between experi-
ments on man and those on animals. Variations in the antiscorbutic
value of meat in man and guinea pigs may be due to just such
procedures.

Another difficulty in the interpretation of results is the lack of
quantitative methods for the estimation of the vitamine; this is most
apparent in the comparison of the results obtained at the outset of
vitamine research, with those of modern investigations carried out
with a better understanding of this new subject. Many of the older
results were obtained without paying any attention to quantitative
relationships, and Chick and Hume (640) correctly noted that in
this regard, we should be constrained to work quantitatively. For
example, if we would wish to study the effect of alkali on vitamines, it
does not suffice merely to add alkali to the foodstuff and feed it to
the animal. We must first determine the minimal protective dose of
the natural preparation, for if too great a dose is used from the start,
the preparation, in spite of an extensive destruction, may still contain
sufficient vitamine to act therapeutically, and we would make the
erroneous conclusion that the treatment to which the foodstuffs had
been subjected was without influence on the vitamine content.

Most of the animal experiments conducted up to the present
suffered because of the fact that with the addition of vitamine in the
form of food, or in the form of extracts, the composition of the basal
diet was at the same time altered, without having the corresponding

236

VITAMINE CONTENT OF FOODSTUFFS

237

controls with no vitamine addition. Most of the investigators also
regarded the vitamine content of any particular diet as an unchange-
able constant, as a mathematical certainty. We shall see in this
chapter that this is not at all the case, and it will become clear how
the various investigators could have obtained dissimilar results with
the same food.

After pointing out the possible relationship between vitamines and
appetite (Mendel 641), and the necessity of new legislation for food-
stuffs with regard to the vitamines (Ranwez 642), we shall take up
the description of the treatment of foodstuffs as related to the vita-
mine content. Here, there are the effect of heating, cooking, drying,
storing, sterilizing, and preserving. Here, too, we shall describe the
more important foodstuffs commonly used in human nutrition. It
will be impossible, however, to go into all of the investigations made
in this respect, and for this reason we shall put all of the data con-
sidered of importance in the form of a table at the end of this chapter,
showing the relative richness of the foodstuffs in the three known
vitamines. The first chart of this kind was made by Cooper (643)
in 1912-14, including but a few foodstuffs and concerning itself only
with the vitamine B content. A similar table was given in the
Report of the Medical Research Committee (I.e. 333) wherein it was
attempted to record quantitatively the values found. The values for
vitamine B are recorded, compared to 100 for wheat germ.

FOODSTUFFS

VALUE

MOISTURE CONTENT

Wheat germ

100

per cent

10-13

Wheat bran

25

10-13

Rice germ

200

10-13

Pressed yeast

60

70

Dried peas

40

12

Lentils

80

Egg volk. .

50

70

Ox liver

50

70

Ox muscle

11

75

Potatoes

4.3

80

In the same report, we see also the first investigation of the con-
tent of other vitamines. All of these values were obtained on
animals and apply strictly only to the species upon which the results

238 THE VITAMINES

were determined. This is true especially as regards quantitative
relationships. Chick and Dalyell (644) prepared another table with
regard to vitamine C, where the values are based on 100 for lemon
juice.

Fresh orange juice or cabbage leaves 110

Fresh raw orange juice 100

Beet juice (swede) 60

Green beans 30

Sprouted fresh peas 30

Carrot juice 7.5

Red beet juice 7.5

Meat juice (ox) 7.5

Potato (cooked 39 minutes) 7.5

Fresh cow's milk 1-1 . 5

In our table, we were not satisfied to give only the vitamine value,
but also the nutritive value of the proteins of various origin. This
rubric will, perhaps, help us to demonstrate the difference between
the nutritive value of plant and animal protein, which may be of
importance in relation to the etiology of pellagra and war edema;
unfortunately, however, the data found on this subject are very
meagre. It appears to us not impossible that some protein substances
may contain adsorbed either a fourth vitamine, or a hitherto unknown
amino acid, essential to life.

INFLUENCE OF HEATING AND COOKING ON THE VITAMINE CONTENT1

We owe to Schuffner and Kuenen (I.e. 56) the first data on this
subject of such practical importance. Studies on the stability of
vitamine B in foodstuffs were undertaken also by Grijns (I.e. 52).
Recently this field was systematically investigated by Chick and Hume
(645), using an improved technique. They studied the influence of
heat on vitamine B of wheat germ and found that a temperature
of 100°C. for two hours has little influence, while a temperature over
120°C. is quickly followed by destruction. Miller (646) found that
cooking and autoclaving (at 115°C. for 45 minutes) had no effect oh
carrots and beans; this was confirmed by Whipple (647) for cab-
bage and onions. In the last two investigations the content of
vitamine B was determined by the yeast method already described

1 A good review of the known data is found in the paper by Emmett and
Luros (I.e. 94).

VITAMINE CONTENT OF FOODSTUFFS 239

by us, but was not controlled by animal experiments. The stability
of vitamine C against heat depends very much on the nature of its
source. Weill and Mouriquand (648) stated that this vitamine is
very labile, but noted that lemon juice may be heated to 110°C. for 1
hour without marked loss in activity. Delf and Tozer (649), co-
workers of Chick, carried out investigations on cabbage : heating for
1 hour at 60°C. decreased the antiscorbutic value about 70 per cent,
at 90°C, about 90 per cent, while on short heating (20 minutes), at
90° or 100°C., the activity is decreased only about 70 per cent; we
see from this that the chief influence is produced by the time of heat-
ing and not by the temperature. Strudwick (650) investigated, in
this respect, the influence of cooking in a fireless cooker in which the
time of cooking was markedly prolonged. The results showed that
a rapid heating is much superior to the above method.

Delf (651 )2 investigated, in the same way, the relationship between
cabbage leaves and cabbage juice, showing that in the natural con-
dition, vitamine C is much more resistant. Turnip juice (swede)
was shown to be far more stable than cabbage juice. Orange juice
heated to 130°C. lost 50 per cent of its activity. Givens and McClug-
age (652) found tomatoes quite resistant to heat. However, if they
are heated for 15 minutes or more at 100°C, a greater amount, in
comparison with the crude material, must be fed in order to protect
guinea pigs against scurvy. Aside from heating, oxidation appar-
ently also plays a role in the inactivation of vitamine C. Rossi (653)
fed guinea pigs on oats and hay sterilized in open kettles at 126°C.,
with the result that the animals died of scurvy. However, a different
result was obtained when the food was heated as above in closed
kettles; on this food, the guinea pigs were still alive after two months.
Delf (I.e. 651) also believed that in the presence of air the inactiva-
tion of vitamine C was hastened, and this condition, according to
Steenbock, Boutwell and Kent (I.e. 594), and others, plays a part
in the destruction of vitamine A.

The destruction of vitamines by heat depends on still another
factor — the chemical reaction of the starting material. In this
connection, the data are not so certain for vitamine B. While McCol-
lum and Simmonds (I.e. 315) reported on the supposed destruction

2 In comparing the temperature coefficient of the heat destruction of
vitamine C with toxins and ferments, Delf concluded that this vitamine
must possess a relatively simple chemical structure.

240 THE VITAMINES

of this vitamine by alkalis, Daniels and McClurg (654) stated that
they observed no destr iction of cabbage and soya beans so treated.
It is possible, however, that the latter investigators fed too much
of the active material. According to Daniels and Heisig (655), the
effect of the addition of soda upon vitamine C was deleterious. In
this connection, Hess and Unger (656) point out that soda is used in
the preparation of proprietary infant foods. This is true also of
commercial cod liver oil emulsions, which give an alkaline reaction,
and to which soda is added in order to obtain a better emulsion. Yet
we have no exact experiments to show whether the activity of the
above emulsions is affected by the manner of preparation. Experi-
ments started by us have not yielded any definite results up to the
present. We must say, however, that the vitamine A of cod liver
oil must be quite resistant in order to withstand the treatment
undergone during the process of manufacture.

It has often been stated that the stability of orange juice is
explained by the acid reaction of the medium. Still, according to
the findings of Delf (I.e. 651), the neutralization of the juice did not
ha.ve any harmful effect.

Summing up the subject matter of this important chapter, we may
say that the stability of the vitamines on heating depends not on
the temperature but on the duration of the heating. In this respect,
the view of Givens and McClugage (657) is of interest; they believed
that when antiscorbutics are heated at a low temperature for a long
time, as in gradual drying, the ferments contained in the tissues
destroy the vitamine. If the ferments are inactivated by rapid
heating at high temperatures, then vitamine C is much more resist-
ant. These statements naturally require further corroboration.3
Aside from this, the vitamines seem to be more stable in the natural
condition than in the form of extracts or pressed-out juices. Oxidation
plays a big part in the destruction of vitamines A and C, while the
chemical reaction of the substrate plays a part in the destruction
of all the vitamines.

3 This behavior may be brought into relationship with the stability in
acid solution, if it is assumed that these ferments act in alkaline solution;
we have in mind here the oxidases, the action of which is delayed in acid
medium.

VITAMINE CONTENT OF FOODSTUFFS 241

The vitamine content of cooking water

That vitamines may be lost by discarding the cooking water, was
first stated by Schuffner and Kuenen (I.e. 56). In addition, vita-
mines are lost in other water-soluble substances and it is therefore not
without reason that Hindhede (658) pointed out the great nutritive
value of soups, especially vegetable soups. Hill (659) stated, for
example, that on cooking potatoes, losses ensued which represented
70 per cent of all water-soluble substances. From the viewpoint of
vitamines, the same thing was pointed out by a number of workers —
Denton (660), Daniels and McClurg (I.e. 654), Miller (I.e. 646) and
Whipple (I.e. 647). The last two investigators, in particular, found
that by discarding the cooking water, 30 to 70 per cent of the sub-
stance necessary for the growth of yeast was lost.

THE INFLUENCE OF DRYING

Since the question of the dehydration of foodstuffs has already
assumed practical importance, it is regrettable that comparatively
little is known of its effect on vitamines. Murlin (661) believed that
the nutritive value of dried foodstuffs is strictly comparable with
that of fresh. Prescott (662) put the practical side of this problem
to the fore, in relation to transportation and storage, particularly for
the provisioning of any army in the field. He stated also that the
loss of vitamine C on drying may be prevented by modifying the
procedure. Givens and Cohen (663) investigated this question, using
cabbage and potatoes. Drying in a stream of air at 40° to 52°C.
resulted in an appreciable loss of activity. With the simple drying,
the preparation of foods is not yet completed, and practically two
other factors are to be considered — the storing and the cooking just
before eating. These two factors have unfortunately not been
considered by most investigators; nevertheless, it was found that
when cabbage or potatoes were first heated or cooked and then dried,
no more vitamine C was left, as shown in guinea pig experiments.4

4 Hoist and Frolich (663a) found that if moisture is excluded as much as
possible to diminish the chances of hydrolysis, a stable cabbage preparation,
as regards vitamine C content, may be obtained. The dehydration was
carried out in vacuum at 37°C. in presence of phosphorus pentoxide, which
is replaced as soon as it becomes moist. This preparation was kept for 10, 18,
and 26 months and found active.

242 THE VITAMINES

Givens and McClugage (664) found tomatoes more resistant in this
respect, at least in relation to storage after drying, since after three
months, the activity was still retained. Subsequent cooking experi-
ments appear to have not been made, but are not particularly impor-
tant inasmuch as tomatoes may also be used raw. Shorten and Ray
(665) dried various kinds of vegetables in the sun, and investigated
their vitamine B and C content. The results showed that vitamine
C of carrots, onions, and cabbage were still present in sufficient
amounts for guinea pigs. Spinach, on the contrary, lost this prop-
erty. We find no statements here to the effect of ageing and subse-
quent cooking. Falk, McGuire and Blount (666) studied the ferments
of dehydrated vegetables. They found them inactivated and pointed
out the similarity to the resistance of the vitamines. Givens and
McClugage (I.e. 657) published a very careful investigation upon the
drying of potatoes. Taking their work as a whole, we find that when
potatoes undergo a series of treatments, the individual effects are
summed up, resulting in a totally inactive product. If the potatoes
are boiled, steamed or baked and then dried, an inactive product
is obtained, as a rule. Drying alone does not have this destructive
effect. The vitamine was least affected when the potato was baked
in the jacket (perhaps on account of absence of air) and then dried.
Addition of acids before treatment did not have the desired effect.
Potatoes dried in vacuum also lost considerable activity. We have
already seen that quick drying at high temperatures is less destructive
than gradual drying at low temperatures. Ageing and subsequent
drying were not investigated, although it is clear from what has
already been said that vitamine C of potatoes is not resistant.

CANNING

This subject too has not yet been extensively investigated. We
believe that for this question, the foodstuffs may be classified in two
groups — those used raw, and those to be cooked before consumption.
We find the conditions here similar to those in drying. Bigelow (667)
described the problems arising in canning. Different sources of
supply give rise to different conditions so that general conclusions
can hardly be drawn as yet. Hess and Unger (668) found that canned
tomatoes still showed a splendid antiscorbutic action in children, and
adopted this antiscorbutic in their children practice; it is specially
suitable for institutions since the expensive orange juice may be

VITAMINE CONTENT OF FOODSTUFFS 243

substituted by something cheaper. This finding is of practical
significance, since in the anticipation of scurvy epidemics, canned
tomatoes, which appear to keep well, may be used, though it should
be remembered (Hess and linger, I.e. 631) that the activity decreases
on cooking. Daniels and McClurg (I.e. 654) believe that the canning
proces does not destroy vitamine B; it seems to us that an absolute
proof of this has not been obtained in canning, although they admit
that changes initiated during canning may progress automatically dur-
ing storage. Campbell and Chick (669) prepared cabbage and beans
in a manner similar to that used in canning, and found that these
substances lost about 66 per cent and 75 to 90 per cent of their
activity, respectively. Vitamines A and B were also affected although
the investigators explained it by losses of vitamine A into the cook-
ing water. When this was given to guinea pigs, it resulted in better
growth. We must assume, therefore, with Campbell and Chick,
that vitamine A went over into the cooking water.
.N

AGEING AND STORING

The last factors to be taken up in this connection are those of ageing
and storage. Hulshoff Pol (670) could cure human beriberi with
puree of peas, although peas, which were three years old, were entirely
inactive. Weill, Mouriquand and Michel (I.e. 392) found the same
to be true of denatured meat, stored for 14 months. For cats, this
meat was much more harmful than freshly denatured meat. Delf
and Skelton (671) dried cabbage and stored it at ordinary tem-
perature; after 2 to 3 weeks, the preparation lost 93 per cent of its
activity and after 3 months, 96 to 97 per cent. Hess and Unger (672)
found no vitamine C in prunes, while orange peel was active even
after 3 months. The storage of orange juice has a definite influence,
since in old juice, only half of the activity remained. Dried carrots,
kept for 3 to 5 weeks and for 7 months, were quite inactive. Hess
and Unger concluded, from this that dried vegetables actually retain
their nutritive value, though as regards vitamine C, they cannot
replace the fresh natural products. The ageing applies not only to
storing and drying, but also to fresh vegetables. In this respect,
Hess and Unger (I.e. 631) investigated carrots of various ages. While
young carrots retained their activity even after cooking and drying
in a current of air, and storing for 3 months, this was not the case
with old carrots.

244 THE VITAMINES

From what has been said, we see that universal rules as to the
stability of the vitamines cannot be formulated. Every single case
must be investigated separately, since we do not yet know of any
rule for determining the stability or lability of these substances.
Evidently, the vitamine content of foodstuffs prepared for ingestion
depends considerably upon its original content in the natural state.
This original content may vary with the country in which it is
produced, with the condition of the soil, the climate and so on.
This is true to our primary sources of vitamines. As for the second-
ary sources, animal products, the question is much more complicated
and depends, for the most part, upon known factors, but partly
upon unknown factors too, which we have already discussed.

CEREALS

Under this heading we shall describe not only the cereals, such as
wheat, barley, rye and maize but also the breadstuffs. Although an
etiological relationship between the refining of cereals for human
consumption and beriberi has been suspected for a long time, and in
fact demonstrated, the economic conditions arising out of war have
stimulated interest in the question and in established knowledge on
the subject. In almost all countries, the investigators have endeav-
ored to utilize the nutritive value of cereals to the best possible
advantage. To the problem of the economic utilization of the grain
kernels, another factor is added which we have already noted in the
etiology of avian beriberi, namely, that in most cases, vitamines A
and B are localized in a certain portion of the kernel. Because of
this, modern refining may result in a more or less complete removal
of these essential factors. It is indeed true that under normal con-
ditions, when the population eats other foodstuffs, in addition to
bread, there is no immediate danger of an avitaminosis, and yet we
must remember that at present bread may constitute a large per-
centage of the diet. Although whole wheat bread is used almost
exclusively in many countries, in more cultured sections, white bread
is used in preference. Aside from the aesthetic factor, some scientific
facts are known which make it desirable to mill the grain. One of
the reasons frequently cited is the fact that flour from whole wheat
keeps with greater difficulty. In fact, the whole grain, very rich in
fat, is very difficult to store. The polishing of rice is often defended
on these grounds, and Ottow (I.e. 518) proposed measures for increas-

VITAMINE CONTENT OF FOODSTUFFS 245

ing its conservability. This is even more applicable to corn, so rich
in fat. Winton, Burnet and Bornmann (673) state that corn may
be stored much better if the germ has been removed. The second
reason why it is difficult to introduce whole grain in practice, is that
many investigators are of the opinion that bread baked with the
chaff in uneconomic. The assimilation of this bread is supposed to
be not only poor, but it occasions losses in other foodstuffs simulta-
neously consumed. Still, the investigators are not agreed upon this
point. While Hindhede (674), Rohmann (675), Eijkman (676),
Hulshoff Pol (677), Butcher (678) and Stoklasa (679) believe that
whole wheat bread is very well utilized; we also encounter views,
such as those of Pugliese (680), van Leersum and Munk (681) and
Rubner (682) who asserted that such breadstuffs are poorly utilized
and in fact provoke losses, as has been noted above.

Since the time of Liebig (683), we have known that milling of
grain decreases its nutritive value; Liebig said: "No single food-
stuff loses its value so readily as whole grain through the modern
process of milling. The whiter the flour the less nutritive value it
possesses." In 1871, Magendie (684) knew that pigeons die when fed
on fine wheat flour, while on rye, they do very well. Dogs on wheat
flour die after 40 days, while on coarse "soldiers' bread" they continue
to live. Oseki (685) investigated the value of various kinds of bread
on mice. He found rye and army bread suitable, while wheat bread
and barley flour were unsuitable. Weill and Mouriquand (686)
chose to add extract of bran, instead of the bran itself, in baking
bread, whereupon the addition of the occasionally objectionable
cellulose was obviated. Sherman, Rouse, Allen and Woods (687)
believed in the greater nutritive value of the whole wheat kernel as
compared with white bread, judging from their rat experiments.

Our knowledge as to the localization of the vitamines in grain
kernels has undergone some modification in recent years. While
the first workers in this field stated that the protective substances
are found in the "silver skin" (a thin membrane surrounding the
rice kernel), Fraser and Stanton thought the protective substances
were to be found in the aleurone layer. Later, McCollum and Davis
(688) showed that vitamine B is found chiefly in the germ. Chick
and Hume (689) found this vitamine in rice, wheat and corn, partially
also in the pericarp. Voegtlin, Lake and Myers (690) believed that
they could show by animal experiments that vitamine B of wheat

246 THE VITAMINES

and maize occurs chiefly in the germ, as well as in the pericarp;
in this they adopted the view of Chick and Hume at first. Later how-
ever, Voegtlin and Myers (I.e. 271) showed that when wheat or corn
kernels (see fig. 53-54) are cut into two portions, and the part without
the germ fed to animals, results are obtained which indicate an
absolute lack of vitamine B; therefore, the vitamine must be in the
germ only. However, it must not be forgotten that by the procedure
followed by Voegtlin and Myers, other constituents are removed (for
example, protein), and that possibly the vitamine is also present in
the remaining part of the grain in sub-optimal amounts. Funk and
Dubin (I.e. 493), showed that the substance promoting the growth
of yeast was found in the germ, but it was likewise present outside
of the germ in appreciable quantities. For practical purposes,
according to Greig and Curjel (691), it seems sufficient when wheat
germ is added to zwieback to the extent of 15 per cent. This measure
was adopted for the English colonial army in order to protect the
personnel against beriberi.

When cereal flours are heated in baking, the vitamine B content is
altered. Weill and Mouriquand (692) heated decorticated barley to
120°C. for 1| hours. When this was fed to pigeons, they died after
30 days with definite beriberi symptoms. On white rice, this disease
appears in 18 to 25 days and on sterilized rice, in 9 to 16 days. They
believed, therefore, that decorticated grain kernels still contained
vitamine, especially since the symptoms appeared sooner when the
food was sterilized. Chick and Hume (I.e. 645) heated wheat germs
to high temperatures, whereupon they observed inactivation at
120°C., but not on heating at 100°C. for 2 hours. From these results,
they believed that baking does not affect vitamine B to any great
extent. If yeast is used in the dough, then a part of the B vitamine
of the bread may arise from this source.

We have frequently touched upon the point that seeds develop
vitamine C upon germination. This fact, first observed by Furst,
was further developed for practical purposes by Chick and Delf
(693), and in addition to cereals, was confirmed in peas and beans.
Weill and Mouriquand (694) conducted similar experiments with
germinating barley and later they showed, in conjunction with
Pdrronet (695), that vitamine C develops in greater amounts only in
the later stages of the germination.

VITAMINE CONTENT OP FOODSTUFFS

247

FIG. 52. 1, UNPOLISHED RICE; 8, PABBOILED RICE; S, POLISHED RICE
(FKASEE AND STANTON)

248 THE VITAMINES

Rice

We have already touched upon most of the data on rice, including
those of Fraser and Stanton (I.e. 49), who showed by means of
illustrations how the loss of substances occurs during the polishing of
rice. From the appended illustration, we see why such great differ-
ences in the number of beriberi cases are apparent with the use of
polished and parboiled rice. The question as to whether polished
rice still contains vitamine B has likewise already been discussed.
It is important now to determine whether this rice contains vitamine
A, but we find no observations on this point. It would be of special
interest to see whether pigeons can be maintained for a long time on
polished rice with the addition of vitamine B. Funk and Dubin
(696) have started experiments in which pigeons are being fed on
rice to which has been added 5 per cent casein and 3 per cent salts,
and the mixture autoclaved at 120°C. for 3 hours. By means of the
prolonged heating, it was expected that the rest of the vitamine A
would be destroyed. The pigeons have gained in weight and have
remained in good health for over six months. Despite the fact that
others have investigated this problem, it is not yet certain whether half
grown pigeons can live without an addition of vitamine A, or whether
it must be concluded that autoclaved, polished rice Still contains traces
of vitamine A. As regards the nutritive value of rice polishings,
Mattei (697) believed that pigeons could not be maintained on rice
polishings alone; he found that an addition of white rice was neces-
sary. Wise and Broomel (698 ) investigated the effect of the modern
milling methods on the composition of the remaining rice kernel. On
polishing, the husk germ, 6 layers of bran and a part of the seventh
layer were removed, so that the total loss in weight was about 10
per cent. On analysis, it was possible to show a loss of 70 per cent
ash, 85 per cent fat, and 10 per cent protein. To prevent these
losses, Guareschi (699) has suggested the use of rice which is not so
highly milled.

Barley

According to the older statements about barley, it seems that its
vitamine B is not localized in the same way as in other cereals. In
the older Japanese data, for example, that of Saneyoshi (700), we find
that barley can cure avian beriberi, whereas Weill and Mouriquand

VITAMINE CONTENT OF FOODSTUFFS

249

(701) used decorticated barley to produce beriberi. Steenbock, Kent
and Gross (702) observed that for rats, barley, like other cereals, was
deficient in vitamine A, protein and salts.

Oats

McCollum, Simmonds and Pitz (I.e. 372) regarded the composition
of oats as unfavorable for the nutrition of animals. Tested on rats,
it showed a lack of vitamine A, protein and salts. This applies much
less to pigeons, as we ourselves have shown.

Wheat

Voegthn and Myers (I.e. 271)
believed that vitamine is found ex-
clusively in the germ; we have
already published our findings in
this respect (I.e. 271). How the
various parts of the wheat kernel
are related to each other, we may
see from the following diagram,
taken from the work of the above
investigators, from which we may
note the method of sectioning used
by them in their experiment on
the localization of vitamine B.
McCollum, Simmonds and Pitz
(703) investigated, in particular,
the vitamine value of wheat germ.
Qualitatively considered, this por-
tion of the wheat has been shown to be adequate, but for practical
reasons, an addition of salts must be given. Vitamine B occurs
in the germ in large quantities; vitamine A, on the other hand,
in smaller amounts. Osborne and Mendel (704) investigated the
nutritive value of various milling products of wheat kernels, and
did not believe in the exclusive localization of vitamine B in the
germ. The contiguous portion was also rich in vitamine B, as was
also the periphery of the kernel, the endosperm and bran not con-
taining very much.

FIG. 53. WHEAT KERNEL SHOWING

DIFFERENT LAYERS (VOEGTLIN-

MYERS)

250 THE VITAMINES

Maize

Because of a possible etiological relationship of corn consumption
to pellagra, the numerous investigations on the nutritive value and
composition of corn and corn products are of special importance.
Bezzola (705) showed that corn does not provide an adequate diet
for the maintenance of guinea pigs in good health; they lose hair,
develop severe diarrhea and subsequently die. A difference between
the use of good and bad corn was not observed. Lucksch (706)
found that guinea pigs, fed on good corn, lost their hair, showed a
hyperemia of the intestinal mucosa and an enlargement of the adre-
nals. On a mixture of corn, flour and greens, they lost hair never-
theless, and developed paralysis of the hind legs and catarrh of the
small intestine. Corn was likewise shown to be inadequate for
rabbits and dogs, the results obtained in spring being more definite
than those in autumn. The blood of the experimental animals was
investigated and found sterile at all times, v. Neusser (707)
reported on a disease of horses, called "emmaisadura" in Mexico and
Colombia supposed to be caused by rotted corn. The symptoms of
this disease are emaciation, dizziness, lethargy, cramps, wildness, loss
of hair and teeth and decay of the hoofs. Hoist (708) remarked on
the frequent occurrence of scorbutic symptoms in pellagra, especially
the porosity of the bones. He repeated Lucksch's experiments with
guinea pigs and came to the conclusion that the disease described by
him could be prevented by the addition of fresh cabbage. In par-
ticular, the constant loss of hair could be overcome although the
experiments were carried out in spring. The whole syndrowe was
thereupon regarded by Hoist as scurvy. Baglioni (709) likewise
carried out some feeding experiments with corn products on guinea
pigs. In the light of modern investigations, Hoist was right when he
characterized as scurvy, the disease developed by guinea pigs fed on
corn. Since in most corn-feeding experiments, no antiscorbutic was
given, the resulting condition was complicated by scorbutic symptoms
to such an extent, that these investigations are of no value in the
explanation of pellagra. The problem appears in a different light
when birds are used as experimental animals. Ohler (I.e. 262)
observed that when decorticated maize is fed to chickens, they

VITAMINE CONTENT OF FOODSTUFFS 251

develop beriberi. Under the same experimental conditions, Driscoll
(710) noted a condition which he thought was similar to pellagra in
man; among other symptoms, he observed an erythema of the legs;
the chickens recovered when given whole corn. Clementi (711) fed
chickens on cooked corn and "polenta" and observed a development
of nervous and digestive disturbances, but only after many months
of feeding. Szalagyi and Kriwuscha (712) also conducted some corn
feeding experiments with chickens, ducks and geese.

Urbeanu (713) carried out some rat and chicken experiments, and
noted some skin symptoms in both. In the former, this was apparent
only after 10 to 12 months of feeding, and was favorably influenced
by the addition of potatoes and soya beans. Suarez (714), experi-
menting with pigeons and mice, observed in both a disease similar
to beriberi after 25 to 35 days, which was favorably affected by
yeast. Experimenting with dogs, Nitzesco (715) stated that corn
was not a complete food. Rondoni (716) attempted to use guinea
pigs in his pellagra work, but recognized that these animals were
unsuited for this purpose, and therefore suggested that the work be
continued on man and monkeys.

Hogan (717) fed corn simultaneously to rats and pigs; in the
former, this diet was very nearly sufficient, but in the latter, a lack
of protein was noted, since on adding egg-albumin, growth was
resumed. This brings us to a consideration of the nutritive value of
corn proteins, a subject which has assumed considerable significance
in the etiology of pellagra. In this connection, Mendel and Fine (718)
carried out a series of dog experiments. According to their results,
corn glutellin, a preparation consisting of zein and glutenin, seemed
to be as well utilized as meat. The relationship between the two
protein components varied within certain limits, but averaged 1:1.
The proteins were separated by means of alcohol, in which zein is
soluble. Osborne and Mendel (719) and Baglioni (720) conducted
experiments on the nutritive value of corn protein. The utilization
was better with the flour than with the proteins isolated. All three
products however gave a nitrogen retention, and it would therefore
be concluded that corn also contains proteins of a higher nutritive
value than that of zein. McCollum and Simmonds (721) found that
when rats are fed on a mixture of 80 per cent corn and 20 per cent
beans, there is still evidence of a lack of vitamine A and protein.

252 THE VITAMINES

The investigations of Johns, Finks and Paul (722) gave the following
percentage composition of corn as regards various protein compounds:

Globulin 21.9

Zein 41.4

Glutellin 30.8

Alcohol-insoluble part 5.9

While it is known that zein is not a complete protein, the com-
position of glutellin shows it to be of good nutritive value. As an
explanation of the poor nutrition resulting from an exclusive corn
diet, these investigators stated that certain flour products lack, first
of all, vitamine B ; second, although they contain complete proteins,
they occur in dilute form, so that the protein must be added as con-
centrates. It is interesting to note that this view corresponds
exactly with our present conception of the matter. Osborne and
Mendel (723) found that the dietary deficiency arising from corn
meal feeding is caused by a deficiency of protein. This deficiency
may be corrected by an addition of skimmed milk powder, which
must amount to 50 per cent of the corn fed, before the nutritive
condition is restored to normal.

We come now to a consideration of the vitamine content of corn,
and how it is affected by milling. It has been known for a long
time that corn germ contains much more fat than the whole corn.
The appended table is taken from the work of Woods (724).
The protein content varies in the same way.

FAT

PROTEIN

Whole corn

per cent

4.3

per cent

12.7

Endosperm

1.5

12.2

Husk

1.6

6.6

Germ

28.6

21.7

We have personally made analyses of various South Africa corn
products (I.e. 525) and concluded that milling causes considerable
losses, which may be prevented. Our figures on milling are recorded
below.

VITAMINE CONTENT OF FOODSTUFFS

253

Data pertaining to milled maize (97 per cent of the total grain)

COLOR RE-

WATER

a

m

NITROGEN

PHOSPHORUS: PaO,

\

FATTY ACIDS

CHOLESTEROL

LIPOID-P2O«

ACTION.
FOLIN AND
MACALLUM

Kjeldahl

Dumas

Melanin

Van Slyke

Alcoholic
extract

Hot

Cold

1. Whole

maize

grain

12.71

1.56

1.73

1.74

0.14

0.99

0.56

4.5

3.88

0.247

0.0154

0.35

0.55

2. Highly

milled

meal 86
per cent
3. First

12 63

1 48

1 67

1 73

0 13

0 %

0 36

3.87

3.36

0.22

0 0098

0.23

0.45

milling

from no. 2

10.48

2.09

1.23

0.14

0.61

0.30

4.66

4.18

0.357

0.0164

1.00

1.00

4. Second

milling

from no. 2

10.71

4.04

2.31

2.34

0.16

1.40

1.43

12.79

11.09

0.438

0.0353

0.70

0.60

5. Slightly

milled

meal, 97

per cent

12.41

1.60

1.73

1.84

0.12

1.00

0.54

4.21

3.63

0.233

0.0153

0.30

0.45

6. Bran

from 5,

above

10.55

1.40

0.65

0.07

0.300.23

1.86

1.58

0.2710.00840.35

0.70

We have found nothing in the literature pertaining to the nutritive
value of the proteins of different corn layers. It is, of course, possible
that the two chief proteins of corn — zein and glutenin — are localized
in various layers, so that milling may deprive the corn of glutenin,
and hence enrich it with the less valuable zein. With the above
analyzed products, we have carried out feeding experiments on
pigeons, but have never observed beriberi. From the diagram of
the corn kernel (fig. 54), it is possible to see the localization of the
germ, explaining the loss of substance and nutritive value during
milling. Analyses of corn milling products were also published by
Juritz (725) and by McCrae (726). Poppe (727) found, besides this,
that by steeping the corn, similar to cooking, the nutritive substances
extracted by the water amount to 36.2 per cent. McCollum, Sim-
monds and Pitz (728) found that corn contains all of the necessary

254

THE VIT AMINES

amino acids, but not in the proper proportion to each other and,
in addition, was poor in vitamine A and salts. While this was found
to be the case in rats, Weill and Mouriquand (729) were able to
maintain pigeons on whole corn for more than 240 days, whereas
decorticated or sterilized corn produced severe disturbances. Voegt-
lin, Lake and Myers (I.e. 690) found that whole corn contains suffi-
cient vitamine B for pigeons. However, if they fed the same corn
with the biggest part of the aleurone layer and germ removed the
pigeons and chickens developed, in about 3 weeks, a disease similar
to beriberi which could be cured by giving vitamine B. Voegtlin,
Sullivan and Myers (730), as well as Hughes (731), also obtained
similar results.

FIG. 54. CROSS-SECTION OF CORN KERNEL
A, endosperm., B, husk, C, germ

Steenbock and Boutwell (I.e. 588) showed, contrary to the ruling
view that corn is poor in vitamine A, that certain strongly yellow
colored varieties are quite rich in this vitamine. In connection with
this, we wish to emphasize once more that corn is a complete diet for
most species of birds. It is different with mammals, for which corn
is not so favorable, and this fact is now generally attributed to the
low biological value of the corn proteins, although, inasmuch as corn
contains a protein that is adequate for birds, it is incorrect to speak
of the low biological value of corn proteins. In experimenting with
mammals, particularly those that can develop scurvy, we must not
forget that corn, at least when dry, contains no C vitamine. This

ViTAMINE CONTENT OF FOODSTUFFS 255

minimal value of corn proteins does not depend so much on the
unfavorable content of amino acids, as that for some animals, the
dilution of the complete proteins by the incomplete, and by large
amounts of starch, has an unfavorable effect on the utilization.

POTATOES

We have already stated that in a population that is nourished
chiefly on potatoes, no avitaminoses appear and pellagra is unknown.
The value of potatoes as a protection against deficiency diseases has,
to a certain extent, been recognized by some investigators. Although
the protein content of potatoes is small, and although large quan-
tities must be used because of the great dilution of the more valuable
components present, it seems to be certain that this dilution is harm-
less to people. The author has often had the opportunity of observing
Polish peasants at their meals. When the men returned from the
fields, an enormous plate of potatoes was set before them, so that a
city dweller could hardly conceive of the possibility of consuming
such a great amount of food. Still, the entire contents of the dish
(not plate), cooked with a trace of lard to give added taste, dis-
appeared in an unbelievably short time. From the many modern
investigations on the nutritive value of potatoes, it is clear to us
why these peasants must consume such an unusual amount of pota-
toes. It is correlated with the small content of protein, together
with the three known vitamines. The splendid utilization of potato
proteins was unquestionably demonstrated by Hindhede (732), and
we shall discuss this later on. His findings were confirmed by
Abderhalden, Fodor and Rose (733) as well as by Rose and Cooper
(734). Despite the work of Rubner (735), we believe that Hind-
hede's results are on a firm foundation.

The experiments with animals, especially rats and guinea pigs,
resulted differently from those on man. We have already seen in
the author's earlier work on guinea pig scurvy, that fresh potato
juice did not possess any great protective action against this disease.
McCollum, Simmonds and Parsons (736), experimenting with rats,
obtained results quite dissimilar to those in man. They found that
potatoes cannot take the place of other grains in their nutritive value,
and are deficient in salts, protein and vitamine A. One would be
tempted to conclude from this that, between rats and man, there are
differences in the utilization of potatoes. Osborne and Mendel (737)

256 THE VITAMINES

found that the vitamine B content of potatoes is sufficient for rats,
and no difference was apparent between new and old potatoes. Abder-
halden and Schaumann (I.e. 499) made the observation that when
pigeons are fed on dried potatoes, they develop edema on the lower
part of the legs. As regards the vitamine content, Chick and
her co-workers (I.e. 333) found that potatoes (cooked and raw)
contain all three vitamines in small quantities. Boruttau (738)
studied the effect of temperature on the nutritive value of potatoes
in mice. Similar experiments were made by Auer (739) in Hof-
meister's laboratory. The effect of drying was studied by Givens and
McClugage (I.e. 657).

MILK

Milk is such an important food for life that we must de-
scribe it more in detail. With regard to the milk question, we
were first to suggest the relationship between the composition of the
milk and some deficiency diseases in children. In 1912, we stated
two factors, without which the explanation of certain avitaminoses
would be impossible. The two essentials are first, that on heating
the milk, certain vitamines become more or less completely destroyed,
and second, that the vitamine content of milk must depend on the
vitamine content of the fodder. Although both are self-evident, they
gained recognition only with difficulty. The following rules, laid
down by us at that time for feeding of milk to children, are still of
value at present : Hygiene of the cow, good fodder, least possible boiling
of the milk, least possible storage after milking and after pasteurization.
Our uncompromising attitude on the milk question was made neces-
sary by the fact that Lane-Claypon (740), in a report to the Local
Government Board in London, came to the conclusion that boiling
does not affect the nutritive value of milk. As a matter of fact,
the same investigator (741), one year later — 1913 — declared that the
question needed further development. The ensuing years have
brought to light a great number of investigations which corroborated
our view on the subject completely. Later, Lane-Claypon (742)
published a book dealing with the question in detail.

Demonstration of vitamines in milk

In 1912 we (I.e. 324), proved the presence of vitamine B in milk,
and Hopkins also discovered the presence of a growth substance.

VITAMINE CONTENT OF FOODSTUFFS 257

The description of infantile beriberi by Hirota (743) and Andrews
(744) showed, of course, that the vitamines in milk must originate in
the food of the mother, since these young ones were nursed by mothers
suffering from beriberi. The question as to the nutritive value of
heated milk, especially in relation to infantile scurvy and without
knowledge of vitamines, was treated by Sir Thomas Barlow, Neu-
mann, Heubner and others ; we shall discuss this work in detail in the
chapter on scurvy.

Influence of food on the vitamine content and composition of milk

Because of the proof of the variation in the composition of
milk, we are in position to explain some of the divergent statements
in the literature. These facts, long known to cattle breeders
and milk producers, of course without any knowledge of vita-
mines, must be proven anew from the viewpoint of the vitamines.
That the milk may possess a different nutritive value according to the
fodder, has been shown by Carr, Spitzer, Caldwell and Anderson
(745), Hart, Nelson and Pitz (746), McCollum and Simmonds (747)
and Drummond (748). A series of interesting experiments were
published by Eckles, Palmer and Swett (749-750) . The latter showed
that the composition of milk is not constant; especially does the
protein and fat content vary considerably. The recognition of the
variation in the protein content of milk bears some relationship to our
modern conception of pellagra — which we shall discuss further under
the subject of pellagra — since under these conditions infantile
pellagra should be more frequently encountered, which is not the
case. The variation in the composition of milk explains the results
of Chick, Hume and Skelton (751), who found that for the protection
of guinea pigs against scurvy, a larger quantity of milk than that
found by us was necessary. The question as to the origin of milk
vitamines from the food has already been treated experimentally.
McCollum, Simmonds and Pitz (752) accepted this view for the vita-
mine A and B content of milk. Osborne and Mendel (I.e. 326) ob-
served no difference in the vitamine B content of summer and winter
milk. Regarding vitamine C, this relationship is very apparent.
Hart, Steenbock and Ellis (753) fed cows on dry and fresh fodder.
While of the milk resulting from the fresh fodder, 15 to 50 cc. daily
were necessary to protect guinea pigs from scurvy, of that arising from
the dry fodder, 50 to 75 cc. were necessary to produce the same result.

258 THE VITAMINES

These findings were confirmed by Hess, Unger and Supplee (754), who
showed further that even a 3 weeks feeding with dry and fresh food
was sufficient to show the above variations. Guinea pigs lived an aver-
age of 56 days on milk produced on the dry fodder, while on the milk
resulting from the fresh fodder the animals lived more than 120 days,
and after this time showed only a mild scurvy. In the milk contain-
ing more vitamine C, more calcium and phosphorus and 50 per cent
more citric acid were found. Butcher and co-workers (754a) cor-
roborated the above findings as to the dependence of the vitamine
content of the milk on the vitamine content of the cow's food.
According to Steenbock, Boutwell and Kent (I.e. 594) the same is
likewise true of vitamine A, after only a 3 weeks' feeding.

The vitamine content of milk

Inasmuch as we have seen that the vitamine content of milk varies
according to the composition of the diet,8 we must add that the
vitamine content even of raw milk is not very high. Therefore it is
obvious that any treatment of milk, or merely storing, may easily
diminish the 'margin of safety. The author (I.e. 325) as well as
Osborne and Mendel (I.e. 326) and Gibson and Conception (755)
showed that the vitamine content of milk is not very high. That this
is also true of vitamine C was demonstrated by Chick, Hume and
Skelton (756). As regards vitamine A, it seems to occur in larger
quantities; still, we are rather uncertain of its stability, since every
manipulation, because of the apparently easy oxidizability of the
substance, may have an effect on the nutritive value of the milk.

We may readily conceive, from the above, that the treatment of
milk may produce a deficiency disease in children whose practically
sole food it is. That this is the case in infantile scurvy, was shown
by Hess (757) using pasteurized milk. The disease developed after
a few months and was only of a mild nature. Hess (758) and Hess
and Unger (759) correctly observed that, aside from heating, storage
plays a great part in the conservation of antiscorbutic power. Miller
(760) was also of the opinion that since raw milk contains only a
small quantity of vitamine C, just within the margin of safety,
further manipulation may be dangerous.

4 In addition, Moore (754b) raised the question as to whether infectious
conditions of the cows may not influence the vitamine content of milk.

VITAMINE CONTENT OF FOODSTUFFS 259

The influence of heat on the vitamine content of milk

Practically all of the studies related to this point were made with
vitamine C. In this respect, most workers agree that the use of
heated milk may result in scurvy. Some investigators, on the con-
trary, believe that the decrease in the nutritive value after heating
cannot be attributed entirely to the vitamines. McCollum and
Davis (I.e. 97) stated that this is related to an injury of the casein
by heat. We have already mentioned this explanation and experi-
mentally disproven it. Daniels and Stuessy (761) showed that rats
fed on milk, cooked for 1 minute, could not grow. This experiment
was repeated, varying the temperature and time of heating. They
found, at the time, that an addition of protein, in the form of casein
or egg-yolk, permitted the rats to resume growth. In a recent paper,
Daniels and Loughlin (762) report that the results of Daniels-
Stuessy with casein and egg-yolk were to be attributed simply to an
addition of calcium salts. They found that when milk is heated
slowly at a lower temperature, as in pasteurization, a deposit con-
taining calcium salts is formed in the kettle. If the milk is quickly
boiled up, or if care is taken to mix the above deposit in with the
milk, then the rats grow just as well as on unheated milk. The same
results were obtained when soluble calcium salts, like calcium glycero-
phosphate, were added to the heated milk. Regarding the signifi-
cance of the results of Daniels and Loughlin critically, they maintain
that the decrease in the nutritive value of milk after heating is due,
not to the destruction of B- and A-vitamines, but to the precipitation
of calcium salts. Although the destruction of vitamine C is not
touched upon in this work, the investigators wished to convey the
impression that the changes of the milk on cooking had nothing to
do with the vitamines. It is not impossible, however, that if they
would repeat their work, using children, as they proposed to do, the
results would be different from those with rats.

The first investigation on the influence of heating on vitamine C
was made by Frolich (763). Milk heated at 98° C. for 10 minutes
lost its protective action against guinea pig scurvy, while heating at
70° C. for 30 minutes gave uncertain results. Whereas the author
found that 50 cc. milk prevented scurvy in guinea pigs, Chick, Hume
and Skelton (I.e. 751) observed that this was not sufficient. Barnes
and Hume (764) investigated the effect of heating, drying and rapid

260 THE VITAMINES

boiling on the vitamine content of milk. Dried milk (dried rapidly
at a high temperature) decreased markedly in its vitamine C content
and was inferior to milk boiled for a short time. The activity of
vitamines A and B seemed to be unaffected by drying. Hart, Steen-
bock and Smith (765) obtained similar results with raw and evapo-
rated milk.

Influence of drying and evaporation on milk

From an early "Report to the Local Government Board" (766),
we see that vigorous objections were raised against the nutritive
value of dried milk. In particular, the dilution necessary before
consumption was regarded as fraught with danger. This important
question was officially investigated in the United States and in
England, and a report on this subject appeared in the United States
Public Health Reports (767). Winfield (768) studied this question,
using 87 children and 40 rats, and came to the conclusion that dried
milk seems to be complete, but only for the first growth period. The
rats did not grow any more after they reached two-thirds of their
size, and the usual mixed diet had to be resorted to at this point.
Naish (769) stated that in the use of dried skimmed milk for feeding
children the danger of scurvy does not exist, even without fruit
addition, while Pritchard (770) recommended the addition of fruit.
Hess and Unger (I.e. 631) are of the opinion that milk, dried according
to the Just-Hatmaker process for a few seconds at 116°C., is protective
against scurvy equally as well as fresh milk. In a few cases, a cure
could even be brought about. Sekine (771) conducted some experi-
ments with condensed milk and showed that young mice remained
normal on this diet for about 100 days, after which they suffered from
beriberi and anemia. On the addition of iron and vitamine B, they
again became normal. Coutts (772) found that dried milk gave
better results in the nutrition of children than did fresh milk. He
believed that 3 months old infants could tolerate dried whole milk.
He recommended also a synthetic milk, in which the cream was
substituted by cheaper vegetable fats which were, however, appar-
ently poor in vitamine A. Gerstenberger (773) and his co-workers
tried to replace the butter-fat by a cheaper fat. They started with
skimmed milk to which were added vegetable fat and cod liver oil.
This milk was fed to 311 children, who subsequently showed no signs
of rickets, spasmophilia or anemia.

VITAMINE CONTENT OF FOODSTUFFS 261

General remarks on the nutritive value of milk

The question of dilution in connection with the use of condensed
and dried milk appears also to be of some practical significance.
Washburn and Jones (774) have shown that the fat content of milk
is likewise of great significance. Working with young pigs, they
demonstrated that skimmed milk, as such, does not permit of growth
in these animals. Milk with a moderate amount of fat (2.5 per cent)
seemed to give the best results, while too much fat (5 per cent)
produced rapid growth, the animals being too fat and apathetic.
Too great a dilution of the milk was followed by a considerable
decrease in its nutritive value, while a very concentrated milk reared
young that were excessively fat and weak, the bones showing only
two-thirds of their normal strength. Lately, views have appeared
which emphasize the exceptional position of milk in nutrition.
Freise (I.e. 342), as a result of some rat experiments, expressed the
view that the milk was either lacking an important constituent or
had lost it during manipulation. He observed, however, that
animals on a deficient diet recovered rapidly when an addition of
10 per cent dried milk was given, so that milk possesses a splendid
supplementary action. The same characteristic is manifested by
barley-malt. Honey, cereals, alcoholic extracts of barley-malt,
carrots, egg-yolk, beans, casein and smaller amounts of dried milk
(less than 10 per cent) no longer show the activity of dried milk.
Mattill and. Conklin (775), working with rats, obtained the same
results. On ordinary milk the animals grew well at first, but later,
especially in 50 to 100 days old females, growth ceased. Autolyzed
and ordinary yeast and wheat germ were added, resulting in only
temporary improvement; addition of iron and casein was without
effect. On the other hand, better results were obtained on adding
dried milk, showing that the dilution of the foodstuff given is of
significance. Dried milk was shown not to be the ideal food for
later stages of life. Here again, a dilution (55 parts dried milk,
40 parts starch and 5 parts butter) was of help resulting in normal
growth and reproduction. The dilution of dried milk with lard gave
the same results as with starch. Addition of 1 per cent yeast
resulted in normal growth. For these reasons, the authors raised
the question, whether the addition of yeast does not introduce some-
thing that is missing in the other combinations, and proposed new

262 THE VITAMINES

experiments to clear up this point. From what has been said in
this chapter, it is evident that we are not yet fully informed as to
the nutritive value of milk, and that this foodstuff still presents some
riddles for us to solve.

MEAT

The significance of meat as a component of our diet is threefold.
First, as an important source of protein; second, as a source of vitam-
ines when it is used as a sole food; and third, for its "vitamine-
sparing" action. The last characteristic will be treated more fully
later. In accord with our own experience, we know of no single food
which requires so little vitamine addition as does protein material,
especially meat.

When we speak of meat in this chapter, we have in mind muscle
tissue only, since glandular organs like liver, kidney, etc., are charac-
terized by a special richness in vitamines. Vitamine B was first
demonstrated in meat by Cooper (I.e. 535). Osborne and Mendel
(776) investigated the value of dried meat and did not find it par-
ticularly rich in vitamine; on successive extraction with water, the
amount of vitamine B decreased considerably. According to Osborne
and Mendel (777), dried meat contains sufficient vitamine A, while
pig's brain and heart has enough of both A- and B-vitamines. Cole
(778) reported that powdered meat contains a sufficient amount of
A- and B-vitamines.

Pertaining to the vitamine C content of meat, practical experience
and laboratory findings do not seem to go parallel. Naturally, the
discrepancies are only apparent, since practice has shown that fresh
meat contains small amounts of vitamine C. This is evident from
the report of Nansen (779) on his polar expedition, as well as from the
report of Stefansson (780). The latter stated that scurvy was cured
by an addition of fresh meat. However, we already knew from the
work of Curran (781) that 2 to 4 pounds of raw meat are necessary,
since he observed cases of scurvy on a daily consumption of 500 grams
cooked meat. Willcox (782) stated that in the English army in
Mesopotamia scurvy developed because the Indian troops refused
to eat meat. He was of the opinion that meat could function as an
antiscorbutic. Pitz (I.e. 365), in his studies on guinea pig scurvy,
found that an increase in the protein proportion of the diet definitely
prolongs the life of the animal, which may perhaps be regarded as

VITAMINE CONTENT OF FOODSTUFFS 263

the "vitamine-sparing action" of protein. Dutcher, Pierson and
Biester (783) found that guinea pigs could not be protected against
scurvy by an addition of fresh meat juice. This was also shown by
Givens and McClugage (784) with dehydrated meat; however, these
results may be criticized on the basis that guinea pigs need more
vitamine C than does man. That the vitamine requirements of
different animals may vary has already been demonstrated by
Harden and Zilva (I.e. 455).

Parsons (I.e. 191a) showed, on the 'contrary, that some organs,
especially the liver, but also spleen and kidney, contain relatively
large amounts of vitamine C — so much so, that 10 grams of pig's
liver per day is sufficient to protect guinea pigs from scurvy. By
the preparation of extracts, instead of juices or organs themselves,
Parsons was able to add larger quantities of extractives from the
muscle, showing that an extract of 55 to 95 grams of muscle suffices to
protect guinea pigs from scurvy. Fish muscle possessed no anti-
scorbutic property.

THE VITAMINE CONTENT OF THE MOST FREQUENTLY USED
FOODSTUFFS

The compilation, which, as we are well aware, cannot be complete,
gives the reader an idea of the number of investigations carried out
for the demonstration of the present known facts. We have not
sought to give the figures for the vitamine content since we believe
that sufficient progress has not yet been made. In the table, a single
plus sign denotes that the respective vitamine is present only in
traces. With regard to protein, the plus sign indicates that it is not
adequate for nutrition. From the table, we may see that those start-
ing materials which are very rich in vitamines contain proteins of
high biological value.

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272

PART III

The Human Avitaminoses — Conditions in
which the Vitamines Play a Role

CHAPTER I
BERIBERI

The first stage of vitamine research, which we shall assign to the
period between 1910 and 1920, served as the instructive period. The
vitamine hypothesis, which can really no longer be regarded as such,
was promulgated during this period, and will survive in a more or
less modified form. The stage of enthusiasm, in which all possible
pathological conditions were regarded as deficiency diseases, has
passed and we shall endeavor now to treat the matter from the
objective point of view. In the study of the known and also the
suspected avitaminoses, we meet with the real difficulty that most
of the medical men are divided into two camps. One champions
the vitamines, while the other will have none of them. The resultant
feud is not very conducive to a speedy solution of the questions that
interest us here, for, if one has a preformed idea, then all facts are
viewed in a light best in accord with this idea. We find, in addition,
little cooperation between experimental investigators and clinicians.
This would be the more desirable, because most investigators have
very little clinical expeiience, while the clinician lacks the broad
scientific basis of the modes of nutrition.

We frequently read in clinical papers, in connection with the
description of a possible avitaminosis, that dietary changes and
vitamine therapy have been resorted to without success. This
whole situation is usually disposed of in a few words and there is
frequently a failure to report important observations. In purely
scientific investigations, as in the field of physiological chemistry,
physiology or pharmacology, it remains for the author of the par-
ticular subject to prove his results. Very frequently, his facts inter-
est us far more than his conclusions. In the clinical work on avitam-
inoses, we often see the conclusions of the individual authors merely
as personal impressions, without giving all the particulars, upon
which basis, perhaps, a different conclusion might be possible. In the
interest of progress, it would be very desirable if in the clinical
investigations in this field, an exact description of the previous diets,
as well as of the vitamine therapy, were given. In this way, there
would be the opportunity, in every individual instance, of forming a

275

276 THE VITAMINES

personal conception of the cases quoted, which is now usually quite
impossible. Since our first edition, no great progress has been
made because of the above reasons, in spite of many publications.
The conditions regarded at that time as avitaminoses are con-
sidered as such to the present day, while the suspected avitaminoses
are still as uncertain as before. The author, as the originator of
this classification would like to see all these conditions demonstrated
as avitaminoses; on the other hand, it would be worth while to elim-
inate as soon as possible all that does not belong to this complex.

Even as regards the definite avitaminoses, we are well aware that
there will always remain a minority who will have a different opinion
from ours. While we recorded all of these divergent opinions in our
first edition, we believed it necessary, from the point of view of uni-
formity of the subject, to avoid this at present. It should, however,
not be thought that we wish to ignore the opponents of the vitamine
subject. This book is intended to treat of vitamines and we shall
not stray from its main purpose.

What has just been said applies even more to the description of
those pathological conditions which cannot possibly be regarded as
avitaminoses, but in which the presence or absence of vitamines may
play a part. The consideration of these conditions in this book
should, however, not be construed to indicate the author's belief
that they may be related to the vitamines directly. It is possible
that some of the diseases to be described here may in the future be
explained differently. This book merely pictures the possible
aspects of future vitamine study, and does not exclude another solu-
tion of the problems.

The reader may ask why an answer to the above problems is not
yet at hand. Why can we not say whether or not a certain condition
can be associated with the vitamines etiologically? Apart from
the fact that the vitamines are indisputably essential for life, we
know very little regarding their physiological significance. To be
sure, we see signs of a generally diminished assimilative ability,
negative balances of some important inorganic and organic constitu-
ents, and decreased resistance to infection. Besides this, we see
changes in most organs, in the glands of internal and external secre-
tion, and trophic changes in the skin. But just how all these con-
ditions can be correlated with the lack of vitamines is at present
unknown to us. The above list of symptoms is so large that we

BERIBERI 277

may associate them with all possible conditions. If now we con-
sider that there is the possibility that new vitamines may yet be
found, we recognize the many-sidedness of the possible vitamine
aspects.

The investigation of the avitaminoses is rendered more difficult,
in that the symptoms characteristic of these conditions may be
attributed to entirely different causes, aside from the deficiency of
vitamines. Thus, we see the development of edema, resembling very
much the hunger edema, which is, however, not of dietetic but of
infectious origin. We note beriberi-like symptoms, not related to
lack of vitamine B but to toxins and poisons; this is- also true of
many trophic appearances. Because of these complications, the
literature on deficiency diseases has become very clouded, since it
frequently happens that the finding of a case explainable by some-
thing other than vitamine deficiency is often followed by the attempt
to discredit the vitamines entirely. For these reasons, we must be
careful in judging the etiology of sporadic and isolated cases; it may
easily be .associated with an error in diagnosis. If such cases occur
in great number (as is usually found in practice) then a mistaken
diagnosis can scarcely be made.

The vitamines have lately been associated with orthopedic surgery,
certain eye troubles, such as hemeralopia and ophthalmia; some
infections such as tuberculosis, pneumonia, leprosy; some metabolic
diseases such as arthritis, diabetes (also cancer) ; and with dental
caries. We know now that these conditions have some relation to
the diet but whether they are specifically associated with vitamines
has not yet been shown, and is in fact difficult to do so. Since many
of these pathological conditions, in spite of tiresome research, have
given no real explanation, we see no harm if vitamine research will
give a new impulse to the study of these questions. We see many
examples of how a misleading hypothesis has led to a favorable
solution of some questions; at any rate, if a subject remains quiescent
it can never lead to progress.

In the description of beriberi, we shall dwell exclusively on
the human type, since many phases of this question have already
been treated in avian beriberi. Above all, we recommend to the
reader who is particularly interested in beriberi the fundamental
work of Vedder (838) as well as to contributions by Castellani and
Chalmers (839) and Schilling (840), who treat the subject from the

278 THE VITAMINES

viewpoint of the modern conception of beriberi. Beriberi is a disease
which, according to Findlay (841), has been known for hundreds, if
not for thousands, of years. Its occurrence was described in the
Roman army which invaded Arabia in 24 B.C. Chinese writers in
the second century referred to this point. N etching, the oldest
medical book (2697 B.C.), speaks definitely of this disease.

East Asia, including the group of Polynesian Islands, is the main
beriberi zone. The second greatest area is in Brazil and the neigh-
boring countries. Furthermore, the African coast shows numerous
foci. It was commonly held that a moist warm climate was especially
conducive for the development of the disease This report is incorrect
as, for example, in the Russo-Japanese War, a severe outbreak of
beriberi was noted in the Japanese army in spite of the severe winter;
beriberi is endemic also in northern Japan. The above mentioned
statement is based largely upon the fact that rice grows well only in
moist heat. Japan is the most afflicted country. Balz and Miura
(842) estimated the number of beriberi patients at about 50,000
yearly. China is very much less afflicted, though in Korea we find
numerous foci. Furthermore, we find the disease quite well spread
over the Malay Peninsula. The number of cases there was estimated
by Fraser (843) in 1911 to be 5540, among which 695 deaths occurred.
This is true also of the islands of the Malay Archipelago, Java,
Borneo, Celebes, the Molucca Islands, New Guinea, and especially
Sumatra. Simpson (844) reported cases in Singapore, where the
disease was often noted among Chinese coolies. In Hawaii, New
Caledonia and North Australia, the disease is mostly seen in Japanese
and Chinese immigrants. More beriberi zones are found in Cochin-
China, Siam (according to Hepburn, 845) and Burma, at the mouth
of the Ganges (according to Mulvany, 846), on the east coast of the
East Indies, Ceylon, Malabar, and less so on the Koromandel Coast.

In Africa, numerous foci exist on the east and west coasts. Thus,
Chevalier (847) reported on an outbreak in Serenli (East Africa) with
112 cases in six months, with 44 deaths. In the Congo, the rice
eaters are often attacked by the disease, while other natives, to whom
rice feeding is unknown, remain free from the disease. Dubois and
Corin (848) describe a number of cases in Bokala (Belgian Congo)
which were caused by the consumption of "maniok." Similarly,
beriberi is known in Madagascar, and in the Mauritius and Reunion
Islands.

BERIBERI 279

The Philippines and Brazil form two important foci. From Brazil,
there is a report of Lovelace (849), who observed 934 cases within
four years; the disease is known there for 250 years. The beriberi
zone broadens out to the north, till Rio de la Plata. Lovelace is
not the only observer in Brazil; we have also the reports of Wolcott
(850) and Fraga (851), the latter having seen cases in Bahia. Riddel,
Smith and Igravidez (852) described cases in the military hospital
at Porto Rico. From the previous chapter, we saw that it did not
require a consumption of white rice to produce the disease. Little.
(853) saw a number of cases in Newfoundland resulting from the
ingestion of white flour. In the United States, cases of beriberi are
very rare occurrences. Parker (854) describes an outbreak of this
disease in the prison at Elizabeth, N. J., as a result of eating white
bread. Reed (855) saw cases in California among Chinese and
Japanese. Travis (856) described cases in Eddyville, Kentucky.

In Europe, beriberi was observed in lunatic asylums in Ireland in
1894, 1896 and 1897; in 1894, there were 147 cases in Richmond
Asylum near Dublin, with 25 deaths. Chantemesse and Ramond
(857) reported an outbreak of beriberi in the lunatic asylum in Saint-
Lemmes (Angers), in 1898, resulting in 40 deaths. Schiiffner (858)
believed, in general, that beriberi cases occurred sporadically from
time to time and were treated under another name. Recently,
cases were observed by Martinez (859) in Spain and Portugal. We
recognize from this survey that beriberi occurs in places, such as
prisons, hospitals, etc., where the food is prepared in quantities in
large kettles. A further opportunity for the occurrence of beriberi
is provided by wars. In the Russo-Japanese war, the Japanese,
according to Schilling (I.e. 840), had about 70,000 to 80,000 cases,
while the Russians, as we shall see later, had only scurvy cases. In
the last war, we have similar observations. Sicard, Roger and
Rimbaud (860), Roger (861) and Leggate (862) reported on numerous
cases among Chinese and Indo-Chinese laborers, who were trans-
ported to France for labor during the war. Massalongo (863)
described cases on the Italian front. Willcox (864), from the
Mediterranean area in the English army in 1916, and later also from
Mesopotamia (865), reported cases which could not be attributed to
rice consumption. Braddon (866) investigated a number of cases in
the Mediterranean war zone. Hehir (867) and Sprawson (868)
observed cases during the siege of Kut-el-Amara.

280

THE VITAMINES

MODE OF OCCURRENCE OF AND DIETS LEADING TO BERIBERI

It is very important for the proper understanding of the nature of
beriberi to know the composition of the diet leading to its occurrence..
This is the more important since a number of investigators have
reached the conclusion that the diet of their patients was completely
satisfactory, from the viewpoint of nutritional studies. As for this,
we may bring to our support the slight amount of exact data available
from two sources. On the one hand, we know the composition of the
diet in a few instances where the disease has been produced experi-
mentally in man; on the other, we have the reports of a few beriberi
outbreaks, in which the composition of the diet was accurately noted.

In the first series, there is the work of Fraser and Stanton (I.e. 47),
who produced beriberi in healthy Japanese coolies on a diet made up
almost exclusively of white rice. Here, 80 to 90 days passed before
the disease made its appearance. Another, and far more important,
investigation was that of Strong and Crowell (869). They conducted
experiments with volunteers from a prison in the Philippines. The
subjects were divided into three groups, each group receiving a
certain amount of fish, lard, bananas, potatoes and sugar. Rice, in
three different forms, was the chief component of the diet.

NUMBER OF

NUMBER OF

VOLUNTEERS

BERIBERI CASES

1

White rice and polishings

8

2

2

White rice

17

13

3

Unpolished rice

7

1 (mild form)

From this, it is evident that unpolished rice can practically prevent
the occurrence of beriberi, though the vitamine B content of this
kind of rice barely exceeds the necessary amount. The first symp-
toms were noted after 60 days. Caspari and Moszkowski (870)
reported an experiment upon Moszkowski himself. He lived 230
days on white rice and other vegetable food, without meat, eggs or
cheese, and developed mild beriberi, but with cardiac symptoms and
considerable nitrogen loss. On the addition of rice polishings, the
symptoms promptly disappeared. It seems likely that that diet
contained small amounts of vitamine B, since the development of the
disease was so long delayed. Fraga (871) conducted an experiment
with nine prisoners. He was unable to produce the disease with

BERIBERI 281

sterilized rice and sterilized beans during a period of 43 days; the sub-
jects objected to the food, and the experiment had to be terminated.

The second group of the accessible data pertains to the composition
of the diet leading to beriberi. Chamberlain (872) investigated the
personnel of the native Philippine Scouts comprising 5200 men. Of
these, 618 developed beriberi in 1908, and 558 in 1909. The food
leading to this outbreak consisted of 340 grams fresh beef or its
equivalent of bacon, canned meat or fish, 560 grams white rice, 225
grams flour or bread and 225 grams potatoes or onions. When the
diet was improved in 1910 by the addition of more beans and unpol-
ished rice, the number of cases decreased to 6. The cases described
by Riddel, Smith and Igravidez (I.e. 852), in Porto Rico, developed
after three months on a diet consisting of rice, vegetables and canned
meat. From the report of Hehir (I.e. 867), we see that there were
numerous cases of beriberi among the English troops at Kut-el-Amara
on a diet of horse-meat, canned meat and white flour. The siege
lasted over 4| months, and this time sufficed for the development of
the disease.

In confirmation of the author's findings (I.e. 559) and those of
Braddon and Cooper (I.e. 561) on pigeons, Braddon (I.e. 866) made
some observations in the Mediterranean war zone on the dependence
of beriberi on the composition of the food. This investigator drew
up a table, which we shall reproduce, giving the composition of two
diets capable of producing beriberi, compared with two other diets
which did not result in beriberi. The content of vitamine B was

V

calculated on the basis of 100 for wheat germ. The quotient ^

-A.

shows the relationship between foodstuffs containing and those lack-
ing vitamine B.

This method of calculation is obviously not very exact, but it
gives us an insight into the relationship between protein and carbo-
hydrates; in beriberi-producing diets the quotient was much smaller.
It is also interesting to note that Braddon, although the individuals
were still in good condition, suspected beriberi, because of the lack
of reflexes, a suspicion which was later confirmed This showed
definitely that beriberi may be recognized prior to the appearance of
characteristic symptoms.

282

THE VITAMINES

FOODSTUFF

VITAMINE
VALUE

BERIBERI-
PRODUCING DIETS

SATISFACTORY DIETS

•Diet Ai

Diet Aj

DietB

DietC

Rice

800
6400

400
400

400
8400

6400

400
400

400
7600

600
3200
200
200
400
400
575
200

5750

800

4800
200

100

5875

White bread

Jam

Sugar

Cheese

Dried fruit

Salt fish

Margarine, butter or oil

X = Total quantity of food lack-
ing in vitamine B

Oatmeal

10
10
50
5

1200
400
1600

1200
800
400

2400

400
850
400
1200

2850

400
1200
600
800

3000

Fresh meat or bacon

Peas, beans or lintels

Potatoes or fresh vegetables

V = Total wgt. of foodstuffs con-
taining vitamine B

V
Ratio — — . .

0.2

0.3

0.5

0.5

X

* Weekly amount of food.

THE SYMPTOMATOLOGY OF BERIBERI

There is no doubt that a latent form of beriberi does exist (Braddon,
I.e. 866). Beriberi symptoms are not infrequently noted, acute
for the most part, after a trauma or over-exertion. After an opera-
tion, severe fatal manifestations occur in persons who, before the
operation, showed no symptoms at all. Acute cases of beriberi occur
in soldiers after a long march; light abortive cases are very common.
The patients complain of a feeling of heaviness in the legs and of
unsteadiness in the knees on walking or standing, so that they
frequently collapse. This occurs chiefly in summer months. Very
often they complain of palpitation of the heart after an exertion, and
of tingling and numbness in the skin of the legs. Skin symptoms in
the form of erythema, petechiae, and exanthema are noted. Hem-
eralopia belongs to the earlier symptoms; the old therapy (liver) of
Hippocrates attains thereby a scientific explanation. Hemeralopia
belongs also to the symptomatology of scurvy and pellagra, and may

BERIBERI 283

be thought of as a symptom of vitamine hunger. A decrease of sexual
ability is frequently observed.

Hepburn (I.e. 845) enumerates, in observations of 100 cases, the
following symptoms: The first symptoms, as a rule, consisted in
one of three manifestations, (1) edema of the feet and legs in 50 per
cent of all cases; (2) numbness, irritability and tingling of the legs
in 42 per cent, and (3) epigastric manifestations in 7 per cent. A soft
mitral systolic murmur was noted in 20 cases in the first investigation.
Cardiac arrhythmia was noted in 5 cases in the first stage. Kato
and Yamada (873) noted arrhythmia in the convalescent stage. The
body temperature was found subnormal by Doyle (874) . As regards
the blood picture, Chun (875) found an increase in the number xof
leucocytes amounting to 100 per cent. The blood pressure was
found to range up to 200 mm. by Yoshikawa, Yano and Nemoto
(876). Jida (877) investigated the cerebrospinal fluid. In a number
of cases he found a large pressure, particularly in the acute stage,
up to 280 mm. with a low blood pressure. This fluid, in cases with
high pressure, injected into the .ear of a rabbit showed a powerful
vaso-constrictor action (which indicates, perhaps, a high adrenaline
content). Sicard and Roger (878), on the contrary, failed to dis-
cover any peculiarities in the cerebrospinal fluid.

The picture of beriberi shows many different forms, according to
Balz and Miura.

1. The light, sensory-motor form.

2. The dry, atrophic form.

3. The wet, atrophic form.

4. The pernicious, acute form.

These forms may also merge into one another. As a rule, the
picture of the disease shows paralysis and muscle atrophy in chronic
cases; in acute cases, however, there are gastro-intestinal disturb-
ances, anasarca, hydropericardium and severe cardiac symptoms.

1. The sensory-motor form

This is the most common form of beriberi. After an exertion,
especially during the summer months, the patient feels unsteady and
weak in the legs; often there is numbness of the arches of the feet
and of the legs; the calf muscles are sensitive to pressure. In addi-
tion, there is palpitation of the heart after exertion, and edema of the

284

THE VITAMINES

FIG. 55. ATROPHIC BERIBERI (BALZ-MITTRA)

BERIBERI

285

FIG. 56. SAME AS FIG. 55, BACK VIEW (BALZ-MITTRA)

286

THE V1TAMINES

BERIBERI 287

legs. The knee reflex is increased at first, and later usually entirely
absent; the temperature remains normal. In this stage a cure may
result but it may go over into the dry atrophic or the wet atrophic
form.

2. The dry atrophic form (figs. 55-56)

In this stage, there is general paralysis and muscle atrophy; at
first the legs, then the thighs, hands, arms, and finally the back muscles
are involved. The patients then show a skeleton-like emaciation.
In this stage, too, improvement may take place under certain con-
ditions, and in the end, after many months, a complete cure may
result. The motility, electrical reaction, the original musculature,
the patellar reflex — all gradually return. In other cases, reoccur-
rences appear, the paralysis and atrophy increase, the foot remains
in the position of pes equinovarus (Fig. 57) ; the hands hang weakly
with half-closed fingers, as in any radial paralysis. After long
prevalence of the disease, permanent contractions of the foot de-
velop, less frequently of the fingers and biceps. The patient can
only walk on tip-toes with the help of crutches. These severe chronic
conditions are incurable.

8. The wet atrophic Jorm

In this condition, besides the above symptoms, the disturbance of
circulation is more sharply prominent. There is palpitation of the
heart, tachycardia, dyspnea, oliguria, pain in the calf muscle and
edema. Over the ankle there is a pale edema and in severe forms
also in the thighs and in the back, especially in the shoulder and
neck region; often also in the arms and on the face. Hydropic
condition of the serous cavities, namely, the pericardial and the
pleural, belongs to the most important symptoms. The elimina-
tion of urine is greatly decreased, often to 200 to 300 cc. or even less.
The urine has a high specific gravity, is protein free, urates are
precipitated out, and it shows a marked indican reaction. If a cure
is initiated, the edema disappears with marked diuresis, whereupon
the great emaciation and muscle atrophy become apparent. In nu-
merous cases, death follows, partly because of the severe hydroperi-
cardium, partly through paralysis of the heart muscle, or, finally,
through a merging into the acute pernicious form. In puerperal
beriberi, the wet form is noted, as a rule. In severe cases, there is

288

THE VITAMIXES

also paralysis of the intercostal muscle, the diaphragm, and less
frequently of the cephalic nerves, optic, abducens, facial and hypo-

FIG. 58. PARALYTIC FORM OF BERIBERI (FRASER)

glossal nerves. Usually, the laryngeal muscles are paralyzed.
Paralysis of the vagus is generally regarded as the cause of death.

Epidemic dropsy. Under this name, Greig (879) described a disease
which appeared to result from rice feeding, and which was regarded

BERIBERI 289

at first as identical with wet beriberi. It is likely now, however, that
epidemic dropsy, sometimes accompanied by fever, is related not to
beriberi but to hunger edema. Greig (880) later investigated the
mode of development of the disease somewhat more closely, and
found that some natives feed on wheat from which the husk and bran
are carefully removed. Addition of meat in these cases exercised a
beneficial influence. The question of the etiology of this disease will
be taken up again later.

4- The acute pernicious or cardio-vascular form

This variety can occur either primarily or secondarily from the
above described forms by over-exertion, after an operation, and also
without apparent causes. Quite suddenly, often in but a few hours,
a severe picture develops : precordial pains, pain in the epigastrium,
dyspnea, tachycardia, nausea, vomiting, diarrhea and staggering.
The temperature remains normal and the mind clear. The heart is
markedly enlarged, especially the right ventricle; the whole heart
region pulsates, due to paresis of the intercostal muscle and the
increased heart labor. The visible pulsation in the epigastrium
belongs to the early symptoms of the disease. Frequently, there
develops a systolic murmur at the apex, and also in the second or
third intercostal space; the second heart sound is strengthened. In
the arteria cruralis, we hear an arterial-diastolic murmur; in severe
cases, even a tone. The pulse is accelerated, 120; in severe cases,
130 and 140, full and soft. The breathing is wild and wheezing,
the whole thorax is raised spasmodically. Hoarseness or aphonia
frequently develops. The quantity of urine is greatly diminished,
100 to 200 cc. per day, and even less. The urine gives a marked
indican reaction; in some cases it contains some albumin, and occa-
sionally it gives a diazo reaction. Death occurs with a small frequent
pulse, cyanosis and edema of the lungs, sometimes in but a few days
or weeks, but sometimes very suddenly in a few hours.

The mortality in beriberi varies within wide limits. In some out-
breaks in Sumatra, Java and Manila, the death rate was frightful —
up to 60 to 70 per cent and even higher, while among the Dutch
troops in Insulinda 2 to 6 per cent, and among the Japanese troops
2 to 4 per cent. The death rate in the wet form is greater than in
the dry, and highest in the acute pernicious form. Formerly, the
average death rate was calculated to be 37 per cent, but in recent

290

THE VITAMINES

years, following a better understanding of the causes and the therapy
of the disease, there has been a marked decrease in the rate. A
portion of the chronic cases die with complications, chiefly tuberculosis.

FIG. 59. HYDKOPIC FORM OF BERIBKHI (PHASER)

Beriberi in infants

This form was first described by Hirota (I.e. 743), and develops in
infants nursed by mothers suffering from beriberi; usually, the latter
exhibit no signs of the disease. The observed symptoms are similar
to those of adults: persistent chronic vomiting, cyanosis around the

BERIBERI 291

mouth and nose, dyspnea, tachycardia, aphonia, edema of the face
and the legs, oliguria, but no fever. In some of the cases, the mother
is still in the latent stage and shows only indicanuria, while more
marked symptoms develop later. These statements were confirmed
by Andrews (I.e. 744) in the Philippines. He found that a great
number of infants died there because of this reason. The milk of
such mothers was shown to be poor in fat (2 per cent instead of 4 per
cent), while the calcium and phosphorus content was increased. If
this milk was given to young dogs, they developed beriberi. The
question as to the mode of development and the nature of infantile
beriberi was also treated by Lagane (881), Weill and Mouriquand
(882) and Walshe (883). In addition, Ogata and Jizuki (884)
investigated the influence of beriberi in the mother on the fetus.
According to Roxas (885), the disease in the Philippines occurs less
frequently in artificially fed children. The great number of cases of
infantile beriberi, compared with infrequency of scurvy and pellagra
in naturally nourished infants, makes it evident that the formation
of vitamine B in the milk proceeds somewhat differently from that
of the other vitamines.

PATHOLOGY OF BERIBERI

General

Diirck (886) has written a good monograph on the subject. For
the most part, sections were made of acute cases. The skin of the
beriberi corpse is quite edematous, cyanotic at the extremities, lips
and ears, often with subcutaneous hemorrhages; the musculature is
also edematous. One of the most common typical findings is hydro-
pericardium, which develops as a result of the degeneration of the
nerves of the pericardium. Hydrothorax, ascites fluid and pin-
point hemorrhages are frequently observed in the serous membranes.
In acute cases, ecchymoses are not infrequently noted in the stomach
and in the duodenum. The heart, especially the right ventricle and
auricle, is markedly dilated and hypertrophied. The heart muscle
itself shows yellowish spots and streaks (of a fatty nature). The
lungs are usually edematous. The kidneys are full of blood, but here,
as in other organs, there are small hemorrhages. Nephritis was not
observed.

292 THE VITAMINES

The most important changes in beriberi were noted in the periph-
eral nerves and muscles. In the peripheral nerves, no macroscopic
changes were apparent, aside from the occasional observation of
minute hemorrhages in the sheath or between the fibers. Micro-
scopically, however, all stages of degeneration are noted. The
changes begin with the nerve; the medulla becomes varicose, espe-
cially near the constrictions; it falls apart in clumps and masses and
finally in a granular detritus. The axis cylinder, on the contrary,
frequently remains intact for a long time, but in the end this too dis-
integrates. The Schwann's sheath then becomes a shapeless mass,
with increase of the nuclei. In fresh acute cases, the number of
degenerated fibers is very slight. At the nerve endings in the
muscles, only those nerve fibers leading to the degenerated muscle
fibers are swollen and granulated. This degenerative neuritis occurs
chiefly in the ulnar, radial and medial nerves in the legs and later in
the phrenic and vagus nerves. Balz (837) found the same to be true
of the kidney nerves. Most apparent, however, was the degeneration
of almost all of the spinal cord nerves. Balz (I.e. 887), as well as
Ellis (888), found changes in the sympathetic and especially in the
cardiac, pulmonary, splanchnic and solar plexi. Rumpf and Luce,
(889) found degenerative processes in the dorsal roots. In the brain
and spinal cord, as well as in other organs, there is venous stenosis
and edematous impregnation. Some investigators have found the
motor ganglia cells of the ventral horn atrophically vacuolized. The
nucleus lies often asymmetrically, pushed to one side by a large
vacuole. Rumpf and Luce found fresh parenchyma and diffused
medullary sheath degeneration in the white substance of the spinal
cord. The pathological-histological changes in the nervous system
will be further described under the subject of pellagra, accompanied
by numerous illustrations, for according to the view of an eminent
neuro-pathologist, Mott (890), these changes belong, in both diseases,
to the same type; however, it should not therefore be said that
both diseases must be etiologically related.

The affected muscles show all signs of degeneration and atrophy.
The first signs of this consists of the dimming of the striations. Fre-
quently, the muscle fibers become waxy, homogeneous and fragile.
For the remaining pathological changes, we may refer to what has
been said under the subject of avian beriberi.

BERIBERI 293

Chemical pathology — Blood

The blood-sugar content in 35 cases of infantile beriberi was
determined by Suzuki (891) to be 0.0808 per cent. This decrease
was attributed to a decreased activity of the adrenals, and is different
from that noted in avian beriberi. DeLangen, Schut, Wechuizen and
Alting (892) found a very low lipoid and fat content in the blood in
beriberi. Arima (893) investigated the non-protein nitrogen in 28
cases and found it increased.

Cerebrospinal fluid

Suzuki (894) found the sugar content of cerebrospinal fluid to be
normal.

Urine

Suzuki (895) found the amino-nitrogen of the urine markedly
increased in the cardiac form of beriberi. Pagniez and Vallery-Radot
(896) studied the elimination of sodium chloride on a diet poor in
salt in cases of wet beriberi. The elimination was very large, indi-
cating a previous marked retention. In the wet form of beriberi,
as in nephritic edema, sodium chloride passes the kidney only with
great difficulty; in beriberi, the water gathers in the muscles, while in
nephritic edema, the fluid collects subcutaneously. These inves-
tigators believed that the development of wet or dry beriberi is
determined by the severity of the kidney lesions.

In confirmation of the adrenaline finding in pigeons, Ono (897)
investigated the adrenals microscopically and found a larger quantity
of adrenaline, as compared with the amount in other pathological
conditions.

THERAPY OF BERIBERI

General

Vitamine therapy in beriberi is at present so well established that
we need not dwell on this point unnecessarily. One of the first
investigations was that of Thompson and Simpson (I.e. 60), who used
yeast for this purpose with good results. The experiments were
thereupon extended by Vedder and Williams (I.e. 471) to include a
preparation of rice polishings. They thought that they had obtained
good results in dry beriberi with a vitamine preparation, made

294 THE V1TAMINES

according to our procedure, while in wet beriberi it was ineffective.
From this, it was concluded that for the therapy of various types of
beriberi, vitamines of various sources were necessary. This observa-
tion may be explained by the severity of the symptoms, without
having recourse to such a hypothesis. Good results with vitamine
preparations were obtained by Williams and Saleeby (I.e. 472).
Later on, Saleeby (898) used autolyzed yeast in adults and children.
The dose used was 15 to 40 cc. three times daily for adults, and 2 to 4
cc. every 3 hours for children, greater doses being of no advantage.
Marked results were noted after 3 days, but the treatment was con-
tinued for 14 days more. In infants, the results were noticeable even
sooner. A standardization of the extract of rice polishings was
recommended by tel Rosario and Maranon (899). In the late war, a
yeast preparation (marmite) was used in the English army with
splendid results (Willcox, I.e. 864). Hepburn (I.e. 845) too reported
results with the vitamine therapy in Siam.

Therapy in infantile beriberi

This consists either of a change of milk, or of the administration of
vitamine. Particularly good results were obtained with an extract
of rice polishings (tiki-tiki). According to Albert (900), the symp-
toms disappear in a few days, sometimes after a few hours. Cox
(901) remarked particularly on the decrease in the number of cases
of infantile beriberi in the Philippines since the introduction of
vitamine therapy.

It need hardly be said that poor results have also been obtained
by the use of vitamine therapy. What we said about avian beriberi
applies here also, namely, that for a successful vitamine therapy it
is necessary throughout that the anatomical changes should not have
progressed too far.

SHIP-BERIBERI

It is theoretically quite possible that vitamines B and C should
both be lacking in the diet, and in this way give rise to a mixed form
of scurvy and beriberi. Such mixed forms have not been described
with great certainty, but it would be possible to regard ship-beriberi
as such. The various investigators are not agreed as to the nature
of this type of beriberi. It occurs on sailing vessels, when the supply

BERIBERI 295

of fresh vegetables approaches the end. Characteristic symptoms
are numbness of the extremities, short breathing, and finally death
from heart failure. The patients recover very rapidly when they
receive fresh provisions, nervous symptoms being noted only seldom;
this is not the case in ordinary beriberi. If the crew is made up of
representatives of rice-eating people, who keep to their dietary
customs, then true rice-beriberi, and not ship-beriberi, develops.
Since long sea voyages are now of rare occurrence, ship-beriberi is
rarely met with. During the World War, the German raider
Kronprinz Wilhelm, with a great number of cases of ship-beriberi,
was interned in New York harbor. The crew lived on frozen meat,
while the officers received a daily portion of fresh vegetables and fruits
in addition, and therefore did not contract this disease. Since fruits
and vegetables contain both vitamines B and C, the above cases
can not give us any clue as to the true nature of the above condition.
Nocht (902) was of the opinion that the disease was not identical
with rice-beriberi, while Vedder (I.e. 838) believed that it was. Hoist
and Frolich (903), on the contrary, saw therein a greater resemblance
to scurvy, although Hoist (904) stated later that on Norwegian ships
the crews, which ate rye bread, were free of this disease. However,
if the rye was mixed with wheat, the disease developed. From this,
it might be concluded that we are dealing with beriberi, in which
view Hoist was strengthened by the results of his pigeon experiments.
Ship-beriberi is apparently a mixed syndrome, differentiated from
scurvy by numbness of the extremities, and from beriberi by scor-
butic gum changes.

RELATIONSHIP BETWEEN BERIBERI AND SCURVY

As we have already seen in the previous chapter, we believe that
under certain conditions mixed avitaminoses of beriberi and scurvy
may arise, though in practice, from a lack of both vitamines, one
condition develops in preference to the other. It is nevertheless
possible that scurvy should develop on an exclusive diet of rice.
During the siege of Paris in 1871, if the observations are correct,
such cases occurred and were reported by Delpech (905) and Bucquoy
(906). Garcia (907) described a mixed form of scurvy and beriberi.
Scherer (908) had the opportunity of observing an outbreak of scurvy
in German Southwest Africa, in which there was a death rate of 30
per cent. The disease developed because of an exclusive rice diet

296 THE VITAMINES

and was accompanied by beriberi symptoms (heart dilatation). In
individual cases, anasarca, serous fluids in the pericardium, pleura and
peritoneum were noted — symptoms which belong sometimes to
both scurvy and beriberi. Greig (I.e. 880) also observed, in cases of
epidemic dropsy, scorbutic symptoms, with bleeding of the gums in
10 per cent of the cases. Gouzien (909) described an interesting
picture of a disease in Hanoi. At first beriberi developed; if white
rice was then replaced by the hand-milled variety, the entire 64
patients promptly recovered, only to develop scurvy somewhat later.
The cases of scurvy which develop on decorticated corn in South
Africa are explained with some difficulty. Fleming, Macaulay and
Clark (910) reported thousands of such cases among miners in South
Rhodesia. The composition of the diet there was as follows : Mealie-
meal (a corn product poor in vitamine B) — 2 pounds daily; meat — 1
pound weekly; beans — 2 pounds weekly; and nuts (monkey-nuts) —
If to 2 pounds weekly. Orenstein (911) is of the opinion that the
differential diagnosis of beriberi and scurvy in these cases was not
made properly; he could, however, firmly establish the point that
these diseases promptly disappear on giving fruit and vegetables.
Darling (912) regarded this South African disease as scurvy, whose
picture was complicated by beriberi. Among the pathological find-
ings, there was noted hypertrophy of the right heart, fatty degenera-
tion of the heart musculature and the vagus, in addition to accen-
tuated reflexes. In a later contribution, Darling (913) stated that
scurvy at the Rand mines can not be so easily cured as pure scurvy,
and it was therefore regarded as a mixed form.

What we wish especially to establish by these statements is the
fact that we encounter in beriberi, as well as in other human avitam-
inoses, a pathological condition which perhaps can not be regarded
as being entirely due to the lack of a single vitamine.

CHAPTER II

SCUEVY

A historical survey of this interesting disease is given by Schroder
(914), Schelenz (915), Hirsch (916) and Autran (917); in a new
monograph by Hess (918), the whole field of scurvy is critically
treated. We have already touched lightly upon the develop-
ment of this subject and have spoken of the accurate conception
of the nature of scurvy by physicians in the eighteenth century —
Kramer (I.e. 32), Bachstrom (I.e. 33), Lind (I.e. 34). From all of
these data, we see that scurvy has been known for hundreds of years,
and makes it appearance particularly on long sea voyages, polar
expeditions, and in time of war. Cook (919), in the report of his
expedition, mentioned the foodstuffs needed for the prevention of
this disease. On one occasion, raw sea-lion meat was used for this
purpose; on a second voyage, he used a malt infusion. Based
upon these and similar experiences, the English navy in 1804, at
the suggestion of Sir Gilbert Blaine (920), introduced the daily portion
of lime juice, whereupon, according to Budd (921), the number of
cases decreased markedly. It appears certain that scurvy was prev-
alent in every war, including the last, in great numbers; however,
in times of peace it was due to bad harvests, especially of potatoes.
MacNab (922) saw numerous cases in the English army at Nassirabad
in Rajputana (India) during 1833-1834. The disease was brought to
an end by using a native sour plant, Phyllanthus emblica. In the
Civil War in the United States (923), there occurred 30,714 cases of
scurvy, because of the use of dried vegetables in winter. In the
Franco-Prussian War of 1870-1871, numerous cases were observed
during the siege of Paris, described by Delpech (I.e. 905), Bucquoy
I.e. 906) and other physicians (924). The disease was at time time
prevalent in prisons and was frequently accompanied by edema. In
the Russo-Japanese War, scurvy was observed among the Russians
and was described in detail by Sato and Nambu (925) and Blau (926).
Sir W. G. MacPherson (927) saw 20,000 cases during the siege of
Port Arthur. As regards the disease among the civilian population,
it made its appearance when there was a scarcity of potatoes and

297

298 THE VITAMINES

fresh vegetables, especially when large amounts of meat were not
available. Curran (I.e. 781) reported an outbreak of scurvy in
Swift Hospital, Dublin, in 1847, among patients eating almost a
pound of meat daily, or drinking a half liter of milk, in addition to
their usual food. Rae (928) reported that in the Hudson Bay Expe-
dition, the natives did not develop scurvy because they ate a large
amount of meat per day — the men, 8 pounds; the women, 6 pounds,
and the children, 2 pounds. Lanceraux (929) reported cases in the
prisons of the Department of the Seine arising from a lack of fresh
vegetables and potatoes. In Russia, the disease has been prevalent
almost at all times, and one of these occurrences has been described
by Berthenson (930). Muller (931) observed numerous cases in
Niirnberg early in the winter of 1911, arising as a result of a poor
harvest. Taussig (932) believed that in Austria, also before the
World War, scurvy was not infrequently noted, especially when it
was difficult to obtain potatoes. The importance of potatoes as a
protective against scurvy is firmly established, and this is also shown
by the newest data. In England, especially in 1917, cases of scurvy
occurred in poorhouses in Glasgow (Pickens, 933) and in Newcastle
(Harlan, 934), which could be attributed to the above cause (lack of
potatoes). In a health report of the city of Manchester (935), cases
of scurvy were described in the spring of the same year. An article
in the British Medical Journal (936) at this time showed the serious
situation, and the danger of the substitution of potatoes in the diet
by rice and bread. The quantity of potatoes necessary to protect
against scurvy, according to the Committee of the English Royal
Society (937), is about one pound per day. Hess (I.e. 819) showed
the etiological relationship between the occurrence of the disease and
the amount of potatoes allotted, in one institution where 200 cases
were involved. Lind (938) describes a number of cases in a hospital
at Kew, Victoria (Australia), while G. R. Hopkins (939) observed
3000 cases in Aruba, a small island in Dutch Guiana, which had a
total population of 10,000 in 1915. The cause of the disease was a
total crop failure during the period 1912-1914. The diet of the
natives consists of corn, corn meal, salt fish and a small amount of
meat, mostly dried or salted. The well-to-do class, which imports
fresh vegetables and fruits, were free from this disease. With the
arrival of the rainy period, and the consequent availability of fresh
vegetables, the disease disappeared completely.

SCURVY 299

During the World War, thousands of cases were observed, and it is
most likely that no single army was entirely immune. Hehir (I.e.
867) described numerous cases among the Indian troops, in Mesopo-
tamia, who refused to eat meat because of religious scruples. Turner

(940) states that 30 to 50 per cent of these troops developed scurvy,
and Wilcox quoted the following figures:

1916 11,445

1917 2,197

1918 • 826

On the other hand, among the English troops, only sporadic cases
were seen. The situation was so serious that a corps of gardeners

(941) was sent to Mesopotamia by the English government to plant
vegetables of every kind and to distribute seeds which could be
sprouted when the need developed. L. Hill (942) described an
occurrence of scurvy among English troops in Scotland, attributed
to excessive cooking of vegetables. The disease seemed to single out
particularly those men who were engaged in hard work.

During the late war, the disease was noted in Russia by Horschelmann
(943) and among the Russian prisoners in Turkestan, by Disque (944).
Borich (945), chief surgeon of a Red Cross Station in Russia, person-
ally observed 1343 cases. Among some troops, 75 per cent of the
men were disabled by this disease.

In France, Harvier (946) reported that in 1917 95 per cent of his
800 troops developed scurvy, while at the same time the disease was
discovered in other detachments. Benoit (947) saw 63 cases out of a
body of 350 men, but only of a transient nature; the patients recovered
without special treatment. Mouriquand (948) too spoke of a number
of scorbutic cases in the army.

For the Italian army, we have the reports of Ferrari (949),
Ramoino (950) Gingui (951), and of Vanutelli (952), who observed
200 cases of infectious purpura with hemorrhagic scurvy. Vallardi
(953), in Macedonia, saw cases among Italian soldiers who mani-
fested icterus and enlargement of the glands.

From Germany and Austria, we have many reports of the army
medical staff, from which it is clear that many of the cases were
diagnosed (954) as purpura or rheumatism. The cases of scurvy
during the war were so numerous that special stations were opened.
Schreiber (955) observed 30 German prisoners suffering from scurvy,

300 THE VITAMINES

which was at first diagnosed as purpural rheumatism. Speyer (956)
was sent by the German government to Bulgaria to make a study
of scurvy. Lobmeyer (957) observed scurvy in the Turkish army, but
it is possible that some of these cases were complicated by war edema
and beriberi. Other cases of scurvy manifested during the war
will be discussed in the course of this chapter. Simultaneously
with scurvy in adults, numerous instances of infantile scurvy were
noted in Central Europe. However, it is no longer necessary to
differentiate between these two types.

MODE OF DEVELOPMENT

It is always instructive and of practical importance to know the
exact diets that lead to the respective avitaminoses. With regard
to scurvy, such data are at hand. Scherer's report (I.e. 908) treat
of more than 862 cases of scurvy in the earlier German colonies,
among miners and railroad laborers, of which 289 died. The diet
there consisted of a kilo of white rice and 500 grams of meat two times
weekly, almost without fruits and vegetables. The first symptoms
appeared in 5 to 6 weeks. In the Bulletin of Tropical Diseases
(958) there is a description of a dietary which led to numerous cases
of scurvy in the prison at Burma:

grams. grams.

Hulled rice 684 Spices 4

Beans 114 Fish paste 14

Vegetables 285 Salt 7

It was shown that an addition of vegetables, milk, meat or fish
had little effect on the disease, but an addition of sweet potatoes
was effective. Dyke (959) described numerous cases among South
African natives, transported to France for war work. The number
of patients amounted to 680 out of a total of 1700. The diet con-
sisted of 450 grams frozen or canned meat and 225 grams fresh
vegetables. In addition, corn or rice, bread and margarine were
provided. Vegetables were cooked for 3 hours, which was obviously
partly responsible for the outbreak of the disease; aside from this,
Dyke noted that the natives drank 3 liters of beer, called "Kaffir,"
prepared in Africa from germinated kernels, but in France from
ungerminated corn. This beer is supposed to protect the natives
from scurvy in their home land. During the war, Comrie (960)

SCURVY 301

was with the English troops in North Russia; in the prisons there,
he observed scurvy. The disease began after 4 to 7 months on a
diet consisting of 313 grams flour or zwieback; 250 grams rice, oat-
meal, peas or beans; 205 grams frozen or canned meat or salted her-
ring; 50 grams bacon or pork; 7 grams tea; 28 grams sugar; 21 grams
salt and 14 grams preserved lime juice. Of the antiscorbutics
investigated, the order of activity was as follows : Sour milk, fresh
meat, fresh lemon juice, germinated peas, canned fruit and germi-
nated beans. Stevenson (961) believes that in adults the disease
develops in 4 to 8 months, and that 200 grams cooked vegetables
daily may prevent the occurrence of the disease. According to Chick
and Dalyell (962), prolonged cooking of vegetables is frequently
responsible for the outbreak of scurvy. Forty such cases were
observed in Pirquet's children's clinic in Vienna; these cases
developed eight weeks after the fresh vegetables were dealt out,
somewhat sparingly. These investigators believed also that rapid
growth on a diet very rich in calories is especially conductive to the
development of scurvy. During and after the war, a number of cases
of scurvy were observed among older children, which was very seldom
the case in times of peace. Tobler (963) commented in particular
upon this, mentioning more than 200 such cases. Erich Miiller
(964) describes a number of these cases in the orphan asylum
(Frederick the Great) in Rummelsburg near Berlin, which were
attributed to the use of dried vegetables. Weill and Dufourt (965)
noted it in children between 2| and 6 years old in a zone formerly
occupied by German troops.

Regarding infantile scurvy, "Barlow's disease" (cf. reviews by
Morse, 966), this condition occurs almost exclusively in artificially
fed children. There are, however, a few investigations, for example,
that of Netter (957), who described the disease in breast-fed infants;
however, from a critical survey of these cases, Hess (I.e. 918) con-
cluded that these data were uncertain, although theroretically not
impossible. The cause of the disease is to be sought in the heating
of the milk (pasteurization or sterilization). The feeding of artificial
milk products such as condensed, homogenized or preserved milk,
or of children's proprietary foods as chief diet (Cheadle and Poynton,
968) brings about the disease. After the introduction of the soxhlet
apparatus, the milk was not infrequently warmed for 45 minutes
and longer. The first extensive investigation of this disease we owe

302 THE VITAMINES

to Sir Thomas Barlow (I.e. 35). He concluded that this type is
identical with scurvy in adults, as did also Hoist and Frolich (I.e.
903) and Looser (969).

Through the work of Neumann (970), we recognized the causative
relationship between heating the milk and the appearance of infantile
scurvy. He observed the development of scurvy on a milk which was
first pasteurized at the dairy farm, and on the next day, before
consumption, heated in a soxhlet for 10 to 15 minutes. Neumann's
conclusions were confirmed by Heubner (971) and by A. Meyer
(972). Brachi and Carr (973), in England, reported some cases of
of scurvy on cooked milk. In 1898 the American Pediatric Society
(974) issued a report on 356 cases, all on artificial feeding. Up to
1908, Cheadle and Poynton (I.e. 968) collected 80 such cases.

More recently, Hess has occupied himself, together with his co-
workers, with the subject of infantile scurvy. Hess and Fish (975)
found that scurvy in children may be brought about by feeding milk
heated for 30 minutes at 145°F. The condition may be prevented
by the addition of antiscorbutics. Sittler (976) stated that milk,
warmed for 5 minutes on the water bath, does not cause scurvy.
Comby (977) described 41 cases,. on proprietary foods, among which
there were several sent in with an incorrect diagnosis. Epstein
(978) reported a number of cases in Prague during the war, at a time
when oranges and fresh fruits were scarce ; the milk was evidently
heated several times before using.

We must also consider the flour diet as an e biological factor.
Bendix (979) described a case, on cow's milk diluted with Kufeke's
flour food, which was cured by raw milk. Even if the carbohydrates,
as such, do not hasten the symptoms, they act unfavorably because
of the milk dilution. Hess and linger (I.e. 656) have shown that
malt soup, prepared with the addition of alkali, is productive of
scurvy, while Gerstenberger (I.e. 829), under certain conditions,
could effect a cure with it. Such cures, however, may also be
attributed to variations in the quality of the milk. Fordyce (980)
believes that the disease can be attributed to the use of proprietary
foods; he rarely saw the disease in children less than six months
and more than one year old. Apart from the facts touched upon
here, the points to be considered in infantile scurvy are those that
have been discussed in the description of the nutritive value of
milk.

SCURVY 303

SYMPTOMATOLOGY AND PROGRESS OF SCURVY

The primary stage in scurvy is characterized by a peculiar pale-
ness of the skin, apathy, melancholy, muscle weakness and slight
dyspnea. The skin becomes yellowish, dry and scaly, and is covered,
especially on the lower extremities, with petechiae and larger livid
spots, due to subcutaneous hemorrhages. In addition, there are
frequently pains in the lumbar region and the legs, often re-
garded as rheumatic. Changes in the skin are frequently noted;
thus Taussig (I.e. 932) reports an exanthema of the legs. AschofT
and Koch (981) describe small hemorrhages near the hair papillae
(piquete scorbutique), and an exanthema localized in certain parts
of the body, mostly observed in the scorbutic cases during the war
in Rumania and in the Carpathians. Similar skin changes were
described by Rheindorf (982). Wiltshire (983) saw among the 3000
cases of scurvy in the Serbian army numerous skin symptoms, which
he called "hyperkeratosis," a condition occurring in 87 per cent
of his cases; later he noted an exanthema of the petechial type,
localized in parts of the body, already mentioned by Aschoff and
Koch. Wiltshire also observed conical swellings around the hair
follicles, v. Niedner (984) observed that infectious exanthema
frequently assumes a hemorrhagic appearance in the presence of
scurvy. According to the paper of Sato and Nambu (I.e. 925), the
skin, as well as the subcutaneous tissue, is often edematous.
According to Wassermann (985), subcutaneous, diffuse, painful swell-
ings appear on the thighs; Speyer (I.e. 956) occasionally noted
abcesses on the calves. Swellings in the knee caps also appear.
The gums become dark red, soft and swollen, especially around
carious teeth, although this characteristic sign may be entirely
lacking. Sometimes the gums swell so that the teeth are entirely
covered. Zlocisti (986) described an ulcerous, gangrenous sto-
matitis. Aschoff and Koch noted a falling out of the teeth with
resultant hemorrhages.

In later stages, the patients suffer very much from dyspnea and
palpitation; they faint easily, and heart weakness progresses. The
sufferers becomes markedly emaciated, the muscles become atrophic,
and contractions are seen at times. Scherer (I.e. 908) described a
tip-toe position, which he believed to be the result of hemorrhage;
Aschoff and Koch described a characteristic position which they

304 THE VITAMINES

called "Seiltaenzerstellung." Richter (987) speaks of tetanus symp-
toms in scurvy. It is not quite clear whether these symptoms
belong to the picture of scurvy or whether caused by complications,
for example, with beriberi.

FIG. 60. SKIN EXANTHEMA IN SCURVY (AscnoFF-Kocn)

The pulse is rapid; the left and right ventricles are frequently
greatly enlarged. Ascites fluid, dystrophic edema, subperiosteal
hemorrhages, bloody-serous fluid in the pericardium, pleura, or in
any of the large joints — all belong to the picture of scurvy. In

SCURVY 305

this stage, we meet with persistent diarrhea with bloody-serous
stools. Sometimes there is icterus, as was found by Scherer (I.e.
908) and by Urizio (988). Death results with complications of an
infectious nature; although it may also be due to heart failure.

Complication with hemeralopia (night blindness) was noted by
Zak (989) among Russian prisoners. Hift (990) was able to avoid

FIG. 61. "SEILTANZER" POSITION IN SCURVY (ASCHOFF-KOCH)

these complications by means of liver therapy, which shows, perhaps,
that the condition of scurvy in these cases was complicated by still
another avitaminosis. O'Shea (991) saw 300 cases of hemeralopia
among English troops, in 22 cases of which the fundus of the eye
was examined. In three of these cases, the only change was a definite
paleness. Changes in the retina in scurvy were investigated by
Kitamura (992).

306 THE VITAMINES

Infantile scurvy

Infantile scurvy develops between the 6th and 18th month, seldom
earlier or later, and Hess (I.e. 918) differentiated between the acute,
sub-acute and latent forms ; this view was accepted by Miller (993) .
The subacute form is not very well defined but growth ceases. Aside
from a somewhat abnormal color of the skin and an occasional edema
of the upper eyelids, no further pathological signs are noted. The
child is capricious, and the lower limbs are somewhat sensitive to
touch. The knee reflexes are almost always markedly accentuated.
All of these symptoms are promptly overcome on the addition of
antiscorbutics, whereupon growth is resumed, according to Hess
(l.c; 461).

The latent form is even less pronounced, and the children recover
in an extraordinary manner after the administration of antiscor-
butics. According to Hess, the latent form occurs especially in
cities, where pasteurized milk is sold. It does not need to be
particularly exphasized that these less important forms of infantile
scurvy occur more frequently than do the acute forms. Franchetti
(994) described a chronic case of scurvy in an older child, with
bone dystrophy, painful paraplegia, cachexia, anemia, absolute
anorexia, but without hemorrhages.

Muscle weakness, anemia, and anorexia, are the initial symptoms
noted. Pain develops in the legs; the child holds them motionless
and cries out when they are moved. This behavior resembles pseudo
paraplegia, according to the statement of Comby (995). Swellings
are frequently noted, sometimes only on one extremity, of a
periosteal nature, especially of the tibia. Besides this, swelling of
the muscles was noted, caused by hemorrhages. The skin on the
arches of the feet is frequently edematous, while the skin over the
swelling appears pale. If teeth are present, the characteristic gum
changes occur. The spongy, bleeding swellings become ulcerous
in some cases and the teeth may loosen and fall out. Oliguria,
described by Gerstenberger (996), is quite characteristic of this
condition, and hematuria less frequently so. The temperature
remains normal, occasionally going up to 39°C., following blood
resorption after hemorrhages (Barlow fever). When no thera-
peutic measures are applied, death may ensue as a result of heart
failure or hemorrhages. Initial phases of scurvy were described by

SCURVY 307

Cozzolino (997). He recommended the frequent inspection of the
gums for the purpose of the speedy recognition of the disease.

A sort of eczema was noted by Hess (I.e. 918) in 8 cases. In one
instance a symmetrical erythema was observed, resembling pellagra
erythema very strongly. Proptosis of the eyeball, following
hemorrhages, was observed by Cheadle and Poynton (I.e. 968) as
well as by Zilva and Still (I.e. 456). It occurs usually in the left eye.
Schodel and Naumwerk (998) observed enlargement of the right
heart; Hess (999) noted the quite common occurrence of polypnea
and tachycardia, apparently because of lesions in the pneumogas-
tricus. Erdheim (1000) also described heart lesions in Barlow's
disease.

DIAGNOSIS

From the foregoing, the clinical picture of scurvy is quite obvious
and becomes even more simple as, a result of the newer possible
differential diagnosis, by the administration of vitamine C. Changes
in the complexion, follicular changes in the skin, petechiae, sub-
cutaneous hemorrhages, hemeralopia, changes in the gums, feeling
of fatigue, shortness of breath, rapid pulse (about 140) — all these
are specially characteristic of scurvy. Some times it may be confused
with rheumatism and perhaps with hemorrhagic diathesis. In
order to differentiate here, the anamnesis is of value as is also the
result of vitamine therapy. Occasionally the hematological investi-
gation may be used for the diagnosis, which, however, according to
Rosin (1001), shows an unspecific picture of anemia. The so-called
"tibialgia" of v. Schrotter was designated as scurvy by Labor
(1002).

In the diagnosis of infantile scurvy, the "white line," described
by Frankel (1003) in the X-ray of the bones, has been regarded as
characteristic. It is a shadow which is apparent at the epiphyseal
ends of the long bones. This indication was observed by Brown
(1004) and by many others, although Hess (I.e. 918) did not believe
that it was significant in the diagnosis. He observed this shadow
also in cases of scurvy, which had been cured for a long time. Hess
and Unger (I.e. 759) have introduced a capillary resistance reaction
for the diagnosis of scurvy, which consists in the compression of the
arm for three minutes with a tourniquet, whereupon petechial spots
appear if scurvy is present.

308 THE VITAMINES

Hematology

A number of investigators have recorded their findings as to the
blood picture, but it does not appear that specific indications were
discovered. On this subject, there is the work of Labor (1005),
Leitner (1006), Brandt (1007), Wassermann (1008) and Benoit
(1009); the last investigator reported findings in 63 adults. Hess
and Fish (I.e. 975) made hematological studies in Barlow's disease,
including the clotting-time of the blood, which was found normal.

As regards the chemistry of the blood, the sugar content was
determined in some cases by Roily and Oppermann (1010) as well
as Schumm (1011), and found normal. Hess and Killian (1012)
determined the calcium, sugar and urea content, as well as the dia-
static and the carbon dioxide combining power of scorbutic blood.
They found a moderate acidosis, and a smaller calcium content
(different from rickets) ; as for the rest, the values were normal.

Metabolism experiments

Bauman and Howard (1013) investigated the inorganic metab-
olism in one case of adult scurvy. The chlorine and sodium balances
were negative and were not much more favorably affected by the
administration of antiscorbutics. The sulphur metabolism in adults
was found abnormal by Labbe, Haguinea and Nepreux (1014).
The case investigated was, perhaps, in the convalescent stage and
was marked by a large urea output, indicative, possibly, of the
disappearance of edema. Experiments in an older child were carried
out by Lust (1015), and especially by Lust and Klocman (1016).
The child showed a retention of all inorganic constituents, which
were subsequently eliminated after a cure; this applies, in particular,
to calcium. These data, which seemed to be somewhat uncertain
at first, appear now to be corroborated. As for the calcium metab-
olism, the above results were confirmed by JVLoll (1017). Frank
(1018), experimenting with two cases, found a calcium retention in
the acute stage; during convalescence, this calcium was again
eliminated.

Pathology of scurvy

A splendid monograph on this subject is that of Aschoff and Koch
(Lc. 981) as well as of v. Samson-Himmelstiern (cited by Aschoff
and Koch, I.e. 981); with the exception of severe cases, only well

SCURVY 309

nourished corpses came to necropsy. The subcutaneous tissue,
especially of the extremities, is infiltrated with bloody-serous fluid.
Widespread blood extravasations, old and new, are noted. The
adductor side of the legs shows more hemorrhages than does the
abductor; hemorrhages in the muscles themselves are rare. Hemor-
rhages occur also in the periosteum, especially in the tibia, and in
younger individuals in the epiphyseal ends of the long bones and
at the cartilage ends of the ribs. Bone investigations in adults
were made infrequently but it appears that here, too, fractures
are noted. Rosary is often observed, as in rickets. Enlargement
of the right and left ventricles, with a possible fatty degenera-
tion of the heart muscles, was also described. Most of the organs
were examined microscopically by Aschoff and Koch. In the bones,
osteoporosis and a disappearance of osteoblasts were found; nothing
of importance was seen in the endocrine glands. Suprarenals showed
only a greater lipoid content. Feigenbaum (1019) found hem-
orrhages in the spinal cord; Scherer (I.e. 908) found hemorrhages
in the dura and pia as well as rank growth of connective tissue in
the liver.

Ecchymoses occur frequently in the pleura, less often in the peri-
toneum, and in severe cases in the lungs. Bloody-serous fluids in
the pericardium and pleura — less frequently ascites fluids — were
found by Johnson Smith (1020).

Barlow's disease

The pathological findings were described by Frankel (1021), the
bones in the infantile form having been frequently investigated. The
periosteum is filled with blood and is thickened, but free from small
cell infiltration; between the periosteum and the bones widespread
hemorrhages are found. At the diaphyseal ends of the long bones,
the bone marrow loses its lymphoid character and forms reticular
tissue poor in cells and blood vessels which, in stained sections, is
characterized by light color, because of "Helles Mark." This
degeneration of the bone-marrow is specific for infantile scurvy and
prevents normal ossification. The new bone formation is delayed
or completely prevented, and the bone tissue present is atrophied,
particularly in the ossification zones. In rare cases, hemorrhages
are found in the dura, in some joints, in the lungs, spleen and kidneys.
As we have already stated in discussing guinea pig scurvy, Hess and

310

THE VITAMINES

Unger (I.e. 378) showed that a scorbutic rosary may be cured by
giving vitamine C. It may seem from this that the complications
of scurvy by rachitic signs frequently mentioned in the literature
may be attributed to the above condition. By investigation of the
bones, however, the scorbutic lesions may be differentiated from
the rachitic. In rickets, we find a broad band of osteoid tissue
formed at the epiphyseal ends; there is a broad band of insufficiently

FIG. 62. SCORBUTIC ROSARY
"Scurvy" Hess, courtesy of J. B. Lippincott

calcified cartilage, which is never observed in scurvy. In scurvy,
the growth of the osteoblasts is markedly inhibited, but when growth
starts, it proceeds in a normal manner. In addition, we find a
difference in the number of capillaries in the cartilagenous tissue and
in the marrow, very rare in scurvy, but much more frequent in rickets.
Hess (1022) saw also a degeneration of the ventral horns in the lumbar
region of the spinal cord.

SCURVY 311

Bahrdt and Edelstein (1023) analyzed the various organs in infan-
tile scurvy. The bone-marrow was poor in solids, calcium and phos-
phorus. This was also true of the muscles; normal figures were
obtained for the other organs.

THERAPY

Since we have already spoken of the antiscorbutic value of various
foodstuffs, we need not go further into the matter at this point.
On using antiscorbutics, the symptoms disappear, in most cases,
after 2 weeks. Even severe symptoms, such as hydropericardium
and hydrothorax are very favorably influenced. Still, when the cases
have progressed too far, curative results may not be obtained.
Paget (1024) treated 133 cases with camel's milk, with good results.
As for lemon juice, 24 cc. daily appear to be sufficient for prophylaxis.
Germinated peas and beans are also used in practice. They were
given to Serbian soldiers, with good results, by Wiltshire (I.e. 806).
The treatment of postscorbutic conditions was described by Schul-
hof (1025).

As for the therapy in Barlow's disease, it consists in the addition
of raw or concentrated milk, orange peel extracts, canned tomatoes
(Hess and linger, I.e. 668), and natural fruit juices (Delille, 1026).
Hess and Unger (I.e. 637) reported successful intravenous therapy
with neutralized orange juice. Harden, Zilva and Still (1027) have
used dry lemon juice preparations with great success.

With regard to blood transfusion therapy, Hess (I.e. 918) did not
find much vitamine C in blood so that a large amount of blood would
have to be used for curative purposes. Similarly, Rueck's (1028)
results did not appear any more certain. With a well chosen
antiscorbutic, the therapy is almost magical. In 24 to 48 hours,
the curative effect is already distinctly obvious.

CHAPTER III

RICKETS

This universally important disease occurs invariably in the first
or second year as a result of a deficient diet. In fact, during the
last war, the disease occurred almost in all ages, so that the boundary
lines between the real rickets, rachitis tarda and osteomalacia have
almost disappeared. As we shall see, the inadequate diet of itself,
even under the best hygienic conditions, suffices to develop the
disease; the role of other factors such as lack of air and light, is not
ruled out. By and large, the number of cases of rickets increases
with the rejection of the natural modes of nutrition and the adoption
of artificial feeding of infants. The further child nutrition strays
from the normal (breast milk), the less it corresponds to the age of
the child, and the more severe are the symptoms of rickets. This
disease is chronic and is not dangerous to life of itself, but it does
predispose towards infections, especially of the respiratory apparatus.

Rickets occurs everywhere, especially in the temperate zone, less
in the north (Iceland, Greenland, Norway), rarer in the tropics and
also southern Italy and Spain. Rickets is seldom noted in countries
where breast feeding is prevalent, as a rule, but increases in the in-
dustrial centers, following artificial infant feeding and lack of sunlight.
A racial influence is not noticeable; for example, rickets frequently
develops among negroes and southern Italians in America, whereas
in their native countries they are unaffected (Cautley, 1029; Strong-
man and Bowditch, 1029a). The frequently observed racial predis-
position may be attributed as well to the dietary customs. Con-
densed milk together with proprietary foods, as well as a one-sided
carbohydrate diet, leads almost invariably to the development of
rickets. According to Dennett (1030), there is no danger of rickets
from cooked milk, as such. However, even breast-milk may be an
insufficient food, especially when the mother's diet is inadequate.
We have noted an analogy to this in other avitaminoses, particularly
in beriberi.

It is evident that rickets arises when the diet either is lacking
in a certain vitamine indispensable for normal metabolism, or contains
it in insufficient amounts. We shall speak of other complicating

312

RICKETS 313

factors in discussing the etiology of this disease. The missing sub-
stance is found in good breast-milk and in cod liver oil, but not in
certain milk preparations and flours. It has nothing to do with a
lack of fat, protein or calcium, per se, as has been often stated, but
with a vitamine which we call "antirachitic vitamine," either
belonging to or identical with the type of A-vitamine. In what
way the lack of vitamine A brings on all of the known symptoms of
rickets, is at present unknown, and we find ourselves upon the same
uncertain foundation characteristic of the explanation of all other
avitaminoses.

OCCURRENCES

According to the newly found facts, it is unlikely that the diseas3
is associated with any definite age, although it seems certain that it
is manifested mostly during the period of active bone growth.
Rickets occurs very frequently and we need mention only two investi-
tigations to illustrate this. Schmorl (1031) stated that on post-
mortem of the cadavers of children under 4 years old, 90 per cent
showed signs of rickets. Schwartz (1032) observed among 4944
cases, 15 per cent craniotabes in the first year; 35 per cent showed
rosary (of which 13 per cent were in the first month). Congenital
rickets too was observed with certainty (Kassowitz, 1032a) . Mery
and Parturier (1033) saw a case in a 6-weeks old child. Wieland
(1034) reported on this too, but he (1035) did not believe in the exis-
tence of congenital rickets. Sinclair (1036) and Carr (1036a), held
that premature children contract rickets very easily, since the neces-
sary reserve material has not yet been prepared. Hamilton (1037)
has described craniotabes in the second month among premature
babies. Retterer and Fisch (1038) describe a case of congenital mi-
cromelia which they regarded as rickets. Since rickets may occur at
any age, the cases of hereditary rickets are not puzzling; the mother
may be suffering from a lack of vitamine A during pregnancy, when
the requirements are even greater than normally. Among the cases
of "osteogenesis imperfecta," there are, to all appearances, those that
may be classified as congenital rickets. On the other hand, some
of them must be regarded as hereditary osteoporosis, with a poor cal-
cium assimilation. Ostheimer (1039) described a condition in chil-
dren, called "fragilitas ossium," resembling osteoporosis (lack of cal-
cium), which in this case could be improved not by a calcium or cod

314 THE VITAMINES

liver oil therapy, but by changing the diet of the mother. He believed
that this disease was associated with a disturbance in the metabo-
lism of the mother. One case of osteogenesis imperfecta has been
described by Bookman (1040). In this instance, calcium lactate and
cod liver oil were used with good results, but it is not quite clear
which of the two was responsible for the curative effect. This child
progressed even better on breast-milk. Another such case was re-
ported by McClanahan and Willard (1041); the child was born with
multiple fractures. Another case was improved with cod liver oil,
according to Schabad (1042).

A logical transitional stage is the rickets in breast-fed children.
Brade-Birks (1043) reported such a case, and in this respect, Hess
and Unger (1044) controlled the diet of negro women in New York
in a district where the children suffered markedly from rickets.
The diet was recognized as being very inadequate from the
dietetic viewpoint.

The deficient nutrition in Europe during the war, especially in
the central European countries, brought with it a great increase in
the number of cases of rickets, according to Japha (1045). The
mortality in rachitic age, due to the pulmonary complications, was
greatly increased according to Engel (1046). Weiss (1047) reported
that 90 per cent of the children born in Vienna after 1917 suffered
from rickets, not excepting the children of well-to-do parents. The
disease appeared mostly among children from 1 to 4 years of age.
Adams and Hamilton (1048) came to the same conclusion as regards
Germany. Rickets in older children was described by Schlee (1049),
Stetter (1050) and Sauer (1050a) in Germany, and by Sutton (1051)
in Australia; the literature on the subject was reviewed by Frangen-
heim (1051a). Schlatter's disease, which consists in a separation of
the epiphyses of the long bones and other similar fractures, was
described in a 13-year old boy by W. Miiller (1052) and in a similar
case by Paus (1052a); these cases were regarded as late rickets.

The effect of rickets on later life has apparently been under-
estimated. Apart from the possibility of permanent bone deformity,
as reported by Park and Rowland (1052b) and by Brusa (1052c),
the disease, according to Engel (1053), exerts an influence on the
growth of the children, perhaps on the mental development. Accord-
ing to Karger (1054), these children are less active and mentally
underdeveloped. In this connection, Leri and Beck (1055) inves-

RICKETS 315

tigated the French soldiers, and noted that those who had suffered
from rickets in childhood showed, aside from anatomical changes,
mental deficiency and little resistance to bodily exertion.

SYMPTOMATOLOGY AND DIAGNOSIS

Rickets is primarily a disease of the bones, although it must be
regarded as a general disturbance of metabolism. As we gain
knowledge of this condition, the latter assertion may perhaps become
more justified. The children show a weak muscle tonus and perspire
profusely, particularly on the scalp. They commence to walk
rather late; the fontanelles remain open longer than usual and the
course of the disease may be controlled to a certain extent by measure-
ment of the opening. Swellings of the cartilage ends of the ribs
(rosary) are easily palpated, and sometimes are apparent even without
this. According to Juaristi (1056) the eyes in rickets are more round
and show more of the sclera, associated with bone changes in the
fundus of the eye; the penis is long and hangs flaccid. Evidently
the delayed development of the teeth is also characteristic of
rickets; the enamel is often lacking, and occasionally is covered with
a greenish deposit which has been emphasized by a number of authors
(1056a). According to Lichtenstein (1056b), rachitic children exhibit
an abnormal sense of taste. Various other symptoms associated
with slight muscle tonus are described. Characteristic skull forma-
tion of rachitic origin is also observed, and as soon as the children
commence to walk, the legs bend in the known way, because of the
weight of the body.

The diagnosis presents no particular difficulty; the differentiation
from scurvy having already been discussed. The differential diag-
nosis between rickets and some pseudo-rachitic appearances, such
as "osteoporosis," is not so easily made. In this respect, the X-ray
may be of some help, in addition to the results of vitamine therapy,
as compared with calcium therapy.

PATHOLOGICAL ANATOMY AND CHEMICAL PATHOLOGY OF RICKETS

We find, in the work of v. Recklinghausen (1057) a careful and
splendid treatise of this subject. The chief sign of rickets is the
persistence of the cartilage in the uncalcified condition. The

316 THE VITAMINES

rachitic bone is recognizable by its abnormally small content of
alkaline earths. The deposit of calcium salts is prevented in that
part of the skeleton which takes those salts during normal growth;
in fact, according to Schmorl (1058), this happens in the entire skele-
ton. A greatly increased resorption of the already calcified bone
is not characteristic for rickets; rather does it characterize pseudo-
rickets. In the investigation of rickety bones, the preponderance
of the cartilage over the bone tissue is especially marked; for in-
stance, Dibbelt (1059) found, in normal bone, 29.4 per cent cartilage
and in rickety bones, 71.3 per cent. According to Schmorl (I.e. 1058),
there is an abnormal formation of osteoid tissue. Important changes
in the bone-marrow were found by Marfan, Bardouin and Feuille"
(1060). Marrow cells were discovered in places where they do not
occur normally. These cells were later substituted by fibroid cells
with outgrowth of cartilage cells; by this means, the disturbance in
the function of the osteoblasts is explained. Hutinel and Tixier
(1061) have confirmed the above findings. Kassowitz (1062) found,
in rapidly growing bones, an abundance of blood in the epiphyseal
ends, causing a proliferation of the cartilage cells and disturbing
the normal ossification. Heubner (1063) and Pommer (1064), how-
ever, found no inflammation but only a seemingly abnormal prolifer-
ation of the cartilage tissue. Ribbert (1065) also noted a marked
breaking down of cartilage cells. Besides the bone findings, muscular
dystrophy was noted in severe cases by Hagenbach and Burckhardt
(1066), Bing (1067) and also by Banu (1087a). According to Mohr
(1068), the central nervous system, besides the muscles, also is
affected. Of the endocrine glands, du Castel (1069) investigated
the thymus and found it hypertrophied. Stolzner (1070) found the
suprarenals small with a slight adrenaline content, while Cattaneo
(1071) was unable to confirm this.

As for the chemical findings, Dibbelt (I.e. 1059) gave the following
composition for normal and rachitic bones:

NORMAL

RACHITIC

Fat

1.89

7.50

Ca3(PO4)2

57.38

15.11

Mg-phosphate

1.72

0.78

CaCO3

8.95

3.15

Other salts

0.83

2.20

RICKETS 317

Gassmann (1072) found the following:

NORMAL

RACHITIC

CaO

24.0

21.0

P2O5

33.0

30.0

CO2

3.0

2.75

MgO

0.10

0.53-0.74

In addition, 5 per cent of organic matter was found in rachitic
bones. Similar analytical results were obtained by Schabad (1073).
Simonini (1074) saw a relationship between the calcium content of
the bones and the teeth. Rost (1075) found the water content of
rachitic bones higher than in the normal; the ash content of the
ribs and the vertebrae, on the contrary, was 20 to 69 per cent lower.
Aschenheim and Kaumheimer (1076) found the calcium content of
the muscles diminished in severe cases. The calcium content of
the blood was found to vary by Aschenheim (1077). Denis and
Talbot (1077a) observed a low calcium content in the serum. How-
land and Kramer (1077b) frequently found the calcium content
of the blood normal, while the phosphorus content was always low.
The sugar content of the cerebrospinal fluid was found diminished
by Suzuki (I.e. 894).

METABOLISM

In rickets, according to Dibbelt (1078), the elimination of salts in
the feces is increased, but decreased in the urine, and sometimes
entirely absent. If a cure is brought about, there is first a hyper-
retention of calcium salts with simultaneous increased elimination in
the urine. Since, in his time, the etiology was associated with the
amount of calcium in the food, Denton (1079) tried to determine the
calcium requirement in children. Aron (1080) tried to prove that
milk, especially mother's milk, contains just sufficient calcium to
cover the minimal needs. % The analyses of mother's milk made by
Schabad (1081) were for the same purpose. Orgler (1082) disputed
the results of Aron, and attributed them to a faulty calculation of the
calcium requirement. Cronheim and Erich Mliller (1083) compared
the mineral metabolism of normal and rachitic children, and found
no marked variations. In Aron's cases, the characteristic metabolic
disturbance had perhaps passed. While the assumption of the lack

318

THE VITAMINES

of calcium in the food could not be confirmed in general, the lesser
utilization of calcium in rachitic children has become a fact which
has gained support from the numerous reports of Schabad (1084)
and others. Schabad (1085) also studied the phosphorus metabolism
in various stages of rickets and convalescence. The children investi-
gated were of different ages, both breast and cow's milk being used ;
normal children were studied as controls. The following figures
were obtained:

DIET

CONDITION

P2Os

EXCRETION
PER KILO
PER DAY

RELATIVE

PzOs

EXCRETION
IN PER CENT
OF AMOCNT
GIVEN

PARTITION
OF P2Oi
EXCRETION

EXCESS IN
FECE8 IN
RELATION TO

CasCPOOt

I

I
•e

p

3

o
H

In urine

J

*c

o

i

9)

0

i
O

1

gram

gram

[

Normal 4-5 months

0.023

.0018

65.3

52.8 80.6

19.4

64.8

Breast-milk {

Progr. rickets,

I

5-13 months

0.034

0.021

122.2

72.2

60.8

39.2

52.8

(

Normal 3-6months

0.214

0.119

70.7

45.1

65.2

34.8

77.8

Cow's milk \

Progr. rickets,

I

5-8| months —

0.186

0.077

94.6

39.3

39.3

60.7

14.1

Mixed diet I

Normal, 4-5 years.
Progr. rickets,
4 years

0.15
0.091

0.102
0.04

80.3
102.1

51.5
44.9

64.4
44.1

35.6
55.9

22.0

39.8

Developed rickets 1 year, 5
months to 2 years, 7 months . . .

0.178

0.069

71.8

27.5

38.2

61.8

(16.5)

23.3

Convalescent 2£ to 8 years

0.103

0.077

65.5

48.7

74.4

25.6

35.6

(64)

Very recently, an important series of metabolism experiments in
rickets was reported by Schloss (1085a) and also by Freise and
Rupprecht (1085b).

According to Flamini (1086) the urinary CaO excretion in normal
children is 0.125 gram per day; in rickets, 0.05 gram on an intake of
3.4 grams per day. Peiser (1087) found the calcium balance negative.
The nitrogen and sulphur metabolism was studied in rachitic dwarfs
by Schwarz (1088), and the mineral metabolism in late rickets, by

RICKETS

319

Schabad (1089). Schabad's findings on the deficient resorption of
calcium were corroborated by Dibbelt (1090).

The study of the influence of the composition of the diet on the
utilization of calcium gives some interesting results. The effect of
protein was investigated by L. F. Meyer (1091) and by Tada (1092).
They found that the nitrogen elimination did not go parallel with the
calcium output. The influence of fats was studied by Meyer (I.e.
1091), Rothberg (1093) and Orgler (1094). They found, surpris-
ingly, that the addition of fats (in the form of whole milk) acted
unfavorably on the calcium utilization, losses occurring in the feces
in the form of soaps. The following figures were obtained on com-
paring the effect of cod liver oil and butter on the calcium balance:

WITHOUT
COD LIVER OIL

WITH
COD LIVER OIL

INSUFFICIENT
BUTTER FAT

EXCESS
BUTTER FAT

gram

gram

gram

gram

1

+0.060

+0.175

6

+0.137

-0.198

+0.141

7

+0.038

-0.034

2

-0.014

+0.143

8

+0.043

-0.120

+0.519

9

+0.037

-0.267

3

+0.073

+0.303

4

-0.038

-0.285

+0.141

+0.108

5

+0.067

+0.465

Hess (1094a) demonstrated the unfavorable influence of butter.
On the other hand, Telfer (1094b) found that fat, whether in the
form of cod liver oil or butter, has no influence on the calcium
metabolism. It should be noted, however, that the duration of his
experiment was too short (4-5 days) ; besides, the child under investi-
gation was not rachitic. Hutchinson (1095) found that the soap
elimination in rachitic stools was somewhat decreased, 2.2 grams
instead of 2.5 grams in normal children. In addition, he (1096)
found that the fat loss can be very large in a voluminous stool, giv-
ing a negative balance, under certain conditions, of 0.6 gram per
day; the fat sometimes constitutes one-third of the total volume
of stools.

Holt, Courtney and Fales (1097) demonstrated a large loss of
fat in the feces. However, the poor utilization of calcium after
the addition of milk fat (butter) could not be corroborated by these
investigators (1098).

320 THE VITAMINES

The addition of carbohydrates appears, according to the prevailing
data, to influence the calcium metabolism favorably, although the
results were not constant. Reports on this subject were made by
Dibbelt (1099), Massaneck (1100), Tada (I.e. 1092) and Rothberg
(I.e. 1093). Rowland and Marriott (1101) have investigated the
action of sugar and cereal addition and found it favorable; but the
effect of large excess of carbohydrates was not investigated.

THERAPY AND THERAPEUTIC EFFECT ON THE METABOLISM.

The therapy of rickets with phospho-cod liver oil was introduced
by Kassowitz, and is still used almost in the original form. Turning
to the findings on the calcium metabolism, the clinical reports on the
therapeutic value of cod liver oil are not particularly uniform, and it
may be concluded that perhaps not all cod liver oils possess the same
therapeutic value. As to this, we have, first of all, the prolific
work of Schabad (1102) and his co-workers, in which the favorable
effect of cod liver oil is demonstrated; the value of cod liver oil
emulsions was also tested. In a later investigation, Schabad (1103)
studied the effect of the Czerny-Keller diet on rickets and found that
it was evidently not adequate, since the addition of cod liver oil still
acted favorably, whereas in a pure milk diet, the oil was unnecessary.
Birk (1104) likewise reported on the results of phospho-cod liver oil
therapy.

It is only questionable whether in the above combination, the
phosphorus addition is necessary. Frank and Schloss (1105) believe
that between the two preparations — cod liver oil and phospho-cod
liver oil — no far-reaching differences exist; however, Schloss (1106)
recommended a simultaneous addition of a calcium preparation.
Grosser (1107) investigated the retention of organic calcium salts.
Pereida y Elardi (1108) did not believe in the favorable effect of the
calcium addition, since a physiologically sufficient amount of calcium
is supposed to be present in the diet. Kurt Meyer (1109) studied
the effect of cod liver oil therapy on the calcium balance, and obtained
good results. Tobler (1110) cured four cases of late rickets with
phospho-cod liver oil, and similar results were obtained by Ricklin
(1111). Phemister(1112)was of the opinion that phosphorus, as such,
possesses no significance in the therapy of rickets, but does stimulate
the formation of osteoid tissue. Phemister, Miller and Bonar
(1112a) believe now that phosphorus, per se, has a curative action.

RICKETS

321

However, the number of cases investigated is insufficient, besides
which there is the possibility of a spontaneous cure to be considered.
As a result of their rat experiments, Sherman -and Pappenheimer
(1112b) and Pappenheimer, McCann, Zucker and Hess (1112c)
are inclined to the belief in the therapeutic influence of phosphorus
alone. They believed they could produce rickets in rats on a
phosphorus-poor diet, and then effect a cure by the addition of
phosphorus. These experiments appeared not very convincing at
first, but have since received additional experimental proof. Lie"naux
and Huynen (1113) do not believe in the therapeutic influence of
calcium in rickets.

In opposition to the above more or less definite results, there are
the findings of Hess and Unge • (1114) on negro children in New York.
In these cases, cod liver oil was used without any phosphorus addi-
tion. In the negro section, about 90 per cent of all the children are
rachitic; the mortality is about 314 per 1000, in which tuberculosis,
pneumonia, and whooping cough play the greatest part, it being
known that rickets predisposes towards disease of the respiratory
organs. Altogether, 40 children were given cod liver oil, while 16
children served as controls. The children were between 4 and 12
months old and the complete results were as follows :

AVERAGE TOTAL
QUANTITY OP
COD LIVEB OIL

TIME OP
THERAPY

NUMBER OP

CHILDREN

KICKETS

NO RICKETS

NO RICKETS

grams

months

percent

1550

6

32

2

30

93

655

6

5

1

4

80

600

4

12

5

7

58

16

15

1

6

The children were mostly breast-fed and the results of the therapy,
especially in cases receiving more cod liver oil, were very marked.
These investigators (I.e. 1044) studied also the diet of the mothers
and found it very deficient. The results of Ferguson (1115) were
about the same as the above. A number of out patient cases of
rickets were treated by Mackay (1115a): Small doses of cod liver
oil, butter or cottonseed oil had no influence, but larger doses of
cod liver oil were effective. According to Aron (1115b) carrot
extract was of no value in rickets.

322

THE VITAMINES

In 1912 Raczynski (1115c) formulated a very interesting theory,
quite in accord with the domestication theory of Kassowitz. He
thought rickets developed because of a lack of sunlight, a view which
seems to have been completely confirmed by the newest experimental
investigations. Huldschinsky (1115d) was able to cure rickets by
means of alpine light, since the X-ray showed a marked deposition
of calcium. Mengert (1115e) used the quartz lamp for prophylactic
purposes and Erlacher (1115f) succeeded by this means, in favor-

k»3*

••^^^ ^(P f. .

FIG. 63. X-RAY DIAGNOSIS OF RICKETS AND THE EFFECT OF COD LIVEII OIL
THERAPY SEEN BY THIS METHOD

Left, May 22, 1920; right, October 31, 1920 (Hess)

ably influencing 46 cases of rickets, without any change in diet.
Hess and linger (1115g) used sunlight for the same purpose, with-
out any dietary changes, and they (1115h) attributed the seasonal
variations in the incidence of rickets to the effect of sunlight. This
explanation, among other factors, sounds quite plausible.

In order to follow the course of the therapeutic measures, it is not
strictly necessary to carry out metabolism experiments. It may be
studied by means of the X-ray, a method utilized by a number of

RICKETS 323

investigators, among whom should be mentioned Phemister (I.e.
1112), Rowland and Park (1116), and Dufour (1116a) and Hess.
The results of cod liver oil therapy are apparent in animals some-
times after only two days; in children, after about three weeks, when
the deposition of calcium may actually be noted. This method is
coming into general use, and will very likely become of great prac-
tical importance.

ETIOLOGY

Although we accept the vitamine etiology in rickets, we are well
aware that still other views for the explanation of the nature of this
disease have been advanced. One of these hypotheses, the causal
relationship with the endocrine glands, we have already discussed in
detail in our first edition, and will, therefore, not go into it again here.
Since the function of the vitamines, especially vitamine A, is entirely
unknown, the vitamins hypothesis in rickets does not exclude the
possible role of the glands in the development of rickets. The
study of the etiology of rickets has been in a state of change during
the past year. According to the results of modern experimental
rickets investigations, this disease seems to' be associated with a
number of factors a fact which might assign to rickets a special
place in pathology. Three such factors have been put in the fore-
ground— lack of vitamine A, lack of phosphorus and lack of sun-
light. Each of these factors alone is able to cure the disease, which
develops as well in rats and dogs as in man, and has a favorable
influence on the calcium metabolism. Similar results have already
been obtained with children. Nevertheless, it is not yet clear as to
just how these three factors are related to each other. They give,
at all events, the same pathological picture. The "domestication,
theory" of v. Hansemann (1117) and also of Kassowitz (1118) has
recently been accentuated by a number of English investigators —
Findlay (1119), Paton, Findlay and Watson (1120) and Ferguson
(I.e. 1115), and by the modern sunlight hypothesis. Dick (1122)
believes that rickets is a disease of great industrial centers, and that
it does not occur in the tropics, Japan or China. However, this
seems to have little foundation in fact. Although we do not wish
to state, at this time, that a poor hygienic condition has nothing
whatever to do with the etiology of rickets, we believe, nevertheless,
that it can not be the only cause. For, as it most frequently hap-

324 THE VITAMINES

pens in practice, the unfavorable hygienic conditions are associated
with a deficient dietary. Mann (1123) investigated statistically
1000 cases of rickets in London, together with 250 controls. His
results speak in favor of the dietetic cause of rickets, inasmuch as
he paid particular attention to the mode of nutrition. A critical
survey of his cases convinced him that:

44 per cent of the cases suffered from a lack of fat and an excess of carbohy-
drates in the diet.

16 per cent were breast fed by a poorly nourished mother.

13 per cent occurred in families in straightened circumstances.

6 per cent exhibited a mis-proportion between the dietary constituents, in
favor of the carbohydrates.

Czerny (1123a) and Siegert (1123b) allot to heredity an im-
portant role in the etiology of rickets. That the cause for all rachitic
manifestations lies in the central nervous system, was assumed by
Pommer (I.e. 1064), and later by Schabad (1124), but this assumption
did not exclude the dietetic factor. Other dietetic theories are as
follows:

1. Lack of fat and excess of carbohydrates; Herter (1124a); Holt
(1125); Cheadle (1126).

2. Lack of a substance of an unknown nature in the diet; Hopkins
(I.e. 23).

3. Lack of calcium in the food; Stolzner (1127).

4. Excessively rich food; Esser (1128).

5. Lack of organic phosphorus combinations; Schaumann (I.e. 2).

6. Nutritional disturbances of every description, acidosis and
excessive nourishment; Pritchard (1129).

7. Lack of antirachitic vitamine; Casimir Funk; Mellanby.

The vitamine etiology of rickets

This theory was first suggested by us and was attributed to the
lack of a specific antirachitic vitamine, in relation to which, the pres-
ence of such a vitamine in cod liver oil has been assumed. These
conceptions were first tested experimentally by Edward Mellanby (I.e.
95). In the discussion as to the vitamine requirements of dogs, we
have already touched upon his work. He divided the foodstuffs
used by him into two groups — one, protective against rickets, the
other, not.

RICKETS 325

Not protective Protective

Bread Whole Milk (500 cc. per day)

Oatmeal Cod liver oil

Rice Butter

Skim-milk Lard

Yeast (10-20 grams per day) Olive oil

Orange juice (5 cc. per day) Arachis oil

Linseed oil Suet

Babassu oil Cottonseed oil

Hydrogenated fat Meat

Ca2 (P O4) 3 Meat extracts
NaCl
Milk protein

At the time of Mellanby's first publication, the view was largely
held that vegetable oils and fats contain no vitamine A and con-
sequently the above table was not so significant as is now the case.
In spite of some differences, hard to explain, Mellanby is of the
opinion that vitamine A is identical with the antirachitic vitamine.
According to his findings, the requirements for this vitamine are
particularly large when growth is most rapid. In a later communica-
tion, Mellanby (1130) stated that the requirement for vitamine A
depends very much upon the composition of the diet. The high
protein content of a diet stimulates the total metabolism; the children
are active and, therefore, require less of this vitamine. On a carbo-
hydrate diet, the metabolism is sluggish and the organism requires
more of this vitamine. However, it seems more logical to regard the
action of protein as "vitamine sparing," judging by our personal
experience. Besides the composition of the diet, age also plays a big
part, according to Mellanby (1131). Rickets is produced in older
dogs only with great difficulty, and spontaneous cures often occur.
That this is not quite in accord with human pathology is evident in
rachitis tarda and osteomalacia. Therefore, the chapter on the
etiology of rickets can not, as yet, be regarded as closed. In spite of
the fruitful investigations of Mellanby, several points are not yet
clear, for example, the prophylactic influence of meat extracts. It is
not impossible that Mellanby's experiments have been rendered
somewhat obscure through the non-observance of the light factor
which will have to be considered in all future research on this subject.

Recently, Noel Paton and Watson (1131a) have subjected the
researches of Mellanby on young dogs to experimental testing and

326 THE VITAMINES

concluded that rickets in dogs is not dependent upbn the lack of
antirachitic vitamine, but upon the energy content of the diet,
exercise and hygiene. However, they admit that dogs on skim-
milk develop rickets more readily.

Hess and Unger (I.e. 96), in particular, have subjected Mellanby's
results to a sharp scrutiny. .They maintained five children, 5 to
12 months old, on a diet composed of 180 grams Krystalak (dried
skim-milk with 0.2 per cent fat), 30 grams cane sugar, 15 to 30 grams
aufcolyzed yeast, 15 cc. orange juice and 30 grams cottonseed oil
(cereals for older children). Some of these children were kept for
several months on this diet, and as they grew older the amount of
cereal was increased, no other change being made. At the time of
publication, the diet was thought to be completely free of vitamine
A. These remarkable experiments showed for the first time that
vitamine A as it occurs in milk plays only a minor part in the develop-
ment of rickets. This finding, which contradicts somewhat the
results obtained by Mellanby in his experiments on dogs, was not
generally accepted. Hopkins (I.e. 595) held it against Hess' experi-
ments that the above diet was not entirely free from vitamine A,
since skim-milk may still contain this substance; cottonseed-oil too
is not free from this vitamine. Although these objections are
partially justified, we can not doubt that the dietary used by Hess
and Unger was indeed very poor in vitamine A. The favorable
result obtained with this diet, extremely poor in vitamine A, suggested
that the diet per se is not the only causative factor in the etiology of
rickets, and that there are still other factors to be taken into consider-
ation. Should they be so interpreted, that rickets has nothing to do
with vitamine A, as Hess and Unger naturally concluded at first?
On the contrary, these important experiments show that under
certain conditions, the vitamine A requirements of children may be
very slight, namely, when the composition of the diet as regards
protein, salts, vitamines B and C leaves nothing to be desired. The
individual differences in children on the same diet may perhaps be
attributed to unequal utilization of dietary constituents, a point
which has not been allowed for in the experiments of Hess and Unger,
or to the influence of light, which was entirely disregarded in
all their earlier experiments. In addition Hess and Unger reported,
that they observed rickets in children fed on milk (600 to 700 cc.
daily) containig 2.5 to 3 per cent fat, and also in breast-fed

RICKETS 327

children, whose mothers apparently had sufficient vitamine A in the
diet. All of these cases were cured with cod-liver oil. Holt,
Courtney and Fales (1132) kept a child for 5 weeks on vegetable
fat, as the sole fat constituent (apparently free from vitamine
A). The child stopped growing but remained in good general
condition. Two children developed styes and two others, facial
eczema, which disappeared when milk fat was given, v. Groer
(1133) conducted an experiment with two children, who grew almost
normally during the first half of the first year on a fat-free diet.
The diet consisted of skim-milk, containing only 0.01 per cent fat,
and cane sugar; later, oatmeal was added. Here we see again the
demonstration that under certain conditions children may thrive
with very little A-vitamine.

Hess (I.e. 636), on the other hand, could observe rickets in about
75 per cent of the children on a diet composed of "Eiweiss" milk
supplemented with cream, dextrin, malt-soup and orange juice. He
admitted, however, that this diet was also poor in vitamine B.

Among the opponents of the vitamine theory cf rickets (lack of
vitamine A), we find at first McCollum, Simmonds and Parsons
(1134), on the basis of their rat experiments, and also Robb (1135).
The latter concluded, from guinea pig experiments, that vitamine C
affects the mineral metabolism, so that the assumption of the existence
of a special antirachitic vitamine does not appear to be necessary.
Later, McCollum and co-workers (1135a), Sherman and Pappenheimer
(I.e. 1112b and c) and Hess, McCann and Pappenheimer (1135b)
conducted some preliminary experiments with rats on diets deficient
in vitamine A and in other respects, and produced a rickets-like
condition suggestive of a causative relationship of this condition to
rickets in man. In 1915, we noted in rats a disease similar to
rickets; a scoliosis-like curvature of the spine with rosary develops
sometimes in these animals on diets very rich in fat and containing
also cod liver oil (I.e. 331 and p. 339). McCollum and his co-workers
fed rats on artificial food mixtures and found that a diet poor in
vitamine A and phosphorus, but rich in calcium, led to rickets.
A simultaneous addition of cod liver oil (but not butter) made the
diet satisfactory for the animals. They found that cod liver oil
specifically influenced the bone development; the deposition of
calcium in the bones could be readily seen in the X-ray in from two
to eight days.

328 THE VITAMINES

Hess, McCann and Pappenheimer (I.e. 1135b) maintain that a
lack of vitamine A is not followed by rickets in rats. This is con-
trary to the findings of Mellanby in dogs, and shows perhaps that
butter is very poor in vitamine A [in agreement with the observa-
tions of Zilva and Miura (I.e. 574a)] and that dogs and rats show
great quantitative differences with respect to their vitamine A
requirements necessary to influence bone growth. In this con-
nection, the vitamine A of butter is less resistant than cod liver oil
towards oxidation. In confirmation of this, there is the work of
Turk (1135c) that melted butter exposed to oxidation does indeed
increase the immunity of children towards infections but does not.
protect against rickets.

In a recent publication, McCollum, Simmonds, Shipley and Park
(1135d) have attributed to cod liver oil a specific role in rickets.
The vitamine contained therein doesn't seem to be identical in its
influence with vitamine A, but perhaps it is a question of quantita-
tive relationships. On the addition of cod liver oil the calcium and
phosphorus requirements of the animals were markedly reduced.
A similar result was obtained on the administration of phosphorus.

On a critical consideration of the vitamine theory of rickets it is
apparent that on a certain diet which must be regarded as leading to
rickets, not all children develop the disease. On the other hand, a
large percentage of the cases may be cured by cod liver oil, but not all
cases. Individual differences arise here which can not be explained
at present, though we firmly believe that we are on the right track.
Recently, of the French investigators, Nathan (1136) has adopted
our view.

Undoubtedly, rickets may be brought into causal relationship with
vitamine A, although the above-mentioned factors are also of signifi-
cance. In accord with this, there is the fact that the manifestations
of rickets in children, in certain periods of the year, are more marked
than in others. Sometimes, as Dr. Hess has told us, spontaneous
cures occur in spring. One might be tempted to associate this with
seasonal variations in the vitamine content of milk, as has already
been partially demonstrated. The influence of light is also to be
considered in this respect. Hess and Unger believe in the specific
action of cod liver oil, but not of milk fat. We cannot agree with
this since it would mean that normal milk contains no substance
protective against rickets, and, therefore, most milk-fed children

RICKETS 329

would suffer from this disease. If we accept the idea, however,
that cod liver oil contains the specific substance, then we admit that
the diet, which brings about rickets, must be lacking in this sub-
stance. But since butter and cod liver oil play the same part in
rat feeding, we have to recognize that rickets is caused by a lack
of vitamine A. We must accept this logical relationship, if we wish
to proceed to the experimental tests of the above conception. In the
meantime, it seems to us that the study of rickets is more likely
to lead to the desired end if dogs are used in preference to rats, for
in the latter only osteoporosis frequently develops. The develop-
ment of this disease in young dogs shows an appreciable similarity
to rickets in small children. With the elucidation of the influence
of light, phosphorus and particularly vitamine A, we may hope for a
solution of the problem of rickets.

OSTEOMALACIA

Late rickets, which we have already mentioned, forms the natural
stepping-stone between rickets in small children and osteomalacia.
The latter seems to be due to an identical metabolic disturbance.
Some demonstration for the genetic relationship is provided by Ogata
(1 137) . In the province of Toyama in Japan, osteomalacia developed
in adults, under conditions apparently similar to those giving rise
to rickets in children. Before the war, the disease developed almost
exclusively in young pregnant women, in which condition it often
assumed a serious aspect. This form of the disease was described by
Liesegang (1138) and also by Scipiades (1139), and was mostly
associated etiologically with the endocrine glands. A relationship
to the pregnancy was often suspected although this condition
undoubtedly was only a secondary factor. It is, however, not
impossible that an interruption of the pregnancy might be accom-
panied by an improvement Koltonski (l!39a). Januszewska (1140)
reported on 3500 cases in Bosnia on an inadequate diet, accompanied
by rheumatic manifestations. The cases appeared between Decem-
ber and April and showed a periodicity similar to that already noted
in rickets, and which was associated with improvement in the diet.
The disease was favorably influenced by cod liver oil.

During the war, central Europe afforded thousands of such cases.
This has been observed in Vienna, especially during the summer of

330 THE VITAMINES

1919 (1141). The frequency of the cases was remarked upon by
Partsch (1142) and by Sauer (1143). The disease did not occur only
in the adolescent period (12 to 20 years), according to Hamel (1144)
and Heyer (1145), but also in men, according to Kopchen (1146)
and in old age, among men and women, according to Curschmann
(1147).

Symptoms

Chelmonski (1147a) describes two forms of alimentary bone
disease which prevailed in Poland in 1917-1918; one form was
accompanied by pain in the joints and brittleness of the bones;
the other, more like the classical osteomalacia, but in the absence
of pregnancy, with severe changes in the pelvis and progressive
paralysis of the lower extremities. As in rickets, the bone changes
are most prominent; without this, the diagnosis is not easy and the
disease is often confused with rheumatism. In women, according to
Croftan (1148), fractures appear chiefly in the pelvic region. The
condition was described by Looser (1149) in young people of both
sexes (17 to 20 years of age). The symptoms consist in an enlarge-
ment of the thyroid, fatigue on walking, pains in the knees and
ankles. Pain in the bones is one of the earliest symptoms. Alwens
(1150) observed 26 cases among women between 19 and 72 years of
age, 65 per cent of which occurred during and after the climacteric.
The weight of the patients in most cases was about 100 pounds;
15 cases showed curvature of the spinal column, and in 10 cases,
spontaneous fractures were noted. Bohme (1151) emphasized, in
particular, the complete analogy between rickets and osteomalacia.
Imhof (1152) believed that osteomalacia is frequently accompanied
by psychoses (dementia praecox). Gould (1153) believed in a
complete analogy between osteomalacia and Recklinghausen's
disease and noted that besides an affection of the peripheral nerves
in the latter, still other factors play a role ; in his opinion, the observed
bone lesions point to this. Cases of Recklinghausen's disease were
described also by Weiss (1153a) and Comby (1153b).

Anatomy

Pathological findings were described by Hanau (1154) and by Wild
(1155). The latter observed changes in the bones, similar to those
in rickets, namely uncalcified osteoid tissue.

RICKETS 331

Metabolism

Scott (1156) studied the calcium metabolism in the puerperal
type of osteomalacia. The amount of calcium in the urine was
increased, while during pregnancy and lactation, the output was
decreased. The figures were as follows: normal CaCl2 per liter of
urine — 1.146 grams; in osteomalacia — 1.36 grams; in pregnancy
and lactation — 0.988 gram. The calcium content of the blood is
increased in osteomalacia; in pregnancy, no variation from the
normal figure was noted; normal — 0.45 gram CaO per liter; osteo-
malacia— 0.58 gram. Leo Zunz (1157) conducted metabolism
experiments in puerperal osteomalacia. He found a negative calcium
balance, with a definite improvement on castration. Elfer and
Kappel (1157a) conducted metabolism experiments with and with-
out the addition of extracts of endocrine glands. The calcium
and magnesium balances were sometimes negative, while the phos-
phorus balance remained positive.

Therapy

The cod liver oil therapy has not been used very much, though
where it has been used, the reports were favorable. Looser, Cursch-
mann, Kopchen, Hamel and many others, have reported on this
subject. In most cases, a change in diet was instituted, together
with the therapy.

Etiology

Hutchinson and Patel (1157b) observed numerous cases among
Mohammedan women in Bombay, and attributed the cause of the
disease not to the diet, early marriage or prolonged lactation, but
to the inertia and lack of air — in accord with the domestication
theory of rickets. The description of cases among men, in middle
and old age, shows that the disease, as such, has nothing to do with
the puerperium. The cases in old age, as well as in middle age, show
that this rachitic-like condition has nothing in common with growth,
contrary to the findings of Mellanby on rickets in young dogs.
Recently, the prevalence of this disease in central Europe was
coincident with a lack of milk, eggs, cheese and butter. The popu-
lation lived on cabbage, turnips, some potatoes and meat. There
was, therefore, a lack of vitamine A, as in rickets.

CHAPTER IV

SOME NUTRITIONAL DISTURBANCES IN CHILDREN — TETANY, CARBO-
HYDRATE DYSTROPHY (MEHLNAHRSCHADEN), ATROPHY

In the description of these conditions, we must state, first of all,
that under this heading, besides conditions that really are avitam-
inoses, others have been included which possess, no doubt, a different
etiology. Unfortunately, at the present status of our knowledge,
this can not be avoided.

TETANY (SPASMOPHILIA)

Taking this disease as an example, the clinical picture of spasmo-
philia is not absolutely denned, and Luzzatti (1158) stated correctly
that it is difficult to differentiate between tetany and convulsions,
especially because of the atypical course of infantile epilepsy.
According to Stheehman and Arntzenius (1159), a poverty of
calcium may be followed by clinical consequences of various kinds.
The diagnosis of spasmophilia may be simplified by testing the
electrical irritability, but this test is not absolutely certain. It is
now generally assumed that the tetanic conditions in children are
brought about by the lack of calcium. That the calcium retention
is impaired in such cases, was demonstrated by Cybulski (1160).
Theoretically, however, it is possible that the calcium-impoverish-
ment of the body may take place in many ways. It may be asso-
ciated with a lack of calcium in the diet, a condition which may be
remedied by means of calcium therapy. The disease may be related
to an affection of the parathyroids, in which event an organo-therapy
would be in order. Finally, it may be associated with an impoverish-
ment of the organism in calcium salts, as a result of rachitic condi-
tions. Pepper (1161) correctly called attention to this possibility.
In practice, apparently all three causes may be accounted for.

Etiology

The lack of calcium as the sole cause of the disease must be quite
rare, since most of the available data do not report favorably on
calcium therapy. When calcium therapy is spoken of, as in the

332

NUTRITIONAL DISTURBANCES IN CHILDREN 333

cases of Rohmer and Vonderweidt (1161a) and Scherer (1161b),
this therapy was for the most part supplemented by cod liver oil
and phosphorus. Thiemich (1162) found the calcium therapy alone
without effect, while Petrone and Vitale (1163) found the parathyroid
therapy ineffective in their cases, though Pincherle and Maggesi
(1163a) reported that their tetany cases showed pathological
changes in the endocrine glands. The third possibility, the calcium
impoverishment as result of rachitic metabolic disease, is strengthened
by the frequent association of this disease with rickets and osteo-
malacia. Thus, Januszewska (I.e. 1140) found 338 cases of tetany
among 3500 cases of osteomalacia. Sauer (I.e. 1143), too, described
this relationship and was strengthened in his view by a favorable
cod liver oil therapy. Takasu (1164) associated spasmophilia with
beriberi. In his cases, there was a possibility of infantile beri-
beri. Since the rachitic disturbance may occur at any age, the
related tetany may appear similarly. Klose (1165) believed that
this disease can not occur under the age of 2 months. Bliihdorn
(1166) on the contrary, found it in children of all ages. Apart from
this, it is not totally excluded that certain forms of tetany of preg-
nancy and lactation are to be attributed to the above causes, although,
this was denied by Faas (1167). Tetany seems to be favorably
affected by a one-sided flour diet, and occurs rarely in breast-fed
infants, according to Guthrie (1168). Lust (1169) stated that tetany
occurs only in 2 per cent of breast-fed and in 55.7 per cent of artifi-
cially-fed children. According to Bossert and Gralka (1169a) true
spasmophilia may be recognized by the fact that it can be influenced
by a change in the diet. v. Meysenbug (1170) did not believe that
spasmophilia had anything to do with the three known vitamines,
still we cannot sae that his results justify such far-reaching conclusions.
Jeppson and af Klercker (1170a) believe that tetany is not due to
the lack of calcium in the diet but to an excess of phosphorus.

Symptoms

The Erb sign (increase of galvanic irritability of the muscles and
the motor nerves) and the Chvostek sign [increase of the irritability
of the nerves especially the facial nerve or the popliteus (Ibrahim,
1170b) to mechanical stimulus] are of value in the diagnosis of
tetany. According to Stheehman and Arntzenius (I.e. 1159), the
Chvostek sign is more sensitive. Bossert (1171) occasionally ob-

334 THE VITAMINES

served edema and carpopedal spasms. The determination of the
calcium content of the blood is of help in the diagnosis. Rowland
and Marriott (1172) found, in normal cases, 10 to 11 mgm. calcium
in the blood; in rickets, somewhat less; in tetany an average of
5.6 mgm. and sometimes only 3.5 mgm. They found that the calcium
therapy, even though gradual, is effective. Eliasand Spiegel (1172a)
found the phosphorus content of the blood very high, while Tisdall,
Kramer and Howland (1172b) found that the ratio of sodium and
potassium to calcium and magnesium is very greatly increased.
Stheehman and Arntzenius (1173) found, on the other hand, that
the calcium content of the blood is no definite indication of tetany.
The calcium in the blood may be high and yet the Erb sign is posi-
tive, which may be associated with a lack of calcium in the tissues,
v. Meysenbug (1173a) states that the Thiemich sign is not associated
with the low calcium content of the blood and that larger doses of
calcium in older children do not affect the anodic reaction. The
determination of salt metabolism in tetany was made by Fletcher
(1174). Liefmann (1175) found the urinary acetone elimination
very high in the acute stage, while Sharpe (1176) found an increased
guanidine elimination in the feces. Hoobler (1177) stated that the
Ca- Mg- and P balances were all negative. Bolten (1177a) found
a decreased blood calcium content with an increase in the electrical
sensitivity in Fragilitas ossium (Osteopsathyrosis).

Therapy

Calcium therapy was described by Howland and Marriott (I.e.
1101). Thiemich (I.e. 1162) recommended breast feeding and
phospho-cod liver oil. Brown and Fletcher (1178) also recom-
mended phospho-cod liver oil. Stheehman (1179) and Brown,
MacLachlan and Simpson (1180) have corroborated the cod liver
oil therapy. The last named workers found the intravenous injec-
tion of calcium lactate without effect if not accompanied by a simul-
taneous phospho-cod liver oil therapy. The clinical manifestations
of tetany disappear in 10 to 17 days while, without the addition of
calcium, the action is very much delayed. Unfortunately, we have
found no data on cod liver oil therapy alone. Recently, Huldschin-
sky (1180a) has used in tetany the sun-ray therapy with which he
was successful in the treatment of rickets. It appears that the sub-
ject requires still further experimental study.

NUTRITIONAL DISTURBANCES IN CHILDREN 335

CARBOHYDRATE DYSTROPHY (MEHLNAHRSCHADEN)

It is evident that under this name conditions are classified, the
etiology of which is fundamentally different. In 1905-1910, par-
ticularly in Germany, but also in other countries, widespread diseases
of children appeared. These arose as the result of proprietary foods
fed over a long period of time. Czerny (1181) was first to recognize
the dangers of this regime, and called the resultant condition,
"Mehlnahrschaden." It need hardly be said that the disease occurred
only in artificially fed babies. Children who could tolerate cow's
milk'poorly or not at all, were fed for weeks, more often for months,
on various proprietary foods. Recently, Bloch (1181a) also warned
against the danger of a carbohydrate-rich diet in infants. At first,
this food seems to be well tolerated; the splendid appearance of the
child permits of no suspicion that the food in inadequate. Later,
however, after a lapse of time which varies within wide limits in
different children, a disease develops which, according to Rietschel
(1182), occurs in three different forms: pure atrophic (hunger),
atrophic-hydremic and hypertonic.

The pure atrophic form occurs in children fed on flour, with the
following symptoms. The stools react acid, with considerable slimy
substance and fermentation. At the same time, there develops a
severe atrophic hypertonicity of the musculature; the muscles are
tightly stretched, which simulates a healthy condition. Electrical
supersensitivity of the peripheral nerves is frequently noted. Here,
there is often a great susceptibility to secondary infections (thrush,
aphthae, abscesses of the skin, otitis, phlegmons, and pneumonia).
In spite of the severe symptoms, the gastro-intestinal tract, on
autopsy, is found almost intact.

The atrophic-hydremic form manifests itself, at the beginning, in
edema of the lower extremeties, which, according to Noeggerath
(1183) is due to heart weakness; later, the children show a blown-up
face, sometimes also scorbutic changes of the gums (Bogen, 1184).

The hypertonic form resembles peripheral nerve affections very
much. In this category, we find cases where the legs are drawn up
to the abdomen, with rigid spinal column, tetany with increased
electrical irritability and carpopedal spasms lasting for weeks. The
prognosis is bad and the children die with neuritis (Griineberg, 1185).

336 THE VITAMINES

Etiology^

Langstein (1186) believed in the analogy of this disease with
beriberi. In one case, he saw such a great hypertonus of the muscles
that he thought it might be a case of Little's disease. Some supposed
cases of carbohydrate dystrophy may really have been beriberi.
According to Hess (I.e. 918), who noted the development of the
disease in the presence of sufficient vitamine C, an analogy with
scurvy is excluded. On the other hand, according to some recent
work, it seems to be similar to hunger edema. L. F. Meyer (1187)
and Rietschel (1188) believed this to be true. Following * this,
Benjamin (1189) reported that in carbohydrate dystrophy, after
feeding "Eiweiss" milk there was high nitrogen retention, pointing
perhaps to a craving of the organism for protein. At first, the reten-
tion amounted to 50 per cent of the intake; later, 15 to 20 per cent.
Since the cause of hunger edema has often been regarded as lack of
protein, this statement is of interest. However, since a di«t com-
posed chiefly of flour may be very poor in vitamines, under certain
conditions, the possibility of dealing with a vitamine deficiency can
not be excluded. Hess (I.e. 918) also came to this conclusion. At
any rate, Keller (1190) is right when he says that an almost exclusive
carbohydrate diet may be used only intermittently. However, there
seem to be cases in the literature, as that of Steinitz and Weigert
(1191), in which, as a result of a poor tolerance for milk, an exclusive
carbohydrate diet was given for a long time. Salge (1192) believed
that in individual children, there are great differences in the tolerance
for starch. He saw a child fed for 2 months on flour exclusively
without any untoward results. The pathological conditions arising
from a diet rich in starch were studied also by Abt (1193).

ATROPHY

In the English pediatric literature the term "Mehlnahrschaden" is
rarely encountered; disturbances of this kind are generally classified
as "atrophy" (Cautley, 1194). The latter described the symptoms
as follows: The disease arises as a result of an inadequate diet,
especially one abundant in flour. There is considerable loss of
weight, diarrhea and vomiting. These symptoms are frequently
accompanied by stomatitis aphthosa, enteritis, bronchitis and furun-
culosis. In severe cases, he found edema of the skin of the extrem-

NUTRITIONAL DISTURBANCES IN CHILDREN 337

ities, later of the face, petechiae and opisthotonos. On post-mortem,
edema of the brain membranes, pleural ecchymoses, erosions in the
stomach and intestine, and subepithelial hemorrhages were found.
In chronic cases, there was fatty degeneration of the liver and kidney
epithelium.

Frank and Stolte (1195) analyzed the organs of children who had
died of carbohydrate dystrophy, special attention being given to
the liver. This was found to be higher in sodium and chlorine and
lower in potassium, while fat, ash, magnesium, calcium, phosphorus
and sulphur showed no great deviations from the normal. Steinitz
and Weigert (I.e. 1191) noticed, in one case, a high fat content of the
organs, and more PaOs.

Noeggerath (I.e. 1183) and Hohlfeld (1196) recommended the
administration of raw milk. Bendix (1197) recommended breast
feeding for atrophy.

OTHER NUTRITIVE DISTURBANCES IN CHILDREN

In many well known nutritional disorders of infants, the present
pediatric literature gives no clear picture of the etiology. Above
all, the individual pictures are not sharply differentiated. Morse
and Talbot (1198) have remarked upon the chaotic condition in the
present conception of atrophy. It is therefore all the more difficult
for one unacquainted with this subject to have a definite conception
as regards the nature of this condition. Of the nutritional disorders
to be treated here, there is exudative diathesis, atrophy (in the sense
of the German pediatric literature), marasmus, milk dystrophy, etc.
Many of these conditions are now thought to be due to excess of fat.
In children practice, cases are known in which every possible dietary
composition has been tested, but without result; there are even
children (though seldom) who can no longer tolerate a completely
normal breast diet. How far this may be due to organic disturb-
ances, or to a previous unsuitable diet, is not always certain. In
some cases, the deficient diet might have caused such great harm,
that the child can no longer be helped, even by the best of diets.
Marriott (1198a) differentiated two forms of nutritive disturbances
in infants, one form arising after severe diarrhoea and another
manifested as chronic malnutrition (marasmus).

For some of the above mentioned nutritional disturbances, Pehu
(1199) introduced, in France, vegetable soups as a therapeutic

338 THE VITAMINES

measure. This therapy was used in Germany by Moro (1200), in
the form of carrot soup, with good results. A similar therapy was
recommended by Stark (1201) in suspected avitaminoses, and
recently, there has been a similar report by Aron and Samelson (1202).
McClendon and Sedgwick (1202a) used spinach powder in infant
nutrition and obtained satisfactory increases in weight. Aron
(1202b) made use of the soluble flour obtained by the action of
ferments and found that such products were well utilized in the
more severe cases.

From time to time, there appear in the literature statements as to
a successful vitamine therapy in certain disturbances. Kohlbrugge
(1203) successfully treated cholera infantum, a disease generally
regarded as an infection or intoxication, with orange juice. Holt,
Courtney and Fales (1204) treated a case of intestinal infantilismus
with cod liver oil, with good results.

It need hardly be said that in the feeding of children, the same
rules are to be observed as have already been formulated for animals.
Besides the already known dietary constituents, we must provide for
the presence of the three known vitamines. It is also essential that
the more important constituents should not occur in the diet in too
great a dilution. Some of the recorded therapeutic results with
vitamines depend upon the fact that either the certain vitamines,
although they were present, were not sufficient, or the relative propor-
tion of the individual food constituents to each other was not quite
correctly chosen. Weill and Mouriquand (1205) have pointed out
such a possibility; as regards vitamine B in this connection, we have
the work of Daniels, Byfield and Loughlin (1206). We may explain,
in this way, certain results of vitamine therapy in an insufficiently
defined condition, usually called "marasmus" or "malnutrition."
Eddy and Roper (1207), and Dubin and Lewi (I.e. 628) obtained
marked results in the conditions just mentioned.

As regards atrophy, it is caused by excess of fat, according to the
present conception. The results of Friedberg and Noeggerath (1208)
are to be regarded in this light; they recommended the use of centri-
fuged breast milk in this condition. This recalls certain investiga-
tions on rats, made by Funk and Dubin (I.e. 331). These animals
received a complete synthetic diet, containing more than 50 per
cent fat. The young rats were able to live on this diet for 2 to 3
months. In spite of the vitamine content of the diet, the animals

NUTRITIONAL DISTURBANCES IN CHILDREN 339

did not grow; they lost about 50 per cent of their weight and their
condition was such as is mostly described in atrophic children. The
animals showed a fatty degeneration of the liver; in one case, there
was edema and scoliosis, with a permanent curvature of the spine.
A physiological explanation of this appearance is still to be found.
Should it appear, however, that the milk dystrophy and the decom-
position of Finkelstein are attributable to the high fat content of the
diet, then the influence of fat may be studied experimentally in the
above manner.

In infantile atrophy, Salge (1209) found the assimilative capacity
satisfactory. The energy and metabolism of these children was
studied by Bahrdt and Edelstein (1210). The assimilation of fat
was found normal by Hutchinson (I.e. 1096) ; fat losses are, however,
quite high as a result of voluminous stools. The protein content of
the blood was found low by Utheim (1211) and he believed that the
protein synthesis proceeds abnormally; in other cases, on the con-
trary, there may have been an excessive carbohydrate nutrition.
Mattill, Mayer and Sauer (1212) state that the glucose tolerance of
atrophic children is higher than normally. Gladstone (1213) recom-
mended a fruit-juice therapy. Cure with breast feeding, and in
one case with cow's milk, was described by Marfan (1213a).

The exudative diathesis of Czerny (1214) is not associated with
lack of vitamines, according to Hess (I.e. 918). The condition,
however, leads frequently to scurvy, eczema and petechial hemor-
rhages. According to Czerny, this disease arises because of the
harmful effect of the dietary fat. Schippers (1215) found, in these
conditions, a low fat content in the blood. Marfan (1215a) regards
athrepsia as a deficiency disease but not as an avitaminosis, and he
accepts as the cause a lack of specific enzymes in breast milk. Be-
cause of the present better understanding of the nutritive require-
ments of infants, a fundamental repetition of these nutritional
problems is greatly to be desired.

CHAPTER V

NUTRITION IN MAN — AN INTRODUCTION TO THE STUDY OF PELLAGRA
AND HUNGER EDEMA

From what has been brought out till now, it is clear that in nutri-
tion, it is not sufficient that all the known dietary constituents be
present; they must also bear the proper relationship to each other.
This idea developed chiefly through observations on rats. If these
animals are given a qualitatively correct diet (ad libitum), dietary
deficiencies develop nevertheless. It would be tempting to assume
that these animals need only ingest more food in order to receive
all of the constituents in correct amounts. In practice, however, it
is evidently not the case. No matter how much food the animal eats,
the relationship of the individual components is not altered. This
excessive dilution of certain components may appear in human nutri-
tion, and yield a rather unclear etiological picture.

Pellagra, sprue, hunger edema, and perhaps carbohydrate dys-
trophy in children, may be associated with the above condition. At
least two of the diseases mentioned have recently been attributed to
the lack of a protein of high biological value. A number of animal
proteins are generally regarded as such. Should this conception
prove to be correct, then these pathological conditions must be
stricken from the list of avitaminoses, which has in fact already
been done by some investigators. We feel, however, that these
diseases have been cleared up so little, etiologically, that this pro-
cedure seems to us altogether unjustified at present. In the last
war, nutrition played a greater role than has been generally assumed
by the laity. At the time of the war, we saw one immense metabolic
experiment, from which a great number of conclusions could have
been drawn, if the serious situation had not prohibited a cold-blooded
survey. The late pharmacologist, Meltzer, remarked at one time
that all scientific data obtained during the war would have to be
corroborated after the war. He may have been correct in this state-
ment, though the convincing power of the large involuntary nutri-
tional experiment remains unquestioned, and we may draw a number
of conclusions for our purpose. Various problems must be taken

into account in this connection, namely:

340

NUTRITION IN MAN 341

1. The question of the protein minimum.

2. Comparison of the nutritive value of vegetable and animal
protein.

3. The chemical groups in protein responsible for its high biological
value.

4. Influence of war food on the health of the people.
Unfortunately, because of lack of space, we can not enter into a

detailed discussion of the above questions. Protein has always been
given an exceptional place in the system of nutrition and correctly
so. Recently, this has been brought out in the work of Gigon (1216)
and Grumme (1217). It is quite important to know the physiological
protein minimum, and in this respect, there are the views held by
two schools — that of Voit-Rubner, which requires a high protein
minimum, and that of Chittenden-Hindhede, which is satisfied with
a considerably smaller figure. The figures of Chittenden were
appreciably reduced by Hindhede (l.c» 732), who showed (1218) in a
great number of extended nutrition experiments, that adults could
maintain themselves and work normally on a daily quota of 20 grams
of assimilated protein and 3900 calories. The investigations were
continued for a longer time than usual for experimental purposes
and therefore are very convincing. In one instance, the experiment
lasted 305 days and the diet consisted of potatoes (cf. p. 255), mar-
garine, and sometimes onions and fruits. Hindhede believed that
when the diet contains sufficient calories, one does not need to worry
about the protein content, taking care only that the diet should
contain sufficient vitamines. The bread experiments of Hindhede
(1219) fall in the same category. Of the bread, made from whole
wheat, 30 to 35 grams assimilated protein were necessary. The rest
of the diet consisted of margarine and fruits, and the longest experi-
ment lasted 242 days. One experiment with coarsely smashed wheat
(I.e. 791) lasted 282 days. In some of these investigations it is even
noteworthy that the persons submitting to the experiment did not
develop scurvy, though it is possible that fruits were used. The
minimum protein figure was very low in experiments with barley-
water gruel (1220), with the addition of margarine and sugar.
Hindhede (1221) also studied the fat minimum in nutrition which is
•likewise of importance in the determination of the value of war diets.
In this case, the experimental diet was composed of potatoes, cabbage,
rhubarb and apples, but without addition of fat. The experimental

342 THE VITAMINES

subjects were in splendid health for 16 months, so that a lack of
animal protein and fat can not be regarded as the cause of pellagra
and war edema, since they were on this diet much longer than is
required for the development of this disease. Bang (1222) believes,
however, that Hindhede's experimental subjects are exceptions and
hence his results can not be used without further confirmation,
especially as regards children and consumptives.

Hindhede assumed that a gram of potato or bread protein when
assimilated, corresponds exactly to a gram of meat or milk protein,
but this conception has failed to gain recognition up to the present.
In a new series of rat experiments, McCollum, Simmonds and
Parsons (1222a) observed that the internal organs, especially the
kidney, can supplement effectively the diet of plant origin. Neu-
mann (1223) believes that it is possible to live on an exclusive bread
diet. Dollner (1224), however, in the description of the etiology of
hunger edema, called it "potato sickness." He saw this condition
develop on large amounts of potatoes and cereals. Kraszewski
(1225) believes that potatoes are poor in calcium and magnesium
and hence must be supplemented with bread and vegetables. Un-
fortunately the individual investigators are not agreed as to the
protein minimum. Jansen (1226) figured that for a man weighing
62 kilos, 2000 calories and 60.5 grams protein are necessary; but the
diets used by Hindhede were much richer in calories. Sherman
(1227) calculated that for an adult weighing 70 kilos, 44.4 grams
protein per day are necessary. Taylor (1228), in his studies on the
nutrition of English prisoners in Germany, concluded that 70 to 90
grams protein must be fed. Benoit (1229) investigated in Germany,
the metabolism of a number of prisoners (Russian officers) and found
that they maintained themselves without complaint for 16 months
on 48 grams protein and 1700 calories, the protein being partly
vegetable and partly animal. Botazzi (1230) was opposed to the
introduction into the Italian army, of Chittenden's figures. Kruse
and Hintze (1231) accepted three-quarters of Voit's food standards.
Praussnitz (1232) also discussed this matter. Stille (1233) supported
the views of Hindhede against those of Rubner. Hindhede recorded
the diet on which he and his family lived during the war, which
consisted of 54.6 grams protein and 2100 calories. Funk, Lyle and
McCaskey (1234) reported a 20 day metabolism experiment on four
men, receiving yeast, rice and white bread. A nitrogen balance was

NUTKITION IN MAN 343

almost obtained with 5 to 7 grams nitrogen. The somewhat unsatis-
factory results were perhaps due to a partial absence of vitamines
(especially during the periods without yeast), or to a relatively low
caloric content. This was one of the few metabolism experiments
where the effect of such a dietary mixture on the composition of the
blood was studied. As to the nutritive value of bakers' yeast,
Hawk, Smith and Holder (1235) reported favorably that yeast can
replace 30 per cent of the protein of the diet.

After this brief consideration of the protein minimum, we come to
the comparison between the nutritive value of vegetable and animal
protein. That there is a great difference in favor of the animal
protein, is now maintained by many, in opposition to the view held
by Hindhede. The present view of Hindhede was advocated many
years ago by Rutgers (1236). Boruttau (1237) showed in metabo-
lism experiments, that vegetable proteins are well utilized; he did
not believe that the biological value was an indication of the vitamine
content, for unpolished rice has a lower biological value than polished
rice. Botazzi (1238), on the contrary, believed in the special nutri-
tive value of animal protein and fat. Rose and Berg (1239) proposed
a view, approaching ours, in that they assumed that the protein
requirements depend on the nature of protein and the composition
of the rest of the diet. Milk protein has mostly been given an
exceptional place in nutrition. Recently, this was again reported
upon by Hart and Steenbock (1240), and we must, therefore, ask
ourselves the question, which chemical group in the protein molecule
plays such an extraordinary role in nutrition.

This problem has been treated experimentally in such great detail,
that a book may well be written on the subject. For this reason,
we must confine ourselves to the chief results. The biological value
of proteins has been determined by Thomas (I.e. 1) on different
products and yielded the following figures:

Beef 104 Rice 88 Peas 56

Cow's milk 100 Potatoes 79 Wheat flour 40

Fish 95 Casein 70 Cornflour 30

This table is based upon the conception of the similarity of animal
proteins with body proteins and is due to their relative content of
essential amino acids. A number of amino acids have been recog-
nized as such. Still, in the study of the nutritive requirements of
the lower organisms (yeast) it has been established that some proteins

344 THE VITAMINES

contain substances which, after hydrolysis, cannot be isolated
and which thus far remain unknown among the cleavage products.
McCollum and Davis (I.e. 97) believed that when casein is
autoclaved, its nutritive value decreases markedly; but, according to
Funk and Macallum (I.e. 98), this deficiency may be overcome by
the addition of vitamine-containing products. Yet, it is not impos-
sible that heating may decrease the nutritive value of some proteins.
Mueller (I.e. 167a), in this connection, made an interesting observa-
tion in the culture of streptococci, namely, that peptone, casein,
edestin, in the tryptophane fraction, contains a substance seemingly
indispensable for the growth of this microorganism. Mueller was of
the opinion, that he was dealing with a new amino acid, in which
belief he was strengthened by the fact that reprecipitated casein
possessed the same ability. Still, this condition does not militate
against the possibility that the substance may be of a vitamine nature,
for in the first place, the active substance is so markedly adsorbable
that it may be completely removed with animal charcoal; second, if
the substance is carried down in the first precipitation of the casein,
it would, in all probability, be carried down on reprecipitation. By
reprecipitation of the casein, therefore, no separation can be obtained.
We saw in the case of other bacteria that the vitamine-like substances
there could not be filtered, even through paper, without being
adsorbed. Mueller found that wool, silk, gelatin and hydrolyzed
yeast do not contain this substance ; it was found in the tryptophane
fraction; but when the known substances in this fraction were
tested for the above action, no result was obtained. If the fraction
is precipitated with phosphotungstic acid, the substance disap-
pears from the precipitate as well as from the filtrate. If the
fraction is crystallized, the substance remains in the filtrate and
appears unstable, de Souza and McCollum (I.e. 529) observed
that hydrolyzed meat exerts some action on the growth of yeast.
Funk and Dubin (unpublished work) noted that a completely
extracted meat (the last extract being no longer active for yeast),
when hydrolyzed, shows definite action. The question whether we
are dealing here with a vitamine or an unknown amino acid must be
left open for the time being, though it seems to us not impossible
that the above finding will give a new stimulus to protein research,
in that we have a biological reaction that may be of value in a frac-
tionation. In addition, the nutritive value of the proteins may
appear to us in a new light, for the experiments show that some

NUTRITION IN MAN

345

proteins, in a more or less impure condition (for the degree of purity
can naturally not be accurately determined), besides the known
cleavage products, still contain something that has been unknown
to us. It is very tempting to associate this substance with the
etiology of pellagra and similar conditions, though we must admit,
that the basis for it is at present hardly at hand. This assumption,
however, explains some of the established facts about pellagra very
well, as we shall presently see.

DIET*

MEAT

SUGAR

STARCH

LARD

SALTS

AUTO-
LTZED

TEA8T

ORANGE

A GAR

COD
LIVER
OIL

Meat

grams

49

grams

12

grams

12

grams

12

grams

3

CC.

4

CC.

3

grams

3

CC.

5

Sugar

12

49

12

12

3

4

3

3

5

Starch

12

12

49

12

3

4

3

3

5

Lard

12

12

12

49

3

4

3

3

5

DIET

WEIGHT — FIRST
25 DAYS

WEIGHT — FOLLOW-
ING 55 DAYS

TOTAL
WEIGHT

INCREASE!

Increase

Decrease

Increase

Decrease

Meat

pe • cent

43
15
15
6
6

per cent

9
9

per cent

86
55
145
62
165

34

per cent

8

per cent

129
70
160
68
171
-17
25

Sugar

Sugar and vitaminet

Starch

Starch and vitaminet

Lard

Lard and vitaminet

* The meat, sugar, starch, and lard were tested and found to be free from
vitamine B.

t In this instance, the figure represents the increase after 60 days, and is
practically the same after 80 days, since most of the animals had already
attained full size.

t Vitamine given during last 55 days.

The rats on the protein diet did not require the addition of extra vitamine
(autolyzed yeast) at all. This may be regarded as the "sparing action of
protein on the vitamine requirement." On the other hand, the rats on the
fat diet took the extra vitamine with great avidity, but showed only a small
advantage over the controls. The replacement of some of the fat by butter
was without any significance, no improvement being noted.

On the starch diet, the rats actually needed extra vitamine (about 2 cc.
per day) in order to resume growth. This was likewise true of the rats on
the sugar diet except that they appeared not to require as much vitamine for
growth as those on the starch diet.

346 THE VITAMINES

That the protein content of a food mixture is of importance, is
apparent from the older work of Osborne and Mendel, who deter-
mined the protein minima of various products on rats. The experi-
ments, however, were not carried out from the viewpoint of vitamines.
In the consideration of avian beriberi we have seen that an excess of
carbohydrates in the diet hastens the outbreak of the disease; an
excess of proteins, on the contrary, delays the onset. Funk and
Dubin (I.e. 331) conducted some experiments with rats, in which it
is easier to control the food intake. The addition of vitamine was
constant in all diets and the dietary composition varied only in its
content of protein, fat, starch and sugar. The composition of the
diets and the results obtained are shown on page 345.

In another series of experiments, rats were kept on a high starch
(curve VIII, p. 347) and high meat (curve VII, p. 347) diet, without
any B-vitamine. The rats on meat lived much longer than those on
starch. The relationship may be seen very well in Fig. 64. Similar
experiments were carried out by Maignon (1241), though he paid
no particular attention to the vitamines. Grafe (1242) likewise
made similar experiments on dogs, and Emmett (1243) took up the
same question, but his detailed report has not yet appeared. Experi-
ments with food constituents mixed in unusual proportions were per-
formed also by Osborne and Mendel (1243a). Tachau (1244) sus-
pected, several years ago, that the utilization of certain dietary
constituents, such as sugar, may depend upon the protein content
of the diet. As for the significance of the above data, they show
that the protein concentration in a given food mixture is of
greatest importance. The dilution of the protein and vitamines by
an excessive carbohydrate addition is not without its harmful
sequelae. In the presence oj considerable protein, the vitamine require-
ments oj the organism are reduced to a minimum. We have no tangible
means of explaining this, as yet. The protein may act as "vitamine-
sparing," so that less vitamine is required in digestion and assimila-
tion, as compared with other dietary constituents. Perhaps the
content of protein in the new substance, mentioned above, is of
significance in this connection. From this, it is clear that the etiology
of pellagra and war edema can not be disposed of merely with the
words, "lack of animal protein." If this were the case, then we
should read of many cases of pellagra in the reports from Central
Europe, which is, however, not the case. Aside from this, it is

NUTRITION IN MAN

347

doubtful that two such different diseases as pellagra and war edema
can arise from the same cause — lack of protein. At least, in one case,
there must logically be an error. Although we believe firmly in the
dietetic cause of both diseases, we cannot accept, for the present the
theory that they arise from a lack of animal protein. If this were so,
then we ought to encounter much more pellagra among the millions
of people that live on an exclusive vegetable diet. In both diseases,
the relationship between the proteins, carbohydrates and vitamines
(perhaps the caloric value of the food is often too low) seems to be
incorrect, and this is all that we know at present.

250
ZSO
2.10
I9O
170

150
130

110

90
70

5

7T7J
O

I

T

7

FIG. 64. THE VITAMINE REQUIREMENTS OF THE RAT ON DIETS RICH IN
PROTEIN, CARBOHYDRATE AND FAT, RESPECTIVELY

I, starch; V, starch + vitamine B after 14 days; II, sugar; VI, sugar +
vitamine B after 20 days; III, fat; IV, meat; VII, meat without vitamine B;
VIII, starch without vitamine B; 0 = died; — > = extra vitamine B given
(Funk-Dubin)

We come now to the consideration of the nutrition in Europe during
the war, and the effect of this diet on the general health of the people.
The mode of nutrition in Germany at this time has been described by
Haupt (1245), Mason (1246), Bornstein (1247) and many others.
Zuntz and Lowy (1248) stated that even at the beginning of the war,
robust men lost about 12 per cent of their body weight, and Rosenthal
(1249) found, usually, a disturbance of the lipoid metabolism. The
influence on the children and the new-born was studied by Hoffmann

348 THE VITAMINES

(1250), Jahreiss (1251) and Bloch (1252) and Kaupe (1252a). The
poor nutrition evidently had no easily recognizable influence on the
intra-uterine life. The size and the weight of the new-born was not
far different from the normal. As regards the children of school age,
there are the reports of Abderhalden (1253), Pfaundler (1254) and
Blanton (1255), all of whom speak of a poor development of the
children, both in body and mind. In Belgium, the same observation
was made by Demoor (1256) and Duthoit (1256a), and in France
by Nobecourt (1257). Fronczak (1258) reported on the effect of
the war in Poland, and Rosenfeld (1259), in Denmark and Switzer-
land.

An exact composition of the diet (per week) was described by
Berg (1260) in 1917 in Dresden:

grams grams

Whole rye bread 1500 Pork 126

Wheat bread 350 Butter or margarine 35

Potatoes 630 Eggs 35

Kohlrabi 3500 Cheese 14

Grits 14 Fruits and vegetables 400

Sugar 21 Coffee substitute 14

This gave 'a daily portion of 33 grams protein (of which 4 grams
were of animal origin), 10 grams fat and 1217 calories. We see from
this that the food was lacking in protein, fat, calories, vitamines A
and (sometimes apparently) C. Many of the observed dietary dis-
turbances in Germany were explained by a poverty of fat, and Thorns
(1261) laid great stress on this dietary component. It is quite true
that a poverty of fat markedly reduces the caloric value of a diet,
but this is frequently confused with a lack of vitamine A. For it is
possible to live very well without fat, as has been shown again by
Osborne and Mendel (1262) on rats. They demonstrated that on a
fat-free mixture, in the presence of vitamine A, rats may grow
splendidly and consume double the quantity of food, as compared
with the usual dietary mixture. Growth in the first stage proceeds
much more favorably, following a greater protein intake. Drum-
mond (1262a) obtained the same results as Osborne and Mendel on
fat-free food mixtures. These results find their replica in experi-
ments on children and adults, which we have already described

If the lack of fat was not responsible for the poor nutrition in
Germany, how did nutritive disturbances develop? In a series of

NUTRITION IN MAN 349

papers, Hindhede (1263) showed how this came about. Although at
the beginning of the war, Germany was in better position than Den-
mark, as regards its food reserves, it tried to conserve its cattle to
the very last, because of the belief in the special nutritive value of
animal protein; for this reason, the cattle consumed a large proportion
of the food which could have been utilized by the population. This
was true particularly of pigs. Kuczynski and Zuntz (1264) laid stress
on this point at the time. In Denmark, on the contrary, cattle con-
servation was prevented by law, and as a result the Danish people
had a dietary composed as follows, according to Hindhede (1265) :

grams per week grama per week

Whole rye bread 1860-2036 Whole milk 700-2100

Wheat bread 420- 700 Skim milk 700-1400

Potatoes 2500-3500 Beef 175- 500

Grits 210- 350 Pork 175- 314

Sugar 448- 467 Butter, margarine 250

Cheese 125-140 Fruits, vegetables 700

Beer 700 Coffee 35-60

This gave a daily ration of 57 to 68 grams protein (of which 17 to
33 grams were of animal origin), 48 to 59 grams fat and 2300 to 2400
calories. This diet differed from the normal in that it contained less
protein, and in the substitution of vegetable for animal protein.
What was the effect of this mode of nutrition on the health of the
population? The mortality dropped to 10.4 per 1000, the lowest
figure of all countries. There was a definite decrease of infections,
including tuberculosis, and a notable drop in cancer mortality, which
fell to 50 per cent of the previous figure. Unfortunately, it is not
certain how much of this improvement was due to the general dietary,
and how much to the abstinence from alcohol, tobacco and coffee.
Of the avitaminoses in Denmark, we have reported only on oph-
thalmia, which we shall speak of later on. In Germany, on the con-
trary, although there was a lack of vitamine A, ophthalmia was
rarely noted.

Besides this, we have observations on the effect of the war diet
on the general health and the various resultant pathological occur-
rences, made by Neuhaus (1266). In women, considerable war-
amenorrhea was observed by Nilsson (1267). Puerperal eclampsia
was markedly decreased according to Gessner (1268). Momm
(1269) and Klotz (1270) found that lactation was unfavorably

350 THE VITAMINES

affected. Zernik (1271) noted a decrease in the resistance to drugs.
The number of tuberculosis cases was considerably increased in
Germany (Kieffer, 1272) and in England (1273). The contrary,
however, was true of cancer (Rumpel, 1274) and also of diabetes,
according to Rumpel, and to Elias and Singer (1275). The effect on
eye diseases was studied by Seefelder (1276).

It is evident from what has been said, that in spite of the small
amount of animal protein in the war diet of Central Europe, no out-
break of pellagra occurred; this was also commented on by S. Harris
(1277). In addition, despite the lack of vitamine A, very little
ophthalmia prevailed. On the other hand, there was considerable
scurvy and hunger edema. We see from the well-controlled diet of
the Danish population during the war, that 68 grams protein (con-
taining 27 gm. animal protein) per day, 50 grams fat and 2500
calories completely sufficed to cover the requirements. Hindhede's
experiments show that the amount of protein might even be decreased
further, and could be substituted by good vegetable protein.

CHAPTER VI

PELLAGRA

Pellagra is a non-contagious, endemic disease of corn-eating popu-
lations, prevalent in northern Italy, Rumania, southern Tyrol and
North America. The disease consists in a specific erythema of the
skin, stomatitis, gastro-enteritis and profound degenerative changes
of the central nervous system. The progress .is acute or chronic,
and the mortality is appreciable. Pellagra has been described by
Bouchard (1278), Roussel (1279), Marie (1280), Roberts (1281),
Schilling (1282), Niles (1283) and Harris (1284).

GEOGRAPHICAL DISTRIBUTION OF PELLAGRA

The greatest prevalence of this disease is in northern Italy, where
it has been known since. 1700, affecting Piemont, Lombardy, Venice
and Emilia. Central Italy is much less affected while the south is
almost free of the disease. It is unknown in Corsica, Sardinia and
Sicily. Pellagra in Italy is evidently decreasing; in 1898, there
were 3987 fatal cases, while in 1905, only 2359. The census shown
as follows:

Year Cases of Pellagra

1879 97,855

1881 104,067

1899 72,603

1905 55,029

1910 33,869

In Austria, especially in southern Tyrol, numerous cases were
recognized between 1875 and 1905; since this time, however, they
have diminshed so rapidly that the number of cases in Rovereto,
according to Weiss (1285), dropped from 8053 in 1904 to 3503 in 1912,
upon the introduction of potatoes. In Roumania, the number of
pellagra patients were calculated at about 75,000 a few years ago;
the disease seems to have increased in Serbia and Bulgaria. In
England, some cases have been described by Sambon and Chalmers
(1286), Box (1287) and Low and Yellowlees (1288). Still, the
diagnosis of sporadic cases seems to be somewhat uncertain, though

351

352 THE VITAMINES

not impossible. In France, the disease is supposed to have been
recognized only since the time of Napoleon I, especially in the
swampy province of Landes, but it has been entirely eradicated by
Roussel (I.e. 1279).

In Egypt, especially in Lower Egypt, we find an insignificant
number of pellagra cases. In Canada, cases were observed by
Pinault (1289) and Rolph (1290). In North America, till a few
years ago, pellagra had been steadily increasing, apparently since
1880; some investigators believe, on the contrary, that it appeared
only in 1900. The progress of the disease here is much more acute
than in Italy, and the mortality markedly greater. The endemia
is most severe in the southern states (especially Texas, Arkansas,
Louisiana, Mississippi, Kentucky, Tennessee, Alabama, Virginia,
North and South Carolina, Georgia) and in Mexico. Lavinder
(1291), between 1907 and 1921, counted 30,000 pellagra cases,
with 40 per cent mortality. Petersen (1292) reported recently that
in the United States, during 1915, there were 10,663 fatal cases;
in 1916, there were 6289. During this time, in the southern states,
the pellagra cases were supposed to represent about 0.5 per cent of
the population, which would make about 165,000 cases. In the
Canal Zone, cases were reported by Decks (1293), arising on a
carbohydrate-rich diet. Tuttle (1294) has described cases in South
America. McDonald (1295) observed pellagra in Antigua; Nicholls
(1296), on the Island of St. Lucia; de Kock and Bonne (1297), in
Surinam.

PROGRESS OF PELLAGRA

Pellagra spreads in rural districts, while the cities remain free
from the disease. In Italy and Egypt, the disease attacked the
poorer population; in North America, however, even well-to-do
farmers who follow the usual mode of country life are not immune.
According to Roberts (1298), the picture of pellagra has changed
in the United States in recent years; here, the acute form resembles
the Italian type, and some forms are so mild that they may be
easily overlooked. The disease occurs more in women than in men;
in fact, the ratio between both sexes is given variously as 2 : 1 or 3 : 1.
According to Siler and Garrison (1299) and Grimm (1300), pellagra
often appears after a birth and in 83 per cent of all cases, it breaks
out in families in straitened circumstances.

PELLAGRA 353

The first attacks and relapses appear in Italy mostly early in the
year — March or April; in North America, beginning in February
and lasting throughout the summer into late autumn. Wellman and
Sparkes (1301) have reported some winter cases.

Pellagra is no respecter of age, but it is seldom seen in infants.
Niles (I.e. 1283), for example, says that he has personally never seen
a case under 5 years and very few under 10 years of age. According
to Snyder (1302), the disease is often seen in older children; thus,
10 per cent of all cases appear before the age of 15. In one case of
a diseased mother who nursed her child for 6 weeks, pellagra
symptoms appeared in the child a few days later on an artificial
diet. Weston (1303) found in Columbia 15 cases among children,
one of which had a sick mother; the child developed pellagra
shortly after being weaned. Voegtlin and Harries (1304) reported
the very interesting case of a 5 months old breast-fed baby, whose
mother showed no symptoms. Byfield (1305) described a pellagra-
like disease in 17 children under 4 years of age, for which, however,
he assumed an infectious and not an alimentary cause. According
to Murphy (1306), the disease is much milder in children and is
without nervous symptoms.

Pellagra is not hereditary, but frequently the children of pellagrans
show stunted growth, mental weakness and other signs of degener-
ation. It is not contagious ; never was there an infection in an orphan
asylum or among the guests of a health resort; according to Gold-
berger (1307), there was never any pellagra among physicans and
nurses of a pellagra hospital.

The course of the disease varies; there are severe acute cases
terminating fatally in a few weeks, and chronic types, with recoveries
and relapses, of many years duration. In an examination of 100
cases, Wood (1308) noted the first symptoms, divided as follows:
skin, 100 per cent; gastro-intestinal disturbance, 77 per cent;
mouth symptoms appearing sometimes before the skin symptoms.
Of the latter, 97 per cent appeared on the hands and the forearms,
and in 39 cases, on the hands only. The uncovered parts of the body
are involved, for the most part, but in one case, the anus, and in
two cases, the back was affected. According to Roberts (I.e. 1281),
there are altogether four different types of the disease.

354 THE VITAMINES

1. Acute, malignant form

Duration is from two weeks to three months. It may appear as the
first attack or during the progress of the disease, or end in chronic
cases with prostration, convulsions, fever and diarrhea. Contin-
uous fever (38 to 40.5°C.); pulse 120 to 130, small and arrythmic;
tongue, deep red, fissured and painful; painful stomatitis and
pharyngitis; nausea; typical dermatitis; skin petechiae. In addi-
tion there are frequently tremors, convulsions, tetanus, delirium and
incontinence. This type, is often called "typhoid pellagra."

2. Light sub-chronic jorm

Prevalent in young individuals; light cases with dermatosis and
dyspepsia, disappearing when cured. Duration, 1 to 2 years; no
fever. This form is very common in Italy and Roumania; less so
in America. The symptoms consist of a light dermatosis on the backs
of the hands and wrists, reddening of the mouth mucosa, consti-
pation or slight diarrhea.

3. Severe, cachectic sub-chronic form

Temperature at the beginning, 37.7°C; pulse, 100. Very severe
symptoms along the digestive tract; the tongue, deep red, eroded;
mouth and pharynx mucosa, deep red and painful; gums, swollen
and bleeding; nausea, vomiting, gastralgia, persistent serous diarrhea,
loss of weight. Later, there are cerebral symptoms, mental weak-
ness and skeleton-like emaciation.

4' Chronic form

Duration, 1 to 20 years. In most cases, three periods are differ-
entiated. A) First period, with dermatosis and dyspepsia, as in
light sub-chronic cases. B) Second period, symptoms along the
digestive tract become more severe; gastralgia, tabes-like stomach
crises, vomiting, diarrhea and tenesmus appears. Pulse, 80 to 100,
with dyspepsia and light dropsy. Typical dermatosis; on recur-
rence the skin of the feet is brownish red and scaly; backs of the hands
are brownish red, wrinkled and aged. To this picture there are
added brain and spinal cord symptoms; uncertain walk, vertigo,
tremor, epileptic and tetanic attacks, contractures, accentuated

PELLAGRA 355

reflexes. C) Sooner or later in the third period, there is cachexia
and mental weakness, with hemiplegia or paraplegia, and finally
with serous diarrhea, petechiae, muscle atrophy and incontinence.
Normal temperature prevails, as a rule; only exceptionally is terminal
fever observed in the last days of the disease. In addition to these
four types, there are light abortive forms, which we shall describe
in the consideration of expermental pellagra in man.

SYMPTOMATOLOGY AND PATHOLOGY OF PELLAGRA

L G astro-intestinal tract

The pellagrous tongue is of diagnostic importance: in the early
stage, coated; later, deep red, fissured in the middle and around
the edge, often painful. The gums, spongy, red, easily bleeding as
in scurvy; sometimes there is alveolar pyorrhea. Swelling and red-
dening of the mouth and pharynx muscosa with aphthae or small
vesiculae, pyrosis, gastralgia, nausea and vomiting occur. In an
investigation of the stomach of 20 cases, Johnson (1309) found a
lack of hydrochloric acid in 16. This finding was confirmed by
Givens (1310), who noted, however, cases in which hydrochloric
acid and pepsin were present, especially in children. Pellagrous
diarrhea is of importance; in a spring attack it may happen that
there will be from 10 to 20 evacuations daily, often slimy and bloody.
In the final stages, the evacuations occur still more frequently, serous
and colorless; indicanuria is often noted. All the symptoms along
the gastro-intestinal tract appear to be of central nervous origin.
Siler (1311) preferred not to regard as pellagra those cases without
gastro-intestinal disturbances, though it seems to us that such a
differentiation would be only artificial.

In acute cases, in the stomach, swelling and reddening with erosions,
especially at the pylorus were found. In chronic cases, the mucous
membrane of the stomach was pale, covered with mucous, and the
muscles were atrophic. In the intestine, in acute cases, there was
enteritis with ulcerations in the small and large intestine, less
frequently in the duodenum; in chronic cases, according to Lynch
(1312), the mucosa was pale, covered with mucus, and atrophic;
the musculosa was thinned. The liver was tough, atrophic, and
occasionally showed fatty degeneration.

356

THE VITAMINES

Fia. 65. TYPICAL PELLAGRA ERYTHEMA

ON THE BACK OF THE NECK (ROBERTS)

FIG. 66. PELLAGRA IN A NEGRESS
(ROBERTS)

Fia. 67. PELLAGRA (ZELLER)

PELLAGRA 357

2. Skin

The pellagrous dermatitis is one of the most important of the early
symptoms. In light forms, this dermatitis is maculo-papular; in
severe acute types, vesicular with intensive reddening, edema and
pain. The dermatitis appears in a typical manner symmetrically
on the backs of the hands, and wrists (the pellagrous glove), then
on the neck, less often on the face, and on the feet (the pellagrous
shoe). This dermatitis was likened by Gurd (1313) to X-ray
dermatitis. The pellagrous glove and shoe extends frequently
one-third of the way up the forearm and leg respectively. On the
face, the dermatitis appears on the nostrils, temples and behind the
ears; sometimes, ecchymoses appear on the eye-lids. On the elbows
and the forearm the skin remains rough and scaly for a long time;
this protracted scaling may appear on the face, shoulders and, in
fact, on the whole body. In chronic cases, the nails sometimes
become grayish white, thickened and brittle. On recovery, the skin
appears brownish red and dark brown on the affected places; in
rare cases, this extends to the entire body.

It is of etiologic importance that in the production of human
experimental pellagra by Goldberger (1314), the first symptoms
appeared on the scrotum. This condition has inclined some investi-
gators to dou^t the validity of Goldberger's findings, pointing out
that it had nothing to do with real pellagra. Because of this, it is
important to state here that Deiaco (1315), Merk (1316), Decks
(I.e. 1293), Crosby (1317) and Wood (I.e. 1308) observed this
localization of the dermatitis as the first sign in their cases.

This specific dermatitis appears to be of trophoneurotic central
origin, but the influence of the sun rays cannot be altogether denied.
The burning of the hands and feet, a very disagreeable occurrence
independent of the dermatitis, belongs to the nervous symptoms.

3. The nervous system

Pain in the back is one of the early symptoms of severe cases. The
reflexes are accentuated. The muscles become atrophic in half of
the chronic cases, with fatty degeneration; contractions in a flexed
position sometimes develop in the hands and feet. The degeneration
reaction is usually lacking. Among the later symptoms, are tremor,
especially of the hands, infrequently of the tongue and lips, and

358

THE VITAMINES

cramps of some muscle groups; in severe cases, there is spastic or
atactic gait. As terminal symptoms, there are tetanic, epileptic
attacks, and paralysis of the sphincters. Mental disorders belong
to the typical picture of severe pellagra. The initial symptoms are
insomnia, quietness, sadness, later hypochondria and finally psychoses

FIG. 68. SPINAL CORD; LUMBAR
REGION (MOTT)

A -

'

4

FIG. 69. CELLS OP VENTRAL
HORN WITH ASYMMETRIC
NUCLEI (MOTT)

FIG. 70. SCIATIC NERVE IN
PELLAGRA; LONGITUDINAL
SECTION (Morr)

FIG. 71. CELL OF BETZ IN
MOTOR CORTEX (MOTT)

of all kinds. The latter develop in 10 per cent of all cases in Italy,
in about 5 per cent in America, apparently more often in the chronic
form. According to Sandy (1319) the number of pellagra psychoses
among the inmates of insane asylums is very large in the southern
states. Miller and Ismail (1320) investigated the various types of
pellagrous lunatics in Egypt, classifying 750 cases as follows:

PELLAGBA 359

Amentia (42 per cent — typical first stage) 320

Mania 115

Melancholia 113

Dementia 103

The mortality was 32 per cent; aside from this, 47 per cent were
discharged, with unknown issue.

In the brain, the following lesions were found : pia and arachnoid
were thickened, with a milky cloudiness and ecchymoses. In the
brain, mostly edema and hyperemia with hydrops of the ventricles
are noted. In chronic cases, the brain and the brain convolutions,
especially the frontal, frequently become atrophic, hard and anemic.
Miscrocopically, cortical nerve cells are degenerated, with swelling,
vacuole formation, displacement of the swollen nucleus to the side,
and later atrophy of the degenerated cells.

In the spinal cord, in acute cases, dilatation of the blood vessels
and edema are found; in chronic cases, chiefly degeneration of the
dorsal fibers and the direct pyramidal tract. Scattered localities
with disappearance of the nerve fibers in the whole white substance
of the spinal cord were also noted. With this, the dorsal roots are
also degenerated with arterial thickening. In the gray substance,
and also in the cells of the ventral and dorsal horns, we find pigmen-
tation, swelling of the cell protoplasm, chromatolysis and displace-
ment of the nucles to one side. In the sympathetic nerve, especially
in the abdominal ganglia, degeneration was observed. It must be
explicitly stated that in the nerve centers, there was not a trace of
inflammatory lesions; all the changes, according to Mott (I.e. 890),
are exclusively of a degenerative nature.

Microscopic hemorrhages were often noted in the nerve centers.
In one of Chalmer's (1321) cases in Roumania, the cause of death
was a marked hemorrhage in the lower cervical and dorsal part of
the spinal cord. In the sciatic nerve, Mott found scattered degener-
ation of nerve fibers.

4- Circulatory system

In the blood, according to Findlay (1318), an increase in lympho-
cytes is mostly found. Hemoglobin, about 80 per cent; pulse, usually
100 and in acute pellagra, up to 120; blood-pressure is mostly low.
Examination of the blood for microorganisms was negative. Bardin
(1322) found an increase in the number of small and large lympho-

360 THE VITAMINES

cytes and a decrease in polymorphonuclear neutrophiles. Micro-
scopic findings were recorded by Nagamatsu (1323). Examination
of the cerebrospinal fluid, according to Lorenz (1324), gave no reason
for regarding pellagra as an infection of the central nervous system.
The heart is mostly atrophic, pigmented, fragile and occasionally
shows fatty degeneration. Sometimes there is lung edema, hyper-
emia and hydrothorax.

FIG. 72. RARIFICATION OF THE PHALYNGEAL ENDS IN PELLAGRA (ROBERTS)

5, Bones

Lombroso (1325) and Babes and Sion (1326) observed, in some
cases, brittleness of the ribs and long bones. Fractures of the long
bones were also noted. Similar lesions are characteristic of human
and experimental scurvy. Roberts (I.e. 1281) gives some X-ray
pictures of pellagrous bones, showing a ratification of the bone ends.

6. Sexual organs

In acute cases, vulvo-vaginitis is not infrequently observed, and
in rare cases, there is gangrene of the labia. In chronic cases, amen-
orrhea frequently occurs. Abortion was noted in 20 per cent of

PELLAGBA 361

pcllagrous pregnant women; hemorrhages following birth are not
uncommon. In latent cases, following pregnancy, an acute condition
is not infrequently brought on; . similar observations have been
made in beriberi.

7. Other organs

Regarding eye symptoms, Marie (I.e. 1280) described conjuncti-
vitis with pterygium and hemeralopia, and sometimes pigmentary
retinitis. Calhoun (1327) found that certain visual disturbances,
especially certain forms of color blindness, precede other symptoms
of pellagra, so that they may be used in the diagnosis.

As regards the endocrine glands, Beeson (1328) observed, out of
316 cases, 25 cases of thyroid complications. Modinos (1329) noted
cases in Egypt with enlarged adrenals, about twice the usual size.
In cases observed by Wilson (1330),, also in Egypt, these glands were
much lighter in weight. Morse (1331) found severe atrophic degen-
erative changes in the thyroid and the adrenal medulla. In the
latest reports, especially from Egypt, special attention was paid to

the insufficiency of the adrenals (Wilson, I.e. 1330).

I

CHEMICAL PATHOLOGY

Koch and Voegtlin (1332) analyzed the spinal cord and brain of
pellagrins and found that the figures agreed, in general, with those
obtained in pigeon beriberi. One of the most important findings
was the loss of lipoids, particularly in the spinal cord.

Indicanuria was found by Ridlon (1333) in 90 per cent of his
cases. Murlin (1334) found high amino nitrogen and hippuric acid
values (two to three times greater than normal), indicating a poorer
nitrogen utilization. If the carbohydrate diet is replaced by a protein
diet, then the values found are smaller. The effect of both of these
diets on the composition of the blood was investigated by Lewis
(1335). The carbohydrate diet gave smaller urea and non-protein
nitrogen figures, but otherwise the findings were normal. Jobling
and Maxwell (1336), as well as Sullivan and Stanton (1337), deter-
mined the alkali reserve of the blood; the figures varied only slightly
from the normal; no acidosis was noted. Sullivan and Jones (1338)
found indican in the saliva in one case; in addition, there was a
thiocyanide reaction, indicating a retardation of protein metabolism.

362 THE VITAMINES

METABOLISM

The first exact metabolism experiments in pellagra were made by
Myers and Fine (1339). The milk-vegetable diet used was well
utilized. Nicolaidi (1340) observed large food losses in the stool.
Albertoni and Tullio (1341) obtained a negative nitrogen balance on
corn; the balance became positive on meat. Hunter, Givens and
Lewis (1342) found in seven cases of pellagra a positive nitrogen
balance on a pellagra-producing diet. Boyd (1343) found, in a
metabolism experiment on Turkish prisoners in Egypt, in confirma-
tion of earlier findings, a decrease of hydrochloric acid in the stomach,
diminished pancreatic secretion, protein and fat loss in the stool
(remarkable in a disease caused by a lack of protein) and a very poor
protein assimilation. The latter is especially emphasized in recent
publications (Sullivan, Stanton and Dawson, 1343a) .

PROGNOSIS

In Italy, 55,029 cases of pellagra were recorded in 1905, with 2359
deaths, equivalent to more than 4 per cent mortality. In America,
on the contrary, the prognosis is much more serious, namely, 50 per
cent death rate in the asylums and 20 to 25 per cent in private
practice. It is obvious that the picture of pellagra, because of the
dietetic factor, frequently varies with the well-being of the popula-
tion, assuming either a mild or more severe type.

THE RELATIONSHIP OF PELLAGRA TO THE ACCEPTED AVITAMINOSES

BERIBERI AND SCURVY

In the short paragraph on the relation of beriberi to scurvy in
South Africa (p. 296), we have seen that scurvy develops on a diet
consisting of very little animal protein and corn, without a sign of
pellagra. This is true also of a widespread occurrence of scurvy,
which G. R. Hopkins (I.e. 939) described in a population which fed
itself almost exclusively on corn. Marie (I.e. 1280) and Viswallingam
(1344) looked upon scurvy as a predisposing cause of pellagra.

As for the relation of pellagra to beriberi, Sheppard (1345) saw
pellagra in Singapore on an exclusive rice diet. Stannus (1346)
stated that he saw a number of cases in Nyasaland on rice (only
partially husked) ; vegetables, fish or meat were eaten only once in
14 days. The disease progressed without any characteristic exan-

PELLAGRA 363

thema, although certain signs of the skin affection were visible
around the mouth. On the other hand, a pellagra-like erythema
was observed in beriberi by Schiiffner and Kuenen (1347). Night-
ingale (1348) saw 1210 pellagra-like cases, in South Rhodesia, which
he called "zeism." The disease developed on milled corn, was
mild and without exanthema, but with light dermatitis and mouth
symtoms. He stated that cod liver oil therapy was quite favorable.
Edwards (1349) designated as "peripheral neuritis" a disease met
with in Jamaica among the poor inhabitants who live on a nitrogen-
poor diet. The symptoms resemble pellagra more than beriberi.
Finally, we wish to comment upon the composition of the diet, which
Braddon (I.e. 866) regarded as productive of beriberi. It was
exceptionally poor in animal protein and must have lead to pellagra,
according to the newest conceptions. This short chapter was
introduced here, not so much to demonstrate the relationship between
pellagra and the avitaminoses, but to show that we may be dealing
with mixed forms under certain conditions, which indicates perhaps
that if the diet is lacking in one constituent, it is likely that others
may be lacking at the same time.

MODE OF DEVELOPMENT OF PELLAGRA

Although theoretically pellagra may also develop without any
corn consumption, it appears and disappears, in practice, with corn
cultivation. Thus, corn was introduced into Egypt in 1840, and in
1847, the first cases of pellagra were noted. In Spain, the disease
has been endemic for two centuries, having appeared there with the
beginning of corn cultivation; at present, considerable rye, wheat and
oats, but very little corn, are grown and the disease has almost been
stamped out there. Weiss (1350) noted that since 1905, after the
exclusion of corn from the diet, cases in the Tyrol appeared more
rarely. Alpago-Novello (1351), in an Italian province, partially sub-
stituted corn growing by turnips and potatoes, whereupon the number
of cases decreased. It is likewise not without significance whether
hand ground or machine milled corn is used. In the United States,
it has often been found that in the mountainous districts where only
the hand milled variety is used no cases of pellagra occur; the con-
trary is true in small industrial cities where the natives use prepared
cornmeal, as brought out by Wood (1352). Blosser (1353) observed

364

THE VITAMINES

130 cases develop on a diet containing a considerable amount of
cane sugar products, especially molasses. By omitting the latter,
which is greatly relished in the southern states, an improvement was
noted. That a diet very rich in carbohydrates is consumed in dis-
tricts where pellagra prevails, is evident from the report of Jobling
and Petersen (1354). Babes (1355) remarked, advisedly, that when
in February 1918, the entire population of Bucharest was placed upon
bread made of corn and wheat, numerous cases of pellagra appeared
as early as in May, so that three months were necessary for the
development of the disease. Lombroso (I.e. 1325) and Camurri (1356)
describe a dietary composition in the Italian pellagra districts:

Lombroso Camurri

Grams daily Grams daily

Corn 1091 Polenta 1500

Rice and barley 67 Rice 100

Beans 60 Beans 100

Potatoes 67 Potatoes 100

Vegetables 250 Vegetables 100

Lard 21 Lard 20

Olive oil 33 Olive oil 10

Fish 67 Milk 100

Poultry 27 Cheese 50

Apart from this, great differences were noted in the summer and
winter diets, which explains very well the periodic occurrence of
pellagra in spring. It seems that the Italian population feeds itself
very poorly in the winter. Lombroso reported on such dietary-
changes in the province of Ferrara :

DIET IN 8
WINTER MONTHS

DIET IN 4
SUMMER MONTHS

Polenta

grams per day
1000

grams per day

160

Milk

Eggs ;

Almost none

Almost none

Onions

One daily

Two daily

Corn bread

50

400

Home-made broad .

50

200

Meat

10

60

Cheese

5

20

Beans

150

40

Fish

20

Very little

PELLAGRA 365

These figures were corroborated by Devoto (1357). Wilson (1358)
saw the development of pellagra on a diet containing 92 grams
protein and 2200 calories. The protein consisted of f wheat and
£ corn; 10 per cent of the war prisoners developed pellagra in one
year, the disease appearing only after hard labor. Boyd and Lelean
(1359) observed pellagra among 6000 Turkish prisoners, to the extent
of 18 per cent, while among German prisoners, there were no cases;
it was only when they were assigned hard labor to perform that 65
cases appeared, according to Lelean (1360). Bouchard (I.e. 1278)
noted that shepherds developed pellagra, while cow attendants
remained free from the- disease. The difference was supposed to lie
in the milk consumed.

The mode of occurrence of pellagra was studied very accurately by
Goldberger and his co-workers. Goldberger, Wheeler and Syden-
stricker (1361) compared the diets of diseased and healthy natives in
a small industrial city in South Carolina. The chief difference was
in the milk, meat, vegetable and fruit consumption. The amount
of protein was about 85 grams, of which one-third was of animal
origin. The reason why other investigators did not observe the above
differences is because the pre-pellagrous diet, and not that which is
consumed at the time of the attack, must be studied ; for in this case,
the disease is already improving. Sydenstricker (1362) could show
by statistical data, that the cases become more numerous, when the
people, because of increased prices, must limit their expenditures.
Apparently, the decrease in the number of pellagra cases in the United
States during the war is due to the great prosperity of the population.
Goldberger and Wheeler (I.e. 109) produced experimental pellagra in
11 prisoners. The diet, accurately controlled, was analyzed by
Sullivan and Jones (1363), and the vitamine content was tested on
animals by Sullivan (1364). The diet consisted of 41 to 54 grams
protein, of which 80 to 97 per cent was of vegetable origin, and it
seemed to be poor in vitamines B and A. The controls received a
better diet. The experiment lasted 6| months, during which time,
out of 11 experimental subjects, 5 developed symptoms, which
must be regarded as somewhat abnormal pellagra symptoms. The
first symptoms appeared on the scrotum, which occurs sometimes,
as we pointed out previously. It is apparent, according to Gold-
berger, that the 11 subjects had pellagra in a mild form, and that
the symptoms may vary with the diet. Goldberger (1365) held
that three months sufficed to develop symptoms of pellagra on a
suitably chosen diet.

366 THE VITAMINES

THERAPY

Roussel (I.e. 1279) stated in 1866 that dietetic measures were of
greatest importance in combating pellagra, a view in which a great
number of investigators are agreed, including Royer (1366) who
expressed himself similarly in 1835. Lombroso (I.e. 1325) saw that
a meat addition produces an extraordinarily favorable effect. Kleim-
inger (1367) cured 12 pellagrous lunatics by means of diet, and
Lorenz (1368) made considerable use of the dieto-therapy. Elebash
(1369) used green vegetables and fruit juices; Allison (1370) used
milk, eggs and fruits; Sylvester (1371) at first used fruit juices, then
skim milk and eggs. Bravetta (1372), Willets (1373) and Ridlon
(1374) proceeded similarly. The latter treated 51 cases with a spe-
cially selected diet, 48 of which recovered. Goldberger, Waring and
Willets (1375) tested, in some asylums, a diet consisting of milk, meat
and vegetables, with a decrease of carbohydrates, and obtained very
good prophylactic and therapeutic results. Voegtlin, Neill and
Hunter (1376) used a vitamine therapy in their practice, with the
following results; with vitamine B, no results were evident, while with
vitamine A, in the form of liver and thymus extracts, some result was
obtained but it was not very marked. Sherman (1377) recommended
milk as a prophylactic.

As we have noted, the best results were obtained by dietary
changes; in fact, the complex products designated as animal protein
proved effective. We have already pointed out that with the addi-
tion of meat, eggs or milk, not animal protein alone is added, but also
food complexes which are, for the most part, unknown. By this
means, a diet rich in carbohydrates and poor in protein is changed
to one poor in carbohydrates and rich in protein, whereupon the
entire nutrition undergoes a radical change. The modern conception
of the etiology of pellagra is based upon the above findings.

ETIOLOGY OF PELLAGRA

Pellagra occurs chiefly on corn. In this connection, a series of
hypotheses were proposed to explain the nature of pellagra, which,
however, we can not discuss here. Even the proponents of the infec-
tion theory, for example, Siler, Garrison and MacNeal (1378) partially
admit that the diet also plays a role in this disease. Tanner and
Echols (1378) point out that it is not enough merely to give a

PELLAGRA 367

satisfactory diet. It must also be noted that the diet is actually
consumed.

As regards the nutritive value of corn, we have already discussed
this matter in detail (p. 250). With the assumption that even the
whole kernel possesses a low biological value, we are not in accord,
since most birds can live on it indefinitely. What happens to the
nutritive value of the protein, after certain parts of the grain are
removed by milling, we do not know exactly. The reports from
Egypt by Wilson (I.e. 1330), Boyd (1379) and Roaf (1380) have shown
that in pellagra, the protein assimilation is disturbed, with simulta-
neous defects of the sympathetic nervous system. Wilson, in par-
ticular, is of the opinion that the biological value of the protein can
not drop below 40 grams casein, without the danger of the occurrence
of pellagra. As a matter of fact, the pellagra-producing diet there
was equal to only 22 grams of casein.

Most of the observers of the Egyptian pellagra came to the con-
clusion that it is related to a lack of an essential aminoacid, probably
tryptophane. Bigland (1381) adopted this view, while Enright
(1382) saw pellagra in German prisoners who, in his opinion, had
sufficient protein in the diet. Based upon Wilson's conception,
Chick and Hume (I.e. 460) undertook to produce experimental
pellagra in monkeys. These monkeys were given a diet containing all
the vitamines, but varying considerably in its protein content in the
form of maize gluten. Three animals were kept on this diet, where-
upon an erythema on the nostrils and other symptoms developed
which were regarded as pellagra, and which were improved by the
administration of casein. Since in this experiment, the food and
vitamine intake were not controlled, and since the study of a disease
supposedly related to a lack of animal protein was, strangely enough,
conducted with plant-eating animals, which normally after the
suckling period never eat animal protein, we must discount the value
of this experiment.

The conception that pellagra is associated with a lack of protein
of high biological value, is in some respects in opposition to Hind-
hede's findings and the experience of specialists connected with the
feeding of the populations of various countries. In Europe too,
during the World War, very little pellagra prevailed despite the
lack of animal protein.

\

368 THE VITAMINES

Infantile pellagra, as for example in the case described by Voegtlin
and Harries (I.e. 1304), might be of help in clearing up the etiology.
It is indeed true, according to Eckles, Palmer and Swett (1383),
that the protein content of the milk can be influenced by nutrition
to a certain extent; still, the possible variations could hardly explain
the occurrence of infantile pellagra, of which some instances have
been observed.

We are inclined to the view of Goldberger (I.e. 109), who con-
sidered the following etiological factors as possible :

1. Partial lack of vitamines.

2. Lack of animal protein.

3. Lack of a still unknown vitamine.

4. The combined influence of all these factors.

We believe that the dilution of protein with carbohydrates increases
the vitamine requirements, so that although the vitamines are
present, they do not suffice. We have mentioned the possible
significance of the important substance, associated with proteins, and
it is not impossible that this factor is lost during the milling of corn.
It would be rather premature to consider the chapter of pellagra as
closed.

CHAPTER VII

SPRUE

This disease, also designated as aphthae tropicae, Ceylon sore
mouth, psilosis linguae et intestini, diarrhea alba, tropical diarrhea
and diarrhee de Cochinchine, has been described in detail by Van der
Sheer (1384), Thin (1385), W. C. Brown (1386), Begg (1387), Schilling
(I.e. 840 and 980) and CasteUani and Chalmers (I.e. 839). This non-
infectious disease, investigated since 1776, is endemic in Asia, espe-
cially in the Malay Archipelago, Siam, Annam, Sumatra, Java, but
also in India, Ceylon, China and Japan, also in Australia and occa-
sionally in Europe. In the United States, cases were reported by
Wood (1388), Boyd (1389) and Sturtevant (1390). These patients
were never in the tropics, but Hiat't and Allan (1391) also observed
cases, which came from the tropics. The symptoms of the disease
consist in painful stomatitis with vesicles and erosions and persistent
diarrhea, with grayish-white foamy stools. The disease is curable by
a dietary therapy.

SYMPTOMATOLOGY

The edge of the tongue and the point are reddened, eroded, with
small vesicles, and covered with ulcers. Similar erosions, vesicles
and ulcers are found on the gums, lips, soft palate, uvula, and
cheeks. The patients complain of marked pain on chewing and
swallowing. According to Bahr (1392), the disease occurs much
more frequently in women, and may be confused with pernicious
anemia.

The abdomen is blown up, especially at the epigastrium, the
patient complains of pressure, fullness and burning in the region of
the stomach, particularly after meals; in addition, there is heart-burn,
flatulency and vomiting (without nausea). On investigation of the
stomach, hyperchlorhydria and achlorhydria are not infrequently
found. In the morning hours, without bodily pain and without
tenesmus, there is a copious, soft, foamy grayish evacuation. The fat
content of the feces is markedly increased, and the findings, according
to Halberkann (1393) are similar to those in a pancreas affection.
According to T. B. Brown (1393a), there is a complete lack of

369

370 THE VITAMINES

pancreatic ferments and the therapeutic administration of such is
recommended. The number of red blood corpuscles is considerably
decreased (1,000,000 to 3,000,000 per cubic millimeter); the hemo-
globin index is 60 to 70 per cent. In some cases, tetany belongs to
the clinical picture of sprue. In this connection, there are the newer
reports of Bassett-Smith (1394) and Barach and Murray (1395), who
investigated the parathyroids and found them normal.

In severe cases, muscle weakness, loss of weight, depression, and
symptoms of cerebral irritation are observed. In the final and fatal
stage, continuous fever is often noted. The skin becomes gray and
scaly; the tongue appears smooth, rimous and atrophic; the ankles
become edematous; the liver becomes atrophic; the diarrhea, more
severe and the pulse, slow and weak.

PATHOLOGICAL ANATOMY

In the intestine, there is a primary congestion of the capillaries of
the sub-mucosa; later, hemoglobin exudation and round cell infiltra-
tion; finally, atrophy of the intestinal mucosa. This constant finding
of primary lesions in the sub-mucosa speaks against a local infectious
cause of the disease. The resophagus and the stomach are similarly
affected, but less intensively. In severe chronic cases, atrophy of
the intestinal wall, liver and pancreas is found.

THEEAPY

Substances like opium, bismuth, etc., are rather harmful. Emetin
was recommended by Schmitter (1396) although, according to
Miihlens (1397), this is indicated only in cases complicated by
dysentery. Simon (1398) used vaccine with good results, although
he stated that a change in diet must be made simultaneously.
Castellani (1398a) recommends sodium bicarbonate administration
in conjunction with the dietetic treatment. According to Wegele
(1399) the dietetic therapy is most productive of results. Among
the best substances to use are, according to Low (1400), milk;
according to Cantlie (1401), fruit, and according to Conran (1402),
meat. These are the same substances used in the treatment of
avitaminoses, but in sprue, special care must be exercised and only
frequent small doses should be administered. Especially to be
recommended, is milk, either raw or cooked for a short time, raw

SPRUE 371

meat juice, meat broth and meat jelly; of the fruits, especially straw-
berries (2 to 3 pounds daily), but also apples; oranges, grapes, goose-
berries, and blackberries. Cantlie (1403) advised beginning carefully
with the meat diet (meat juice, meat jelly) and giving milk every 3
or 4 days. An exclusive milk diet does not suffice to bring about a
cure, so that in certain cases it must be combined with a fruit diet.
Later, light mixed food is gradually and carefully given. Bovaird
(1403a) suggests as therapy the replacing of fats and carbohydrates
by proteins and fruits.

PATHOGENESIS

Since sprue occurs less frequently than pellagra the number of
published communications on the subject is not very large. Al-
though some investigators like Cantlie (I.e. 1401) and Michael (1404)
believe in an infectious cause (a fungus, Monilia psilosis), neverthe-
less the following circumstances point to an avitaminosis : Ashf ord
found (1405) that in Porto Rico only the city dwellers, who eat
much bread, develop sprue, but not the farmers, who eat bananas.
According to W. C. Brown (I.e. 1386), the Chinese coolies in East
India develop beriberi, whereas the Europeans, on a practically similar
diet, develop sprue. Stewart (1406) is of the opinion that no real
basis exists for regarding pellagra and sprue as two different diseases.
Heaton (1406a) believes rather in the dietetic than in the infectious
origin of sprue. Werner (1407) observed scorbutic symptoms
(petechiae on the calves of the legs) in sprue. In the early stage of
sprue, there is often an aversion for meat and this feeling, together
with the known fear of fruit in diarrhea, leads to the further develop-
ment of the disease. Still, the most important reason for considering
sprue here is the influence of dietary changes on the course of the
disease, and the therapeutic results following the administration of
vitamine-containing foodstuffs. In particular, the fact that emphasis
is placed upon the raw state of the foodstuffs in the treatment gives
further support to our conception. Especially in the use of straw-
berries, does Leede (1408) emphatically state that preserved straw-
berries are far less effective than fresh berries, and that the active
substance contained therein is destroyed on heating. Whether we
have, in sprue, a disease similar to pellagra or scurvy, cannot be
stated with certainty at present; we must await further work on
the subject.

CHAPTER VIII
HUNGER EDEMA

Hunger edema had been observed, prior to the late war, and
Sticker (1409) and Prinzing (1410) occupied themselves particularly
with the history of this disease. Prinzing, in his historical compi-
lation, records no cases between the time of the Peloponesian War
(430 to 425 B.C.) and the siege of Port Arthur in 1904. Maliwa
(1411) believed that hunger edema was observed in the Napoleonic
Wars, at the siege of Paris, and in the Boer War. Wheeler (1412)
described actual cases in the Boer War, in the concentration camps.
Digby (1413) and McLeod (1413a) reported on this disease during
famines in India and Ceylon in 1876 and 1877. Patterson (1414)
saw numerous cases in Chinkiang (China) in 1899, during the time
that the population was living on greens. Landa (1415) counted
hundreds of cases in Mexico during the war in 1915, which developed
on a diet of turnips and spinach.

In the last war, the first report on the subject was made by Rumpel
(1416). Then, in rapid succession there came numerous reports of
the disease among prisoners by Jtirgens (1417), Bonheim (1418) in
Bonn and Lange (1419) in East Prussia. From Austria, among
other reports, were those of Knack (1420), Schiff (1421) and Jaksche
(1422) ; indeed, in one part of Bohemia alone, 22,000 cases were noted,
with 4 per cent mortality.

Vandervelde and Cantineau (1423) and Breuer (1424) reported on
cases in Belgium. Beyerman (1425) observed cases in the insane
asylum in Medemblik (Holland) in 1917 and 1918, which were cured
by fresh vegetables; in these cases there was perhaps a complication
with scurvy. Strauss (1426), Guillemin and Guyot (1427) and, at
the beginning of the war, Budzynski and Chelchowski (1428) de-
scribed the disease in Poland. Wells (1429) saw numerous cases
in 1917 in Roumania; Tonin (1430) observed the disease among
prisoners in Italy; Enright (1431), among Turkish prisoners in
Egypt.

Mann, Helm and Brown (1432) noted 3000 cases in Haiti, out of
which 200 came to necropsy. The disease has prevailed there for

372

HUNGER EDEMA 373

years among the prisoners, in fact, it was noted that the disease
broke out in less than three months after incarceration.

In infants, on a carbohydrate-rich diet, there develops a condition
called "Mehlnahrschaden." of which we have already spoken.
Many of these cases, if not all, can be considered analogous to hunger
edema. Vacher (1433) reported such cases during the siege of Paris;
later, similar conditions were described by de Wolf (1434) and
Potter (1435) in children suffering from marasmus. Potter believed
that lack of protein was the cause of the disease. The disease was
also noted by Chapin (1436), Waterman (1437), Hume (1438), Ashby
(1439) and Klose (1440). Many of these investigators studied the
effect of a change in the diet, especially an addition of protein.
In the American literature, the disease is often designated as acro-
dynia and has been described by Weston (1440a) and by Cartin
(1440b).

Of additional interest, is the relationship between hunger edema
and "epidemic dropsy" in India. This disease, described by
Greig (I.e. 880), was first regarded as wet beriberi. Although
scorbutic symptoms were also observed, the condition of the patients
could be markedly improved by an addition of meat. Most of these
cases developed on a rice diet, and yet they could be identified as
hunger edema. McCay (1441) believed that the cause of epidemic
dropsy lay in the lack of protein. In a compilation on this subject,
Maver (1442) believed that hunger edema, edema of children and
epidemic dropsy have the same etiology.

SYMPTOMATOLOGY AND MODE OF DEVELOPMENT

A splendid description of the symptomatology, pathology and
pathogenesis is given by Schittenhelm and Schlecht (1443) and by
Schiff (1444). The most prominent sign is the edema, localized
mostly on the legs and on the face, near the eyes. According to
Hiilse (1445) pains in the calves are very characteristic. Polyuria
is very pronounced, according to Rumpel and Knack (1446) and
Zondek (1447). Jansen (1448) found decreased blood pressure and
subnormal temperatures. Jess (1449) called attention to the eye
symptoms, hemeralopia and peripapillary edema of the retina. The
marked bradycardia is especially characteristic of hunger edema,
according to Rumpel and Knack (I.e. 1446). This is not to be at-

374 THE VITAMINES

tributed to heart weakness, according to Schittenhelm and Schlecht
(I.e. 1443), but to the same causes that bring about the edema. In
these cases, adrenaline dose not raise the blood pressure, and the usual
heart stimulants, such as digitalis and caffeine, have little effect,
while atropine has no effect on the bradycardia. The pulse is
extraordinarily small, slow (32 to 36) and soft; reflexes and normal;
constipation was noted more frequently than diarrhea, though the
latter progressed as the edema disappeared. The disease appeared
mostly in hard-working men; the over-exertion seemed to hasten
the onset of the disease. Frequently, rest in bed sufficed to dissi-
pate the edema and to bring about an improvement. The disease
is often found associated with various infections, such as lobar
pneumonia, bronchitis and furunculosis.

Various investigators offer different explanations of the mode of
development of the disease, though there is no doubt that the poor
diet must be the primary cause; the predisposing conditions may be
severe manual labor and exposure. Knack and Neumann (1450)
stated that the cause was inadequate nutrition with turnips and a
large water intake. Falta (1451) thought that the faulty diet was
bread, cabbage, dried vegetables and turnips. The total diet con-
tained 30 to 59 grams protein and 1200 to 1400 calories; the vege-
tables were cooked in large quantities for 4 to 7 hours. The cases
reported by Falta occurred among war prisoners. Burger (1452)
gave the exact composition of the faulty diet; it consisted of 55.9
grams of protein, 8.5 grams fat and 284.6 grams carbohydrate (1478
calories). Kraus (1453) observed cases developed after hard labor,
on a diet containing 15 per cent indigestible carbohydrates, little
fat and a maximum of 50 grams protein, and 800 to 1300 calories.

PATHOLOGY

On autopsy, a complete disappearance of subcutaneous fat tissue
was found, although well-nourished bodies also came to necropsy.
Atrophy of all parenchymatous organs was observed; the liver and
muscles were glycogen-free. Hiilse (1454) found atrophy of the
heart; in a few cases the adrenals were enlarged; however, according
to Lippmann (1455), no pathological changes in the nerves were
observed. According to Paltauf (1456) the anemia is very insignifi-
cant. In the pre-edematous stage, according to Jacobsthal (1457)

HUNGER EDEMA 375

high hemoglobin and erythrocyte values were found. Woltmann
(1458) found anisocytosis, polychromatophilia, high leucopenia,
lymphocytosis and hydration of the blood. In epidemic dropsy,
Maynard (1459) found increased intra-ocular pressure.

METABOLISM

Schittenhelm and Schlecht (I.e. 1443) conducted metabolism
experiments with a diet regarded as productive of edema (45 to 55
grams protein and 1100 to 1800 calories). It was soon established
that a characteristic assimilative failure was not shown by the
patients, since they gained weight on more protein and fat and re-
tained nitrogen. Zondek (I.e. 1447) had some patients who excreted
six liters of urine daily, with 40 grams, and more, of sodium chlo-
ride, accompanied by a normal nitrogen content. Phosphoric acid
excretion was high (4 to 5 grams); Hiilse (I.e. 1454) was able to
demonstrate a good protein assimilation, but believed that the
de-aminization of the amino acids was faulty. Franke and Gottes-
mann (1460) studied the rate of elimination of urea, sodium
chloride and other salts, and found a retardation. The pathological
chemistry of hunger edema was considered by Feigl (1461), who
studied the cases of Rumpel. Schittenhelm and Schlecht (I.e. 1443)
found the protein content of the blood low, 4.5 to 5.7 per cent instead
of 7 to 9 per cent. The erythrocytes showed a lowered lipoid con-
tent. In the opinion of these investigators, the low figures are not
to be attributed merely to the dilution of the blood, for as the edema
disappears, the blood values drop still more. The blood sugar is
either normal or increased. The composition of the edematous
fluid resembles that of exudates. At the height of the edema, a
retention of water and chlorides is noted. On the disappearance of
the edema, considerable urea and sodium chloride excretion is found,
which is increased still more by thyroidin and decreased by adrenaline.
Pituitary extracts are effective, but only on the sodium chloride
excretion.

THERAPY

Here too, there is no uniformity in the literature. Zak (1462)
believed that hunger edema could be cured not only through the
addition of animal products (eggs, raw calves' liver), but also by
means of carrot juice. Maase and Zondek (1463) were of the opinion
that the addition of 100 grams fat could arrest the progress of the

376 THE VITAMINES

disease. Schiff (I.e. 1444) sought the cause of the disease not in
the lack of calories, per se, but of potatoes, fresh vegetables and
milk; addition of yeast, beer and fat had no therapeutic effect.
Knack and Neumann (I.e. 1450) obtained good results with potatoes.
Reach (1464) made use of organotherapy, in the form of testicles,
with good results, v. Hosslin (1465) and Schittenhelm (1466)
believed mainly in the effect of protein therapy. Isenschmid (1467)
attributed certain edemas, like those occurring after dysentery and
other protracted diseases, to the same causes that bring about
hunger edema, and in his opinion, they should be treated in a similar
manner. Most investigators manifest a particular belief in protein
of animal origin as a therapeutic measure. The above therapy con-
stitutes the sum and substance of the leading ideas as regards the
etiology of hunger edema.

ETIOLOGY

Because of the great lack of fat during the war, some investigators
associated the etiology with this factor. We have just seen that
this is not likely and that it has to do chiefly with a lack of vitamine
A, and most animal products used contain more or less of this sub-
stance. Aron (1467a) considers that hunger edema is due to an
insufficient amount of calories in the diet. Park (1468) was of the
opinion that the edema was associated with a lack of protein and fat,
and an excessive water intake. Rubner (1469) and Determann (1470)
attributed it to the lack of protein. The latter conception is also in
accord with the experience gained in the studies on parenchymatous
nephritis by Epstein (1471) and on renal edema by Allbutt (1472).
Further confirmation is given by the study of the development of
experimental edema in animals, especially of rats and mice. Tachau
(I.e. 1244) stated that when the correct relationship between proteins
and carbohydrates is disturbed by the addition of the latter, edema
develops. Denton and Kohman (1473) observed that when rats
are fed on carrots, casein, starch, butter or lard and salts, this diet
is adequate only when the carrots provide half of the total calories.
However, if this relationship is changed by the addition of starch or
fat, then edema develops in a great percentage of the animals.
Kohman (1474) carried this idea further, in that by means of the
above diet, especially with the addition of water, the animals
developed edema, which was cured by the administration of casein.
No exact figures are given as to the food intake, but in any event.

HUNGER EDEMA 377

we have already voiced our opinion, in the introduction to this chap-
ter, against the conception of a lack of protein as an etiological
factor in the development of hunger edema. By the addition of
casein, it is possible, first of all, that there might have been an
increase in appetite, with a consequently greater food and vitamine
intake. The considerable quantity of water not only diluted the
vitamine and protein content of this diet, but also caused great loss
by washing out. Kohman tried to get around these difficulties by
showing that on a synthetic diet prepared in the usual manner, a
small food intake, per se. is not followed by edema. However, since
the diet was properly constituted, in this case, the results cannot
be applied to the carrot diet.

Another hypothesis deals with the role of the adrenals in the
development of edemas; this was formulated by McCarrison (I.e.
290 and 608). He showed that in beriberi, there was an increased
adrenaline output which was supposed to be responsible for the
development of edema. McCarrison held that butter, but not all
fats, contains a substance (apparently vitamine A) which is protec-
tive against edema. This idea was adopted by Bigland (1475), in
a practically unchanged form, who regarded an increased or de-
creased suprarenal activity as the causative factor in the explanation
of hunger edema and pellagra, respectively. Nixon (1476) assumed,
as the cause of edema, insufficient calories (fat?) and protein and
excess of water; adrenaline was also thought to play a part. In
conclusion, we shall mention the pathogenesis assumed by Schitten-
helm and Schlecht (I.e. 1443). They ascribed the cause to the
lack of protein and fat, the effect of cold and hard labor helping to
bring about the disease. They recognized that hunger edema occurs
on a diet rich in carbohydrates. The differentiation from beriberi
lies in the bradycardia; on the other hand, scurvy-like symptoms,
such as stomatitis and bleeding of the gums were observed.
Hemeralopia and xerosis of the eyes were also noted quite frequently.
Administration of vitamine B exerted no therapeutic effect. There
is a great analogy to "Mehlnahrschaden."

Our presumption is that we are dealing here with a complication
of causes, although the disproportion between the individual dietary
constituents, as well as an inadequate diet in relationship with the
hard labor stands in the foreground. However, in this case too,
further work is necessary in order to arrive at an explanation of the
problem.

CHAPTER IX

PATHOLOGICAL CONDITIONS IN WHICH THE LACK OF VITAMINES
MAY BE SUSPECTED

It is difficult, if not impossible, to determine in the conditions
mentioned below, at the present time, if the described effect is due
to the vitamines, per se, or to the other nutritive constituents admin-
istered simultaneously. The subject may be cleared up only when
we shall have the vitamines available in larger amounts in the pure or
almost pure state.

KALLAK

Little (1477) reported, in 1908, on a disease called "kallak,"
occurring among the Eskimos in New Foundland. The disease
consisted in a marked pustulous dermatitis on the hands, elbows
and other parts of the body, with persistent itching. The disease
is usually curable. The natives already know, from experience,
that the disease makes its appearance when only a small quantity
of seal meat is available. It occurs in autumn, when they live
mostly on fish and when wild berries become scarce. Their diet
consists of seal meat, carribou, birds, fish and berries, and is very
rich in protein. The disease is very predisposing to scurvy, and
Little believed that it might be an avitaminosis.

TRENCH SICKNESS

Bruntz and Spillmann (1478) observed the disease among the
French soldiers in the trenches. It consists partly of neuritic
symptoms (parasthesias, pains, motor disturbances, asthenia), and
scorbutic symptoms in the mouth, especially the gums. Following
a better diet (fresh vegetables and fruits), the number of cases
decreased markedly. It is related to scurvy, with a partial lack of
other vitamines, and is made more severe by cold and over-exertion.
Psychic phenomena and uncleanliness apparently also play a great
role in the pathogenesis. All the extremities were affected. This
disease was also studied by Mercier (1479) and Chauvin (1480).

378

PATHOLOGICAL CONDITIONS DUE TO LACK OF VITAMINES 379
INTESTINAL STASIS, ETC.

Following his interesting findings in the intestines in avian beriberi,
McCarrison (1481) sought to apply his results to an explanation
of some obscure intestinal disorders. He thought for example, that
properly nourished children showed less tendency towards intussus-
ception. Among the diseases which McCarrison associated with
the vitamines is intestinal stasis, which was treated by Sir Arbuthnot
Lane (1482), and later, intestinal toxemia and the so-called "coeliac
disease," described by Still (1483). The latter observed, among
41 cases, 4 that had scurvy previous to developing coeliac disease.
As an indication of the lack of vitamines, McCarrison pointed out
dilatation of the stomach, air pockets in the intestine, impairment
of the neuro-vascular control of the gastro-intestinal tract, formation
of stomach ulcer and especially colitis. Mackenzie Wallis (1484)
adopted the view of McCarrison in regard to coeliac disease, in that
he made use of vitamine therapy in this condition among children,
with good results.

STERILITY

Reynolds and Macomber (1485) found that when rats are fed
on inadequate diets (also in the case of vitamine deficiency) they
exhibit a marked diminution in fertility. This viewpoint which is
well borne out by laboratory experience has been considered in the
treatment of analogous conditions in man and favorable results have
been reported.

HEMERALOPIA

We have already seen that in the various human avitaminoses,
hemeralopia was frequently observed. This is not associated with
any particular vitamine, but arises unspecifically, perhaps on any
kind of inadequate diet. We have nevertheless found data in the
literature pointing to a widespread occurrence of this eye affection,
which was favorably influenced by cod liver oil. Ishiwara (1486)
saw such cases in Japan. He studied the lipoid content of the blood
and found it decreased. When the disease was cured, the figures
returned to normal. Tricoire (1487) observed among French soldiers,
320 such cases which were promptly cured by cod liver oil.
H. Smith (1487a) is of the opinion that hemeralopia has mostly been
simulated during the war since the visual field was not diminished as

380 THE VITAMINES

in true cases. McCollum, Simmonds and Parsons (I.e. 1222a) re-
gard hemeralopia as of dietetic origin, and curable with cod liver
oil. Appletpn (1487b) considers hemeralopia and snow blindness,
as it occurs in polar regions (Labrador), due to the strong sun rays
on the basis of a nervous disturbance and supposed to occur only
in men who work in the open. Pick (1488) and Feilchenfeld (1489)
reported on the influence of war diets upon various eye affections.

EXOPHTHALMIC GOITRE (GRAVES DISEASE)

In the study of experimental rickets in dogs, Edward and May
Mellanby (I.e. 408a and b) made the interesting observation that
on feeding certain fats, like butter, a hyperplasia of the thyroid
develops which can be cured by cod liver oil. This observation has
been made use of in human cases and good results have been obtained,
though the number of cases up to the present have been insufficient
to permit of definite conclusions being made.

THE SIGNIFICANCE OF VITAMINES IN INFECTIONS

Since an inadequate diet, especially the deficiency in vitamines,
lowers the resistance towards infections, an insufficient production
of antibodies was first thought of as an explanation. The question
has been studied experimentally by some investigators. Hektoen
(1490), in rats on artificial diet, and Zilva (1491), in rats and guinea
pigs on a vitamine-poor diet, observed a normal formation of anti-
bodies. On the other hand, according to Kleinschmidt (1492) the
hemolysin formation does not attain its normal value on an un-
suitable diet. It is well known that the method of feeding plays an
important part in various infections, as has been shown by E. Thomas
(1493), Valagussa (1494) and Peiser (1495). The latter reported
especially on the influence of the dietary fat on the resistance to
infection. During the course of our discussion, we have already
pointed out the possible relationship between the vitamines and
some infections, and we must call attention to the fact that since the
recognition of the nature of ophthalmia, this matter has assumed
greater significance. The known data on ophthalmia indicate
perhaps, that they may be partially applied to other infectious
conditions. For here, we have a local infection that may be
influenced specifically by a vitamine therapy.

PATHOLOGICAL CONDITIONS DUE TO LACK OF VITAMINES 381

A further field in which the vitamines may be of significance, is
the diet in various acute, and especially chronic infections. During
convalescence, after acute infections, the diet is of greatest impor-
tance. It happens occasionally that in all these conditions, quite
a one-sided diet is given for weeks at a time, for example, flour
preparations, because of which recovery is impeded, and the resis-
tance of the patients is further decreased. These conditions occur
especially after typhoid fever, as Morrison (1496) showed. The
question of the diet in typhoid was studied by Barker (1497), while
Coleman (1498) Walton (1499), Carter, Howe and Mason (1500)
laid emphasis on the caloric value of the diet in convalescence.
Coleman spoke of the necessity of a well-balanced diet, with which
he treated 222 patients, with the result that the mortality was 50
per cent less. Combe (1501) described the influence of the war
diet on the course and convalescence of typhoid in Germany.

Isenschmid (I.e. 1447) believed that some edemas, which occur
after dysentery or other protracted diseases, are of dietary origin and
belong to the type of hunger edema. In all these cases, it is important
that the diet, even though small in quantity, in accord with the
appetite of the patient, must be complete in its composition. It is
not without reason that milk is so often used for the above purpose.
If the disease or convalescence is too long drawn out simultaneously
with an unsuitable diet, then it is possible for a real avitaminosis
to develop.

OPHTHALMIA

The disease, likewise called "xerophthalmia," seems to occur also
endemicalry. Thus, Hirdlika (1502) described a similar disease
among the Indians in the southwestern part of the United States,
while McCarrison (1503) stated that the disease occurs not infre-
quently in India, on a diet of rice and vegetable oils, and is curable
by cod liver oil. The disease consists in an infection of the con-
junctiva, which, under certain conditions, may lead to a destruction
of the cornea, falling of the lens and the iris, and total blindness.
•Mori (1504) was one of the first to describe the condition somewhat
more in detail. An outbreak of this disease, called "hikan," which
developed at a time of partial famine, was mentioned by Mori, who
investigated 1400 cases in detail. It rarely occurred among fisher-
men, and could be cured with chicken liver. Czerny and Keller

382 THE VITAMINES

(1505) observed the development of the disease on a carbohydrate-
rich diet. Falta and Noeggerath (I.e. 18) were perhaps the first to
produce this disease experimentally in animals. Later, this con-
dition was observed in rats by Knapp (1506). Already at that time,
he believed that a similar condition could occur in children. Since
1913, there have appeared in rapid succession, the papers of Osborne
and Mendel, as well as those of McCollum, already mentioned, the
cause of the disease being associated with the absence of certain fats
in the diet. In the end, this disease was regarded as an avitaminosis,
caused by the lack of vitamine A. This eye affection was also studied
in Germany by Freise, Goldschmidt and Frank (1507), working on
rats. Goldschmidt (1508) stated that the disease could be cured by
a small amount of skim milk. Later it was also produced in mice
and rabbits and although Nelson and Lamb (1509) stated that they
could not produce it in guinea pigs and chickens, we have information
to the effect that ophthalmia has been produced in guinea pigs.
We have personally seen numerous cases in chickens, terminating in
total blindness. Recently, we have been corroborated by Guerrero
and Conception (1510), who observed the disease in chickens fed on
white rice.

In recent years, ophthalmia has been frequently noted in
children. A particularly careful investigation was made by Bloch
(1511) in Copenhagen. He observed, during a period of 5 years in
the children's clinic, 40 cases among children fed on a highly centri-
fuged milk, fat being given in the form of vegetable margarine. A
cure was obtained with whole milk or cod liver oil. The diet in
these cases appeared to be inadequate also in other respects. Bloch
(151 la) believes that ophthalmia occurs quite frequently in Den-
mark and may often be the cause of blindness. Monrad (1512)
and R0nne (1513) reported on the same condition in Denmark.
In addition, one case was reported in France by Sztark (1514) and
in the United States by Parker (1515). Sztark's case was a 27
months old baby fed on vegetable soups, without milk; cod liver oil
effected a cure in about 10 days.

The nature of the disease

Lately, the investigators have been more and more inclined to
assume that ophthalmia is associated with a lack of vitamine A.
The pathogenesis of this disease is not so very simple, since we are

PATHOLOGICAL CONDITIONS DUE TO LACK OF VITAMINES 383

dealing here with a new conception in pathology. According to
Macfie (1516), it is a non-contagious infection which may be pre-
vented and cured by diet. Funk arid Macallum (I.e. 86) reported
that they could prevent this eye disease by antiseptic treatment, and
this was concurred in by Bulley (1517). Later, however, from the
same laboratory in which Bulley worked, the report of Stephenson
and Clark (1518) appeared refuting Bulley's findings. They believed
that Bulley's results were due to the contamination of the casein by
vitamine A. In these cases, the eyes were also examined bacteri-
ologically. The infection was shown to be unspecific, and it is of
interest that before the development of the disease, no histological
changes were found to account for the slight resistance towards the
infection. In addition, Stephenson and Clark found that not all
animals lacking vitamine A develop ophthalmia. Wason (1518a),
who called the disease "ophthalmia," arrived at the same conclusion
on the basis of her findings. The specificity of ophthalmia, as
being due to lack of vitamine A, has recently been studied by Emmett
(1519). His statistical data are as follows:

1. Without vitamine A — 122 rats; ophthalmia in 120 = 98.3 per cent.

2. Without vitamine B — 103 rats; ophthalmia in 0 = 0.

3. Normal diet — 216 rats; ophthalmia in 0 = 0.

This disease could not be transmitted from one animal to another.
Mendel (1520) observed no ophthalmia among 7000 rats fed on
insufficient protein, and in 225 rats which were given no B-vitamine.
However, out of 136 rats receiving no A-vitamine, 69 developed the
disease. The author has personally never worked with great num-
bers of rats; we have had usually about 30 rats at one time under
observation, but on several occasions we have seen ophthalmia in
the presence of sufficient butter or cod liver oil. Thus, Funk and
Dubin (I.e. 331) have recently noted two such cases out of 30 rats.
These two manifested intermittent ophthalmia, lasting for a short
time, disappearing, and re-appearing in a few days, but permanent
lesions were recognizable. This form was favorably influenced by
yeast. It is worthy of note that Guiral (1521) reported ophthalmia
in children, who were completely cured by orange juice in 8 to 10
days. We are in possession of the private data of specialists who
observed the eye condition in the presence of vitamine A, which
was favorably affected by orange juice. Still, it must certainly be

384 THE VITAMINES

admitted that the absence of vitamine A favors the occurrence of
ophthalmia in quite a specific manner, while contrary findings require
further explanation. It is possible, for example, that in the presence
of cod liver oil, but with a small food intake there might be insuffi-
cient vitamine A supplied to the animal. All substances containing
vitamine A may be used in the therapy of ophthalmia.

TUBERCULOSIS

That diet plays an important part in the therapy of tuberculosis,
is too well known to require further discussion. An added confirma-
tion of this is found in the reports on the influence of the war diets,
of which we have already spoken. However, we may mention once
more the communication of Adams and Hamilton (I.e. 1048), dealing
with the increase in tuberculosis in Germany during the war. Ham-
burger (1522) also, in his report on the war diet of children, pointed
out the dependence of the number of cases of tuberculosis on the diet,
stating that it was due to the lack of fat. Stolzner (1523) likewise
reported on the effect of the diet. Geoghegan (1524) reported on
some cases in the West Indies, which, in his opinion, are due to the
prevailing poverty. On these islands, with good climatic conditions,
there should be very little tuberculosis. Richet (1525) believed that
meat juice contains ferments which act favorably on tuberculosis.
Woodcock and Rustin (1526) stated that the diet in tuberculosis
must be rich in protein and fat (in the form of milk or milk products) .
If margarine is used, it must be oleo-margarine which contains vita-
mine A; the diet must also contain plenty of fruits and vegetables.
Gardey (1526a) finds that certain symptoms of pulmonary tuber-
culosis resemble those of a deficiency disease. He suggests that
collecting of statistical data on the relationship between the con-
sumption and the composition of the diet in various countries would
be helpful in the study of this condition.

Just how important the diet is in tuberculosis, is shown by the
experimental investigations of Weigert (1527). He showed in young
pigs that on a constant protein content, tuberculosis develops more
frequently in the presence of carbohydrates than in that of fats.
Similar investigations were made by E. Thomas and Hornemann
(1528), who showed, in pigs infected with tuberculosis, that the
animals manifested tuberculosis less on protein and much more on
carbohydrates, while on fat, varying results were obtained. In

PATHOLOGICAL CONDITIONS DUE TO LACK OF VITAMINES 385

some communications on tuberculosis, particular attention was called
to the significance of the vitamines. Re"non (1529) said that on
vitamine-poor diets, a decreased immunity towards tuberculosis is
evident, and Muthu (1530) likewise believed in the important role
of the vitamines in the development of this disease.

LEPROSY

Button (1531) recently stated that in the development of leprosy
the diet, especially the vitamines, plays an important role. Sir
Jonathan Hutchinson (1532) believed that leprosy was associated in
some way with fish consumption. On perusing the literature, we
see in an article by Deycke (1533), that there is no basis for this
.assumption except, perhaps, the statement that in the treatment, a
satisfactory diet is of significance. Among other known facts, we
see in a paper by Underbill, Honeij,, Bogert and Aldrich (1534) that
in leprosy, certain chemical changes in the bones are apparent,
indicating, perhaps, a disturbance in the calcium and magnesium
metabolism. The atrophy of the bones in leprosy is associated
with the large requirements of these patients for calcium. If calcium
is added, marked retention results. We may, perhaps, associate this
with the action of vitamine A. Vokurka (1534a) observed an
increase in the number of lepers in Bosnia and Herzogowina after
the war, and attributed it to malnutrition. It is obvious that
leprosy occurs largely among the poorer people, but any possible
relationship with the vitamines has not been demonstrated. Never-
theless, it would be worth while to go further into the question
of diet.

PNEUMONIA

The many investigators in the field of experimental vitamine
research have quite regularly reported the frequent occurrence of
infections of the respiratory apparatus on diets poor in vitamines.
Similar observations have been made in man by Fleming, Macaulay
and Clark (I.e. 910) in South Rhodesia. Great epidemics were noted
there on a diet, previously described, which apparently leads to a
mixed avitaminosis. Vaccine therapy was used without success, and
there were 686 deaths out of 2251 cases in 1908, with considerable
meningitis and tuberculosis. At the same time, the avitaminosis was
evident in 100 out of 700 laborers. Here, when a change was made
in the diet, splendid results were obtained. Not only did the avitam-
inosis disappear but the pneumonia decreased also.

LIBRARY

CHAPTER X

INFLUENCE OF NUTRITION (VITAMINES) ON THE ACTION OF SOME

POISONS AND UPON PATHOLOGICAL CONDITIONS OF

NON-INFECTIOUS ORIGIN

The older papers of Reid Hunt have already shown that changes
in the diet are not without effect on the toxicity of different drugs.
Similarly Reach (1535) has shown that the resistance towards the
cramps-producing poison, picrotoxin, is greater in mice on a meat
diet than on a bread diet and he assumed that he probably had to
deal with the action of some unknown substances. Salant (1536)
and Salant and Swanson (1537) observed that young carrots had a
far greater protective action against the poisonous effect of sodium
tartrate in rabbits, than did old carrots. The same results were
obtained with sweet potatoes and carrot leaves. On oats and sugar,
the toxicity was markedly increased, and the differences found were
not due to increased diuresis. Cats behave otherwise, no difference
being apparent; here too, however, the effect of the poison was
increased about 40 per cent in starvation.

ANEMIA, ETC.

Here we have the work of Pearce, Austin and Pepper (1538), who
studied the influence of the diet in anemia, after splenectomy. They
observed in dogs that raw meat acts much better than cooked, and
they thought this difference was due to the vitamine content. Mad-
sen (1539) is of the opinion that the vitamines play a part in chlorosis
and anemia. Geiling and Green (1539a) subjected this question
to experimental test. If rats were given a diet poor in protein vita-
mines or salts, then the blood regeneration after a hemorrhage was
markedly retarded. Davis, Hall and Whipple (1540) produced
necrosis of the liver by chloroform and studied the effect of different
diets on the regeneration of liver cells. Muscle tissue and fats of
various sources were without effect but feeding of liver, kidney and
brain resulted favorably.

Campbell (1541) and van der Bogert (1541a) thought that the
occurrence of adenoids in children is of dietetic origin, but we do not

386

INFLUENCE OF NUTRITION ON POISONS 387

know how correct this assumption may be. The significance of
vitamines in certain orthopedic cases is based on a better foundation.
Schodel and Naumwerk (I.e. 998) believed that scurvy predisposes
to coxa vara and other conditions, diagnosed as congenital hip-
joint dislocation. Hess (I.e. 1022) saw, in one case of scurvy, a bone
affection similar to coxa vara. During the war, Hammer (1542)
observed that some fractures healed with great difficulty, a dis-
turbance in metabolism being justifiably suspected. Peckam (1543)
believed that some deformities like scoliosis, bow-legs, flat feet,
etc., are of dietetic origin. He thought that most orthopedists are
so occupied with the purely orthopedic aspect of their cases that
they pay absolutely no attention to the practical important etio-
logical factors. The presumptions of Peckam are certainly partly
justified because of the relationship between rickets and bone
formation.

We shall now discuss two diseases which are related more to a
luxury consumption than to an insufficient dietary, namely, diabetes
and cancer.

DIABETES

We see already the justification for the conception that this disease
is related to luxury consumption in the papers on the effect of the war
diet. Magnus-Levy (1544) observed that during the war, there was
a marked decrease in the number of diabetes cases, whereas, before
the war, they had been constantly increasing. Gerhardt (1545) had
the same experience in Wiirzburg. Two phases of the diabetes
question interest us here. First, the possibility of an antidiabetic
substance of a vitamine type in the food, and second, the danger of
an altogether rigorous dietary restriction in the usual therapy. The
first possibility was justified, to a certain extent, for a specific action
on diabetes was ascribed to v. Noorden's oatmeal cure. Even
Magnus-Levy (1546) tried to concentrate this hypothetical substance
by alcoholic extraction, but without success. Boruttau's (1547)
experiments resulted differently. He found that pancreas extracts,
yeast and yeast extracts inhibit the cleavage of glycogen in the iso-
lated heart, while extracts from the peripheral oat layers decreases the
sugar elimination in diabetic dogs and in man. Similar results were
obtained by Rose (1548) with one of the substances isolated from the
pancreas. The composition of the diet, especially its carbohydrates,

388 THE VITAMINES

influences the blood sugar content not only of diabetics but also of
normal subjects. This was reported on by Jacobson (1549), v.
Moraczewski (1550) and McCay, Banerjee, Ghostal, Dutta and Ray
(1551). The latter described conditions in India which develop
there on a diet rich in carbohydrates, and which gradually lead to
diabetes. These conditions strongly resemble experimental gly-
cosuria, which we have already described in pigeon beriberi.

Considering the question of therapy in diabetes, we see the pos-
sibility of producing an avitaminosis by adhering too closely to a
strict diet. Burger (I.e. 1452) and Schittenhelm and Schlecht (I.e.
1443) state that in diabetes, on an oatmeal diet, they observed
edema resembling hunger edema. A similar observation was made
by Wilder and Beeler (155 la). An editorial in the Journal of the
American Medical Association (1552) warned of the dangers of a
decrease in the protein of the diet with a simultaneous increase of the
carbohydrates, a regime which produces real diabetics from latent
cases. In addition, there is the actual danger of an avitaminosis.
As regards the starvation therapy in diabetes, we have asked our-
selves the question as to whether the organism, in starvation, suffers
first from a lack of vitamine or of the usual dietary constituents.
The author and Dubin (unpublished data) undertook to test this
question experimentally. However, no difference was noticeable in
the time of survival between the pigeons receiving either vitamine A
or B, or both, and those that were starving. In addition, no beriberi
was observed in the starving pigeons.

CANCER

Cancer is a disease of excessive nutrition, although McCarrison
(1553) believed that under certain conditions, a lack of vitamines
may lead to cancer of the stomach. He saw one such case among
monkeys. That this disease is associated with luxury consumption,
is best recognizable from the statistical figures compiled during the
war and from insurance statistics. Hoffmann (1554) said that cancer
occurs more frequently in cities than in the country, and more among
the well-to-do than among poor people, the reverse being the case in
tuberculosis.

The cancer question is of interest from two points of view, first, the
possibility of a chemical substance as an etiological factor and second,

INFLUENCE OF NUTRITION ON POISONS 389

the influence of the diet on the growth of neoplasms. As for the
first factor, we can not take it up in such great detail as we should
like to, unfortunately, since it does not fall in the sphere of this book.
We were one of the first to point out this possibility, but we must
admit that aside from the influence of the diet on the growth of
experimental tumors and a possible chemical cause of certain neo-
plasms, such as the chicken sarcoma of Rous and analogous tumors,
we have no firm basis for our conception. If we (1555) choose this
line of thought in preference to the other known ones, it is because
cancer research, in the hands of pathologists, has" yielded very little
tangible results. Some of the known pathologists, like Ewing (1556)
and Leo Loeb (1557) were inclined to a similar view at one time.
Certain experimental papers like those of Calkins (1558), who
reported on the stimulation of protozoa (Didinium nasutum) by
tumor extracts, may be interpreted in the above manner; still, it is
possible that it may be associated with the influence of a dietary or
vitamine addition. The various investigations dealing with the
effect of pregnancy on the growth of neoplasms may also be inter-
preted from the viewpoint of the chemical theory. From these
investigations, we may see that the embryo contains a substance
that stimulates the growth of neoplasms. The growing embryo,
however, according to v. Graff (1559) and Slye (1560), needs this
substance for its own growth, thereby inhibiting considerably the
tumor growth.

That the diet exerts some influence on tumor growth, was suspected
by Ehrlich in his athreptic theory. This theory was tested experi-
mentally by Jansen (1561) and Haaland (1562) who established the
inhibiting influence of under-feeding. Similar experiments were
also undertaken by Cramer and Pringle (1563), Rous (1564) and
v. Jaworski (1565). The latter believed that under-feeding inhibits
the growth of tumors. In his opinion, carcinomas were an exception
since, under such conditions, they grew even more rapidly. The
dietetic aspect of cancer research was stimulated by vitamine studies.
Sweet, Corson-White and Saxon (1566), in their rat and mouse experi-
ments, used the Osborne-Mendel diet and saw that the growth of
tumors on a vitamine-poor diet was markedly arrested. In quick
succession, there appeared papers on the same subject by Hopkins
(1567), Centanni (1568), Rous (1569), Drummond (1570) (this work
being very exhaustive), Benedict and Rahe (1571) and Sugiura and

390 THE VITAMINES

Benedict (1572). Van Alstyne and Beebe (1573) believed that they
had stimulated tumor growth by the addition of lactose to an artificial
diet, but this was evidently due to a contamination of the lactose
with vitamine B. In a similar manner, the author (I.e. 244, 245, and
246) showed in 1913 on chickens, that the chicken sarcoma of Rous
grows much slower on a vitamine-poor diet than on one rich in vita-
mines, and these results were confirmed by Drummond (I.e. 247).
Levin (1574), however, could not corroborate these findings and
Frankel and Fiirer (1575) expressed themselves against this phase of
cancer research. The few negative results, naturally, cannot have
any effect on our conclusions in comparison with the conformity of
the conclusions of a great number of investigators in this field.
Undoubtedly, the restriction of the vitamines in the diet markedly
inhibits the growth of neoplasms. This measure would have been
of practical value, if it had not been evident that the tumor tissue
shows a greater affinity for the vitamines than does the somatic
tissue. This indicates that in order to obtain a complete success,
the vitamines would have to be so far curtailed that the organism
itself would die. That certain substances govern tumor growth, is
evident from the paper by Rondoni (1576), who showed that the
sarcoma tissue could favorably influence the growth of experimental
tumors. The same conclusions were obvious from the work of
Murphy (1577), who allowed rat tumors to grow in chicken embryo,
it being impossible to do so in grown chickens. From this, it is
evident that in rapidly growing embryonic tissue, a substance is
present which stimulates the growth of neoplasms. That this view
is justified, is clear from the work of Funk (1578), who showed that
when the mouse chondroma of Ehrlich is implanted in the rat, the
tumor tissue is resorbed in a few days. However, if the mouse tumor
is fed to the rat in large quantities, then not only does the tumor
develop but it may be transplanted in three generations of rats so
treated. Here, in our opinion, it cannot be denied that the mouse
chondroma contains a substance which acts specifically on the
growth of a similar tumor. At the same time, the influence of the
diet is shown very well. The induced tumor possessed, as the
illustration shows, for the most part, the same histological structure.
The dietetic restriction for the purpose of treating cancer has also
been used in practice. Bulkley (1579) used, for this purpose, a diet
so chosen that it contained almost no animal protein. Since this

INFLUENCE OF NUTRITION ON POISONS

391

diet could be regarded for many reasons as pellagra-producing, we
inquired as to the health of the patients and the duration of the
therapy. Dr. Bulkley replied that a number of patients lived
on the diet without showing any signs of pellagra. Copeman (1580)
tried a similar therapy with a diet that was almost vitamine-free.
At the time of his preliminary communication, his experiments had
lasted from three to four months, and a detailed report was still
to be made. His patients increased in weight. The dietetic treat-
ment of cancer is at present not in favor with the medical profession.
We believe that it would be of value to cancer research if this
disease were regarded more as a metabolic disturbance.

'3

.<* -

>•-"• rW '&*'«• V*?G
* (ty \^ • ( ^ ^ ?-•> - jj,_ .,>.•..

^Sa^^^'"'*J

; ",»W '^

•^,2^1

• , y

A B

FIG. 73. A, EHRLICH'S MOUSE CHONDROMA; B, THE SAME TUMOR GROWING IN
A RAT AFTER TUMOR FEEDING

We shall now consider two problems in which the vitamines are
of significance with greater certainty — the influence of the diet in
the development of teeth, and the cause of cystic calculi and similar
formations.

THE DEVELOPMENT OF TEETH

It was suspected by Kunert (1581) that the diet exerted some
influence on the condition of the teeth. Durand (1582) saw a greater
percentage of carious teeth in children who had been fed on condensed
milk than in those fed on breast-milk. A carbohydrate-rich diet
in particular, was held to be responsible for caries. Black and

392 THE VITAMINES

McKay (1583) frequently observed a defective enamel in certain
districts in the Rocky Mountains, where the diet was inadequate.
Castilla (1584) often observed caries and defective enamel after
colitis and other disturbances of the stomach and intestinal tract
in children from 1| to 4 years old. Considerable progress has been
made in this field since the problem has been undertaken experi-
mentally.

Miller and Gies (1585) studied the question of teeth formation in
rats from the standpoint of the calcium and magnesium metabolism.
M. Mellanby (1586) studied the condition of the teeth in young
rachitic dogs. On diets that were poor in vitamine A, the following
changes were visible :

1. Delayed falling out of the milk-teeth.

2. Delayed appearance of permanent teeth.

3. Disarrangement in the position of the teeth

4. Lack of or defects in the enamel.

5. Diminished calcium content.

In quickly growing teeth, the defects were even more pronounced.
Vitamine A, in the form of cod liver oil or butter., exerted a very
favorable influence. Zilva and Wells (1587) observed, in guinea
pigs and monkeys on a scurvy-producing diet, a fibroid degeneration
of the tooth pulp. These changes occur very early, often before all
other scorbutic symptoms. The findings on scorbutic guinea pigs
were confirmed by Percy R. Howe (1588), Robb, Medes, McClendon,
Graham and Murphy (1588a) and developed further experimentally
by P. R. Howe (1588b). The latter managed, by means of a partial
lack of vitamine C, to keep the animals (guinea pigs and rabbits)
alive for about one year, so that the results were more definite.
The teeth were carious, poor in calcium and bent, with
Pyorrhoea alveolaris. There was also observed a disease of the
joints, resembling rheumatism or Arthritis deformans. Ballantyne
(1589) found that 98 per cent of pregnant women show carious teeth.
This may well be explained by the greater requirements for vita-
mines, calcium salts, etc. Sinclair (1590) rightly emphasized that
precautions for the good condition of the teeth of the child must
begin with the pregnant mother, who should receive the best possible
nourishment. The effect of the diet on the teeth does not appear
to be related to any one specific vitamine, but vitamine A, because

INFLUENCE OF NUTRITION ON POISONS 393

of its relationship to calcium metabolism seems to play the most
important part. There is no doubt that the significance of the
factors mentioned will soon receive greater attention in practice.

CALCULI

Osborne and Mendel (1591) found phosphatic calculi in the
bladder in 857 rats kept on a diet poor in vitamine A. Padua (1592)
saw a definite relationship between beriberi and calculi in the
Philippines. Out of 58 cases of phosphatic calculi, 11 had beriberi;
in 18 cases, under-nutrition was observed, but without any beriberi
manifestations; 27 cases occurred in well-nourished patients. The
phosphatic calculi were found mostly in children and young people;
while urate calculi were observed later in life (after 50 years).
Kirschner (1593) and Clemm (1594) observed that in Germany, on
diets poor in fat, gall-stones are found more commonly. All these
cases are associated with the indirect influence of the vitamines,
presumably decreasing the resistance to infections, in which connec-
tion vitamine A is of special significance. For it is apparent that a
local infection is the real cause of this calculus formation.

This concludes our data on the subject. We are well aware that
despite the ten years of experimental and clinical research, most of
the problems discussed by us have only been scratched on the surface.
This is due largely to the fact that such slow progress has been made
in the chemistry of the vitamines. In this connection, the frequent
modifications of the nomenclature is not of the slightest value, and we
must still await the investigator who, through tedious and patient
work, will make the decisive step forward — the recognition of the
exact nature of the vitamines.

Before closing, we shall add a few words touching upon the prac-
tical aspect of the vitamines. From what has already been said,
it is evident that in a properly constituted diet there is no danger
of an avitaminosis. In England and in the United States recently,
there have appeared on the market a number of vitamine prepara-
tions whose purpose was to exploit the present popularity of the
vitamines. There seems to be no doubt that most of these prepara-
tions cannot produce the effect claimed for them. Many investi-
gators, among them Drummond (1595), have expressed the

.06*

394 THE VITAMINES

as being opposed to such preparations, maintaining that the natural
foodstuffs may be used in the therapy and the prevention of avitam-
inoses. In the light of our present knowledge, this appears indeed
to be the case. Still, we do not know how this matter will develop
in the future, and the question can be studied only then when the
vitamines will be obtained in the pure or highly concentrated state.
It is possible that such pure products may manifest properties at
present neither known nor. even suspected. For example, on com-
paring the influence on rickets of vitamine A of butter and cod liver
oil, it was found that only the latter can prevent the occurrence of
the disease; one might therefore be tempted to regard the two sub-
stances as individual chemical entities. As a matter of fact, if
it is true that cod liver oil is about 250 times richer in vitamine A
than is butter, the above difference may perhaps be only
one of concentration. Although most commercial products cannot
withstand sharp scrutiny, it would be desirable not to decry the
present concentration and purification experiments, since they may
perhaps lead to interesting and important results.

GENERAL LITERATURE

In order to give the reader a broader survey of the field, there is
appended a chronological list of some of the work done on vitamines
during the last six years, insofar as the records were available and
insofar as they will not be referred to again in the text. These
references to the existing literature will enable the interested reader
to make himself familiar with the views of other workers. Reports
appearing from time to time in the daily newspapers have not been
considered at all.

1914

FUNK, CASIMIR: Die Vitaminlehre, ihre wissenschaftliche und praktische
Bedeutung. Naturwiss. 2, 121.

HOPKINS, F. G. : Progress in physiological chemistry. Ann. Rep. Chem.
Soc. 11, 188.

HUSSY, P. : Zur klinischen Bedeutung der Vitamine. M. med. W. 61, No. 18.

MELOCCHI: Nuovo orizzonti sull' alimentazione dell' uomo. Napoli.

MENDEL, LAFAYETTE B. : Newer points of view regarding the part played by
different food substances in nutrition. J. A. M. A. 63, 819.

SCHAEFFER, G. : Le be'ribe'ri et les vitamines de Funk. Unprobleme de physio-
logic gene'rale. Biologica 15 Avr., 108.

STERNBERG, W. : Diat und diatetische Behandlung von Standpunkte der
Vitaminlehre und von Standpunkte der diatetischen Kiiche. Arch.
Verdauungsk. 20, 200.

1915

BORUTTAU, H. : Vitamine und akzessorische Nahrungsbestandteile. D. med.

W. 41, 1208.
OSBORNE, T.B., AND MENDEL: Report on Nutrition. Washington. Year Book

Cam. Inst. 14, 378. Also next year 15, 365.
STILES, PERCY G. : The vitamines. The recognition of essential constituents

of the diet hitherto unclassified; deficiency diseases. Am. J. Pharm.

86, 237.
SCHAUMANN: Neue Ergebnisse der Beriberiforschung. Arch. f. Schiffs.

Tropenhyg., 19, 425, 1915; Therap. monatsh., 29, 152, 1915.
WATSON-WEMYSS, H. L. : Recent work on vitamines. Edinb. Med. J., 14,

186, 1914; J. Comp. Path. Ther., 28, 53.

1916

ALSBERG, CARL L. : Biochemical analysis of nutrition. J. Wash. Acad. Sci.,

6, 269.
FISCHER, Louis : Notes on the r61e of vitamines in deficiency diseases. Amer.

Med., 11, 774.

395

396 THE VITAMINES

FUNK, CASIMIR: The vitamines, their chemical nature, their importance in

metabolism and their function in the animal organism. Amer.

Med., 11, 751.
JACOBS, MERKEL H. : Some biological and practical aspects of growth. Univers.

Penn. Bull., 17, Ser. No. 1, Part 2, 131.
McCoLLtJM, E. V. : Malnutrition through errors in the combination of foods.

Amer. Food J., 11, 404.

Mosc, G. : Deficiency diseases. L'ospedale maggiore 4, II, 735.
VEDDER, E. B. : Relation of diet to beriberi, and the present status of our

knowledge of the vitamines. J. A. M. A., 67, 1494, 1916.
VOEGTLIN, C. : Importance of vitamines in relation to nutrition in health and

disease. J. Wash. Acad. Sci., 6, 575; The chemical nature and

physiological significance of so-called vitamines. Science Monthly,

2, 289.

1917

ALB, B. : Les vitamines et les maladies par carence. Revue. Sci., 16 et 23 Juin.
CHRISTENSEN, F. W. : The so-called "vitamines" or "food accessories" and

their importance in the nutrition of animals. Special Bull. N.

Dakota Agr. Exp. Sta., 4, 331.
CORNALBA, G. : Vitamines and new ideas regarding human nutrition. Boll.

chim. farm., 56, 577.
McCoLLUM, E. V. : Supplementary dietary relationship among our natural

foodstuffs. J. A. M. A., 68, 1379. Some essentials to a safe diet.

Ann. Amer. Acad. Polit. and Soc. Sci., 74, 95.

POL, D. J. H. : Deficiency diseases. Nederl. Tijdsr. v. Geneesk., 2, 1212.
STEPP, WILHELM : Einseitige Ernahrung und ihre Bedeutung f iir die Pathologic.

Erg. d. inn. Med. u. Kinderh., 15, 257.

1918

ARON, H. : Akzessorische Nahrungsbestandteile. Berl. klin. W., 55, 546.
EIJKMAN, C. : The importance of vitamines in human nutrition. Chem.

Weekbl., 55, 765.

KLEISSEL, R.: Vitamine. Wien. klin. W., 68, 601; Wien. med. W., p. 547.
McCoLLUM, E. V. : What to teach the public regarding food values. J. Home

Econ., 10, 195; Influence of heat on growth-promoting properties

of food. Amer. J. Publ. Health., 8, 191.
RAMSDEN, W. : Vitamines. J. Soc. Chem. Ind., 37, 53.
SCHAEFFER, G. : Les rccents travaux sur les besoins qualitatifs d'azote chez

les mammife'res et les vitamines. Bull. soc. sci. d'hyg. aliment.

6, No. 5 and 6.

STEENBOCK, H. : Vitamines and nutrition. Science Monthly, 7, 179.
WEILL, E., AND MOURIQUAND, G. : A propos des vitamines et des maladies par

carence. Rev. Sci., 5-12 Juin.

1919

Azzi, A.: Vitamines and oxymones. Riv. Med., 35, 878.
BUHRER, C. : Vitamine. Schweiz. Apoth. Ztg., 57, 377.

GENERAL LITERATURE 397

COMBY, J. : Deficiency diseases. Arch, de me'd. d. enfants., 22, 659.

EDDY, WALTER H. : Bibliographic Review. The Vitamines. Abstr. Bact.,

3, 313.
FORBES, E. B. : Vitamines in human nutrition. Monthl. Bull. Ohio Agr.

Exp. Sta., 4, 299.
G6MEZ, A. : Hormones, vitamines and symbiotes. Repert. de med. y cirurgia,

Bogotd, 10, 463.

GOODBY, SIR K. : Vitamines. Practit., 103, 6.

GUIDI, G. : Recent literature on vitamines. Riv. clin. pediatr, 17, 243.
HALLIBURTON, W. D. : Vitamines. B. Med. J., May 10, 583.
HOPKINS, F. G. : Practical importance of vitamines. Ibid. Apr. 26th, 507.
JACOBY, MARTIN: Die Bedeutung der Qualitat der Emahrung. Ther. d..

Gegenw., 60, 401, 1919.
McCoLLUM, E. V. : Food control from the standpoint of nutrition. Amer.

Food J., 14, 27, 30. The newer knowledge of nutrition. New York,

Macmillan.
PUGLIESE, A. : The vitamine problem. Rend. Reale. Inst. Lomb. sci. lett.

52, 723.

RISQUEZ: Vitamines and symbiotes. Gaceta me'd. de Caracas., 26, 29, 82.
SCHAEFFER, G. : Facteurs accessoires de la croissance et de 1'equilibre. Vita-
mines, auximones. Bull. Inst. Past., No. 1-2, 17, 1, 41.
WEILL, E., AND MOURIQUAND. G. : L'alimentation et les maladies par carence.

J. B. Bailliere, Paris.
WOLFF, L. K. : A study of the inorganic constituents of the animal body and

their significance as a part of the diet. Chem. Weekbl., 16, 1083.

1990

AULDE, J. : Applied calcium therapy. Study of deficiency diseases. Med.

Rec., 97, 257.
BLUNT, K., AND CHI CHE WANG: The present status of vitamines. J. Home

Econ., 12, No. 1, Jan.
VAN DRIEL, B. : Present status of vitamine question. Nederl. Tijdsr. v.

Geneesk k, 1350, Apr. 17.
FORBES, E. B. : Mineral nutrients and vitamines in diet. Ohio State Med.

J., 16, 419.
HAAS, P.: Recent advances in science — organic chemistry. Sci. Progr., 14,

378.

HOJER, A. : Recent research on the vitamines. Hygiea, 82, 449.
LECOQ, R. : Les nouvelles theories de la nutrition. Bull. Sci. pharm., 27, 139.
LEMAIRE, H. : Nourisson, 8, 289.
LINOSSIER, G. : Role of different food elements in nutrition. Paris Me'd.,

10, 349.

LINTON, R. J. : Vitamines. Science and Industry (Australia), 2, 438.
McCLENDON, J. F. : Nutrition and public health, with special reference to

vitamines. Am. J. Med. Sci., 159, 477.
McCoLLUM, E. V.: Nutrition and physical efficiency. J. Franklin Inst.,

189, 421.

398 THE VITAMINES

MOLINAKI, G. : Deficiency phenomena. Riv. med., 36, 182.

POL, D. J. HULSHOFF: One-sided diet and the vitamines. Ned. Tijdsr. v.

Gen., 1, 1625.
PORTIER, P., Vitamines and their mode of action. Bull. Soc. sci. hyg. 8,

521, and 603.

RAMOINO : Policlin. 28, 819.

SEIDELL, A.: Chemistry of the vitamines. J. Ind. Eng. Chem., 13, 72, 1921.
TSCHIKCH, A. : What are vitamines? Schweiz. med. W., 50, 21.
CHICK, HARRIETTS: Role of vitamines in nutrition. Wien med. W., 70, 411,

1920.

NEPPI, BICE: Vitamines. Giorn. chim. ind. appl., 2, 573, 1920.
HALLIBURTON, W. D. : Vitamines. Scientia, 27, 55, 1920.
RAMSDEN, W.: Vitamines. Dent. Rec., 40, 281, 1920.

ASHER, L.: Review of the vitamine theory to date. Deutsch.med.W.47, 510.
BIDATJLT: Vitamines and the preservation of food. Revue sci. 59, 13.
BOTAZZI, FIL.: The problem of vitamines and avitaminoses. Bull. soc. hyg.

9,1.
DALYELL, E. : Accessory food factors and their influence on nutrition. Med.

J. of Australia. 1, 61.

EDDY, W. H. : The Vitamine Manual. Williams and Wilkins, Baltimore.
EMMETT, A. D.: Physiological value of the vitamines. J. Amer. Pharm.

Assocn. 10, 176.
FITCH, W. E.: The necessity for vitamines in the dietary and the role they

enact in constructive metabolism. Amer. Med. 27, 368.
FUNK, CASIMIR: Vitamines and the avitaminoses. Proc. New York Path.

Soc. 20, 119.
HAAS, P.: Recent advances in science. Organic chemistry. Sci. Progr.

15, 377.
HALLIBURTON, W. D.: Importance of the infinitely little. Brit. Med. J. I.,

449; Lane. 1,627.
HARDEN, A.: Vitamine. Chem. and Drugs. 94, 202; Vitamines and the food

supply. J. Soc. Chem. Ind. 40, 79.
HARROW, B.: Vitamines. Essential food factors. Dutton and Co., New

York.
HOPKINS, F. G.: Recent advances in science in their relation to practical

medicine and the nutritional requirements of the body. Lane. 1, 1.
VANLEERSUM, E. C.: Vitamines. Ned. Tijdsr. v. Gen. I, 2137.
LEWI, M. J., AND DUBIN, H. E.: Vitamines and nutrition. N. Y. State J.

Med. 21, 268.

McCARRisoN, R. : Studies in deficiency diseases. Oxford Univer. Press.
QUEAL, E. B. : Vitamines and their relation to deficiency diseases. Colorado

Med., 18, 195.
SHERMAN, H. C.: Vitamines. Physiol. Reviews, October. Vitamines.

Chemical Monographs.: Amer. Chem. Soc.
SPERONI, C. E. : Vitamines, Semana med. 28, 679.

LITERATURE TO THE TEXT1

1. THOMAS, K.: Pfl. Arch. 291, 1909.

2. SCHAUMANN, H. : Die Aetiologie der Beriberi unter Beriicksichtigung

des gesamten Stoffwechsels. Arch. f. Schiffs- u. Tropenhyg. 14,
Beih. 8, 1910.

3. McCoLLUM, E. V., HALPIN, I. G. AND DRESCHER, A. H. : Synthesis of

lecithin in a hen and the character of the lecithin formed. J. Biol.
Chem. 13, 219, 1921.

4. FINGERLING: Die Bildung von organischen Phosphorverbindungen aus

anorganischenPhosphaten. Biochem.Z. 38,448, 1912;Einflussorgan-
ischer und anorganischer Phosphorverbindungen auf die Milchsekre-
tion. Ibid. 39, 239, 1921.

5. ABDERHALDEN, E. : Synthese der Zellbausteine in Pflanze und Tier.

Berlin 1912. Springer.

6. STEPP, W. : Experimentelle Untersuchungen iiber die Bedeutung der

Lipoide fur die Ernahrung. Biochem. Z. 22, 452, 1909.

7. STEPP, W. : Z. f. Biol. 57, 135, 1911.

8. FORSTER, J. : Ueber die Bedeutung der Aschenbestandteile in der Nahrung.

Z. f. Biol. 9, 297, 369, 1873.

9. BUNGE, G. : Lehrbuch der physiologischen und pathologischen Chemie.

10. LTJNIN, N. : Uber die Bedeutung der anorganischen Salze fur die

Ernahrung des Tieres. H. 5, 31, 1881.

11. SOCIN, C. A. : In welcher Form wird das Eisen resorbiert? H. 15, 93, 1891.

12. BUNGE, G. : Weitere Untersuchungen iiber die Aufnahme des Eisens

in Organismus des Sauglings. H. 16, 173, 1892.

13. HAUSERMANN: Die Assimilation des Eisens. H. 23, 555, 1897.

14. HALL, WINFRED S. : Einige Bemerkungen iiber die Herstellung eines

kiinstlichen Futters. Arch. f. (Anat) Physiol. p. 49, 142, 1896;
Ueber die Resorption des Carniferrins. Ibid. 455, 1894.

15. COPPOLA, F. : Sul valore fisiologico e terapeutico del ferro inorganico.

Rendic. R. Acad. d. Line. 6, I, 362, 1890.

16. PASQUALIS, G. : Studi sulla nutritione animale. Atti. d. R. Inst. Veneto

d. Scienze. (7), 7, 1895-6.

17. HENRI QUES AND HANSEN: Ueber Eiweisssynthese im Tierkorper.

H. 43, 417, 1905; Ueber Fiitterungsversuche mit Zein und Gliadin.
H. 60, 105, 1909.

18. FALTA, W. AND NOEGGERATH, C. F. : Fiitterungsversuche mit kiinstlicher

Nahrung. Hofm. B. 7, 313, 1905.

19. JACOB, LUDWIG: Fiitterungsversuche mit einer aus einfachen Nahr-

ungsstoffen zusammengesetzter Nahrung an Tauben und Ratten.
Z. f. Biol. 48, 19, 1906.

20. MCCOLLUM, E. V. : Am. J. Physiol. 25, 120, 1909.

xFor Index to Sectional Divisions of the Literature, see page 476.

399

400 THE VITAMINES

21. WATSON, CHALMERS D. : Influence of excessive meat diet on growth, on

nutrition and on the osseous system. Effect of a meat diet on the
animals and their progeny. Lane. II. 145, 1585, 1778, 1906.

22. WILCOCK, E. G., AND HOPKINS, F. G. : The importance of individual

aminoacids in metabolism. J. Physiol. 35, 88, 1906-07.

23. HOPKINS, F. G.: Analyst 31, 395, 1906.

24. ROHMANN, F. : Ueber ktinstliche Ernahrung und Vitamine. Borntrager

Berlin, 1916.

25. ABDERHALDEN AND LAMPE: Gibt es lebenswichtige, bisher unbekannte

Nahrungstoffe? Z. ges. exp. Med. 1, 296, 1913.

26. ABDERHALDEN, EMIL: Lehrbuch der physiologichen Chemie. Urban

und Schwarzenberg.

27. ABDERHALDEN, EMIL AND FUNK, CASIMIR: Zur Frage nach der Neubil-

dung von Aminosauren im tierischen Organismus. H. 60, 418, 1909.

28. CHITTENDEN, R. H. AND UNDERBILL, F. P.: Production in dogs of a

pathological condition which closely resembles human pellagra.
Proc. Nat. Acad. Sci. 3, 195, 1917; Am. J. Physiol. 44, 13, 1917.

29. ABDERHALDEN, EMIL: Weitere Studien iiber Stickstoffwechsel. H. 96,

1, 1915.

30. GRAFE, E., SCHLAPFER V., TTJRBAN, R. AND WINTZ, H. : H. 77, 1, 1912;

78, 485, 1912; 83, 25, 1913; 84, 69, 1913; 86, 283, 1913.

31. ROHMANN, F. : Allg. med. Central Ztg. 1, 1903; 9, 1908.

32. KRAMER: Medicina castrensis. 1720.

33. BACHSTROM: Observationes circa scorbutum. eiusque indolem, causas,

signa et curam. 1734.

34. LIND: A treatise on scurvy. London 2nd Edit., 1757.

35. BARLOW, T. : Infantile scurvy. Brit. Med. J. Nov. 10th, 1894.

36. HOLST, AXEL AND FROLICH, T. : Experimental studies relating to

ship-beriberi and scurvy. J. of Hyg. 7, 634, 1907.

37. MILLER, R. : The medical diseases of children. New York, 1911.

38. SCHABAD, J. A. AND SoROCHOWETz, R. F. : Lipanin, als Ersatzmittel

des Lebertrans bei Rachitis. Sein Einfluss auf den Stoffwechsel.
Monats. f. Kinderh. 9, Orig. 659, 1911.

39. SCHABAD AND SOROCHOWETZ: Zur Frage von Wesen der gunstigen

Wirkung des Lebertrans bei Rachitis. 1st das wirksame Agens
des Lebertrans ein Ferment? Ibid. 11, Orig. 3, 4, 1912.

40. WERNICH: Geographisch-medizinische Studien. Berlin, 1878.

41. VAN LEENT: Gen. Tijdsr. voor Ned. Indis., 1880.

42. TAKAKI: Sei-i-kwai 1885, 1886, 1887.

43. VORDERMANN: Gen. Tijdsr. voor Ned. Indie, 48, 1898.

44. BRADDON, L. : The cause and prevention of beriberi. London, 1907.

45. FLETCHER, W.: Rice and beri-beri. Lane. 29th June, 1907; J. Trop.

Med. and Hyg. 12, No. 9, p. 127.

46. ELLIS, W. GILMORE: Uncured rice as a cause of beri-beri. Brit. Med.

J. p. 935, 1909.

47. FRASER, HENRY AND STANTON, H. T. : An inquiry concerning the etiology

of beri-beri. Lane. 76, 451, 1909.

LITERATURE TO THE TEXT 401

48. EIJKMAN, C. : Ein Versuch zur Bekampfung der Beriberi. Virch.

Arch. 149, 187, 1897.

49. FRASEE, H., AND STANTON: An inquiry concerning the etiology of

beriberi. Studies from the Inst. for Med. Res., Feder. Malay
States, No. 10, 1909; The etiology of beriberi. Ibid. No. 12, 1911.

50. EIJKMAN, C. : Polyneuritis bij hoenders, nieuwe tot de aetiologie der

ziekte. Gen. Tijdsr. voor Ned. Indie 1896; Eine beriberiahnliche
Krankheit der Huhner. Virch. Arch. 148, 523, 1897.

51. EIJKMAN, C. : Ernahrungspolyneuritis. Arch. d. Hyg., 58, 150, 1906.

52. GRIJNS: Gen. Tijdsr. voor Ned. Indie 41, 1901; 49, 1909.

53. EIJKMAN, C. : Virch. Arch. 222, 301, 1916.

54. BREATTDAT: Origine alimentaire et traitement du be'ribe'ri. Bull, de la

soc. de path. exot. p. 13, 1901.

55. FRASER AND STANTON: Lane. p. 4515, 1900.

56. SCHUFFNER AND KUENEN: Ueber den Einfluss der Behandlung des

Reises auf die Beriberi und die daraus entstehenden Fehlerquellen
bei der Beobachtung Arch. f. Schiffs. u. Tropenhyg. Beih. 7,
1912.

57. SCHATJMANN, H. : Beriberi und Nukleinphosphorsaure in der Nahrung.

Arch. f. Schiffs. u. Tropenhyg., 12, Beih. 5, 1908.

58. SIMPSON AND EDIE: On the relation of the organic phosphorus content

of various diets to diseases of nutrition, particularly beriberi.
Ann. Trop. Med. a. Parasit. 5, 313, 1911.

59. POL, HULSHOFF: Beriberiforschungen in den niederlandisch-ostin-

dischen Kolonien, besonders in Bezug auf Prophylaxis und Heilung
Arch. f. Schiffs- u. Tropenhyg. 14. Beih. 3, 1910.

60. THOMPSON AND SIMPSON: Treatment of beriberi. Ann. Trop. Med. a.

Parasit. 6, 53, 1921.

61. EIJKMAN, C. : Polyneuritis gallinarum and Beriberi. Arch. f. Schiffs- u.

Tropenhyg. 15, 698, 1911.

62. FUNK, CASIMIR: The etiology of the deficiency diseases. J. State Med.

June, 1912.

63. TERUTJCHI: Ueber die Aetiologie der Beriberi unter besonderer Beriick-

sichtigung der Schaumann'schen Phosphormangel theorie. Saiking-
akuzashi. Tokyo No. 79, 1910.

64. CHAMBERLAIN AND VEDDER: A contribution to the etiology of beriberi.

Philipp J. of Sci. 6 B, 251, 1911; 6, 395, 1911; with Williams, R. R.,
Ibid. 7, 39, 1912.

65. SHIGA AND KUSAMA: Arch. f. Schiffs- u. Tropenhyg. Beih. 3, May, 1911;

Shiga: Experimentelle Studien iiber Kakke. C. f. Bakt. I Abt.
Beih. 6, Tagung d. Vereins f. Mikrobiologie p. 158.

66. TSUZUKI : Antiberiberintherapie der Beriberikrankheit 1912.

67. ARON AND HOCSON: Phosphorus starvation with special reference to

beriberi. II. Philip. J. of Sci. 5, February, 1910.

68. COOPER AND FUNK, CASIMIR: Experiments on the causation of beriberi.

Lane. p. 1266, 1911.

402 THE VITAMINES

69. McCoLLUM, E. V. AND PITZ, W. : The "vitamine" hypothesis and defi-

ciency diseases. A study of experimental scurvy. J. Biol. Chem.,
31, 229, 1917.

70. HOPKINS, F. G. : Feeding experiments illustrating the importance of

accessory factors in normal dietaries. J. Physiol. 44, 425, 1912.

71. OSBORNE, T. B. AND MENDEL, L. B.: Feeding experiments with isolated

food-substances. Publ. Cam. Inst., Wash., No. 156, Part I and II.

72. The same. Feeding experiments with fat-free food mixtures. J. Biol.

Chem. 12, 81, 1912; H. 80, 307, 1912.

73. The same. Maintenance experiments with isolated proteins. J. Biol.

Chem. 13, 233, 1912.

74. HOPKINS AND NEVILLE: A note concerning the influence of diets upon

growth. Biochem. J. 7, 97, 1913.

75. McCoLLUM AND DAVIS, M. : The necessity of certain lipins in the diet

during growth. J. Biol. Chem. 15, 67, 1913.

76. OSBORNE AND MENDEL: The relation of growth to the chemical constit-

uents of the diet. J. Biol. Chem. 15, 311, 1913.

77. The same. The influence of butter-fat on growth. Ibid. 16, 423, 1913.

78. McCoLLUM AND KENNEDY. Ibid. 24, 491, 1916.

79. DRUMMOND, J. C. : Note on the role of the antiscorbutic factor in nutri-

tion. Biochem. J. 13, 77, 1919.

80. FUNK. CASIMIR: What is a vitamine? J. A. M. A. 66, 1650, 1916.

80a. DRUMMOND, J. C. : The nomenclature of the so-called accessory food
factors (vitamines). Biochem. J. 14, 660, 1920.

81. FUNK, CASIMIR AND MACALLUM, A. B., JR.: Die chemischen Deter-

minanten des Wachtsums. Ztschr. f. Physiol. Chem. 92, 13, 1914.

82. McCoLLUM AND DAVIS : Observations on the isolation of the substance

in butter-fat which exerts a stimulating influence on growth. J.
Biol. Chem. 19, 245, 1914.

83. OSBORNE AND WAKEMAN: Does butter-fat contain nitrogen and phos-

phorus? Ibid. 21, 91, 1915.

84. WRAMPELMEYER, E. : Der Lecithingehalt der Butter. Landw. Vers.

Sta. 42, 437, 1892.

85. SUPPLEE, G. C. : The lecithin content of butter and its possible relation-

ship to the fishy flavor. Corn. Univers. Agr. Exp. Stat. 29, 101,
1919.

86. FUNK, CASIMIR AND MACALLUM, A. B., JR.: Studies on growth. II.

On the probable nature of the substance promoting growth in
animals. "J. Biol. Chem. 23, 413, 1915.

87. DANIELS, AMY L. AND LOUGHLIN, ROSEMARY: Note on the fat-soluble

growth-promoting substance in lard and cotton-seed oil. Ibid.
42, 359, 1920.

88. SEIDELL, ATHERTON: Vitamines and nutritional diseases; a stable

form of vitamine. U. S. Publ. Health Rep., Repr., 325, 1916.

89. HARDEN, A. AND ZILVA, S. S. : Differential behavior of the antineuritic

and antiscorbutic factors towards adsorbents. Biochem. J. 12, 93,
1918.

LITERATURE TO THE TEXT 403

90. BYFIELD, A. H., DANIELS, A. L. AND LOUGHLIN, R. : The antineuritic

and growth-stimulating properties of orange juice. Am. J. Dis.
Childr. 19, 349, 1920.

91. FURST, VALENTIN: Weitere Beitrage zur Aetiologie des experimentellen

Skorbuts des Meerschweinchens. Z. f. Hyg. u. Infektionsk. 72,
121, 1912.

92. OSBORNE AND MENDEL: Do fruits contain water-soluble vitamine?

Soc. Exp. Biol. Med. 17, 46, 1919; J. Biol. Chem. 42, 465, 1920.

93. MITCHELL, H. H. : On the identity of the water-soluble growth-pro-

moting vitamine and the antineuritic vitamine. J. Biol. Chem.
40, 399, 1919.

94. EMMETT, A. D. AND LTJROS, G. O. : Are the antineuritic and the water-

soluble vitamines the same? Proc. Soc. Biol. Chem., J. Biol.
Chem. 41, VII, 1920; J. Biol. Chem. 43, 265, 1920.

95. MELLANBY, EDWARD: A further demonstration of the part played by

accessory food factors in the etiology of rickets. Proc. Physiol.
Soc. Dec. 14th, 1918; J. Physiol. 52, LIII, 1919; An experimental
investigation on rickets. Lane. March 15, 1919.

96. HESS, ALFRED F. AND UNGER, L. J. : The role of fat-soluble vitamine

in the dietary of infants. Proc. Soc. Exp. Biol. Med. 17, 49,1919;
The clinical role of the fat-soluble vitamine; its relation to rickets.
J. A. M. A. 74, 217, 1920.

97. McCoLLUM AND DAVIS : Essential factors in the diet during growth.

J. Biol Chem. 23, 231, 1915.

98. FUNK, CASIMIR AND MACALLUM, A. B. : Studies on growth. III. The

comparative value of lard and butter fat. Ibid. 27, 51, 1916.

99. HOGAN, ALBERT G. : The effect of high temperature on the nutritive

value of foods. Ibid. 30, 115, 1917.

100. HARDEN, A. AND ZILVA, S. S. : Accessory factors in the nutrition of the

rat. Bio. J. 12, 408, 1918.

101. FUNK, CASIMIR: The nitrogenous constituents of lime-juice. Ibid.

7, 81, 1913.

102. OSBORNE, T. B., WAKEMAN, A. J. AND FERRY, E. A.: Preparation of

protein free from water-soluble vitamine. J. Biol. Chem. 39, 35,
1919.

103. OSBORNE AND MENDEL: Protein minima for maintenance. J. Biol.

Chem. 22, 241, 1915; 20, 351, 1915; 26, 1, 1916; 37, 223, 1919.

104. EDELSTEIN, F. AND LANGSTEIN, L. : Das Eiweissproblem im Sauglings

alter, experimentelle Untersuchungen liber ' die Wertigkeit der
Milcheiweisskorper fur das Wachstum. Z. f. Kinderh. 20, 112,
1919.

105. EMMETT, A. D. AND LUROS, G. O. : Is lactalbumin a complete protein

for growth? J. Biol Chem. 38, 147, 1919; The stability of lactal-
bumin toward heat. Ibid. 38, 257, 1919.

106. McCoLLUM, SIMMONDS AND PARSONS: Ibid. 37, 287, 1919.

107. EMMETT, A. D. AND McKiM, L. H. : The value of the yeast vitamine as

a supplement to a rice diet. Ibid. 409, 1917.

404 THE V1TAMINES

108. SUGIURA, K. AND BENEDICT, S. R. : The nutritive value of the banana.

J. Biol. Chem. 36, 171, 1918; 40, 449, 1919.

109. GOLDBEEGER, JOSEPH AND WHEELER, G. A. : The experimental produc-

tion of pellagra in human subjects by means of diet. Hyg. Labor.
Wash. Bull. 120, Febr., p. 7, 1920.

110. CLARK, GUY W. : The properties and composition of oocytin. J. Biol.

Chem. 35, 253, 1918.

111. ROBERTSON, BRAILSFORD T. : On the extraction of a substance from the

sperm of a sea-urchin, which will fertilize eggs. Ibid. 12, 1, 1912.

112. PASTEUR, Louis: Note sur un memoir de M. Liebig, relatif aux fer-

mentations.

113. WILDIERS, E. : Nouvelle substance indispensable au deVeloppement de

la levure. La Cellule. 18, 313, 1901.

114. AMAND, ABEL: Le "Bios" de Wildiers ne joue pas le role d'un contre-

poison. Ibid. 20, 225, 1902; 21, 324, 1904.

115. DEVLOO, RENE: Ibid. 23, 36, 1906.

116. PRINGSHEIM, H. H.: Centr. f. Bakt. II Abt. 16, 111, 1916.

117. RUBNER, MAX: Die Ernahrungsphysiologie der Hefezelle bei der alko-

holischen Gahrung. Leipzig, 1913.

118. VLAHUTA, E. : Preparation of a peptone by the decomposition of the

cells of beer yeast and the role of this peptone in fermentation.
Bull. acad. Sci. Roumanie. 3, 123, 1914-15.

119. KURONO, K. : Significance of oryzanin for the nutrition of fermentative

organisms. J. Coll. Agr. Univers. Tokyo. 5, 305, 1915.

120. BRILL, H. C. AND THURLOW, L. W. : Alcohol from discarded molasses

in the Philippine Islands. Philipp. J. Sci. 12, 267, 1917.

121. MOUFANG, E. : Beschleunigung der Gahrung (durch getotete Hefe)

Allg. Brau. Hopfen. Ztg. 55, 605, 1915.

122. SAITO, K. : Chemical conditions for the development of the reproductive

organs of some yeasts. J. Coll. Sci. Univ. Tokyo, 39, 1, 1916.

123. BOKORNY, TH. : Bio. Z. 82, 359, 1917.

124. VANSTEENBERGE, P. : The autolysis of yeast and the influence of prote-

olysis on the development of yeast and lactic bacteria. Ann.
Inst. Past., 31, 601, 1917.

125. LAMPITT, L. H. : Nitrogen metabolism in Saccharomyces cerevisiae.

Biochem. J. 13, 459, 1919.

126. ABDERHALDEN, E. AND SCHAUMANN, H. : Die Wirkung einiger aus der

Hefe mit Alkohol extrahierten Substanzen auf die Aktivitat der
Hefefermente. Fermentf. 2, 120, 1918.

127. ABDERHALDEN AND KOHLER, A. : Die Wirkungsweise einer in Alkohol

loslichen Substanz, wahrend der alkoholischen Gahrung durch
Hefe gebildet, auf niedrigere Organismen. Arch. ges. Physiol.
176, 209, 1919.

128. WILLIAMS, ROGER J. : The vitamine requirements of yeast. A simple

biological test for vitamine. J. Biol. Chem. 38, 465, 1919.

129. BACHMANN, F. M. : Vitamine requirements of certain yeasts. J. Biol.

Chem. 39, 235, 1919.

LITERATURE TO THE TEXT 405

129a. FULMER, ELLIS I., NELSON, VICTOR E. AND SHERWOOD, F. F. : The
nutritional requirements of yeast. I. The role of vitamines in
the growth of yeast. II. The effect of the composition of the
medium on the growth of yeast. J. Amer. Chem. Soc. 43, 186-
191, 1921.

129b. NELSON, V. E., FULMER, E. I. AND CESSNA, RUTH: Nutritional re-
quirements of yeast. III. Synthesis of Vitamine B by yeast.
J. Biol. Chem. 46, 77, 1921.

129c. HARDEN, A. AND ZILVA, S. S. : The synthesis of vitamine B by yeasts
(preliminary note). Biochem. J. 15, 438, 1921.

129d. FUNK, CASIMIR AND DUBIN, HARRY E.: Vitamine requirements of
certain yeast and bacteria. J. Biol. Chem. 48, 437, 1921.

129e. EDDY, WALTER H., HEFT, HATTIE L., STEVENSON, HELEN C., AND
JOHNSON, RUTH: Proc. Soc. Exp. Biol. Med. 18, 138, 1921.

129f. MACDONALD, MARGARET B. AND McCoLLUM, E. V.: The cultivation
of yeast in solutions of purified nutrients. J. Biol. Chem. 45, 307,
1921.

129g. IDE, M.: The "Bios" of Wildiers and the cultivation of yeast. J.
Biol. Chem. 46, 521, 1921.

129h. MACDONALD, M. B. AND McCoLLUM, E. V.: The "Bios" of Wildiers
and the cultivation of yeast. Ibid. 46, 525, 1921.

129i. WILLIAMS, R. J.: Vitamines and yeast growth. Ibid. 46, 113, 1921.

130. BERTRAND, GABRIEL: Etudes biologiques sur la bacterie du sorbose.

Ann. de chim. et de phys. (8), 3, 121, 1904.

131. NOGUCHI, H. : A method for the pure cultivation of pathogenic Trepo-

nema pallidum. (Spirochaeta pallida). J. Exp. Med. 14, 99,
1911.

132. VEDDER: J. Infect. Dis. 16, 385, 1915.

133. DORYLAND, C. J. T. : J. Bact. 1, 135, 1916.

134. PIEPER, E. J., HUMPHREY, C. J.AND AGREE, S. F. : Phytopath. 7, 214, 1917.

135. LOCKEMAN, GEORG: Welche Substanzen sind zum Wachstum des Tuber-

kulose Bacillus absolut notig. Centr. Bakt. u. Parasitenk. Abt.
I, 83, 420, 1919.

136. BAINBRIDGE. J. Hyg. 11, 341, 1911.

137. SPERRY AND RETTGER: J. Biol. Chem. 20, 445, 1915.

138. ROBINSON, HAROLD C. AND RETTGER, LEO F. : The growth of bacterias

in protein-free enzyme- and acid-digestion products. J. Bact.
3, 209, 1918.

139. PACINI, J. P. AND RUSSELL, D. W. : The presence of growth-producing

substances in cultures of typhoid bacilli. J. Biol. Chem. 34,
43, 1918.

140. MACKENZIE WALLIS, H. L. : Improvements in bacteriological media.

I. A new and efficient substitute for "Nutrose". Agr. J. India
12, 621, 1917.

141. HUNTOON, F. M. : Hormone medium, a simple medium employable as a

substitute for serum medium. J. Infect. Dis. 23, 169, 1918.

142. AYERS, S. H. AND RUPP, P. : Extracts of pure dry yeast for culture

medium. J. Bact. 5, 89, 1920.

406 THE VITAMINES

143. LLOYD, DOROTHY J. : Vitamines, amino acids, and other chemical

factors involved in the growth of the Meningococcus. J. Path.
Bact. 21, 113, 1916.

144. The same. Brit. Med. J. II, 143, 1916.

145. The same. Chemical factors involved in the growth of the Meningo-

coccus. Ibid. I, 11, 1917.

146. GORDON, M. H. AND HINE, T. G. M. ; Experimental study of the cultural

requirements of the Meningococcus. Ibid. II, 678, 1916.

147. FLACK, MARTIN: Note on the constituent of pea-flour augmenting

growth of the Meningococcus on the trypagar. medium. Ibid. II,
682, 1916.

148. SHEARER, C. : The presence of an accessory food factor in the nasal

secretion and its action on the growth of the Meningococcus and
other pathogenic bacteria. Lane. I, 59, 1917.

149. The same. The action of spinal fluid in stimulating the growth of the

Meningococcus. Ibid. II, 714, 1917.

150. EBERSON, FREDERICK: Yeast mycelium for prolonging the viability of

Meningococcus. J. A. M. A. 72, 852, 1919.

151. COLE, SYDNEY W. AND LLOYD, DOROTHY J. : The preparation of solid

and liquid media for the cultivation of the Gonococcus. J. Path.
Bact. 21, 267, 1917.

152. MORINI, L. : Cultivation of Gonococcus in gelatin with beer yeast.

Rif. med. Napoli. 36, 2, 1920.

153. DOUGLAS, S. R., FLEMING, A. AND COLEBROOK, L. : Studies in wound

infections. The growth of anaerobic bacilli in fluid media under
apparently aerobic conditions. Lane. II. 530, 1917.

153a. SAZERAC, ROBERT: C. r. 171-, 278, 1920.

153b. MASUCCI, PETER: A note on the effect of amino acids on the growth
of tubercle bacilli. J. Lab. Clin. Med. 6, 96, 1920.

154. DAVIS, DAVID JOHN: Food accessory factors (vitamines) in bacterial

culture, with special reference to hemophilic bacilli. J. Inf. Dis.

21, 392, 1917.

155. The same. II. Ibid. 23, 248, 1918.

156. WOLF, C. G. L. AND HARRIS, J. E. G. : Contribution to the biochemistry

of pathogenic anaerobes. IV. The biochemistry of Bacillus
histolyticus. J. Path. Bact. 22. 1, 1918.

157. WOLF, C. G. L. : V. The biochemistry of Vibrion septique. Ibid.

22, 115, 1918.

158. The same. VI. The biochemistry of Bacillus sporogenes and Bacillus

Welchii. VII. The biochemistry of Bacillus proteus. Ibid. 22,
270, 1919.

159. BOYER, Louis : Un nouveau milieu de culture, particulierement adapte"

au deVeloppement du streptococce pyog^ne. C. r. soc. de biol.
81, 229, 1918.

160. OTABE, SHORABURO : Wheat culture-media. Lane. 197, 576, 1919.

161. DIENERT, F. AND GUILLERD, A.: Milieu de culture d'un extrait de la

leVure autolyse"e pour les cultures de B. coli. C. r. 168, 256, 1919.

LITERATURE TO THE TEXT 407

162. BUNKER, JOHN W. M. : Studies on the diphtheria bacillus in culture.

J. Bact. 4, 379, 1919.

163. DAVIS, L. AND FERRY, N. S. : Studies on diphtheria toxin II. The role

of the aminoacids in the metabolism of B diphtheriae. Ibid.
4, 217, 1919.

164. AGULHON, H. AND LEGROUX, R. : Contribution a 1'etude des vitamines

utilisables. a la culture des microorganismes. Application au ba-
cille de F influenza (B. de Pfeiffer) C. r. 167, 597, 1918.

165. LEGROUX, REN£ AND MESNARD, JOSEPH : Les vitamines pour les cultures

des bacteries. Ibid. 170, 901, 1920.
165a. DAVIS, D. J.: Food accessory factors in bacterial growth. Ill, IV,

V. J. Infect. Dis. 29, 171, 1921.
165b. RIVERS, T. M. AND POOLE, A. K.: Growth requirements of influenza

bacilli. Johns Hopk. Hosp. Bull. 32, 202, 1921.
165c. THJOTTA, TH.: Studies in bacterial nutrition. J. Exp. Med. 33,

763, 1921.
165d. THJOTTA, TH. AND AVERT, O. T.: Growth accessory substances in

nutrition of bacteria. Proc. Soc. Exp. Biol. Med. 18, 197, 1921;

J. Exp. Med. 34, 97, 1921.

166. KLIGLER, I. J. : Yeast autolysate as a culture medium for bacteria. J.

Bact. 4, 183, 1919.

167. The same. Growth accessory substances for pathogenic bacteria in

animal tissues. J. Exp. Med. 30, 31, 1919.

167a. MUELLER, J. HOWARD: Observations of bacterial metabolism. Proc.
Soci Exp. Biol. Med. Oct. 20, 1920.

167b. MUELLER, J. HOWARD,: Growth determining substances in bac-
teriological culture media. Soc. Exp. Biol. Med. Apr. 10, 1921.

167c. FREEDMAN, L.: Comparison of yeast and bacteria growth-promoting
vitamines. Amer. Chem. Soc. Sept. 1921.

167d. NORRIS, D. : Preparation of a simplified culture medium for field
work. Ind. J. Med. Res. 7, 704, 1920.

168. LUTZ, C.: Ann. Mycol. von Sydow. 7, 91, 1909.

169. CURRIE, JAMES N. : The citric acid fermentation of Aspergillus niger.

J. Biol. Chem. 31, 15, 1917.

170. WILLAMAN, J. J. : J. Ind. Eng. Chem. 10, 863, 1918; The function of

vitamines in the metabolism of Sclerotinia cinerea. J. Amer.
Chem. Soc. 42, 549, 1920.
170a. LINOSSIER, G. : C. r. soc. biol. 82, 381, 1919.

171. CLINTON, G. P.: Conn. Agr. Exp. Sta. Rep. 1909-10, p. 753-

172. HABERLANDT, G. : Zur Physiologic der Zellteilung. Sitzungsb. d.

Preuss. Akad. Wiss. Berlin. 318, 1913.

173. BOTTOMLEY, W. B. : The bacterial treatment of peat. J. R. Soc. Arts.

72, 372; Some effects of humates on plant growth. Rep. Br. Ass.
Adv. of Sciences. 680, 1912.

174. The same. Some accessory factors in plant growth and nutrition.

Proc. R. Soc. 88 B, 237, 1914; The significance of certain food
substances for plant growth. Ann. Bot. 28, 531, 1914.

408 THE VITAMINES

175. Anonymous: Experiments with bacterized peat. Agr. News. 13, 263,

1914.

176. BOTTOMLEY, W. B. : A bacterial test for plant food accessories (Auxi-

mones). Proc. Roy. Soc. 89 B, 102, 1915.

177. CHITTENDEN, F. I. : Report on experiments with bacterized peat or

humogen. J. Roy. Hort. Soc. 41, 305, 1915.

178. BOTTOMLEY: Some effect of organic growth-promoting substances

(auximones), on the growth of Lemna minor in mineral culture
solution. Proc. R. Soc. 89 B, 481, 1917.

179. The same. Ann. Bot. 34, 1920.

180. ROSENHEIM, OTTO: Accessory factors for plant growth. Biochem. J.

11, 7, 1917.

181. BOTTOMLEY: Isolation from peat of certain nucleic acid derivatives.

Proc. R. Soc. 90 B, 39, 1917.

182. APPLEMAN, C. O.: Maryland Agr. Exp. Sta. Bull. 212, 1918; Science

N. S. 48, 319, 1918.

183. BOTTOMLEY: Effect of nitrogen-fixing organisms and nucleic acid

derivatives on plant growth. Proc. R. Soc. 91 B, 83, 1919.

184. JONES, D. H. Growth of plants. Abstr. Bact. 1, 43, 1917.

185. STEAD, AKTHUK. Notes on humus, humogen, and its accessory plant

food substances. S. Afr. J. Sci. 13, 239, 1917.

186. MOCKERIDGE, FLORENCE A. : Some effects of organic growth-promoting

substances (auximones) on the soil organisms concerned in the
nitrogen cycle. Proc. R. Soc. 89 B, 508, 1917.

187. SCHREINER, REED AND SKINNER: U. S. Dept. Agr. Bureau of Soils. Bull.

47, 1907; Schreiner and Skinner: Ibid. Bull. 87, 1912; Schreiner and
Shorey: Ibid. Bull. 74, 1910; Shorey Bull. 88, 1913.

188. MACALISTER: The action of Symphytum officinale and allantoin.

Brit. Med. J. Sept. 21, 1912.

189. COPPIN: The effect of purine derivatives and other organic compounds

on growth and cell-division in plants. Biochem. J. 6, 416, 1912.

190. MOCKERIDGE, FLORENCE ANNIE: The occurrence and nature of the

plant-producing substances in various manurial imposts. Ibid.
14, 432, 1920.

191. GRAFE, VICTOR: Sterilisieren lebender Pflanzen. Abderhaldens Handb.

d. biochem. Arbeitsm. 6, 139, 1912; 8, 178, 1915.

191a. PARSONS, HELEN T. : The antiscorbutic content of certain body tissues
of the rat. The persistence of the antiscorbutic substance of the
rat after long intervals on a scorbutic diet. J. Biol. Chem. 44,
587, 1920.

192. SPRIGGS, E. J. : The effect of a meat diet upon the coats of rats with and

without the supply of lime. J. Physiol. 36, xvii, 1907.

193. MAIGNON, F. : L'influence des saisons sur la toxicite" du blanc d'oeuf,

chez les rats blancs. C. r. de la ass. fran9. avanc. sci. 43, 553; C. r.
166, 1008, 1918; 167, 91, 1918.

194. The same. Toxicit6 du blanc d'oeuf; 1'influence des saisons sur la

sensibility de 1'organisme a 1'intoxication azotee. C. r. 166, 919,
1918.

LITERATURE TO THE TEXT 409

195. DAVIS, W. A.: The phosphate depletion of the soils of Bihar; its effect

on the quality and yield of crops and the contingent risks of malnu-
trition and endemic diseases in cattle and man. Agr. J. India
(Spec. Ind. Sci. Congr. No.) 77, 1917.

196. SCHOTTELIUS: Arch. f. Hyg. 34, 1899; 42, 1902; 47, 1908.

197. BELONOWSKY: Sur 1'alimentation sterilisee. Z. f. Bakt. 44, 322, 1907.

198. BOGDANOW, E. A. : Ueber das Ztichten der Larven der gewohnlichen

Hausfliege (Calliphora vomitoria) in sterilisierten Nahrmitteln.
Arch. f. ges. Physiol. 113, 97, 1906; Arch. Physiol. Suppl. 173, 1908.

199. DELCOURT, A. AND GUYENOT, E. : Bull. Sci. France et Belg. 14, 249,

1911; Guyenot: C. r. soc. biol. 65, Part i, 97, 178, 223, 270, 1914;
Part i, 66, 483, 548, 1914; Recherches expe'rimentales sur la vie
aseptique et le developement d'un organisme enfonctiondumillieu.
These Doct. Sci. Paris, 1917.

200. WOLLMANN, N. : Sur la vie sans microbes. Ann. Inst. Past. 25, 79, 1911;

La vie aseptique. Bull. Inst. Past. 15 et 30 dec. 1914; Elevage
aseptique des larves de la mouche a viande (Calliphora vomitoria)
sue millieu sterilise a haute temperature. C. r. soc. biol. 31 mai
1919; Richet, Ch. : L'alimentation avec les aliments ste>ilise"s remarque
a propos de la note de M. Wollmann. C. r. soc. biol. 7 juin, 1919.

201. METCHNIKOFF, MME. : cited by Cohendy. Ann. Inst. Past. 27, 106,1912.

202. Mono: Das Leben ohne Bakterien. J. f. Kinderh. 62, H. 5, 1905.

203. COHENDY: Experiences sur la vie sans microbes. Ann. Inst. Past. 26,

106, 1912; II. Experiences sur la vie en cultures pures succe'dant a
la vie sans microbes. C. r. 154, 670, 1912.

204. The same. La vie aseptique. C. r. 158, 1283, 1914.

205. SCHOTTELIUS: Die Bedeutung der Darmbakterien fur die Ernahrung,

IV. Arch. f. Hyg. 79, 1913.

206. KUSTER: Leben onhe Bakterien. Arb. a. d. Kais. Gesundheitamte.

48, 1, 1914.

207. CHARRIN AND GUILLEMONAT: cited by Weill-Mouriquand. Maladies

par carence. Bailliere, Paris, 1919.

208. KIANIZIN, J. : Effect on higher animals of the sterilization of the inhab-

ited medium, the air and the food. J. Physiol. 50, 391, 1916.

209. NUTTAL AND TmERFELDER : cited by Kuster. Die keimfreie Ziichtung

von Saugetieren. Abderhaldens H. d. Biochem. Arbeitsm. 8, 311,
1915.

210. WEILL, E. AND MOURIQTTAND, G. : C. r. soc. biol. 81, 1253, 1918.

211. SCHAEFFER, G. : Conception de la carence dans 1'interpretation des

resultats obtenus dans les travaux sur 1'alimentation artificielle et
la vie aseptique. Ibid. 82, 2, 1919.

212. PORTIER, PAUL. : Les symbiotes. Masson & Cie, Paris, 1918.

213. BIERRY, H. AND PORTIER, P. : Vitamines et symbiotes. C. r. 166, 963,

1918; Action des symbiotes sur les constituants des graisses. C. r.
166, 1055, 1918; L'importance de la function cetonique dans la
nutrition, sa creation par les symbiotes. Ibid. 167, 94, 1918.

214. DELAGE, YVES: Ibid. 166, 966, 1918.

410 THE VITAMINES

215. LUMIERE, AUGUSTE : Le mythe des symbiotes. Masson, Paris, 1919.

216. RANG, A. : Bioenergesis et la the"orie des symbiotes de Paul Portier.

L'ind. chim. 6, 136, 1919.

217. POBTIER, P. AND RANooiN, LuciE : La production des vitamines dans

les intestins des lapins, relevant une nourriture sterilised a haute
temperature. C. r. 170, 478, 1920.

218. LUND: Am. J. Physiol. 47, 167, 1918.

219. CALKINS, G. W. AND EDDY, W. H. : The action of pancreatic vitamine

upon the metabolic activity of Paramecium. Proc. Soc. Exp.
Biol. Med. 14, 162, 1917.

220. FLATHER, MARY DRUSILLA: Effects of polished and of unpolished rice

upon the metabolic activity of^ Paramecium. Biol. Bull. 36, 54,
1919.

221. CHAMBERS, MARY H. : Effect of some food hormones and glandular

products on the rate of growth of Paramecium caudatum. Ibid.
36, 82, 1919.

222. PETERS, R. A. : Nutrition of the protozoa. The growth of Paramecium

in sterile culture medium (Prelim. Comm.). Proc. Physiol. Soc.
Febr. 21, 1920, J. Physiol. 53, CVIII, 1920; 54, L, 1920; The sub-
stances needed for the growth of a pure culture of Colpidium
colpoda. J. Physiol. 55, 1, 1921.

223. GOLDFARB, A. J. : A study of the influence of lecithin on growth. Proc.

Soc. Exp. Biol. Med. 4, 159, 1907.

224. The same. Does lecithin influence growth? Arch. Entw. Mech. 29,

255, 1910.

225. CARREL, A. : Technique for cultivating a large quantity of tissue. J.

Exp. Med. 15, 393, 1912.

226. The same. On the permanent life of tissues outside of the organism.

Ibid. 15, 516, 1912.

227. The same. Handb. der biochem. Arbeitsm. 6, 519, 1912.

228. CARREL AND BURROWS : An addition to the technique of the cultivation

of tissues in vitro. J. Exp. Med. 14, 244, 1911.

229. CARREL, A. : Artificial activation of the growth in vitro of connective

tissue. Ibid. 17, 14.

230. WALTON, A. J. : The influence of internal secretion glands on growth.

Ibid. 20, 554, 1914.

231. LOEB, JACQUES: The simplest constituent required for growth and the

completion of the life cycle in an insect. Science 41, 169, 1915.

232. The same. The salts required for the development of insects. J.

Biol. Chem. 23, 431, 1915.

233. LOEB, JACQUES AND NORTHROP, J. H. : Nutrition and evolution. J.

Biol. Chem. 27, 309, 1916.

234. The same. The influence of food and temperature upon the duration

of life. Ibid. 32, 103, 1917.

235. NORTHROP, JOHN H. : The effect of prolongation of the period of growth

on the total duration of life. Ibid. 32, 123, 1917.

236. The same. The role of yeast in the nutrition of an insect (Drosophils) .

Ibid. 30, 181, 1917.

LITERATURE TO THE TEXT 411

237. GUYENOT, EMILE: Bull. biol. de France et de la Belg. 51, 14 aout et 15

d<5c. 1917.

238. BAUMBERGER, J. PERCY: A nutritional study of insects, with special

reference to microorganisms and the substrate. J. Exp. Zool.
28, 1, 1919.

239. MORGULIS, SERGIUS : Studies on the nutrition of the fish. Experiments

on brook trout. J. Biol. Chem. 36, 391, 1918.

240. ALMY, L. H. AND ROBINSON, R. K. : Toxic action of ingested linseed oil

meal on trout. J. Biol. Chem. 43, 97, 1920.

241. EMMETT, A. D. AND ALLEN, FLOYD P. : Nutritional studies on the growth

of frog larvae (Rana pipiens). Ibid. 38, 325, 1919.

242. EMMETT, ALLEN AND STTTRTEVANT, MARGUERITE : Relation of vitamines

and iodine to the size and development of the tadpoles. Proc.
Soc. Amer. Biol. Chem. J. Biol. Chem. 41, LIV, 1920.

243. HARDEN, ARTHUR AND ZILVA, SOLOMON SYLVESTER: Dietetic experi-

ments with frogs. Biochem. J. 14, 263, 1920.

244. FUNK, CASIMIR: Studien iiber das Wachstum. I. Das Wachstum auf

vitaminhaltiger und vitaminfreier Nahrung. Ztschr. f. physiol.
Chem. 86, 352, 1913.

245. FUNK, CASIMIR AND MACALLUM, A. B. : Die chemischen Determinanten

des Wachstums. Ibid. 92, 13, 1914.

246. FUNK, CASIMIR: Studies on growth: the influence of diet on growth,

normal and malignant. Lane. 186, 89, 1914.

247. DRUMMOND, J. C. : Observations upon the growth of young chickens

under laboratory conditions. Biochem. J. 10, 77, 1916.

248. FUNK, CASIMIR: J. Biol Chem. 27, 1, 1916 (Footnote p. 4).

249. BUCKNER, G. D., NOLLAU, E. H. AND KASTLE, J. H. : Feeding of young

chicks on grain mixture of high and low lysine content. Am. J.
Physiol. 39, 162, 1915.

250. OSBORNE AND MENDEL: The effect of the amino-acid content of the diet

on the growth of chicken. J. Biol. Chem. 26, 293, 1916.

251. The same. The growth of chickens in confinement. Ibid. 33, 433,

1918; J. Pharmac. 11, 170, 1918.

252. HART, E. B., HALPIN, J. C. AND STEENBOCK, H. : Use of synthetic diets

in the growth of baby chicks. A study of leg weakness in chickens.
J. Biol. Chem. 43, 421, 1920.

253. PALMER, LEROY S. : The physiological relation of plant carotiniods to

the carotinoids of the cow, horse, sheep, goat, pig and hen. Ibid.
27, 27, 1916.

254. PALMER AND KEMPSTER, HARRY L. : Relation of plant carotinoids to

growth, fecundity, and reproduction of fowls. Ibid. 39, 299, 313,
333, 1919.

255. HART, E. B., HALPIN, J. G. AND McCoLLUM, E. V.: Ibid. 29, 57, 1917.

256. HART, HALPIN AND STEENBOCK: The behavior of chickens restricted to

the wheat and maize kernel. II. Ibid. 31, 415, 1917.

257. HARNEY, R. N. : Egg-producing values of some Texas feeding stuffs.

Texas Agr. Exp. Sta. Bull. 220, 11, 1917.

412 THE VITAMINES

258. KAUPP, B. F. : Mineral content of Southern poultry feeds and mineral

requirement of growing fowls. J. Agr. Res. 14, 125, 1918.

259. McCoLLUM AND DAVIS, MARGUERITE: The nature of the dietary defi-

ciency in rice. J. Biol. Chem. 23, 181, 1915.

260. HOULBERT: Vitamines et la croissance. Paris me'd. 9, 473, 1919.

261. HILL, L. AND FLACK, M. : Brit. Med. J. Sept. 16th, 311, 1911.

262. OHLER, RICHARD W. : Experimental polyneuritis — Effects of exclusive

diet of wheat flour, in form of ordinary bread, in fowls. J. Med.
Res. 31, 239, 1914.

263. WEILL, E. AND MOURIQUAND, G. : Troubles provoqu6s par une alimen-

tation exclusive. Ann. de me'd. et chir. infant. 18, mai 15, 1914.

264. WELLMAN, BASS AND EUSTIS: Investigations of Louisiana rice, with

reference to the etiology of beriberi. New Orl. Med Surg. J. 65,
191, 1912. Wellmann: South. Med. J. 6, 516, 1913..

265. WELLMAN, C., AND BASS, C. C. : Polyneuritis gallinarum caused by

different foodstuffs. Am. J. Trop. Dis. and Prev. Med. 1, 129, 1913.

266. VEDDER AND CLARK: A study of polyneuritis gallinarum. A fifth contri-

bution to the etiology of beriberi. Philipp. J. Sci. 7 B, 423, 1912.

267. SEGAWA: Ingestion of polished rice by chickens and pigeons. Mitt.

med. Ges. Tokyo 27 No 7, 1914; Virch. Arch. 215, 404, 1914..

268. TASAWA: Experimentelle Polyneuritis, besonders bei Vogeln, im Ver-

gleich zur Beriberi des Menschen. Z. f. exp. Path. Ther. 17, 27,
1915.

269. POL, HULSHOFF: Geneesk. Tijdsr. v. Ned. Indie. 52, 11, 245, 1912.

270. WILLIAMS, R. R. AND JOHNSTON, J. A. : Miscellaneous notes and com-

ments on beriberi. Philipp. J. Sci. 10B, 337, 1915.

271. VOEGTLIN, CARL AND MYERS, C. N. : Distribution of the antineuritic

vitamine in the wheat and corn kernel. A contribution to the
biology of the antineuritic vitamine. Am. J. Physiol. 48, 504, 1919.

272. The same. Growth-promoting properties of foods derived from corn

and wheat. U. S. Public Health Serv. Repr. 471, 23, 1918.

273. FUNK, CASIMIR: Is polished rice plus vitamine a complete food? J.

Physiol. 48, 228, 1914.

274. STEPP, WILHELM: Zur Frage der synthetischen Fahigkeiten des Tier-

korpers. Z. f. Biol. 66, 300, 1916.

275. FUNK, CASIMIR: Studien iiber Beriberi XI. Die Rolle der Vitamine

beim Kohlenhydratstoffwechsel. Ztschr. f. physiol. Chem. 89,
378, 1914.

276. DUTCHER, R. ADAMS: Vitamine Studies IV. Antineuritic properties of

certain physiological stimulants. Prelim. Paper. J. Biol. Chem.
39, 63, 1919.

277. COOPER, E. A. AND FUNK, CASIMIR: Experiments on causation of beri-

beri. Lane. Nov. 4th, p. 1266, 1911.

278. THEILER, A. GREEN, H. H., AND VILJOEN, P. R. : Contribution to

the study of deficiency diseases, with special reference to the
lamziekte problem im South Africa. 3rd and 4th Director Veter.
Res. Rep. Dept. Agr. Union S. Africa 9, 1915.

LITERATURE TO THE TEXT 413

279. LUMIERE, A. : Bull de 1'acad. de meU 83, 96, 1920.

280. MERKLEN, P. : Modification of diet saves ducklings from epidemic

diseases. Bull, de la soc. de p6d. 16, April, 1914.

281. KtiLZ, L.: Ueber Beriberi derEnten. Arch. Schiffs- und Tropenh. 16, 163,

1912.

282. FUJITANI, J. : Beitrage zur aetiologischen Kenntnis der bei Reisfiitterung

auftretenden Krankheit der Vogel. Mitt. Berib. Stud. Komm.
Tokyo. 306, 1911.

283. TOYAMA, C.: Ueber eine kakkeahnliche Krankheit der Vogel. Ibid.

274, 1911.

284. FINK, G. L. : Beriberi und weisser Reis, cin Versuch mit Papageien. J.

Trop. Med. Hyg. 15, VIII, 1910; Arch. Schiffs u. Tropenhyg. 15,
270, 1911.

284a. JANSEN, B. C. P.: Vitamine content of extract of rice bran. Medel.
v. I. Burg. Dienst. Ned. Indie. No. 1, 23, 1920.

285. KIMURA, ONARI: Degeneration and regeneration in peripheral nerves.

Mitt. Path. Inst. Univers. Sendai, Japan, March 24th, 1919.

286. SCHNTDER, K. : Pathologisch-anatomische Untersuchungen bei experi-

mentellen Beriberi (Reispolyneuritis) Arch. f. Verdauungskrankh.
20, 147, 1914.

287. WEILL AND MOURIQUAND: Neurologic expe"rimentale. Les paralysies

par carence. Soc. de neurol. Mai, 1917.

288. KATO, G. AND SHIZUME, S. : Physiologic and pathologic examination of

nerves and muscles of domestic fowl suffering from so-called
"polished rice disease". Japan Med. World. Nov. 23, No. 310, 1919.

289. PAGUCHI, K. : Ibid. Dec. 14th, No. 313, 1919.

290. McCARRisoN, R. : Genesis of edema in beriberi. Proc. R. Soc. 91 B, 103,

1920; Pathogenesis of deficiency diseases II. Effects of deprivation
of "B" accessory food factor. Ind. J. Med. Res. 6, 550, 1919.

291. FUNK, CASIMIR AND DOUGLAS, MACKENZIE: Studies on beriberi VIII.

The relationship of beriberi to glands of internal secretion. J.
Physiol. 47, 475, 1914.

292. WILLIAMS, R. R. AND CROWELL, B. C. : Thymus gland in beriberi.

Philipp. J. Sci. 10 B, 121, 1915.

293. MCCARRISON, R. : Involution of thymus in birds. Ind. J. Med. Res.

6, 557, 1919.

294. DOUGLAS, MACKENZIE: The histology of the thyroid in animals fed on

various diets. J. Path. Bact. 19, 341, 1915.

295. MCCARRISON, R. : Pathogenesis of deficiency diseases. X. Effect of

some food deficiencies and excesses in thyroid gland. Ind. J. Med.
Res. 7. 633, 1920.

296. MCCARRISON: Ibid. 5, 2755, 1919.

297. MCCARRISON: Ibid. 2, 369, 1914.

298. MCCARRISON: Pathogenesis of deficiency diseases. V. Histopathology.

Ibid. 7, 269, 1919.

299. MCCARRISON: The influence of deficiency of accessory food factors on

the intestine. Brit. Med. J. July 12th, 1919.

414 THE VITAMINES

300. McCARRisoN: The pathogenesis of deficiency diseases. III. The

influence of dietaries deficient in accessory food factors on the
intestine. Ind. J. Med. Res. 7, 167,4919.

301. FUNK, CASIMIR: The effect of a diet of polished rice on the nitrogen and

phosphorus of the brain. J. Physiol. 44, 51, 1912.

302. WIELAND, HERMANN: Analytische Untersuchungen tiber den P-gehalt

der ernahrungskranken Tieren. Arch. exp. Path. Pharm. 69, 93,
1912.

303. KOCH, MATHILDE AND VOEGTLIN, CARL: Chemical changes in the central

nervous system as a result of restricted vegetable diet. II. S. Publ.
Health Serv. Hyg. Lab. Bull. 103, 1916.

304. FUNK, CASIMIR AND v. SCHONBORN: The influence of vitamine-free

diet on the carbohydrate metabolism. J. Physiol. 48, 328, 1914.

305. FUGI, S. : Diastase in blood of chickens fed on polished rice. Jikwa

Zasshi No. 203, 36, 1917.

305a. FINDLAY, GEORGE MARSHALL: Glyoxylase in beriberi. Biochem.
J. 15, 104, 1921.

306. FUNK, CASIMIR: Ueber die physiologische Bedeutung gewisser unbisher

bekannter Nahrungsbestandteile, der Vitamine. Erg. d. Physiol.
13, 125, 1913.

307. BENEDICT, STANLEY R. : Uric acid in its relation to metabolism. J.

Lab. Clin. Med. 2, No. 1, 1916.

308. HUNTER, ANDREW AND WARD, F. W. : Purine metabolism in various

representative mammals. Trans. R. Soc. Canada. 13, Section IV,
7, 1919.

309. McCoLLUM, E. V., SIMMONDS, N. AND PITZ, W. : The effects of feeding

the proteins of the wheat kernel at different planes of intake. J.
Biol. Chem. 28, 211, 1916.

310. JACKSON, C. M. ; Biol. Bull. 23, 171, 1917.

311. MACALLUM, ARCHIBALD BRUCE: The relation of vitamines to the growth

of young animals. Trans. R. Can. Inst. p. 175, 1919.

312. FERRY, EDNA L. : Nutrition experiments with rats. J. Lab. Clin. Med.

5, 735, 1910.

313. DRUMMOND, J. C. : The growth of rats upon artificial diet containing

lactose. Biochem. J. 10, 89, 1916.

314. OSBORNE AND MENDEL: J. Biol. Chem. 34, 309, 1918.

315. McCoLLUM AND SIMMONDS : A study of the dietary essential water-soluble

B, in relation to its solubility and stability towards reagents.
Ibid. 33, 55, 1918.

316. OSBORNE AND MENDEL: Ibid. 18, 95, 1914; The suppression of growth

and the process of growth. Ibid. 23, 439, 1915.

317. JACKSON, C. M. AND STEWART, C. A. : The effects of underfeeding and

refeeding upon the growth of various systems and organs of the
body. Minn. Med. Nov., 1918; J. of Zo6l. 30, 97, 1920.

318. BRUNING, H. : Experimentelle Studien iiber die Entwickelung neuge-

borener Tiere bei langerdauernden Trennung von der saugenden
Mutter und nachheriger verschiedenartiger kiinstlichen Ernahrung.
J. f. Kinderh. 80, 65, 1914.

LITERATURE TO THE TEXT 415

319. ARON, HANS: Untersuchungen iiber die Beeinflussung des Wachsturns

durch Ernahrung. Berl. klin. W. 51, 972, 1914.

320. SLONACKER, J. R. : Leland Stanford, Jun. Univers. Public. 1912.

321. OSBORNE AND MENDEL: The choice between adequate and inadequate

diets, as made by rats. J. Biol Chem. 35, 19, 1918.

322. SURE, BARNETT: The nutritive value of lactalbumin; cystine and tyro-

sine as growth-limiting factors in that protein. Ibid. 43, 457, 1920.

323. SUZUKI, UMETARO, OKUDA, YUZURU, MATSUYAMA, YOSHIHIKO, OKIMOTO,

TAMAO, KATAKURA, KEI AND IWATA, MOTOE: Nutritive value of
various proteins and fats. J. Tokyo Chem. Soc. 41, 381, 1920.

324. FUNK, CASIMIR : The preparation from yeast and certain foodstuffs of

the substance, the deficiency of which occasions polyneuritis in
birds. J. Physiol. 45, 75, 1912.

325. The same. The study of certain dietary conditions bearing on the

problem of growth in rats. J. Biol. Chem. 27, 1, 1916.

326. OSBORNE AND MENDEL: Milk as a source of water-soluble vitamine.

Ibid. 34, 537, 1918; II 41, 515, 1920.

327. STEPP, W. : Weitere Untersuchungen liber die Unentbehrlichkeit der

Lipoide fur das Leben. Ueber die Hitzezerstorbarkeit lebens-
wichtiger Lipoide der Nahrung. Z. f. Biol. 59, 336, 1912.

328. LANDER, PERCY E. : On the cholesterol content of the tissues of growing

rats when under various diets. Biochem. J. 9, 78, 1915.

329. ARON, H. : Die Bedeutung von Extraktstoffen fur die Ernahrung.

Monatsh. Kinderh. 13, 359, 1915.

330. FUNK, CASIMIR AND MACALLUM, A. B., JR.: Studies on growth. III.

The comparative value of lard and butter fat in growth. J. Biol.
Chem. 27, 51, 1916.

331. FUNK, CASIMIR AND DUBIN, HARRY E. : The vitamine requirements of

the rat on diets rich in protein, carbohydrate- and fat respectively.
Science, N. S., 52, 447, 1920.

332. DESGREZ, A. AND BIERRY, H. : L'equilibre d'azote et 1'absence des

vitamines. C. r. 170, 1209, 1920.

333. Report of Medical Research Committee on the present state of knowledge

concerning accessory food factors (vitamines). No. 38, 1919.

334. VOEGTLIN, Carl AND LAKE, G. C. : Experimental mammalian polyneuritis

produced by a deficient diet. Am. J. Physiol. 48, 558. 1919.

335. OSBORNE, T. B. : The water-soluble vitamine. N. Y. State Med. J.

20, 217, 1920.

336. OSBORNE AND MENDEL. The role of vitamines in the diet. J. Biol.

Chem. 31, 149, 1917.

337. DRUMMOND, J. C. : A study of the water-soluble accessory growth-

promoting substance. II. Its influence upon the nutrition and
nitrogen metabolism of the rat. Biochem. J. 12, 25, 1918.

338. OSBORNE AND MENDEL: The nutritive value of yeast protein. J. Biol.

Chem. 38, 223, 1919.

339. EMMETT, A. D. AND ALLEN, F. P.: Pathogenesis due to vitamine defi-

ciency in the rat. Proc. Soc. Amer. Biol. Chem. XIV, J. Biol.
Chem. 41, LIII, 1920.

416 THE VITAMINES

339a. TSULI, K.: Effect of diet on thyroid functioning. Acta Scholae
Med. Kyoto, 3, 713, 1920.

340. ARON, HANS: fieri, klin. W. No. 23, 546, 1918.

340a. MORGULIS, S. AND GIES, W. J. . Calcium content of bones and teeth

from normal and thymectomized albino rats. J. Exp. Med. 20,

499, 1914.
340b. MATTILL, H. A.: Influence of fasting and vitamine B deprivation on

the non-protein nitrogen of rat's blood. Science, 54, 176, 1921.
340c. BRUNING, H. : Untersuchungen iiber das Wachstum von Tieren jenseits

der Sanglingsperiode by verschiedenartiger kunstlichen Ernahrung.

J. f. Kinderh. 79, 305, 1914.

341. MACALLUM, ARCHIBALD BRUCE: The relationship of vitamines to animal

growth. Amer. Med. 11, 782, 1916.

341a. HOPKINS, F. G.: Note on vitamine content of milk. Biochem J.
14, 721, 1920.

342. FREISE, E. : Nahrungsbestandteile die das Wachstum erregen. J. f.

Kinderh. 91, 79, 1920.

342a. FUNK, CASIMIR AND DUBIN, HARRY E.: Vitamines of autolyzed
yeast in nutrition. Soc. Exp. Biol. Med. Oct. 19, 1921.

343. ROBERTSON, T. BRAILSPORD: The normal growth of the white mouse.

J. Biol. Chem. 24, 363, 1916.

344. THOMPSON, HELEN B., AND MENDEL, L. B. : Alternating growth and

suppression of growth in the albino mouse, with special reference
to the economy of food consumption. Am. J. Physiol. 45, 431, 1918.

345. MENDEL AND JUDSON, S. E. : Some interrelations between diet, growth

and chemical composition of the body. Proc. Nat. Acad. Sci. 2,
692, 1916.

346. SCHMIDT, M. B. : Eisenstoffwechsel und Blutbildung. Mitt. d. Physik.

Med. Ges. Wurzburg. Ref. D. med. W. 40, 205, 1914; Verh. d. deutsch.
path. Ges. 17 Tagung, 156, 1914.

347. MORPURGO, B. AND SATTA, G. : Sugli scambi de sostanze nutrienti fra

topi in parabiosi. Arch, di fisiol. 11, 360, 1914,

348. STEPP, W. : Untersuchungen tiber die Unentbehrlichkeit der Lipoide

fur das Leben. Z. f. Biol. 62, 405, 1913.

349. ROHL, W. : Experimentelle Untersuchungen iiber den Aufbau von

Lipoiden im Tierkorper. Verh. des 29en deutsch. Kongr. f. inn.
Med. Wiesbaden, 1912.

350. STEPP, W. : Ueber lipoidfreie Ernahrung und ihre Beziehungen zur

Beriberi und Skorbut. D. med. W. 40, 892, 1914.

351. STEPP: Lipoid Hunger und Beriberi. Z. f. Biol. 66, 339, 1916.

352. STEPP: Die Unentbehtlichkeit der Lipoide fur das Leben. Ibid. 66,

365, 1916.

353. DEZANI, SERAFINO: Untersuchungen tiber die Genese des Cholesterins.

Arch. farm. 17, 4, 1914.

354. DEZANI : Ricerche sulla nutrizione alipoidea. Biochimica. 4, 475, 1914.

355. ROHMANN, F. : Ueber die Ernahrung von Mausen mit einer aus einfachen

Nahrungstoffen zusammengesetzter Nahrung. Bio. Z. 64, 30, 1914.

LITERATURE TO THE TEXT 417

356. WHEELER, RUTH: Feeding experiments with mice. J. Exp. Zool, 15,

209, 1913.

357. MACARTHUR, C. G. AND LTJCKETT, C. L. : Lipins in nutrition. J. Biol.

Chem. 30, 161, 1915.

358. MITCHELL, H. H. AND NELSON, B. A. : The preparation of protein-free

milk. J. Biol. Chem. 23, 459, 1915.
358a. HUME, E. M.: Comparison of the growth-promoting properties for

guinea pigs of certain diets consisting of natural foodstuffs. Bio-

chem. J. 15,30, 1921.,
358b. TOZER, F. M. : Effect on guinea pigs of deprivation of vitamine A and

of antiscorbutic factor. J. Path. Bact. 24, 306, 1921 .

359. SMITH, THEOBALD: Bacilli in swine disease. Bur. of Anim. Ind. Wash.

172, 1895-6.

360. HOLST, AXEL AND FROLICH, TH. : Ueber experimentellen Skorbut. Z.

f. Hyg. u. Infektionsk. 72, 1, 1912.

361. JACKSON AND MOORE: J. Inf. Dis. 19, 510, 1916.

362. JACKSON AND MOODY: Ibid. 19, 511, 1916.

363. MOORE, J. J. AND JACKSON, L. : Experimental scurvy produced in guinea

pigs by milk and milk products. J. A. M. A. 67, 1931, 1916.

364. GIVENS, MAURICE H. AND HOFFMAN, GEORGE L. : Preliminary obser-

vations on the relation of bacteria to experimental scurvy in guinea
pigs. Proc. Soc. Amer. Biol Chem. XIV. J. Biol. Chem. 41,
XXXIII, 1920.

365. PITZ, W. : Studies on experimental scurvy. J. Biol. Chem. 33, 471, 1918;

36, 439, 1918.

366. TORREY, J. C. AND HESS, A. F. : The relation of the intestinal flora to

the scurvy of guinea pigs and of infants. Proc. Soc. Exp. Biol.
15, 74, 1918.

367. HARDEN AND ZILVA: Note on the etiology of scurvy in guinea pigs.

Biochem. J. 12, 270, 1918.

368. FUNK, CASIMIR: Nature of the disease due to the exclusive diet of oats

in guinea pigs and rabbits. J. Biol. Chem. 25, 409, 1916.

369. HEIM, P.: Die Bedeutung der Extrakstoffe fur die Ernahrung. Mon.

f. Kinderh. 13, 367, 1914-6; 13, 477, 1916.

370. RONDONI, PIETRO AND MoNTAGNANi, MARIO : Lesioni istologiche nel

maidismo nel digiuno e nello scorbuto sperimentale. H Speriment.
69, 659, 1915.

371. INGIER, A.: Ueber experimentell hervorgerufenen Morbus Barlow an

Foten und Neugeborenen. Nord. Med. Ark. 48, 1, 1915; Abt. II;
J. Exp. Med. 21, 525, 1915.

372. McCoLLUM, SIMMONDS AND PITZ : The nature of dietary deficiencies

of the oat kernel. J. Biol. Chem. 29, 341, 1917.

373. CHICK, HARRIETTS AND HUME, E. MARGARET: Trans. Soc. Trop. Med. &

Hyg. 10, 152, 1917.

374. COHEN, BARNETT AND MENDEL, L. B. : Experimental scurvy of the

guinea pig in relation to the diet. J. Biol. Chem. 35, 425, 1918.

375. HESS, A. F. AND UNGER, L. J. : Experiments on scurvy of guinea pig.

Proc. Soc. Exp. Biol. Med. 15, 82, 1918; J. Biol. Chem. 35, 479, 1918.

418 THE VITAMINES

376. CHICK, HARRIETTS AND co-workers: Cited according to Med. Res.

Comm. Rep. No. 38. p. 43, 1919.

377. HESS, A. F. : The antiscorbutic vitamine. N. Y. State J. of Med. 20,

209, 1920.

378. HESS AND UNGER. Scorbutic beading of ribs. Amer. J. Dis. Childr.

19, 331, 1920; Br. M. Assocn. Meeting June 29 Cambridge, Br.
Med. J. July 31, 154, 1920.

379. RONDONI, P. : Pathogenesis of deficiency diseases and pellagra. Br.

Med. J. I, 542, 1919.

380. McCARRisoN, R. : Influence af scorbutic diet on suprarenals. Ind. J.

Med. Res. 7, 188, 1919; Br. Med. J. Aug. 16th, 200, 1919.

381. LAMER, V. K. AND CAMPBELL, L. H. : Changes in organ weight produced

by deficient diet in antiscorbutic vitamine. Proc. Soc. Exp. Biol.
Med. 1920.

382. MCCARRISON, R. : Pathogenesis of deficiency diseases. VI. Influence

of scorbutic diet on bladder. Ind. J. Med. Res. 7, 279, 1919.

383. BATJMAN, L. AND HOWARD, C. P. : The mineral metabolism of experi-

mental scurvy in the guinea pig. Amer. J. Med. Sci. 153, 650, 1917.

384. SMITH, MILLARD L. AND LEWIS, HOWARD B. : Study of the normal

metabolism of the guinea-pig. J. Amer. Chem. Soc. 39, 2231, 1917.

385. McCLENDON, J. F., COLE, W. C. C., ENGSTRAND, O. AND MIDDLEKAUFF,

J. E. : The effect of malt and malt extracts and the alkaline reserve
of the blood. J. Biol. Chem. 40, 243, 1919.

386. LEWIS, HOWARD B. AND KARR, WALTER G.: Changes in the urea content

of the blood and tissues of guinea pigs maintained on an exclusive
oat diet. J. Biol. Chem. 28, 17, 1916.

387. KARR, W. G. AND LEWIS, H. B. : The phenol excretion of guinea pigs

maintained on an exclusive oat diet. Amer. J. Physiol. 44, 586,
1917.

387a. McCoLLtJM AND PARSONS; The antiscorbutic requirements of the
prairie dog. J. Biol. Chem. 44, 603, 1920.

388. MORGEN, A. AND BEGER, C. : Ueber den schadlichen, auf einer Saure-

vergiftung zurtickftihrenden Einfluss einer ausschiesslichen Ernah-
rung. H. 94, 324, 1915.

389. KURIJAMA, S. : J. Biol. Chem. 33, 215, 1918.

390. MCCLENDON, J. F., v. MEYSENBUG, L., ENGSTRAND, O. J. AND KING,

FRANCIS : Effect of diet on the alcaline reserve of the blood. Ibid.
38, 539, 1919.

391. NELSON, V. E. AND LAMB, A. R. : The effect of vitamine deficiency in

various species of animals. I. The production of xerophtalmia in
the rabbit. Amer. J. Physiol. 51, 530, 1920.

392. WEILL, E., MOTJRIQTJAND, G. AND MICHEL, P.: Recherches sur les

maladies par carence; effets compare's d'alimentation exclusive des
chats, avec la viande crue, conge!4e, salee, buillie et sterilised. C.
r. soc. biol. 79, 189, 1916.

393. VOEGTLIN, CARL AND LAKE, G. C. : Experimental mammalian polyneu-

ritis produced by a diet deficient in antineutiric "vitamine." J.
Pharmac. 11, 167, 1918; Amer. J. Physiol. 48, 558, 1919.

LITERATURE TO THE TEXT 419

393a. MACKAY, HELEN MARION MACPHERSON: The effect on kittens of a
diet deficient in animal fat. Biochem. J. 15, 19, 1921.

394. BLAND-SUTTON: Cited according to Cheadle und Poynton. Allbutt's

System of Medicine, 3, 85.

395. CAHN, A.: Die Magenverdauung im Chlorhunger. Ztschr. f. Physiol.

Chem. 10, 517, 1886.

396. TRAPBE, K. : Ueber Saurebildung im Magen. Diss. Halle. 1892.

397. ROSEMANN, R. : Ueber den Chlorgehalt des tierischen Korpers. Pfl.

Arch. 135, 177, 1910; 142, 208, 1911.

398. BONNIGER, M. : Die Substituirung des Chlors durch Brom im Tier-

korper. Z. f. exp. Path. u. Ther. 4, 414, 1907; 7, 556, 1911; 14, 452,
1913.

399. PFLUGER: Cited according to Hofmeister. Erg. d. Physiol. 16, 540, 1918.

400. HEUBNER, WOFLGANG: M. med. W. 2543, 1911; Lipschiitz, Alexander.

Untersuchung iiber den Phosphorhaushalt des wachsenden Hundes.
Arch. f. exp. Path. Pharm. 62, 210, 1910; Arch. f. ges. Physiol.
143, 91, 1911; Durlach, E. Arch. f. exp. Path. Pharm. 71, 210, 1913;
Heubner. Ueber den P-gehalt tierischer Organe nach verschieden-
artiger Fattening. Ibid. 78, 22, 1915.

401. SCHMORL, C. : Ueber die Beeinflussung des Knochenwachstums bei phos-

phorarmer Ernahrung. Ibid. 73, 313, 1913.

402. MASSLOW: Ueber die biologische Bedeutung des Phosphors fur den

wachsenden Organismus. Bio. Z. 55, 45, 1913; 56, 174, 1913; 64, 106,
1914.

403. SCHAUMANN, H. : Arch, f . Schiff. u. Tropenhyg. Beih. 6. July, 1914.

404. KARR, W. G. : The influence of water-soluble vitamine on the nutrition

of dogs. Proc. Soc. Exp. Biol. Med. 17, 84, 1920; J. Biol Chem.
44, 255, 1920.

405. STILLING AND v. MERING: Ueber experimentelles Erzeugen der Osteo-

malacie. Z. f. d. med. Wiss. 803, 1889.

406. HEBRANT, G. AND ANTOINE, G. : Sur 1'osteomalacie du chien. Acad.

de me"d. Beige. 28, 213, 1914.

407. GUERIN: Cited according to Cautley. Rickets. Garrod, Batten und

Thursfield. Diseases of Children, p. 107. London, 1913.
407a. ROLOFF, F. : Ueber Osteomalazie und Rachitis. Virch. Arch. 37,

1866; Arch. f. wiss. Thierh. 1, 189, 1875; 5, 152, 1879.
407b. REIMERS AND BOYE: Ein Beitrag zur Lehre von der Rachitis. Z. f.

innere Med. 26, 953, 1905.

408. BULL, : J. Comp. Path. 31, 1918.

408a. MELLANBY, E. AND MELLANBY, M.: The experimental production ot
thyroid hyperplasia in dogs. Proc. Physiol. Soc.; J. Physiol. 55,
VII, 1921.

408b. MELLANBY, E. AND MELLANBY, M.: The application of results ob-
tained in experiments on the hyperplasia of dogs' thyroids to the
treatment of exophthalmic goiter (Graves' Disease) . Ibid. 55, X,
1921.

408c. HENDERSON, P. S. : On the creatine content of muscles in rickets in
dogs. Proc. Physiol. Soc. Oct. 19, 1918 ; J. Physiol. Vol. 82.

420 THE VITAMINES

408d. FINDLAY, L., PATON, D. N. AND SHARPE, J. S. : Metabolism of rickets.
Quart. J. Med. 14, 352, 1921.

409. HENRI QUES AND ANDERSEN : Ueber parenterale Ernahrung durch intra-

venose Injektionen. H. 88, 357, 1913.

410. FINGERLING, G. : Landw. Vers. Stat. 86, 75, 1915.

411. HOARE, E. W. : System of veterinary medicine. Chicago 2, 1290, 1915.

412. JONES, F. S. AND ARNOLD, J. F. : Staggers in sheep in Patagonia. J.

Exp. Med. 26, No. 6, Dec., 1917.

413. REID AND ASTON: J. New Zealand. Dept. Agr. 15th Nov., 1910.

414. IBELE, J. : Die Kontrolle von "Lecksucht" und falscher Lecksucht

beim Renntier. Mitt, der Vereins z. Ford. d. Moorkultur im
Deutsch. Reiche. 36, 14, 1918.

415. EVVARD, J. M. : Proc. Iowa Acad. Sci. 22, 375, 1915.

416. HART, E. B., McCoLLUM, E. V. AND FULLER, J. G. : The role of inorganic

phosphorus in the nutrition of animals. Amer. J. Physiol. 33,
246, 1908.

417. HART AND MCCOLLUM: The influence of restricted rations on growth.

Proc. Amer. Soc. Biol. Chem. J. Biol. Chem. 17, II, 1914.

418. McCoLLtTM: The value of the proteins of the cereal grains and of milk

for growth of the pig, and the influence of the plane of intake on
growth. J. Biol. Chem. 19, 323, 1914.

419. HART AND McCoLLtnvi: Influence on growth of rations restricted to the

corn or wheat grain. Ibid. 19, 373, 1914.

420. HART AND STEENBOCK, H. : Maintenance and production value of some

protein mixtures. Ibid. 38, 267, 1919.

421. The same. Maintenance and reproduction with grains and grain

products as the sole dietary. Ibid. 39, 209, 1919.

422. KLEIN: Milchw. Zentralh. 43, 452, 1914; 44, 81, 1915; 44, 97, 1915.

422a. PLIMMER, R. H. ADERS: Note on "scurvy" in pigs. Biochem. J. 14, 570,
1920.

423. GREEN, H. H. : Dietetic deficiency. S. Afr. J. Sci. 12, 289, 1916.

424. HART, MILLER, W. S. AND MCCOLLTJM: Further studies on the nutritive

deficiencies of wheat and grain mixtures and the pathological
conditions produced in swine by their use. J. Biol. Chem. 25, 239,
1916.

425. ROBERTS, G. A. : Effect of feeding cottonseed and its products to swine.

J. Amer. Vet. Med. Ass. 49, 12, 1916.

426. ROMMEL, G. M. AND VEDDER, E. B. : Beriberi and cottonseed poisoning.

in pigs. Prelim. Comm. J. Agr. Res. 5, 489, 1915.

426a. ZILVA, S. S., GOLDING, J., DRUMMOND, J. C. AND COWARD, K. H.:
Relation of the fat-soluble factor to rickets and growth in pigs.
Biochem. J. 15, 427, 1921.

427. FRIEDBERGER, FRANZ AND FROEHNER, EUGEN: Veter. Path. 6th Ed.

Ed. Chicago 2, 1908.

428. HtiTYRA, FRANCIS AND MAREK, JOSEF: Pathology and Therapeutics of

the Diseases of Domestic Animals. Chicago 1, 1916.

LITERATURE TO THE TEXT 421

429. SCHEUNEBT, A., SCHATTKE, A. AND LOTSCH, E. : CaO, MgO, PzOs

gehalt von Heu und Hafer, nach deren Verfiitterung Pferde an
Osteomalacie erkrankten. Bio. Z. 36, 240, 1911.

430. KAWAKAMI, Z. : Disease similar to beriberi, produced in the horse by

food-foundering. Kyoto Igaku Zasshi. 14, 1, 1917.

431. STEWART: A report on the disease affecting cattle in the Moruya District.

New S. Wales Agr. J., 1899.

432. LOTSCH, E. : Ueber den Stallmangel, eine eigenartige Rindererkran-

kung in sachsischen Erzgebirge. Zut Kenntnis des Mineralstoff-
wechsels. Z. f. Inf. u. Hyg. d. Haustiere. 12, 205, 1912.

433. ASTON, B. C. AND REAKES, C. J. : J. New Zealand Dep. Agr. Nov., 1911;

Aug., 1912; Apr., 1913; Febr., 1914.

434. RUSSELL, H. L. AND MORRISON, F. B. : Univers. Wise. Agr. Exp. Sta.

Bull., 302, 55, 1919.

435. FLEISCHMANN, FRITZ: Veranderungen welche bei der Dtirrheubereitung

im Grase vor sich gehen. Landw. Vers. Stat. 76, 237, 1912.

436. HENRY, MAX: Mortality of cattle in the Bega District of New

South Wales. Veter. J. 71, No. 47S, Febr., 1915.

437. FORBES, E. B.: Mineral feeds for farm animals. Annual Meet. Amer.

Feed Manuf. Ass. Chicago, June 17-8, 1920.

438. PLACE : Diet deficiency and disease in live stock. The Farmers Weekly

5, No. 125, July, 1913.

439. THEILER, A. : Facts and theories about Stijfziekte and Lamziekte Agr.

J. Union of S. Africa, 1912.

440. HEDINGER, E. : Pathological investigation into Lamziekte. Dep. Agr.

Un. S. Africa, 1915.

441. VILJOEN, P. R. : Investigation into Lamziekte in cattle. V and VI.

Dir. Rep. Veter. Res. Un. S. Africa 257, 1918.

442. SPRUELL: Lamziekte on the Cap Plateau. Agr. J. May, 1908.

443. MITCHELL, D. F. : "Lamziekte" II Rep. Dir. Veter. Res. Dep. Agr.

Un. S. Africa, 1912.

444. WALKER, JAMES: Investigations into the disease Lamziekte in cattle.

Ibid.

445. FUNK, CASIMIR: Results of studies in vitamines and deficiency diseases

during the years 1913-5. Biochem. Bull. 4, 304, 1915.

445a. HART, E. B., STEENBOCK, H. AND HOPPERT, C. A.: The comparative
influence of green and dried plant tissue, cabbage, orange juice and
cod liver oil on calcium metabolism. Science 52, 318, 1920; J.
Biol.Chem.48,33, 1921.

446. STEAD: Some clinical reflections concerning Lamziekte. Agr. J. Un.

S. Africa 5, 386, 1913; Ibid. 725, 1914; July, 9, 1914.

447. THEILER, A., GREEN, H. H., DU TOIT, P. J., MEIER, H. AND VILJOEN, P.

R. : The cause and prevention of Lamziekte. Agr. J. Un. S. Africa
Reprint 13, July, 1920.

448. SHIGA, K. AND KUSAMA, SH. : Arch, f . Schiffs- u. Tropenhyg. 15, Beih.

3, 1911.

449. NOE: Bull soc.. path, exotique. p. 315, 1911.

422 THE VITAMINE6

450. MCCARRISON: Pathogenesis of deficiency diseases. VII. Effects of

autoclaved rice dietaries on gastro-intestinal tract of monkeys.
Ind. J. Med. Res. 7, 283, 1919; VIII. 7, 308, 1919; Br. Med. J. I.
249, Febr. 21st, 1920.

451. Hart, Carl: Ueber die experimentelle Erzeugung der Moller-Barlowscher

Krankheit und ihre endgiiltige Identifizierung mit dem klassischen
Skorbut. Virch. Arch. 208, 367, 1912.

452. HART, C. AND LESSING, OSCAR : Der Skorbut der kleinen Kinder. Ferd.

Enke. Stuttgart 1913.

453. TALBOT, TODD AND PETERSON: Boston Med. & Surg. J. 169, 232, 1913.

454. HARDEN AND ZILVA: Experimental scurvy in monkeys. J. Path. Bact.

22, 246, 1919.

455. HARDEN AND ZILVA: The antiscorbutic requirement of the monkey.

Biochem. J. 14, 131, 1920.

456. ZILVA, S. S. AND STILL, G. F. : Orbital hemorrhage with proptosis in

experimental scurvy. Lane. I. May 8th, 1008, 1920.

457. HOWARD, C. P. AND INGVALDSEN, F. : The mineral metabolism of experi-

perimental scurvy of the monkey. Bull. Johns Hopk. Hosp. 28,
222, 1917.

458. HARDEN AND ZILVA: Oedema observed in a monkey. Lane. II, 197,

780, 1919.

459. HEWLETT, TANNER R. AND DE KORTE: Oedema in a monkey. Br.

Med. J. II, 201, 1907.

460. CHICK, HARRIETTS AND HTJME, ELEANOR MARGARET: The production

in monkeys of symptoms closely resembling those of pellagra by
prolonged feeding on a diet of low protein content. Biochem. J. 14,
135, 1920.

461. HESS, ALFRED F. : Infantile scurvy. Its influence on growth (length

and weight). Amer. J. Dis. Childr. 12, 152, 1916.

462. CHICK, HARRIETTE AND DALYELL, ELSIE J. : 88th Annual Meeting.

Br. Med. A. Cambridge, June 30- July 2nd, 1920, Br. Med. J. July
31st, 147, 1920.

462a. SEIDELL, ATHERTON: Chemistry of vitamines. J. Ind. Eng. Chem.
13,72, 1921.

463. FUNK, CASIMIR: On the chemical nature of the substance which cures

polyneuritis in birds induced by a diet of polished rice. J.
Physiol. 43, 395, 1911.

464. SCHAUMANN, H. : Zu dem Problem der Beriberiatiologie. Arch, f .

Schiffs- u. Tropenhyg. 16, 825, 1912.

465. FUNK, CASIMIR: Studies on beriberi. VII. Further facts on the chem-

istry of the vitamine-fraction from yeast and rice-polishings. J.
Physiol. 46, 173, 1913.

466. SUZUKI, SHIMAMURA AND ODAKE: Ueber Oryzanin, ein Bestandteil der

Reiskleie und seine physiologische Bede*utung. Bio. Z. 43, 89, 1912.

467. SUZUKI AND MATSUNAGA: On the occurrence of nicotinic acid in rice

bran. Biochem. Bull. 2, 228, 1913.

LITERATURE TO THE TEXT 423

468. SCHAUMANN, H. : Ueber die Darstellung und Wirkungsweise einer der

in der Reiskleie enthaltenen gegen experimentelle Polyneuritis
wirksamen Substanz (Vorl. Mitt). Arch. f. Schiffs- u. Tropenhyg.
16, 349, 1912; D. med. W. No. 26, 1255, 1912.

469. WELLMAN, C., EUSTIS, A. C. AND SCOTT, L. C. : Rapid cure of polyneu-

ritis gallinarum by intramuscular injection of a substance isolated
from rice. Amer. J. Trop. Dis. & Prev. Med. 1, 295, 1913.

470. EUSTIS, A. C. AND SCOTT, L. C. : Isolation of vitamine from rice-polish-

ings. Biochem. Bull. 3, 466, 1914.

471. VEDDER, E. B. AND WILLIAMS, R. R. : Beriberi-preventing substances

or vitamines contained in rice polishings. VI. Philipp. J. Sci. 8
(B), 175, 1913.

472. WILLIAMS, R. R. AND SALEEBY, N. B. : Experimental treatment of

human beriberi with constituents of rice polishings. Ibid. 10(B),
99, 1915.

473. WILLIAMS, R. R. : Chemistry of the vitamines. Ibid. 11 (A), 49, 1916.

474. ISSOGLIO, GIOVANNI: Chemical composition of the by-products of the

working up of the rice. Atti acad. sci. Torino, 54, 980, 1919.

475. KONDO, H. AND GOMI, N. : Efficacious constituents of rice bran. J.

Pharm. Soc. Japan No. 391, 1013, 1913.

476. MTJRAI, T. : A new method for the extraction of the efficacious constit-

uents of the rice bran. Ibid. No. 386, 1914.

477. TSTJZXJKI, J. : Remedy for beri-beri. Brit. 25,322, Nov. 5th, 1912.

478. GAMS, A. AND SCHREIBER, B.: Vitamines. U. S. P. 1,235,198, July 31,

1917; Soc. Anon, pour 1'Ind. chim. a Bale. Schweiz. 76, 494, Apr.
1st, 1918.

479. BRILL, HARVEY C. : The antineuritic properties of the infusorial earth

extract of the hydrolyzed extract of rice polishings. Philipp. J.
Sci. 12, 199, 1917.

480. FRASER, H. AND STANTON, A. T. : The chemistry of rice-polishings.

Lane. I, 1021, 1915.

481. DRUMMOND, J. C. AND FUNK, CASIMIR: Chemical investigation of the

phosphotungstic acid precipitate fron rice polishings. Biochem.
J. 8, 598, 1914.

482. BARGER: Simpler Natural Bases. Longmans, Green & Co., London,

1914.

483. HOFMEISTER, FRANZ AND TANAKA, M. i cited by Hofmeister, Erg. d.

Physiol. 16, 510, 1918.

484. HOFMEISTER, FR. : Zur Kenntnis der alkaloidischen Bestandteile der

Reiskleie. Bio. Z. 103, 218, 1920.

485. FUNK, CASIMIR: Further facts concerning the chemistry of the vitamine-

fraction from yeast. Brit. Med. J. Apr. 19th, 1913.

486. FOLIN, O. AND MACALLUM, A. B., JR. : J. Biol. Chem. 11, 265, 1912; 13,

363, 1912.

487. BARSICKOW: Experimentelle Untersuchungen liber die therapeutische

Wirkung der Hefe bei der alimentaren, multiplen Polyneuritis der
Meerschweinchen und Tauben. Bio. Z. 48, 418, 1913.

424 THE VITAMINES

488. EDIE, EVANS, MOORE, SIMPSON AND WEBSTER: The antineuritic bases

of vegetable origin in relationship to beriberi, with a method of
isolation of torulin, the antineuritic base of yeast. Biochem. J. 6,
234, 1913.

489. FUNK, CASIMIR: Fractionation of phosphotungstic acid precipitate

with acetone as a useful method for the preparation of the vitamine-
fraction from yeast. Biochem. Bull. 5, 1, 1916; U. S. P. 1,162,908,
Dec. 7th, 1915.

490. VAN SLYKE, D.D .: J. Biol. Chem., 30, 115, 1917.

491. BOHRINGER, C. F. AND SONS: Separating active substances from food

and organ extracts. Holl. 2600, Oct. 15, 1918.

492. COOPER, E. A. : The curative action of autolyzed yeast against avian

polyneuritis. Biochem. J. 8, 250, 1914.

493. FUNK, CASIMIR AND DUBIN, HARRY E. : A test for antiberiberi vitamine

and its practical application. J. Biol Chem. 44, 487, 1920.

494. DRUMMOND, J. C. : Observations on the phosphotungstates of certain

bases and aminoacids. Biochem. J. 12, 5, 1918.

495. SEIDELL, ATHERTON: Vitamines and nutritional diseases. A stable

form of vitamine, efficient in the prevention and cure of certain nu-
tritional deficiency diseases. Repr. 325, U. S. P. H. S. Rep. Febr.
18th, 1916; Seidell A., Vitamines, U. S. Pat. 1,173,317, Febr. 28th,
1916.

496. WILLIAMS, R. R. AND SEIDELL, A. : The chemical nature of the "vita-

mines". II. Isomerism in natural antineuritic substances. J.

Biol. Chem. 26, 431, 1916.
496a. OSBORNE, T. B. AND LEAVENWORTH, C. L.: Effect of alkali on the

efficiency of water-soluble vitamine B. J. Biol. Chem. 45, 423, 1921.
496b. RHODEHAMEL, H. W. AND STUART, A. H.: Atropine sulphate from

Datura stramonium. J. Ind. Eng. Chem. 13, 48, 1921.
496c . SEIDELL, A . : A stable silver vitamine compound obtained from brewer's

yeast. Publ. Health Rep. 36, 665, 1921.

497. VOEGTLIN, C. AND WHITE, G. F. : Can adenine acquire antineuritic

properties? J. Pharmacol. 9, 155, 1916.

498. DRUMMOND, J. C. : A study of water-soluble accessory growth promoting

substance in yeast. I. Biochem. J. 11, 255, 1917.

499. ABDERHALDEN, E. AND SCHAUMANN, H. : Beitrag zur Kenntnis von

organischen Nahrungsstoffen mit spezifischer Wirkung. Arch. f.
Physiol. (Pfl. Arch.) 172, 1, 1918.

500. ABDERHALDEN, E. : Weitere Beitrage zur Kenntnis von organischen

Nahrungsstoffen mit spezifischer Wirkung. Ibid. 178, 260, 1920.

501. SUGIURA, KANEMATSU: A preliminary report on the preparation of

antipolyneuritic substances from carrots and yeast. J. Biol. Chem.
36, 191, 1918.

502. OSBORNE, T. B. AND WAKEMAN, A. J. : Extraction and concentration of

Vhe water-soluble vitamine from brewer's yeast. Ibid. 40, 383,
1919.

503. MYERS, C. N. AND VOEGTLIN, C. : The chemical isolation of vitamine.

Ibid. 42, 199, 1920.

LITERATURE TO THE TEXT 425

504. POL, D. J. HULSHOFF : X-acid as a remedy in polyneuritis and beriberi.

J. Physiol. 51, 432, 1917; Ned. Tijdsr. v. Gen. 11, 806, 1917.

505. VOEGTLIN, C. AND TOWLES : The treatment of experimental beriberi

with extracts of spinal cord. J. Pharmacol. 5, 67, 1913.

506. SULLIVAN, M. X. AND VOEGTLIN, C. : Distribution in foods of the so-called

vitamines and their isolation. Proc. Soc. Biol Chem., Dec. 27th,
1915; J. Biol. Chem. 24, XVI, 1956.

507. STEENBOCK, H. : Antineuritic substance from egg yolk. Proc. Amer.

Soc. Biol. Chem. J. Biol. Chem., 29, XXVII, 1917; J. Biol. Chem.
29, 495, 1917.

508. WILLIAMS, R. R. : The chemical nature of the "vitamines." I. Anti-

neuritic properties of the hydroxypyrimidines. J. Biol. Chem. 25,
437, 1916.

509. The same. Structure of antineuritic hydroxy-pyridines. Proc. Soc.

Exp. Biol. Med. 14, 25, 1916.

510. HARDEN, A. AND ZILVA, S. S. : Alleged antineuritic properties of

o-hydroxypyridines and adenine. Biochem. J. 11, 172, 1917.

511. DUTCHER, R. ADAMS, HOLM, C. E. AND BIERMAN, HARLOW: Further

observations on the antineuritic properties of chemical substances.
Science 52, 589, 1920.

512. FUNK, CASIMIR: The influence of radium emanation on the activity of

vitamines. Proc. Soc. Exp. Biol. Med. 14, 9, 1916.

513. ZILVA, SOLOMON SYLVESTER: The action of ultra-violet rays on the

accessory food factors. Biochem. J. 13, 164, 1919.

514. SUGIURA, KANEMATSU AND BENEDICT, STANLEY R. : The action of radium

emanation on the vitamines of yeast. J. Biol Chem. 39, 421, 1919.

515. WEILL, E. AND MOURIQUAND, G. : Action des rayons X sur grains d'orge

et carence. Soc. biol. 9 Oct., 1918.

516. VOEGTLIN, C. AND MYERS, C. N. : Phosphorus as an indicator of the

vitamine content of corn and wheat products. U. S. Publ. Health
Serv. Repr. No. 471, 49, 1918.

517. GREEN, HENRY H. : The vitamine content of maize and maize-milling

products, and the ambiguity of its correlation with the phosphoric
acid content. S. Afr. J. Sci. 14, 519, 1918.

518. OTTOW, W. M. : Examination, preservation and treatment of the silver

skin of rice. Gen. Tijdsr. v. Ned. Indie, 55, 75, 1915.

519. FUNK, CASIMIR: An attempt to estimate the vitamine-fraction in milk.

Biochem. J. 7, 211, 1913.

520. BRILL, HARVEY C. AND ALINCASTRE, CECILIO: The possible maximum

vitamine content of some Philippine vegetables. Philipp. J. Sci.
12 A, 127, 1917.

521. SEIDELL, A. : The vitamine content of brewers, yeast. J. Biol. Chem.

29, 145, 1917.

522. EDDY, W. H. : Further observations on pancreatic vitamine. Proc.

Soc. Exp. Biol. Med. 14, 164, 1917.

426 THE VITAMINES

523. FUNK, CASIMIR AND MACALLUM, A. B., JR.: On the chemical nature of

substances from alcoholic extract of various foodstuffs which give
a color reaction with phosphotungstic and phosphomolybdic acids.
Biochem. J. 7, 356, 1913.

524. LEWIS, HOWARD B. AND NICOLET, BEN H. : The reaction of some purine,

pyrimidine and hydantoin derivatives with the uric acid and phenol
reagents of Folin and Denis. J. Biol. Chem. 16, 369, 1913.

525. FUNK, CASIMIR: The influence of the milling of maize on the chemical

composition and the nutritive value of maize-meal. J. Physiol.
47, 389, 1913.

526. GREEN, HENRY, H. : Experimental expression of the relationship between

the content of foodstuff in antineuritic hormone and the period of
healthy survival of animals upon it. S. Afr. J. Sci. 14, 483, 1918.

527. EDDY AND STEVENSON, H. C. : Further studies in the measurement of

the vitamine content. Proc. Soc. Exp. Biol. Med. 17, 52, 122, 1920;
J. Biol. Chem. 43, 295, 1920.

528. WILLIAMS, R. J. : A quantitative method for determination of vitamine.

J. Biol Chem. 42, 259, 1920.

529. DE SOUZA, GERALDO PAULA AND McCoLLUM, E. V. : A study of the

factors which interfere with the use of yeast as test organism for
the antineuritic substance. Ibid. 44, 113, 1920.

530. FUNK, CASIMIR AND MACALLUM, A. B., JR. : The action of yeast fractions

on the growth of rats. J. Biol. Chem. 27, 63, 1916.

531. EMMETT, A. D. AND STOCKHOLM, MABEL: Water-soluble vitamine. II.

The relation of the antineuritic and water-soluble B-vitamines to
the yeast growth-promoting stimulus. Ibid. 43, 287, 1920.

531a. GOY, PIERRE.: Les v6g6taux inferieurs et les facteurs assessoires de
la croissance. C. r. 172, 242, 1921.

531b. FRANKEL, S. AND SCHWARTZ, E.: Biochem. Z. 112, 203, 1920.

532. EIJKMAN, C. : Ueber die Natur und Wirkungsweise der gegen die

experimentelle Beriberi wirksamen Substanzen. Arch, f . Schiff- u.
Tropenhyg 17, 328, 1913.

533. FUNK, CASIMIR: Fortschritte der experimentellen Beriberiforschung

in den Jahren 1911-1913. M. med. W. No. 36, 1913.

534. FUNK: Further experimental studies on beri-beri. The action of certain

purine- and pyrymidine-derivatives. J. Physiol. 45, 489, 1913.

535. COOPER, E. A. : The preparation from animal tissues of a substance

which cures polyneuritis in birds induced by diets of polished rice.
Biochem. J. 268, 1913.

536. COOPER: On the protective and curative properties of certain foodstuffs

against polyneuritis induced in birds by a diet of polished rice.
Part I, J. of Hyg. 12, 436, 1913.

537. ABDERHALDEN, E. AND EWALD, G. : Gibt es lebenswichtige unbekannte

Nahrungstoffe? Z. f. ges. exp. Med. 5, 1, 1916.

537a. SEAMAN, EMILY C. : Influence of alcoholic extract of thyroid gland
upon polyneuritic pigeons and the metamorphosis of tadpoles.
Amer. J. Physiol. 53, 101, 1920.

LITERATURE TO THE TEXT 427

538. RAMOINO, P. : Incomplete diets. Researches on gaseous exchange on

subjects receiving a rice diet. Arch. ital. biol. 65, 1, 1916.

539. JANSEN, B. C. P., AND MANGKOEWINOTO, R. M. M. : Medel v. d. Burg.

Gen. Dienst. Ned. Indie, No. 31, 51, 1920.

540. MASSALONGO, R. : Acute polyneuritis, a form of beriberi. Contribution

to a theory of vegetable hormones. Riv. med. 32, 1316, 1916.

541. LTJMIERE, A.: Bull, de 1'Acad. de He'd. 83, 310, 1920; Presse meU 28, 29,

1920.

541a. DAMIANOVICH, H. L. AND PILADO, MATHAU C.: Biochemical and
clinical research on vitamines. Rev. de la Ass. med. Arg. 34, 279,
286,303,1921.

542. DUTCHER, R. ADAMS : Vitamine studies I. Observations on the catalase

activity of tissues in avian polyneuritis. J. Biol. Chem. 36, 63,
1918.

543. DUTCHER AND COLLATZ: II. Ibid. 36, 547, 1918.

544. BURGE, W. E. AND NEILL, A. J. : Effect of starvation on the catalase

content of the tissues. Amer. J. Physiol. 43, 58, 1917.

545. JANSEN, B. C. P. : Is vitamine identical with secretin? Gen. Tijdsr.

v. Ned. Indie. 58, 191, 1918.

546. VOEGTLIN, C. AND MYERS, C. N. : Comparison of influence of secretin

and antineuritic vitamine on pancreatic secretion and bile flow.
J. Pharmacol. 13, 301, 1919.*

546a. COWGILL, G. R.: Studies in the physiology of vitamines. Is water-
soluble vitamine identical with secretin? Proc. Soc. Exp. Biol.
Med. 18, 148, 1921. Does vitamine B stimulate glands in a manner
similar to the alkaloid pilocarpine. Ibid. 18, 290, 1921.

546b. ANREP, G. V. AND DRTJMMOND, J. C.: Note on supposed identity of
vitamine B and secretin. J. Physiol. 54, 349, 1921.

547. BICKEL, A. : Ein neues Pflanzensekretin. Berl. klin. W. 54, 74, 552,

1917. Eisenhardt, W., 54, 553, 1917. Djenab, K, 54, 624, 1917.

548. VAN EWEYK: Die Wirkung des Spinatextraktes auf die innere Pankreas-

sekretion. Virch. Arch. 227, 113, 1920.

549. BoRUTTAtr, H. : Das Verhalten der Erganzungsstoffe. II. Spezifische

antidiabetische Substanzen. Bio. Z. 88, 420, 1918.

550. UHLMANN, F. : Beitrag zu der Pharmakologie der Vitamine. Z. f . Biol.

68, 419, 1918; 68, 457, 1918.

551. BRADDON, L., AND COOPER, E. A.: J. Hyg. 14, 331, 1914.

552. MTJCKENFUSS, A..M. : The presence of food accessories in urine, bile, and

saliva. J. Amer. Chem. Soc. 40, 1606, 1918. The excretion of
vitamines. Arch of Pediatr. 36, 80, 1919.

553. GAGLIO, G. : Vitamine in urine. Policlinico. 26, 1381, 1919.

554. OSBORNE, T. B. AND MENDEL, L. B. : The role of vitamines in the diet.

J. Biol. Chem. 31, 149, 1917.

555. MATJRER: Polyneuritis der Hiihner und Beriberi, eine chronische Oxal-

saurevergiftung. M. med. W. 54, 731, 1907.

428 THE VITAMINES

556. CHAMBERLAIN, BLOOMBERGH AND KILBOURNE: A study of the influence

of the rice diet and of inanition on the production of multiple
neuritis of fowls and the bearing thereof to the etiology of beri-
beri. Philipp. J. Sci. 6 (B), 177, 1911.

557. EIJKMAN, C. AND VAN HooGENHUYZE, C. J. C. : Influence of feeding and

of starvation on the development of polyneuritis gallinarum. Proc.
Akad. Wetterschap. 18, 1467, 1916; Virch. Arch. 222, 301, 1916.

558. WALSHE, F. M. R. : The deficiency theory of the origin of beriberi in

the light of clinical and experimental observations on the disease,
with an account of a series of 40 cases. Quart. J. Med. 11, 320,
1918.

559. FUNK, CASIMIR: Studies on beri-beri. The probable role of vitamines

in the process of digestion and utilization of food. Proc. Physiol.
Soc. Dec. 13th, 1913; J. Physiol. Vol. 47.

560. FUNK: Studien iiber Beriberi. XI. Die Rolle der Vitamine bein Kohlen-

hydratstoffwechsel. Ztschr. f . physiol. Chem. 89, 378, 1914.

561. BRADDON, L. AND COOPER, E. A.: The influence of the total fuel-value

of a dietary upon the quantity of vitamine required to prevent
beriberi. Brit. Med. J. June 20, 1914; J. Hyg. 14, 331, 1914.

562. WEILL, E. AND MOURIQUAND, G. : Les maladies par carence (carence

experimentale, carence clinique). Rev. de me"d. No. 1 et 2, 1916.

563. VEDDER, E. B. : Is the neuritis-preventing vitamine concerned in carbo-

hydrate metabolism? J. Hyg. 17, 1, 1918.

564. MAIGNON, F. : Etudes comparers sur 1'influence des hydrates de carbone

et des graisses sur le pouvoir nutritif des albumines alimentaires.
C. r. 167, 172, 1919.

565. BIERRY, H. AND PORTIER, P. : C. r. soc. biol. 81, 574, 1918.

566. BIERRY, H.: C. r. 169, 197, 1919.

567. JOHNS, CARL O. AND FINKS, A. J. : Studies in nutrition IV. The nutri-

tive value of peanut flour as a supplement to wheat flour. J. Biol.
Chem. 42, 569, 1920.

567a. SCOTT, E. L., AND HONEYWELL, H. E.: Sugar in blood of normal
pigeons. Amer. J. Physiol. 54, 349, 1921 .

568. FUNK, CASIMIR: Action of substances, influencing the carbohydrate

metabolism, in experimental beriberi. J. Physiol. 53, 247, 1919.

569. ELIAS, H. AND KOLB, L. : Die Rolle der Saure in Kohlenhydratstoff-

wechsel. II. Hungerdiabetes. Bio. Z. 52, 330.

570. OSBORNE AND MENDEL: The influence of cod liver oil and some other

fats in growth. J. Biol. Chem. 17, 401, 1914.

571. Anon. : Cod liver oil industry. Pharm. Era 48, 255, 1915.

572. ISCOVESCO: Lecithide contenue dans 1'huile de foie de morue. C. r.

soc. biol. 76, I, 34; II, 74, II, 117, 1913; 10 et 17 Janv. 1914.

573. GAUTIER, ARMAND AND MOURGUES: C. r. 107, 110 et 626, 1888.

574. FUNK, CASIMIR : Biochemistry of cod-liver oil. Biochem. Bull. 4, 365, 1915.
574a. ZILVA, S. S. AND MIURA, M.: A note on the activity of fat-soluble

accessory factors in cod liver oil and butter. Lane. I. 323, 1921.

575. PAAL, C., AND ROTH, K. : Reduktion der Fette. B. 42, 1541, 1909.

LITERATURE TO THE TEXT 429

576. CHAPMAN, A. C. : Spinacene. A new hydrocarbon from certain fish oils.

J. Chem. Soc. Ill, 56, 1917.

577. TSUJIMOTO, M. : Squalene : a highly unsaturated hydrocarbon in shark

liver oil. J. Ind. and Eng. Chem. 12, 63, 1919; 12, 73, 1919.

578. KUBOTA, B. : The chemical constitution of squalene. Tokyo Kwagaku

Kwaishi. 39, 879, 1918.

579. BULL, HENRIK: The composition of cod liver oil. Tidskr. Kemi Farm.

Terapi 14, 1916.

580. ROGERS, L. : Preparation of sodium morrhuate. Brit. Med. J. Sept.

27th, 1919.

581. OSBORNE AND MENDEL: Further observations of the influence of natural

fats upon the growth. J. Biol. Chem. 20, 379, 1915.

582. DRUMMOND, J. C. : Researches on the fat-soluble accessory substance.

I. Observations upon its nature and properties. Biochem. J. 13,
81, 1919.

582a. STEENBOCK, H., SELL, MARIANA T. AND BUELL, MARY B.: Fat-
soluble vitamine. VII. The fat-soluble vitamine and yellow
pigmentation in animal fats, with some observations on its stability
to saponification. J. Biol. Chem. 47, 89, 1921 .

583. McCoLLUM, SIMMONDS AND PITZ : Distribution of the fat-soluble A, the

dietary essential of butter fat. Amer. J. Physiol. 41, 361, 1916.

584. OSBORNE AND MENDEL: The extraction of "fat-soluble" vitamine from

green foods. Proc. Soc. Exp. Biol. Med. 16, 98, 1919.

585. ZILVA, S. S. : The extraction of the fat-soluble factor of cabbage and

carrot by solvents. Biochem. J. 14, 494, 1920.

586. STEENBOCK, H. AND BOUTWELL, P. W. : Fat-soluble vitamine VI. The

extractability of the fat-soluble vitamine from carrots, alfalfa,
and yellow corn by fat solvents. J. Biol. Chem. 42, 131, 1920.

587. STEENBOCK, H. : White corn vs. yellow corn and a probable relation

between the fat-soluble vitamine and yellow plant pigments.
Science 50, 352, 1919.

588. STEENBOCK AND BOUTWELL: Fat-soluble vitamine. III. The compara-

tive nutritional value of white and yellow maizes. J. Biol. Chem.
41, 81, 1920.

589. PALMER, LEROY S. : Carotinoids as fat-soluble vitamine. Science, 50,

501, 1919.

589a. PALMER, LEROY S. AND KENNEDY, CORNELIA: The relation of plant
carotinoids to growth and reproduction of albino rats. J. Biol.
Chem. 46, 559, 1921.

590. ROSENHEIM, OTTO AND DRUMMOND, J. C. : Relation of lipochrome

pigments to fat-soluble accessory food factors. Lane. I, 862, 1920.

590a. STEPHENSON, M.: Note on differentiation of yellow pigments from
fat-soluble vitamine. Biochem. J. 14, 715, 1920.

590b. STEENBOCK, H., SELL, M. T., NELSON, E. M. AND BUELL, M. V. : Fat-
soluble vitamine. Proc. Soc. Biol. Chem. J. Biol. Chem. 46,
XXXII, 1921.

591. VAN DEN BERGH, A. A. HUMANS AND MULLER, P.: Serum-lipochrome.

I. Proc. A'cad. Sci. Amsterdam 22, 748, 1920. '

430 THE VITAMINES

592. DRUMMOND, J. C. AND COWARD, KATHARINE HOPE: Researches on the

fat-soluble accessory substance. V. The nutritive value of animal
and vegetable oils and fats considered in relation to their color.
Biochem. J. 14, 668, 1920.

593. McCoLLUM, E. V., SIMMONDS, N. AND STEENBOCK, H. : A method for the

separation of the dietary essential "fat-soluble A", from butter
fat. Proc. Amer. Soc. Biol. Chem. ; J. Biol. Chem. 29, XXVI, 1917.

594. STEENBOCK, H., BOUTWELL, P. W. AND KENT, HAZEL E. : Fat-soluble

vitamine. I. J. Biol. Chem. 35, 517, 1918.

595. HOPKINS, F. G. : Discussion on the present condition of vitamines in

clinical medicine. Brit. Med. J. July 31st, 147, 1920.,

596. McCoLLtrM, E. V. AND DAVIS, M.: Proc. Soc. Exp. Biol. Med. 11, 101,

1913-1914.

597. DRUMMOND, J. C. : The fat-soluble accessory factor. J. Physiol. 52,

344, 1919.

598. STEENBOCK AND BOUTWELL: Fat-soluble vitamine. V. Thermostability

of the fat-soluble vitamine in plant materials. J. Biol. Chem. 41,
163, 1920.

599. OSBORNE AND MENDEL: The stability of the growth-promoting substance

in butter fat. Ibid. 24, 37, 1916.
599a. HOPKINS, F. G. : Effects of heat and aeration on fat-soluble vitamine.

Biochem. J. 14, 725, 1920.
599b. DRUMMOND, J. C. AND COWARD, K. H.: Researches on fat-soluble

accessory factors (vitamin A). VI. Effect of heat and oxygen on

vitamin value of butter. Ibid. 14, 734, 1920.

600. The same. Nutritive factors in plant tissues. IV. Fat-soluble vita-

mines. Ibid. 14, 549, 1920.

600a. ZILVA, S. S. : Action of ozone on fat-soluble factor in fats. Biochem.
J. 14, 740, 1920.

601. FAHRION, W. : Fett und Vitamine. Chem. Umschau 97, 109, 1920.

602. FREUDENBERG, ERNST AND KLOCMAN, LUDWIG: Spasmophilia. II. J. f.

Kinderh. 79, 700.

603. HESS, A. F. AND UNGER, LESTER J. : The deleterious effect of the alkalin-

ization of infants' foods. J. A. M. A. 73, 1353, 1919.

603a. DRUMMOND, J. C., COWARD, K. H. AND WATSON, A. F. : Notes on the
factors influencing the value of milk and butter sources of vitamin
A. Biochem. J. 15, 540, 1921.

604. DRUMMOND, J. C. AND COWARD, K. H. : Researches on the fat-soluble

accessory substance. III. Technique for carrying out feeding
tests for vitamine A (fat-soluble A). Biochem. J. 14, 661, 1920.

604a. STAMMERS, A. D.: The value of steam distilled palm kernel oil as a
control fat. Biochem. J. 15, 489, 1921.

605 COWARD AND DRUMMOND: Researches on the fat-soluble accessory
substance. IV. Nuts as a source of vitamin A. Biochem. J. 14,
665, 1920.

605a. DELF, E. MARION: The distribution of accessory food factors (vita-
mins) in plants. S. Afr. J. Sci. 17, 121, 1920. -

LITERATURE TO THE TEXT 431

605b. COWARD, K. H. AND DRUMMOND, J. C.: Formation of vitamin A in
living plant tissues. Biochem. J. 15, 530, 1921.

605c. HUGHES J. S.,: Effect of fat-soluble vitamine content of the feed on
the fat-soluble vitamine content of adipose tissue. Science 52,
565, 1920.

605d. DRUMMOND, J. C., GOLDING, J., ZILVA, S. S. AND COWARD, K. H: Nu-
tritive value of lard. Biochem. J. 14, 742, 1920.

606. DRUMMOND, J. C. : Researches on the fat-soluble accessory substance.

II. Observations on its role in nutrition and influence on fat metab-
olism. Ibid. 13, 95, 1919.

607. MENDEL, LAFAYETTE B. : The fat-soluble vitamine. N. Y. State Med.

J. 20, 212, 1920.

608. McCARRisoN, R. : Dietetic deficiency and endocrine activity with special

reference to deficiency edema. Brit. Med. J. Aug. 14, 236, 1920.

609. CRONHEIM: Die Rolle des Lecithins im Stoffwechsel der Erwachsene.

Z. physikal. u. diat. Ther. 16, 262.

610. ROBERTSON, T. BRAILSFORD: Experimental studies on growth. VI.

The influence of lecithin upon the growth of the white mouse. J.
Biol. Chem. 25, 647, 1916.

611. McLEAN, HUGH: A simple method for the preparation of lecithin. J.

Path. Bact. 18, 490, 1914; The composition of "lecithin" together
with observations on the distribution of phosphatides in the tissues
and methods for their extraction and purification. Biochem. J. 9,
351, 1915

612. ISCOVESCO: Les proprietes physiologiques des lecitides du foie. C. r.

soc. biol. 76, 74, 117, 1914..

613. ROBERTSON, T. BRAILSFORD : On the isolation and properties of tethelin,

the growth-controlling principle of the anterior lobe of the pituitary
body. J. Biol. Chem. 24, 409, 1916.

614. COOPER, EVELYN ASHLEY: The relation of vitamines to lipoids. Biochem.

J. 8. 347, 1914.

615. SULLIVAN, M. X. AND VOEGTLIN, CARL: The relation of lipoids to

vitamines. Proc. Amer. Soc. Biol. Chem., J. Biol. Chem; 24, XVII,
1916.

616. ARON, HANS: Nahrwerte. Bio. Z. 92, 221, 1918; Nahrungswert und die

Bedeutung von nahrenden Fett. Ibid. 103, 172, 1920.

617. STEPP, W. : Die Lipoide in ihrer Bedeutung als akzessorische Nahrstoffe.

Med. Klin. No. 3, 1920.

618. The Same: Ueber Versuche mit lipoidfreier Ernahrung an Ratten und

Hunden. Z. f. Biol. 69, 495, 1919.

619. CRAMER, W. : Vitamines and lipoid metabolism. Proc. Physiol. Soc.

May 15, 1920; J. Physiol. 54, III, 1929; Glandular adipose tissue;
relation to other endocrine organs and to vitamine problem. Brit.
J. Exp. Path. 1, 184, 1920.

620. HOLST, AXEL AND FROLICH, TH. : Experimenteller Skorbut. Weitere

Untersuchungen tiber die Konservierung und Extraktion der spezi-
fischen Bestanteile der antiskorbutischen Nahrungsmittel. Z. f.
Hyg. u. Infektionsk. 75, 334, 1913.

432 THE VITAMINES

621. SMITH, ALICE HENDERSON: A historical inquiry into the efficiency of

lime-juice for the prevention and cure of scurvy. J. Roy. Army
Med. C. Feb.-March, 1919; Lane. II, 725, 1918.

622. FREISE, E. : Der Alkoholextrakt aus Vegetabilien als Trager barlowheil-

ender Stoffe. M. f. Kinderh. 12, 687, 1914.

623 FREUDENBERG, ERNST. : Beitrag zur Frage des Barlow Schutzstoffes.
Ibid. 13, 141, 1914-6.

624. HARDEN AND ZILVA: Antiscorbutic factor in lemon juice. Biochem. J.

12, 259, 1918.

625. HARDEN AND ROBISON, ROBERT: The antiscorbutic properties of

concentrated fruit juices. J. Roy. Army Med. C., Jan., 1919.

626. The same. The antiscorbutic properties of concentrated fruit juices.

III. Biochem. J. 14, 171, 1920.
626a. VEDDER, E.B.: Etiology of scurvy. Milit. Surg.49, 133, 1921.

627. GIVENS, M. H. AND MCCLUGAGE, H. B. : Antiscorbutic property of

fruits. I. An experimental study of dried orange juices. Amer.
J. Dis. Childr. 18, 30, 1919.

628. DUBIN, HARRY E. AND LEWI, MAURICE J. : Amer. J. Med. Sci. 159, 264,

1920.

629. BASSETT-SMITH : Scurvy. With special reference to prophylaxis in the

Royal Navy. Lane. May 22nd, 1102, 1920.

630. HARDEN AND ZILVA: A note on the susceptibility of the antiscorbutic

principle to alkalinity. Lane. II, 320, 1918.

631. HESS AND UNGER: The scurvy of guinea pigs. III. The effect of age,

heat, and reaction on antiscorbutic foods. J. Biol .Chem. 38, 293,
1919.

632. McCLENDON, J. F. AND SHARP, PAUL F. : The hydrogen ion concentra-

tion of foods. J. Biol. Chem. 38, 531, 1919.

633. SOMMER, H. H., AND HART, E. B. : The effect of heat on the citric acid

of milk. Ibid. 35, 313, 1918.

634. FABER, HAROLD K. : Sodium citrate and scurvy. Proc. Soc. Exp. Biol.

Med. 17, 140, 1920.

635. FOWLER, G. S. : Edinb. Med. J. Jan. 2nd, 1914.

636. HESS, A. F. : Brit. Med. J. July 31st, 154, 1920.

636a. ZILVA, S. S. AND MIURA, M.: The differential dialysis of the anti-
neuritic and the antiscorbutic factors. Biochem. J. 15, 422, 1921.

637. HESS AND UNGER: Experiments on antiscorbutics. Report of an anti-

scorbutic for intravenous use. Proc. Soc. Exp. Biol. Med. 15, 141,
1918.

638. HESS, A. F. : Infantile scurvy. Proc. II, Pan-Amer. Sci. Congr. Wash.

X, 48, 1917.

639. HESS, A. F. : Scurvy. Past and present. Lippincott, Philadelphia &

London, 1920.

640. CHICK, HARRIETTE AND HUME, E. MARGARET: Note on the importance

of accurate and quantitative measurements in experimental work
on nutrition and accessory food factors. J. Biol. Chem. 39, 203,
1919.

LITERATURE TO THE TEXT 433

•641. MENDEL, L. B.: Food factors in gastro-enterology. Amer. J. Med.
Sci. 158, 297, 1919.

642. RANWEZ, F. : Les nouvelles theories sur la valeur des produits alimen-

taires en relation avec la suppression des fraudes. J. pharm. beige.
2, 537, 1920.

643. COOPER, E. A. : On the protective and curative properties of certain

foodstuffs against polyneuritis induced in birds by a diet of polished
rice. J. Hyg. 12, 436, 1912; 14, 12, 1914.

644. CHICK, H. AND DALYELL, E. : Skorbutgefahr in Wien. Wien. klin. W.

32, 1219, 1919.

645. CHICK, H. AND HUME, E. M. : Effect of exposure to temperature at or

above 100° upon the substance (vitamine) whose deficiency in a
diet causes polyneuritis in birds and beriberi in man. Proc. R.
Soc. 90B, 60, 1917.

646. MILLER, ELIZABETH W. : The effect of cooking on the water-soluble

vitamine in carrots and navy beans. J. Biol. Chem. 44, 159, 1920.

647. WHIFFLE, BERTHA E. : Water-soluble B in cabbage and onion. Ibid.

44, 175, 1920.

648. WEILL AND MOURIQUAND : Recherches sur le scorbut experimental

Arch, physiol. path. gen. 17, 849, 1918.

649. DELF, E. M. AND TOZER, F. M. : Antiscorbutic value of cabbage. I.

The antiscorbutic and growth-promoting properties of raw and
heated cabbage. Biochem. J. 12, 416, 1918.

650. STRUDWICK, F. : Scurvy. Cited by Med. Res. Comm. No. 38, p. 65, 1919.

651. DELF, MARION ELLEN: Effect of heat on the antiscorbutic accessory

factor of vegetables and fruit juices. Biochem. J. 14, 211, 1920.

652. GIVENS, MAURICE H. AND MCCLUGAGE, HARRY B. : Influence of tem-

perature on the antiscorbutic vitamine in tomatoes. Proc. Soc.
Biol. Chem.; J. Biol. Chem. 41, XXIV, 1920.

653. Rossi, G. : Experimental scurvy: the possibility of sterilizing at high

temperature without altering the alimentary value of the sterilized
substances. Arch, fisiol. 16, 125, 1918.

654. DANIELS, AMY L. AND McCuuRG, NELLIE I. : Influence of high tem-

peratures and dilute alkalies on the antineuritic property of food.
J. Biol. Chem. 37, 201, 1919.

655. DANIELS AND HEISIG, ESTHER: The acidity of various sirups used in

cooking. J. Home Econ. 11, 193, 1919.

656. HESS AND UNGER: The deleterious effect of the alkalinisation of infants

food. J. A. M. A. 73, 1353, 1919.

657. GIVENS AND MCCLUGAGE: Antiscorbutic property of vegetables. II.

An experimental study of raw and dried potatoes. J. Biol. Chem.
42, 491, 1920.

658. HINDHEDE, M. : Dytidskost samt Kortfattet Ernaeringslaere. Smaas-

kriften ud givne af det af Indenrigsministeriet nedsatte Hushold-
nigsudvaig, No. 10, 1918.

659. HILL, J. R. : Food wastage on potato cooking. Pharm. J. 100, 149, 1918.

660. DENTON, MINNA: Change in food value of vegetables due to cooking.

J. Home Econ. 11, 143, 1919.

434 THE VITAMINES

661. MTJELIN, J. R. : Boston Med. & Surg. J. 179, 395, 1918.

662. PRESCOTT, S. C. : Dried vegetables for army use. Amer. J. Physiol.

49, 578, 1919.

663. GIVENS, MAURICE H. AND COHEN, BARNETT : The antiscorbutic property

of dessicated and cooked vegetables. J. Biol Chem. 36, 127, 1918.
663a. HOLST AND FroucH : Scorbutics and its prophylaxis. Norsk Mag. f.
Laegerldensk. 77, 989, 1916; J. Trop. Med. Hyg. 23, 261, 1920.

664. GIVENS AND MCCLUGAGE: The antiscorbutic property of vegetables.

I. An experimental study of raw and dried tomatoes. Ibid. 37,
253, 1919.

665. SHORTEN, J. A. AND RAY, C. : Report on the antiberiberi vitamine

and antiscorbutic property of sun dried vegetables. Proc. Asiat.
Soc. Beng. 15, 226, 1919; Ind. J. Med. Res. Spec. No. 60, 1919.

666. TALK, K. GEORGE, McGuiRE, GRACE AND BLOUNT, EUGENIE : Studies on

enzyme action. XVII. The oxidase, peroxidase, catalase and
amylase of fresh and dehydrated vegetables. J. Biol. Chem. 38,
229, 1919.

667. BIGELOW, W. D. : Problems of canning operations. Amer. J. Publ.

Health 8, 212, 1918.

668. HESS AND UNGER: Canned tomatoes as an antiscorbutic. Proc. Soc.

Exp. Biol. Med. 16, 1, 1918.

669. CAMPBELL, MABEL E. D. AND CHICK, H. : The antiscorbutic and growth-

promoting value of canned vegetables. Lane. Aug. 23, 1919.

670. POL, D. J. HULSHOFF : Dietetic treatment of beriberi in the Dutch Indies.

Norsk Mag. Laegeridenskaben. 77, No. 1, 1916.

671. DELF, E. M. AND SKELTON, R. F. : Effect of drying on the antiscorbutic

and growth-promoting properties of cabbage. Biochem. J. 12, 448,
1918.

672. HESS AND UNGER: The scurvy of guinea pigs. II. Experiments on the

effect of the addition of fruits and vegetables to the dietary. J.
Biol. Chem. 35, 487, 1918.

673. WINTON, A. L., BURNET, W. C. AND BoRNMANN, J. H. : Composition of

corn (maize) meal manufactured by different processes and the
influence of composition on the keeping qualities. Bur. of Chem.
Dept. of Agr. Bull. 215, 1915.

674. HINDHEDE, M. : Das Ganzkornbrot. Z. physik. u. diatet. Ther. 18,

1914; Skand. Arch. Physiol. 28, 165; 32, 59, 1915. Ned. Tijdsr. v.
Gen. 1, 2115, 1919.

675. ROHMANN, F. : Der Nahrwert des Ganzkornbrotes. Berl. klin. W. 53,

105, 1916.

676. EIJKMAN, C. : White bread or dark bread. Arch. ne"erl. physiol. 1, 766.

1917

677. EIJKMAN AND POL, HXJLSHOFF: Experiments on the nutritive value of

standard white bread and white bread. Prod. Acad. Sci. Amsterd.
21, 48, 1918.

678. DUTCHER, R. A. : Shall we eat whole wheat bread? Science 47, 228, 1918.

679. STOKLASA, J. : Das Brot der Zukunft. Gust. Fischer, Jena 1917.

LITERATURE TO THE TEXT 435

680. PUGLIESE, A. : Valeur nutritive de different genres de pain. Rev. ge"n.

sci. 6, 612, 1915.

681. VAN LEERSXJM, R. L. AND MUNK, J. : Nutritive value of bread from

whole wheat flour. Arch, neerl. de physiol. 1, 446, 1917.

682. RUBNER, M. : Ganzweizenbrot. Arch. Physiol. 245, 1918.

683. v. LIEBIG, JUSTUS: Ein Mittel zur Verbesserung und Entsauerung des

Roggenbrotes. Lieb. Ann. 1854.

684. MAGENDIE: Cited according to Gustav Meyer. Z. f. Biol. 7, 1, 1871.

685. OSEKI, S. : Untersuchungen iiber qualitativ unzureichende Ernahrung.

Bio. Z. 65, 158, 1914..

686. WEILL AND MOURIQUAND: Note sur la valeur du pain de guerre. Soc.

me"d. milit. de la XIV region. 7 nov. 1915; Acad. de me'd. 26 juin
1917; Paris med. 8, 258, 1918.

687. SHERMAN, H. C., ROUSE, M. E. ALLEN, B. and WOODS E. : Growth and

reproduction upon simplified food supply. Proc. Soc. Exp. Biol.
Med. 17, 9, 1919.

688. McCoLLUM AND DAVIS : The influence of certain vegetables fat in growth.

J. Biol Chem. 21, 179, 1915.

689. CHICK AND HUME: Distribution in wheat, rice and maize grains of the

substance, the deficiency of which in diet causes polyneuritis in
birds and beriberi in man. Proc. R. Soc. 90B, 44, 1917.

690. VOEGTLIN, LAKE AND MYERS: The dietary deficiency of cereal foods

with reference to their content in antineuritic vitamine. IT. S.
Publ. Health Serv. Repr. 33, 647, 1918.

691. GREIG, E. D. W. AND CURJEL, D. F. : Antiberiberi vitamine in atta

biscuits. Ind. J. Med. Res. 6, 56, 1918.

692. WEILL AND MOURIQUAND: C. r. soc. biol. 79, 194, 1916.

693. CHICK, H. AND DELF, ELLEN MARION : The anti-scorbutic value of dry

and germinated seeds. Biochem. J. 13, 199, 1919.

694. WEILL AND MOURIQUAND: Des resultats compares sur la cobaye de

1'orge complete, dans 1'etat de reposet dans 1'etat de germination.
C. r. soc. biol. 80, 33, 1917.

695. The same and PERRONET, MLLE. : Apparition d'une substance anti-

scorbutique pendant la germination des grains de ble. Ibid. 81,
608, 678, 1918.

696. FUNK, CASIMIR AND DUBIN, HARRY E.: Unpublished data.

697. MATTEI, P. D. : Coffee and the vitamines. Policlin. 27, 1011, 1920.

698. WISE, F. B. AND BROOMELL, A. W. : The milling of rice and its mechanical

and chemical effect upon the grain. U. S. Dep. Agr. Bull. 330, 31 pp.
1915.

699. GUARESCHI, T. : Use of rice as food. Giorn. farm. chim. 66, 161, 1917;

Ann. acad. d'agr. Torino. 60, 41..

700. SANEYOSHI : On kakke. Proc. XIII. Intern. Congr. Med. Paris. 17, 78,

1900.

701. WEILL AND MOURIQUAND : Beribe'ri experimental provoque" par 1'alimen-

tation exclusive par 1'orge decortique"e ou sterilise'e. C. r. soc.
biol. 78, 649, 1918.

436 THE VITAMINES

702. STEENBOCK, H., KENT, HAZEL E. AND GROSS, E. G. : The dietary quali-

ties of barley. J. Biol. Chem. 35, 61, 1918.

703. McCoLi/UM, SIMMONDS AND PITZ : The nature of the dietary deficiencies

of the wheat embryo. Ibid. 25, 105, 1916.

704. OSBORNE AND MENDEL: The nutritive value of the wheat kernel and its

milling products. Ibid. 37, 557, 1919.

705. BEZZOLA: Beitrag zur Kenntnis der Ernahrung mit Mais. I. Einwir-

kung der Maisfiitterung auf Meerschweinchen. Z. f. Hyg. 56, 75,
1907.

706. LUCKSCH: Untersuchungen zur Pellagrafrage. Z. f. Hyg. 58, 474, 1908.

707. v. NEUSSER: Munch, med. W. 1887.

708. HOLST, AXEL: Ueber das Vorkommen skorbutischer Symptome bei

Pellagra und ihre Erklarung. Medicinsk Revu's festskrift. Juli
1911.

709. BAGLIONI. S: Feeding of corn. Arch. ital. de biol. 64, 45, 1915; Atti

accad. Lincei. 24, II, 213, 254, 1915.

710. DRISCOLL: A theory of the etiology of pellagra. South. Med. J. 6, 400,

1913.

711. CLEMENTI, A. : Observations of the effect of exclusive maize and exclusive

rice diet in relation to so-called vitamines. Rass. clin. terap. sci.
affini 16, 121, 1917; Arch. farm. sper. 2, 441, 1916.

712. SZALAGYI, K. AND KRiwuscHA, A. : Ausnutzung des Maises von Hennen,

Enten und Gansen. Bio. Z. 88, 286, 1918.

713. URBEANU, A. : Die Gefahr einer an Kaliumverbindungen zu armen

Ernahrungsweise und ihre Beziehung zu Ernahrungskrankheiten.
Berlin 1916.

714. STJAREZ, P. : Maisfiitterung in Beziehung zu der Pellagrafrage. Bio.

Z. 77, 17, 1916.

715. NITZESCO, J. J. : Nutrition exclusive des animaux avec le mais. C. r.

soc. biol 78, 222, 1915.

716. RONDONI, P. : Maize diet from the point of view of the etiology of pellagra

Speriment. 69, 723, 1915.

717. HOGAN, ALBERT G. : The nutritive properties of corn. J. Biol. Chem.

27, 193, 1916.

718. MENDEL, L. B. AND FINE, M. S. : The utilization of the proteins of the

corn. Ibid. 10, 1911.

719. OSBORNE AND MENDEL: Feeding experiments relative to the nutritive

value of the proteins of maize. Proc. Soc. Biol. Chem. ; J. Biol.
Chem. 14, XXXI, 1913.

720. BAGLIONI, S. : The effects of corn feeding. VI. Nutritive value of flour

of wheat, corn and egg in white rats. Atti accad. Lincei 24, 1. 1158,
1915.

721. McCoLLtJM AND SIMMONDS : A biological analysis of pellagra producing

diets. I. The dietary properties of mixtures of maize kernel and
bean. J. Biol. Chem. 32, 29, 1017.

722. JOHNS, CARL O., FINKS, A. J. AND PAUL, MABEL S. : Studies in nutrition.

III. The nutritive value of commercial corn gluten meal. Ibid.
41, 391, 1920.

LITERATURE TO THE TEXT 437

723. OSBORNE AND MENDEL : Skimmed milk as a supplement to corn in feeding.

Ibid. 44, 1, 1920.

724. WOODS, CHARLES D. : Food value of corn and corn products. U. S.

Dep. Agr. Bull. 298.

725. JURITZ: The chemical composition of South-African maize and other

cereals. Union S. Afr. Agr. J. Sept. 495, 1913.

726. MACCRAE: The phosphoric oxide content of maize flours. J. Hyg. 14,

395, 1914.

727. POPPE, E. : Wanderung der Bestandteile des Maiskernes in Wasser und

in wasserige Losungen. Bull. soc. chim. belg. 27, 103, 1913.

728. McCoLLUM, SIMMONDS AND PITZ : Dietary deficiencies of the maize

kernel. J. Biol. Chem. 28, 153, 1916.

729. WEILL AND MOURIQUAND. : Pellagre. Progres med. mai 1917 ; C. r. soc.

biol. 80, 372, 1917.

730. VOEGTLIN, SULLIVAN AND MYERS: Bread as a food. Change in its

vitamine content and nutritive value with reference to the occur-
rence of pellagra. U. S. Publ. Health Serv. Repr. 333, Apr. 14th,
1916.

731. HUGHES, J. S. : Some nutritive properties of corn. Kansas Agr. Exp.

Star, Bull. 5, 39, 1918.

732. HINDHEDE, M. : Skand. Arch. Physiol. 27, 277, 1912; 31, 259, 1914; Studien

iiber das Eiweissminimum. Deutsch. Arch. klin. Med. Ill, 366,
1913.

733. ABDERHALDEN, E. FODOR, A., AND ROSE, C. : Arch. ges. Physiol. 160,

511, 1915.

734. ROSE, MARY S. AND COOPER, LENA F. : The biological efficiency of

potato nitrogen. J. Biol. Chem. 30, 201, 1917.

735. RUBNER, MAX: Untersuchungen von Hindhede tiber die Verdaubarkeit

der Kartoffel. Arch. (Anat.) Physiol. 16, 1918.

736. McCoLLUM, SIMMONDS AND PARSONS: The dietary properties of the

potato. J. Biol. Chem. 36, 197, 1918.

737. OSBORNE AND MENDEL: Nutritive factor in plant tissue. III. Further

observations on the distribution of water-soluble vitamine. Ibid.
41, 451, 1920.

738. BORUTTAU, H. : TJeber das Verhalten von Erganzungstoffen. I. Bio. Z.

82, 103, 1917.

739. AUER, ALOYS: Ueber qualitativ unzureichende Ernahrung. Ibid. 93,

1, 1919.

740. LANE-CLAYPON, JANET: Value of boiled milk as a food for infants and

young animals. Rep. Loc. Gov. Board New Ser. No. 63, 1912.

741. The same. "The biological properties of milk" both of the human

species, and of cows, considered in special relation to the feeding of
infants. Ibid. No. 76, 1913.

742. The same. Milk and the Public Health. Longmans, Green & Co.,

London.

743. HIROTA: Ueber die durch die Milch, der an Beriberi leidenden Frauen,

verursachte Krankheit der Sauglinge. Z. f. inn. Med. No 16, 385,
1898.

438 THE VITAMINES

744. ANDREWS: Infantile beriberi. Philipp. J. Sci. 7, 67, 1912.

745. CARR, R. H., SPITZER, GEORGE, CALDWELL R. E..AND ANDERSON C. H.

The efficiency of certain milk substitutes in calf feeding. J. Biol.
Chem. 28,501, 1917.

746. HART, E. B., NELSON V. E. and PITZ, W. : The synthetic capacity of

the mammary gland. I. Can this gland synthetize lysine. Ibid. 36,
291, 1918.

747. McCoLLUM AND SiMMONDs: The nursing mother as a factor of safety

in the nutrition of the young. Amer. J. Physiol. 46, 275, 1918.

748. DRUMMOND, J. C. : Some aspects of infant feeding. Lane. II, 482, 1918.

749. ECKLES, C. H. AND PALMER L. S. : Influence of plane of nutrition of

the cow upon the composition and properties of milk and butter
fat. Influence of underfeeding. Miss. Exp. Agr. Sta. Bull. 25, 107,
1916.

750. ECKLES, PALMER AND SWETT W. W. : Factors influencing the compo-

sition of milk. The influence of condition at parturition on com-
position of milk and butter-fat. Ibid. 163, 33, 1919.

751. CHICK, HUME AND SKELTON R. F. : Antiscorbutic value of cow milk.

Biochem. J. 12, 131, 1918.

752. McCoLLtrM, SIMMONDS AND PITZ: The relation of the unidentified

dietary factors, the fat-soluble A and water-soluble B, of the diet
to the growth promoting properties of milk. J. Biol. Chem. 27,
33, 1916.

753. HART, STEENBOCK AND ELLIS N. R. : Influence of diet on the antiscor-

butic potency of milk. Ibid. 42, 383, 1920.

754. HESS, UNGER AND SUPPLEE G. C. : The relation of fodder to the anti-

scorbutic potency and salt content of milk. Proc. Soc. Exp. Biol.

Med. Oct. 20th, 1920.
754a. DUTCHER. ECKLES, DAHLE, MEAD AND SCHAEFER: Influence of diet

of the cow upon the nutritive and antiscorbutic properties of cow's

milk; J. Biol. Chem. 45, 119, 1920.
754b. MOORE J. J. : Experimental studies in diet deficiency diseases. Proc.

Inst. of Med. Chicago, 254. 1918.

755. GIBSON, B. B. AND CONCEPTION, J.: Philipp. J. Sci. 11B, 119, 1916.

756. CHICK, HUME AND SKELTON: The antiscorbutic value of milk in infant

feeding. Lane. I. 1, 1918.

757. HESS, A. F. : Scurvy and pasteurized milk. Amer. J. Obstetr. and

Dis. Childr. July 1916.

758. HESS, A. F. : Infantile Scurvy. A study of its pathogenesis. Amer.

J. Dis. Childr. 14, 337, 1917.

759. HESS AND UNGER: Scurvy VIII. Factors affecting the antiscorbutic

value of foods. Ibid. 17, 221, 1919.

760. MILLER: CLEVELAND. Med. J. Aug. 1917.

761. DANIELS, AMY L. AND STUESSY SYLVIA: The nutritive value of boiled

milk. Amer. J. Dis. Childr. 11, 45, 1916.

762. DANIELS AND LOUGHLIN. : A deficiency in heat-treated milks. J.

Biol. Chem. 44, 381, 1920.

LITERATURE TO THE TEXT 439

763. FROLICH, TH. : Experimentelle Untersuchugen iiber den infantilen

Skorbut. Z. f. Hyg. u. Infektionskr. 72, 155, 1912.

764. BARNES, ROSAMUND E. AND HUME E. M. : Relative anti-scorbutic

value of fresh, dried and heated cow's milk. Lane. Aug. 23rd
1919; Biochem. J. 13, 306, 1919.

765. HART, STEENBOCK AND SMITH D. W. : Studies in experimental scurvy.

Effect of heat on the antiscorbutic properties of some milk products.
J. Biol. Chem. 38, 305, 1919.

766. Food Rep. to Loc. Gov. Board No. 15,1911.

767. Anon: Dried milk powder. U. S. Publ. Health Rep. No 473, 1052, 1918;

Food Rep. No 24.

768. WINFIELD, G. : Some investigations bearing on the nutritive value

of dried milk. Loc. Gov. Board Food Rep. 24, 139, 1918.

769. NAISH: PEDIATRICS. 26, 247, 1914.

770. PRITCHARD. Ibid. 26, 300, 1914.

771. SEKINE, HIDESABURO: Nutritive defect of condensed milks. J. Tokyo

Chem. Soc. 41, 439, 1920.

772. COUTTS, F. J. H. : Upon an inquiry as to the dried milks with special

reference to their use in infant 'feeding. Loc. Gov. Board Food
Rep. 24, 1, 1918.

773. GERSTENBERGER, H. J. : Studies in adaptation of artificial food to

human milk. Amer. J. Dis. Childr. 10, No. 4, 1915; GERSTEN-
BERGER, Run, H. O., BRICKMAN, M. J., LESLIE, H. J., AND OCHSNER,
R. J. : Studies on the adaptation of an artificial food to human
milk. II. A report of three years' clinical experience with the
feeding. S. M. A. (synthetic milk adapted) Ibid. 17, 1, 1919.

774. WASHBURN, R. M. AND JONES, C. H. : Studies of the values of diff-

erent grades of milk in infant feeding. Vermont Agr. Exp. Sta.
Bull. 195, 6, 1916.

775. MATTILL, H. A. AND CONKLIN, RUTH E. : The nutritive properties

of milk, with special reference to reproduction in the albino rat.
J. Biol. Chem. 44, 137, 1920.

776. OSBORNE AND MENDEL: Nutritive factors in animal tissues. I. J. Biol.

Chem. 32, 309, 1917.

777. OSBORNE AND MENDEL: Nutritive factors in animal tissues II. Ibid.

34, 17, 1918; Proc. Soc. Exp Biol. Med. 15, 71, 1918.

778. COLE, S. W. : Report on a dried meat powder. Food (War) Comm.

R. Soc Lond. 6, 1917.

779. NANSEN, FRIDJOP. : Voyage on Fram.

780. STEFANSSON, V.: J. A. M. A.. 71, 1715, 1918.

781. CURRAN: DUBLIN J. Med. Sci. 7, 83, 1847.

782. WILLCOX, W. H. : Rations in relation to disease in Mesopotamia. Lane.

II. 677, 1917.

783. DUTCHER, R. A., PIERSON EDITH M. AND BIESTER, ALICE: The anti-

scorbutic properties of raw beef. Science. 50, 184, 1919; J. Biol.
Chem. 42, 301, 1920.

784. GIVENS, M. H. AND MCCLUGAGE H. B. : The antiscorbutic property

of dehydrated meat. Science 51, 273, 1920.

440 THE VITAMINES

785. McCoLLUM AND DAVIS: The influence of certain vegetable fats on-

growth. J. Biol. Chem. 21, 179, 1915.

786. WHEELER, RUTH AND BIESTER, ALICE: A study of the nutritive value

of some proprietery infant foods. Amer. J. Dis. Childr. 7, 169,
1914.

787. WHEELER, R. : A study of the nutritive value of some proprietary infant

foods. II. As food modifiers. Ibid. 9, 300, 1915.

788. OSBORNE AND MENDEL: The vitamines in green foods. J. Biol. Chem.

37, 187, 1919.

789. ABDERHALDEN, E. : Studien iiber den Einfluss der Art der Nahrung

auf das Wohlbefinden des einzelnen Individuums, seine Lebens-
dauer, seine Fortpflanzungsfahigkeit und das Schicksal der Nach-
kommen. Arch. ges. Physiol. 175, 187, 1919.

790. SWOBODA, FREDERICK H. : A quantitative method for the determina-

tion of vitamine in glandular and other tissue. J. Biol. Chem.
44, S31, 1920.

791. HINDHEDE, M. : Ernahrungsversuche mit grob zerquetschtem Weizen.

Skand. Arch. Physiol. 33, 263, 1916.

792. HESS, A. F. : The therapeutic effect of wheat embryo and of yeast in

infantile scurvy. Proc. Soc. Exp. Biol. Med. 13, 145, 1916; Amer.
J. Dis. Childr. 13, 98, 1917.

793. SHERMAN, H. C. : Protein requirement of maintenance in man and

nutritive efficiency of bread protein. J. Biol. Chem. 41, 97, 1920.

794. OSBORNE AND MENDEL: The food value of soy bean products. Proc.

Soc. Exp. Biol. Med. 14, 174, 1917.

795. DANIELS, AMY L. AND NICHOLS, NELL B. : The nutritive value of the

soy bean. J. Biol. Chem. 32, 91, 1917.

796. OSBORNE AND MENDEL. : The use of soy bean as food. Ibid. 32, 369,

1917.

797. WALLIS, K. L. MACKENZIE: Food value of the ground nut (arachis)

Ind. J. Med. Res. 6, 45, 1918.

798. DANIELS, AMY L. AND LOUGHLIN, ROSEMARY: Feeding experiments

with peanuts. J. Biol. Chem. 33, 295, 1918.

799. GREIG, E. D. W. : Report on antiberiberi vitamine content of ground

nut (peanut) meal biscuits. Ind. J. Med. Res. 6, 143, 1919.

800. McCoLLUM, SIMMONDS AND PITZ : The dietary deficiencies of the white

bean, Phaseolus vulgaris. J. Biol. Chem. 29, 521, 1917.

801. RICHARDSON, ANNA AND GREEN, HELENS.: Nutrition investigations

upon cottonseed meal. III. Cottonseed flour. The nature of
its growth-promoting substances, and a study in protein minimum.
J. Biol. Chem. 31, 379, 1917.

802. RICHARDSON. I & II. Ibid., 25, 307, 1916; 30, 243, 1917.

803. HOLMES, ARTHUR D. : Digestibility of steam cooked soy bean and pea-

nuts. J. A. M. A. 74, 798, 1920.

804. JOHNS, C. O. : The proteins of the peanut. Cotton Oil Press. 2, No.

12, 41, 1919.

805. McCoLLUM, SIMMONDS AND PARSONS: The dietary properties of the

pea (Vicia sativa) J. Biol. Chem. 37, 287, 1919.

LITERATURE TO THE TEXT 441

806. WILTSHIRE, H. W. : The value of germinated beans in the treatment

of scurvy. Lane. II. 811, 1918.

807. WELLS, C. A. AND EWING P. V: Cottonseed meal as an incomplete food.

J. Biol. Chem. 27, 15, 1916.

808. OSBOENE AND MENDEL: The nutritive value of some cottonseed products

in growth. Proc. Soc. Exp. Biol. Med. 13, 147, 1916.

809. STEENBOCK AND GROSS: Fat-soluble vitamine. IV. The fat-soluble

vitamine content of green plant tissues together with some obser-
vations on their water-soluble vitamine content. J. Biol. Chem. 41,
149, 1920.

810. STEENBOCK AND GROSS: Fat-soluble vitamine. II. The fat-soluble vita-

mine content of roots, together with some observations on their
water-soluble vitamine content. Ibid., 40, 501, 1919.

811. OSBORNE AND MENDEL: Nutritive factors in plant tissues. II. The

distribution of water-soluble vitamine. Ibid., 39, 29, 1919.

812. CHICK, H. AND RHODES, MABEL: The antiscorbutic value of the raw

juice of root vegetables with a view of their adoption as an adjunct
to the dietary of infants. Lane. II, 774, 1918.

813. JOHNS, C. O., FINKS, A. J. AND PAUL, M. S. : Studies in nutrition. I.

The nutritive value of coconut globulin and coconut press cake.
J. Biol. Chem. 37, 497, 1919.

814. JANSEN, B. C. P.: Fat-soluble vitamine in coconut oil. Med. v. d.

Burg. Gen. Dienst. Ned. Indie. No 6, 78, 1918; Gen. Tijdsr. v.
Ned. Indie. 58, 173.

815. PICKARD, GLENN N. : The manufacture of nut margarine. Amer. Food

J. 13, 16, 1918.

816. CAJORI, F. A.: Some nutritive properties of nuts; their proteins and

content of water-soluble vitamine. Proc. Soc. Exp. Biol. Med.
17, 78, 1920; J. Biol. Chem. 43, 583, 1920.

817. MCLEAN, STAFFORD: Case report of scurvy with a summary of fifty

other cases. Arch. Pediatr. 35, 477, 1918.

818. CHICK, HUME AND SKELTON: The relative content of antiscorbutic

principle in limes and lemons. Lane. II, 735, 1918.

819. HESS, A. F. : The role of antiscorbutics in our dietary. J. A. M. A.

71, 941, 1918.

820. LEWIS, HOWARD B. : The antiscorbutic value of the banana. J. Biol.

Chem. 40, 91, 1919.

821. CHICK, HUME AND SKELTON. The antiscorbutic value of some Indian

dried fruits. Lane. Aug. 23rd 1919.

822. JANSEN, B. C. P. : Coconut press cake as food for man. Med. v. d. Burg

Gen. Dienst. Ned. Indie No. 1, 3, 1920.

823. VAN DER WIELEN, P. The preparation of malt extract containing vita-

mine. Pharm Weekblad. 52, 673, 1915.

824. HARDEN AND ZILVA: Investigation of beer for antineuritic and anti-

scorbutic potency. J. Inst. Brewing. 24, 197, 1918.

825. DUTCHER, A. R. : Vitamine studies. III. Observations on the cura-

tive properties of honey, nectar and corn pollen in avian polyneu-
ritis. J. Biol. Chem. 36, 551, 1918.

442 THE VITAMINES

826. CHICK AND HUME: Distribution among foodstuffs (especially those

suitable for the rationing of the armies) of the substances required
for. the prevention of beriberi and scurvy. J. R. Army Med. C.
29, 121, 1917.

827. FABER, HAROLD K. : A study on the antiscorbutic value of honey. J.

Biol. Chem. 43, 113, 1920.

828. McCLENDON, J. F. AND COLE, W. C. C. : The antiscorbutic properties-

of green malt. Proc. Amer. Physiol. Soc. Amer. J. Physiol. 49,
145, 1919.

829. GERSTENBERGER, HENRY J. : Has malt-soup extract an antiscorbutic

value? Amer. Pediatr. Soc. May 31-June 2nd 1920; J. A. M. A. 75,.
99, 1920.

830. OSBORNE AND MENDEL: The influence of beef fat on growth. Proc.

Soc. Exp. Biol. Med. 12, 92, 1915.

831. EDDY, WALTER H. : The isolation of a growth-producing substance from

sheep pancreas. J. Biol. Chem. 27, 113, 1916.

832. SEKINE, HIDESABXJRO: Vitamine, fat-soluble A, in fish oils. J. Tokyo-

Chem. Soc. 41, 426, 1920.

833. EMMETT, A. D. ANT>LUROS, G. O.: The absence of fat-soluble A vitamine

in certain ductless glands. J. Biol. Chem. 38, 441, 1919.

834. HALLIBURTON, W. D. AND DRUMMOND, J. C. : The nutritive value of

margarine and butter substitutes with reference to their content
of the fat-soluble accessory growth substance. J. Physiol. 51,
235, 1917.

835. DRUMMOND, J. C: The nutritive value of certain fish. J. Physiol,

52, 95, 1918.

836. SUZUKI, UMETARO, OKUDA, SUZURU, OKIMOTO, TAMARO, AND NAGASAWA,

TAMARY: Nutritive value of various proteins. I. Nutritive
value of muscle proteins of marine animals. J. Tokyo Chem.
Soc. 40, 385, 1919.

837. McCoLLUM AND DAVIS : Nutrition with purified food substances. J.

Biol. Chem. 20, 641, 1915.

838. VEDDER, EDWARD B. : Beriberi. John Bale Sons and Danielson, Lon-

don 1913.

839. CASTELLANI AND CHALMERS: Manual of Tropical Medicine. 3d Ed.

840. SCHILLING, V. : Tropische Stoffwechselkrankheiten. Beriberi Spezielle

Path. u. Ther. d. inn. Krankh. Kraus-Brugsch. II a, 949, 1915.

841. FINDLAY: Beriberi. Practitioner. 98, 69, 1917.

842. BALZ AND MIURA: Beriberi. Mense's Handbuch der Tropenkrankhei-

ten. II. Bd. S. 140.

843. FRASER, HENRY: Rep. from the Inst. of Med. Res. Fed. Malay States

Apr. Ist-Sept. 30th, 1912.

844. SIMPSON, K. : A note on the environmental factor in the causation of

beriberi. Lane. 197, 1027, 1919.

845. HEPBURN, H. H. : Analysis of early cases of beriberi. Br. Med. J.

Apr. 3rd, 466, 1920.

846. MULVANY: Beriberi. Ind. Med. Gaz. 52, 98, 1917.

LITERATURE TO THE TEXT 443

847. CHEVALIER: Report on beriberi in Serenli. East Africa Protect. Trop.

Dis. Bull. 1, 487, 1913.

848. DUBOIS AND CORIN: Rapport sur une petite e'pidemie de be'ribe'ri

a Bokala (Congo Beige) Bull. soc. path. exot. 7, 402, 1914.

849. LOVELACE, CARL. The etiology of beriberi, J. A. M. A. 59, 2134, 1912.

850. WOLCOTT, A. M: J. A. M. A. 65, 2145, 1915.

851. FRAGA, C. : Brazil Med. 31, 20th-27th Jan. ; Feb. 3rd, 1917.

852. RIDDELL, J. D., SMITH, CHAS. H. AND IGRAVIDEZ P. G. : Beriberi at

U. S. Army Base Hospital, San Juan, Porto Rico. J. A. M. A. 72,
569, 1919.

853. LITTLE: Beriberi caused by white flour. J. A. M. A. 58, 2029, 1912.

854. PARKER: A report on beriberi in the county jail at Elizabeth, N. J.

Publ. Health Rep. 29, 339, 1914.

855. REED: Beriberi. California State J. Med. 15, 158, 1917.

856. TRAVIS: Beriberi. Kentucky Med. J. 15, 476, 1917.

857. CHANTEMESSE AND RAMOND: Cited by Schiiffner. M. med. W. 642, 1913.

858. SCHUFFNER, W. : 1st Beriberi eine in Europa endemische Krankheit.

M. med. W. No. 12, 642, 1913. ,

859. MARTINEZ, F. F. : Beriberi en Espagne et Portugal. Paris me'd. 9,

54, 1919; Arch. m<§d. beige. 73, 299, 1920.

860. SICARD, ROGER AND RIMBAUD: Paris me'd. 1 dec. 1917.

861. ROGER, H: Le be'ribe'ri en France. Med. 1, 151, 1919.

862. LEGGATE, A. R. : Observations on beriberi among the Chinese in France.

Edinb. Med. J. 24. 32, 1920.

863. MASSALONGO: Boll, delle cliniche. 31, marz. 1917.

864. WILLCOX, W. H. : Beri-beri, with special reference to prophylaxis and

treatment. Lane. 190, 553, 1916.

865. WILLCOX, W. H. Treatment and management of diseases due to

deficiency of diet: scurvy and beriberi. Brit. Med. J. I, 73, 1920.

866. BRADDON, LEONARD: Investigation as to occurrence of beriberi in

Mediterranean area. Med. Res. Comm. Rep. No 38, p. 55.

867. HEHIR: Beri-beri. Proc. Asiat. Soc. Bengal 15, 212, 1919; Mesopot.

Comm. Rep. Appendix. Ill, 1917.

868. SPRAWSON, C. A. : Beri-beri in Mesopotamian Force. Quart. J. Med.

13, 337, 1920.

869. STRONG, RICH/.RD P. AND CROWELL, B. C. : The etiology of beriberi.

Philipp. J. Sci. 7B, 271, 1912.

870. CASPARI AND MOSZKOWSKI: Weiteres zur Beriberifrage. Berl. klin.

W. 50, 1515, 1913; D. med. W. 39, 1479, 1529, 1913.

871. FRAGA, C.: Beriberi. Brazil med. 33, 49, 1919.

872. CHAMBERLAIN, WESTON P.: Prevention of beriberi among "Philippine

Scouts," by means of modification in the diet. J. A. M. A. 64,
1215, 1915.

873. KATO, S. AND YAMADA S. : Arrythmia with beriberi. Mitt. d. med. Fak.

Tokyo Univers. 19, 229, 1918.

874. DOYLE.: The clinical manifestations of beriberi. N. Y. Med. J. Apr.

8th, 1916.

444 THE VITAMINES

875. CHUN: Beriberi. Nation. Med. J. Shanghai. 3, 113, 1917.

876. YOSHIKAWA, I, YANO K, AND NEMOTO, T. : Studies of the blood in beri-

beri. Arch. Internal Med. 20, 103, 1917.

877. JIDA, H. : Examination of the cerebrospinal fluid in beriberi. Chagai

Iji Shimpo 895, 812, 1917.

878. SICABD, J. A. AND ROGEB H. : Le liquid cerebrospinal dans le be'ribe'ri.

Bull, et mem. soc. me'd. hopitaux de Paris. 34, 137, 1918.

879. GKEIG: Epidemic dropsy. Scient. memoirs by officers of the medical

and sanit. dep. of the Gov. of India. New Ser. No. 45, 1911.

880. Greig: Note on an investigation on the causation of epidemic scurvy

in Calcutta. Trans. Soc. Trop. Med. Hyg. 5, 106, 1912.

881. LAGANE: Le nourisson. No. 5, Sept. 1913.

882. WEILL, E. AND MOURIQUAND G. : La notion de la "carence" en patho-

logie infantile. Paris me'd. 3 fevr. 1917.

883. WALSHE, F. M. R. : "Food deficiency or "vitamine." theory in its

application to infantile beri-beri. Br. J. Childr. Dis. 15, 258, 1918.

884. OGATA, J. AND JIZUKI S. : Ueber den Einfluss der miitterlichen Kakke

auf den Fotus. Beitr. z. Geburtsh u. Gynak. 17, 196, 1912-13.

885. ROXAS, MANUEL: Cited according to McCollum. Newest Knowledge

of Nutrition, p. 127.

886. DUKCK, H. : Pathologische Anatomic der Beriberi. Jena 1908.

887. BALZ: Z. f. klin. Med. 4, 616, 1882.

888. ELLIS, W. G. : Lane. II, 985, 1898.

889. RUMPF AND LUCE: Deutsch. Z. f. Nervenheilk. 18, 63, 1900.

890. MOTT: The histological changes in the nervous system of Dr. Box case

of pellagra, compared with changes found in a case of pellagra
dying in Abbasieh Asylum, Cairo. Trans. Soc. Trop. Med. Hyg.
6, 156, 1913.

891. SUZUKI, T. The amount of sugar in the blood of nurslings suffering

from beriberi. Tokyo Igakukai Zasshi. (19) 30, 29, 1916; Jikwa
Zasshi No. 199, 41, 1916.

892. DE LANGEN, SCHUT, WECHUIZEN AND ALTING: Fat and lipoid content

of blood in the tropics. Med. Gen. Labor, te Welteverden, Java,
No 2-3, 44, 1919.

893. ABIMA, E. : The non-protein nitrogen blood content in cases of beriberi.

Tokyo Igakukai Zasshi. 30, 1, 1916.

894. SUZUKI, T. : Sugar content of cerebrospinal fluid in various diseases of

children. Tokyo Igaku Zasshi. 30, 231, 1916.

895. SUZUKI : Amino-acid content of the urine in beriberi. Kyoto Igaku

Zasshi. 14, 249, 1917.

896. PAGNIEZ AND VALLERY-RADOT: Be'ribe'ri. Ann. de me'd. 4, 45, 1917.

897. ONO, S. : Adrenaline content of the suprarenal glands of cadavers of pa-

tients dying from beriberi. Jikwa Zasshi. Dec. 20th, 1014, 1916;
Tokyo Igakukai Zasshi 31, 1, 1917.

898. SALEEBY, N. M. : Treatment of human beriberi with autolyzed yeast

extract. Philipp. J. Sci. 14, 11, 1919.

LITERATURE TO THE TEXT 445

899. TEL ROSARIO, M. V., AND MARANON, JOAQUIN. : Physico-chemical valua-

tion of tiki-tiki (rice polishings) extract. Philipp. J. Sci. 15,
221, 1919.

900. ALBERT, JOSE: Treatment of infantile beriberi with the extract of

tiqui-tiqui. Ibid. 10B, 81,1915.

901. Cox, A. J. : Annual Rep. Philipp. Bur. of Sci. 1918.

902. NOCHT: Festschrift zum 60 Geburtstag von R. Koch. p. 203, 1903.

903. HOLST AND FROLICH. : Experimentelle Unterschungen viber die Ur-

sache des Skorbuts. Verh. d. 6 Norddeutsch. Kongr. f. inn. Med.
p. 328, 1909.

904. HOLST, AXEL: The etiology of beriberi. Trans. Soc. Trop. Med.

Hyg. 5, 76, 1911.

905. DELPECH: Le scorbut pendant le siege de Paris. Ann d' hyg. publ.

et me'd. 16g. II ser. janv. 35, 1871.

906. BUCQTJOY: Union m6d. Sept. Oct. 1871.

907. GARCIA: Case of associated scurvy and beriberi. Siglomed. 67, 361, 1920.

908. SCHERER: Ueber Skorbut in Deutsch Siidwestafrika. Arch. f. Schiffs-

u. Tropenhyg. 17, 191, 1913.

909. GOTJZIEN: Le beriberi au Tonkin. Ann. d'hyg., et m6d. colon.

15, 445, 1912.

910. FLEMING, MACAULAY AND CLARK: Report on the prevalence and pre-

vention of scurvy and pneumonia in Southern Rhodesia among
native laborers, 1910.

911. ORENSTEIN, A. J. : Annual Rep. Sanit. Rand Mines Ltd. 1915-16.

912. DARLING: The pathological affinities of beriberi and scurvy. J. A.

M. A. 63, 1290, 1914.
913 DARLING: Trop. Dis. Bull. 5, 1915.

914. SCHRODER: Ein Beitrag zu Geschichte des Skorbuts. Arch, f . Schiff s-

u. Tropenhyg. 17, 263, 1913.

915. SCHELENZ: Scharbock: Ibid. 16, 113, 1912.

916. HIRSCH, AUGUST: Handbook of geographical and historical pathology.

London 1885. .New Sydenham Society.

917. AUTRAN : Essai historique sur le scorbut et le be'ribe'ri. These de Lyon.

1916.

918. HESS, ALFRED F. : Scurvy, Past and Present. J. B. Lippincott Co.

Philadelphia 1920.

919. Captain Cook's Voyages. Everyman Ed.

920. ELAINE, SIR GILBERT: Observations on the diseases incident to seamen.

London J. Murray 1789.

921. BUDD: Scurvy. Tweedie's System of Practical Medicine. London 1840.

922. MACNAB: Quart. J. Calcutta Med. Phys. Soc. 1,306, 1837.

923. Medical and Surgical History of the War of the Rebellion. 1888 Wash.

I, Part 3, Chapter 8.

924. GRENET, A. L. Z. : Le scorbut au Fort de Bic6tre pendant le si4ge de

Paris par les prussiens. L'hiver de 1870-71. Ann d'hyg. publ.,
36, II ser. 1871; Hayem, M. G. : Gaz. m<§d. No. 13, 186, 1871; Lasegne,
Ch. and Legroux, A.: Arch. g<5n. de m6d. 2, 680, 1871; Lev6n, M.:
Gaz. m<§d. de Paris p. 469, 1871; Dechambre: Ibid. p. 147, 1871.

446 THE VITAMINES

925. SATO, T., AND NAMBU, K. : Zur Pathologie and Anatomie des Skorbuts

Virch. Arch. 194, 151, 1908.

926. BLAU: Der Skorbut in der russischen Armee und Marine. Deutsch.

militararztl. Z. 38, 657, 1909.

927.MACPHERSON, SIB W. G.: R. Soc: Med. May 10th 1920; Lane. May
22nd, p. 1112, 1920.

928. RAE: Rep. to Scurvy Commission in 1876 about the Hudson Bay Co.

cited by Med. Res. Comm. No. 38, p. 63, 1919.

929. LANCERAUX: Le scorbut des prisons du department de la Seine. Ann.

d'hyg. publ. et med. leg. 13, III, Ser. 1885.

930. BEETHENSON: Zur Statistik und Aetiologie des Skorbuts. Die Skor-

butepidemie von 1889 nach Beobachtungen in St. Petersburger Mili-
tarhospital. Arch. f. klin. Med. 49, 127, 1892.

931. MULLER: Munch, med. W. No. 35, 1894, 1911.

932. TAUSSIG, SIGISMUND: Zur Epidemiologie des Skorbuts, Militarmed.

und arztl. Kriegswiss. Heft IV, 1913, Safar Wien.

933. PICKENS, R. M. : A death from scurvy. Lane. II. 21,1917.

934. HARLAN, G. P. : Land scurvy in England. Brit. Med. J. II, 46, 1917.

935. Report on the Health of the City of Manchester 49, 1917.

936. Edit. Recent cases of scurvy in Glasgow. Brit. Med. J. II, Jly 7, 1917.

937. Memorandum on food and scurvy. Food (War) Comm. of the Roy.

Soc. Lane. Nov. 40, p. 756, 1919.

938. LIND, W. A. T. : Outbreak of scorbutus at Kew Victoria. Med. J. of

Austr. 2, 107, 1919,

939. HOPKINS, G. R. : The etiology of scurvy. J. A. M. A. 69, 1641, 1917.

940. TURNER: Scurvy. Brit. Med. J. II. July 14, 33, 1917.

941. Trop. Dis. Bull. 12, 257, 1918.

942. HILL, LEONARD: Cited by Med. Res. Comm. No. 38, p. 65.

943. HORSCHELMANN, E. : Zur Klinik des Skorbuts in der russischen Armee.

Deutsch. med. W. 52, 1617, 1917.

944. DISQUE: Entstehung und Verlauf des Skorbuts im Jahre 1916 unter

den deutsch-osterr. Kriegsgefangenen in Taschkent (Turkestan)
Med. Klin. No 1, 1918.

945. BORICH, R. : Skorbut. D. Arch. f. klin. Med. 130, 151, 1919.

946. HARVIER, P.: Epidemic de scorbut. Paris m6d. 7, 394, 1917.

947. BENOIT, A. : Une Spidemie de scorbut. Ibid. 9, 469, 1919.

948. MOURIQUAND, G. : Role de 1'aliment frais dans la nutrition. Imprim.

milit. Paris 1915; La die"tetique sur le front. Arch, de m6d. et
de pharm. milit. sept. 1915. L'alimentation aux armies. Lyon me"d.
oct. 1915.

949. FERRARI: Gazz. d. osp. 38, 778, 1917.

950. RAMOINO: Policlin. 24, 616, 1917.

951. GINGUI, F. : Rif . med. 34, 22, 1918.

952. VANUTELLI, F. : Un epidemia di porpora infettiva con manifestazio-

ne emorragico-scorbutiche tra le truppe combattenti. Ibid
24, 873, 1919.

953. VALLARDI, C.: Rif. med. 34, 793, 1918.

LITERATURE TO THE TEXT 447

954. MUCH, HANS: Ueber den Skorbut. Munch, med. W. 64, 854, 1917; Much

and Baumbach, K: Ibid. No. 26, 1917; Saxl, P. and Melka, J. :
Ueber den Skorbut und seine Beziehungen zu den hamorrhagischen
Diathesen. Med. Klin. No. 37, 986, 1917; Feig, S.: Beobachtungen
iiber Skorbut in Kriege. Ibid. No. 31, 832, 1917; Schneider, E. :
Skorbut in Felde. Munch. Med. W. No. 44, 1939, 1917; Hertz, A.:
Wien. klin. W. No. 22, 1917; v. Jaksch, R. : Z. f. inn. Med. No.
32-33, 1917; Hannemann, K. : Munch, med. W. p. 665, 1918; Morawitz :
Ibid. 65, 349, 1918; Tiichler: Med. Klin. No. 5, 112, 1918; Blatt,
N.: Wien. klin. W. 31, 942, 1918; Arneth: D. med. W. 64, 509, 1918.

955. SCHREIBER, G. : Scorbut et dysenteric. Paris med. p. 508, 1918.

956. SPEYER: Wadenabszesse bei Skorbut. D. med. W. 544, 626, 1918.

957. LOBMEYER, G. : Kriegschirurgische Bedeutung des Skorbuts. D. med.

W. 44, 557, 1918.

958. Edit. Fixed diet in prisons. Trop. Dis. Bull. 4, 446, 1914.

959. DYKE, H. W. : Scurvy. Lane. II, 513, 1917.

960. COMRIE, J. D. : Scurvy in North Russia. Edinb. Med. J. 24, 207, 1920.

961. STEVENSON: J. Roy. Army Med. C. 35, 218, 1920.

962. CHICK, H. AND DALYELL, E. J. : Influence of overcooking vegetables

in causing scurvy among children. Brit. Med. J. Oct. 9, 546, 1920.
Epidemic des Skorbuts bei Schulkindern. Z. f. Kinderh. 26, 257, 1920.

963. TOBLER, W. : Skorbut im Kindesalter. Z. f. Kinderh. 18, 63, 1918.

964. MULLER, ERICH : Ueber ein gehauftes Auftreten von Skorbut bei Kindern.

Berl. klin. W. 55, 1024, 1918.

965. WEILL AND DUFOURT A. : Le scorbut tardif chez les enfants. Arch.

meM. d. enfants. 22, 561, 1919.

966. MORSE, J. L. : Re'sume' of literature of infantile scurvy during the past

5 years. Boston Med. Surg. J. 178, 160, 1918; for 1918-9. Ibid.
182, 428, 1910

967. NETTER: Le scorbut infantile. La semaine med. 22 fevr. No. 8, 1899.

968. CHEADLE, W. B. AND POYNTON F. J. : Infantile scurvy. System of Med.

Allbutt-Rolleston 5, 898, 1909.

969. LOOSER, E. : Ueber die Knochenveranderungen beim Skorbut. J. Kin-

derh. 62, 743, 1905.

970. NEUMANN, H. : Bemerkungen zur Barlowschen Krankheit. D. med.

W. 28, 628, 647, 1902; Sauglingsskorbut. Deutsch. Klin. 7, 341, 1904.

971. HEUBNER, O.: D. med. W. 29 Vereinsbeil. 109, 110, 117, 1903; Ueber die

Barlowsche Krankheit. Berl. klin. W. 40, 285, 1903.

972. MEYER, A.: Barlows sygdom. Kj0benhaven 1901.

973. BRACHI AND CARR: Infantile scurvy in a child fed on sterilized milk.

Lane. 662, 1911.

974. Amer. Pediatr. Soc., Collective investigation on infantile scurvy in

North America. Arch, of Pediatr. 15, 481, 1898.

975. HESS, A. F. AND FISH M. : Infantile scurvy. The blood, the blood ves-

sels and the diet. Amer. J. Dis. Childr. 8, 386, 1914.

976. SITTLER: Das Sauglingsheim der med. Universitatsklin. zu Marburg

wahrend der ersten Jahre seines Bestehens 1905-8 Z. f. soc. Med.
4, 1, 1909.

448 THE VITAMINES

977. COMBT, J. : Le scorbut infantile. Arch, de me"d. d. enfants. 20, 337,

1917; 22, 281, 1919.

978. EPSTEIN, A. : Ueber eine auffallige Haufung der Barlow'schen Krank-

heit in den Kriegsjahren 1917-18 J. f. Kinderh. 88, 237, 1918.

979. BENDIX: Barlow'sche Krankheit. D. med. W. 38, 242, 1912.

980. FORDYCE, A. D. : Scurvy in infants. Edinb. Med. J. 20, 110, 1918.

981. ASCHOFP, L. AND KOCH. W. : Zur Epidemiologie des Skorbuts. Eine

pathologisch-anatomische Studie. Fischer, Jena, 1919.

982. RHEINDORF: Med. Klin. No. 5, 1918.

983. WILTSHIRE, HAROLD: Hyperkeratosis of the hair follicles in scurvy.

Lane. 197, 564, 1919.

984. v. NIEDNER: Hamorrhagische Exantheme. Med. Klin. 14, 333, 1918.

985. WASSERMANN, S. : Ueber eine mit Schwellung und Rotung der Beine

verbundene Knochenschmerzhaftigkeit bei Kriegern. Wien. klin.
W. 68, 968, 1918.

986. ZLOCISTI, TH.: Ueber Skorbut. Med. Klin. No 25, 659, 1916; Die ulzero-

gangrenosen Stomatitisformen des Skorbuts, Ibid. No. 46, 1200,
1916.

987. RICHTER, A. : Ueber das Vorkommen von Tetanussymptomen bei Skor-

but. Wien. med. W. 338, 1917.

988. URIZIO, L. : Skorbut und eine nicht spirochatogene Ikterusepidemie.

Wien. klin. W. No 46, 1449, 1917.

989. ZAK, E. : Wien. klin. W. 30, 592, 1917.

990. HIFT: Beobachtungen tiber Skorbut und Hemeralopie. Wien. klin.

W. No. 34, 1918.

991. O'SHEA, H. V.: Scurvy. Practitioner. Oct-Nov. 1918.

992. KITAMURA, S. Ein Beitrag zur Kenntnis der Netzhautveranderungen

beim Skorbut. D. med. W. No. 9, 1910.

993. MILLER: Infantile scurvy. Cleveland Med. J. 16,541, 1917.

994. FRANCHETTI, U. : Riv. di clin. pediatr. 18, 193, 1920

995. COMBT, J. : Le scorbut chez le nourisson. M6d. 1, 673, 1920.

996. GERSTENBERGER, H. J. : Amer. J. Med. Sci. 155, 253, 1918.

997. COZZOLINO, O. : Pediatria 27, 321, 407, 477, 1919.

998. SCHODEL, J. AND NAUMWERK, C. : Untersuchungen iiber die Moller-

Barlow'sche Krankheit. Jena 1900.

999. HESS, A. F. : Cardio-respiratory involvement in infantile scurvy. Proc.

Soc. Exp. Biol. Med., 14, 4, 1916, J. A. M. A. 68, 235, 1917.

1000. ERDHEIM, J. : Ueber das Barlowherz. Wien. klin. W. No 49, 1293, 1918.

1001. ROSIN: Hamorrhagischen Diathesen. Spez. Path. Ther. inn. Krankh.

Kraus-Brugsch 8, 911, 1920.

1002. LABOR, M. : Ueber das Leukozytenbild bei Skorbut und Tibialgia.

Wien. klin. W. 29, 1069, 1916.

1003. FRANKEL, E.: Fortschr. Geb. d. Rontgenstr. 7, No 5-6, 1904; 10,

No. 1, 1906; Erganzungsb 18.

1004. BROWN, ALAN: Some new features in the diagnosis of scurvy with

brief reference to metabolic changes. Arch. Pediatr. 32, No.
10, 1915.

LITERATURE TO THE TEXT 449

1005. LABOR, M: Eine Beobachtung liber das Blutbild des Skorbuts.

Wien. klin. W., No. 29, 912, 1916.

1006. LEITISTER, PH.: Beitrage zur Hamatologie und Klinik des Skorbuts.

Wien. klin. W., No. 31, 978, 1917.

1007. BRANDT, HEDWIG: Das Blut im Skorbut. Arch. f. Kinderh. 67,395,

1919.

1008. WASSERMANN, S. : Ueber hochwertige Erythrozyten und Hamoglobin-

befunde bei Kriegern. M. med. W., 65, 927, 1918.

1009. BENOIT, A. : Le sang dans le scorbut. Arch d. malad. du coeur. 12,

241, 1919.

1010. ROLLY AND OPPERMANN: Blutzucker. Bio. Z. 48,472,1913.

1011. SCHUMM, O. : Untersuchungen iiber den Zuckergehalt des Blutes

unter physiologischen und pathologischen Verhaltnissen. III.
Mitt. H. 96, 204, 1915.

1012. HESS, A. F. AND KILLIAN. J. A. : Chemistry of blood in scurvy. Proc.

Soc. Exp. Biol. Med. 16,43, 1918; Hess: Chemistry of blood in scurvy,
especially its Ca-content. Ass. Amer. Phys. June 16th 1919; J. A.
M. A. Aug. 23rd 1919.

1013. BAUMAN, Louis AND HOWARD, C. P. : Metabolism of scurvy in an

adult. Arch. Internal Med. 9, 665, 1912.

1014. LABBE, HAGUINEA AND NEPREUX: Biochimie du scorbut. Bull. soc.

meU d. hop. 44, 1094, 1920.

1015. LUST, F. : Stoffwechselversuch bei Barlowschen Krankheit. Berl.

klin. W. 49, 862, 1912; Munch, med. W. 59, 785, 1912.

1016. LUST, F. AND KLOCMAN L. : Stoffwechsel bei Barlowschen Krankheit

J. f. Kinderh. 75, 663, 1914.

1017. MOLL, L. : Stoffwechsel in Barlowschen Krankheit. Mitt. d. Ges.

f. inn. Med. u. Kinderh. Wien 18, 1919.

1018. FRANK, M. : Stoffwechsel in Moller-Barlowschen Krankheit. J. f.

Kinderh. 91, 21, 1920.

1019. FEIGENBAUM, D. : Ein Beitrag zur Kenntnis der Riickenmarkblutun-

gen beim Skorbut. Wien. kliri. W. No. 46, 1455, 1917.

1020. SMITH, W. JOHNSON: Scurvy. System of Medicine. Allbutt-Rolleston,

5, 879, 1909.

1021. FRA'NKEL, E. : Archiv und Atlas der normalen u. pathol. Anatomie

in typischen Rontgenbildern. Erganzungsband 17, 35, 1908.

1022. HESS, A. F. : Focal degeneration of the lumbar cord in a case of in-

fantile scurvy. J. Inf. Dis. 23, 438, 1918.

1023. BAHRDT, H. AND EDELSTEIN, F. : Organanalyse bei Barlowschen Krank-

heit. Z. f. Kinderh, 9, 415, 1914.

1024. PAGET: Annual Med. Rep. for year 1912, Somaliland.

1025. SCHULHOF, W. : Ueber die postskorbutische Zustande und deren Be-

handlung. Wien. klin. W. No 25-26, 1918.

1026. DELILLE, ARMAND: Le jus d'orange dans le traitement et la prophy-

laxe du scorbut infantile, d'apres les recherches r^centes. J. des
pratic. No. 15, 225, 1919.

450 THE VITAMINES

1027. HARDEN, A., ZILVA, S. S., AND STILL, G. F. : Infantile scurvy; the

antiscorbutic factor of lemon juice in treatment. Lane. Jan. 4th,
1919.

1028. RUECK: Scurvy. Med. Rec. 91, 152, 1917.

1029. CAUTLEY, E. : Rickets; GARBOD, BATTEN AND THURSFIELD: Diseases

of children. Arnold, London. 1913.

1029a. STRONGMAN, B. I., AND BOWDITCH, H. I.: Impressions Concerning
etiology of rickets among Italians. Bost. Med. Surg. J. 184, 443,
1921.

1030. DENNETT, R. H. : The use of boiled milk in infant feeding. J. A.

M. A. 63, 1991, 1914.

1031. SCHMORL.: Verh. dental-path. Ges. 58, 1909.

1032. SCHWARTZ, H. : Craniotabes and beading of ribs as signs of rickets.

Amer. J. Dis. Childr. 19, 384, 1920.

1032a. KASSOWITZ, M.: Ueber Rachitis bei Neugeborenen. J.f.Kinderh.
76,369,1912..

1033. MERY AND PARTURIER: Le rachitisme congenital. Bull. soc. de peel.

Paris, f<§vr. 18, 1919.

1034. WIELAND, E. : Klinische und anatomische Untersuchungen iiber so

genannte angeborene und liber friihzeitig erworbene Rachitis,
J. f. Kinderh. 70, 539, 1909.

1035. WIELAND, E.: Ueber das physiologische Osteoid bei Foten und Neuge-

borenen und deren Bedeutung fur die histologische Diagnose der s. g.
angeborenen Rachitis. Deutsch. med. W. 35, 1520, 1909.

1036. SINCLAIR, JOHN F. : Problem of the premature infant. Arch. Ped.

37, 139, 1920.

I036a. CARR, W. L.: Care of premature babies. Amer. Ped. Soc. Meeting,
June 2-4, 1921.

1037. HAMILTON, B. : Calcium metabolism of premature infants. Amer.,

J. Dis. Childr. 20 316, 1920.

1038. RETTERER, E. AND FISCH J. : C.r. soc. biol. 80, 182, 1917.

1039. OSTHEIMER, M.: Fragilitas ossium. J. A. M. A. 63, 1997, 1914.

1040. BOOKMAN, A. : The metabolism in osteogenesis imperfecta with special

reference to calcium. Amer. J. Dis. Childr. 7, 436, 1914.

1041. MCCLANAHAN, H. M. AND WILLARD, W. W, : Osteogenesis imperfecta

congenita. Amer. Ped. Soc. June 16th, 1919; Amer. J. Dis. Childr.
19, 181, 1920.

1042. SCHABAD, J. A.: Der Stoffwechsel bei angeborener Knochenbriichig-

keit. (Osteogenesis imperfecta) Z. f. Kinderh. 11, 230, 1914.

1043. BRADE-BIRKS H. K. : Bone condition analogous to rickets in a child

of five months. Lane. March 27, 1920.

1044. HESS AND UNGER : The diet of the negro mother in New York City.

J. A. M. A. 70, 900, 1918.

1045. JAPHA, A.: Kriegsernahrung in Beziehung zur Rachitis. Berl. klin.

W. 56, 921, 1919.

1046. ENGEL: Rickets in Germany; a study of effect of war in children.

Lane. I, 188, 1920.

LITERATUBE TO THE TEXT 451

1047. WEISS, T. : Cited according to J. A. M. A. 74, 262, 1920.

1048. ADAMS, JANE AND HAMILTON, ALICE: A visit to Germany. Brit.

J. Dis Childr. 16, July-Sept. 1919.

1049. SCHLEE: Spatrachitis. Munch, med. W. 66, 1349, 1919.

1050. STETTER, K. : Spatrachitis. Deutsch. med. W. 46, 520, 1919.

1050a. SATJER, H.: Spatraohitis. Deutsch. Zenti. f. Chirurg. 162, 356, 1921.

1051. STJTTON, H. : Manifestation of rickets in school children. Med. J.

of Austr. 2, 190, 1920.

1051a. FRANGENHEIM, P. : Die Krankheiten des Knochensystems in Kindes-
alter. F. Enke. 1913.

1052. MtiLLER, W.: Schlatter' sche Krankheit. Beitr. z. klin. Chir. 110,

389, 1920.

1052a. PAUS, N.: Schlatter's disease. Norsk. Mag. vor Laegeridenskaben
82, 320, 1921.

1052b. PARK, E. A., AND ROWLAND, J.: Dangers to life of severe involve-
ment of thorax in rickets. Johns Hopk. Hosp. Bull. 32, 101, 1921.

1052c. BRUSA, P.: Rachitic deformity of thorax. Riv. clin. pediatr. 19,
210, 1921.

1053. ENGEL, S. : Rachitis in Beziehung zum Wachstumstillstand. Med.

Klin. 16, 383, 1920.

1054. KARGER, C.:M. f. Kinderh. 18,21,1920.

1055. LERI AND BECK: Les consequences du rachitisme chez les adultes.

Ann. de me'd. 6, 449, 1920.

1056. JtTARisTi, V. : The rachitic penis. Arch, espanol. de. pediatr. 3, 286, 1919.
1056a. NEUMANN, H. : Ueber die Beziehungen der Krankheiten des Kindes-

alters zu den Zahnkrankheiten. S. Volkmann, Leipzig, no. 172;
1897. Heubner, Otto: Lehrb. der Kinderkrankh. I, 674, 1903;
Birkenthal: Berl. Verlagsanst. Zweite Aufl. p. 48, 1912; Fleisch-
mann, L.: Cited according to Biedl. Innere Secretion, 2nd
Edition, I, 98, 1913.

1056b. LICHTENSTEIN, ALFRED: Ueber die Geschmacksempfindung gesunder
und rachitischer Kinder. J. f . Kinderh. 37, 76, 1894.

1057. v. RECKLINGHAUSEN: Untersuchungen iiber Rachitis und Osteoma-

lacie.

1058. SCHMORL. : Die pathologische Anatomic der rachitischen Knochen-

erkrankung mit besonderer Berticksichtigung iher Histologie
und Pathogenese. Erg. d. inn. Med. u. Kinderh. 4, 403, 1909.

1059. DIBBELT: Die Bedeutung der Kalkstoffwechselstorung fur die Entsteh-

ung der Rachitis. Munch, rued. W. 57, 2121, 2186, 1910.

1060. MARPAN, BARDOUIN AND FETJILL£ : Lesions de la moelle osseuse dans

le rachitisme. C. r. soc. biol. 66, 862, 1909.

1061. HUTINEL AND TixiER: Modifications de la moelle osseuse des rachi-

tiques. Ibid. 66, 946, 1909.

1062. KASSOWITZ: Ueber Rachitis. J. f. Kinderh. 69, 251, 1909; II. Osteo-

chondritis rachitica. Ibid. 75, 194, 334, 489, 581, 1912.

1063. HEtTBNER: Lehrb. der Kinderkrankheiten. Leipzig, 1906.

1064. POMMER: Untersuchungen liber Osteomalacie und Rachitis. 1885.

452 THE VITAMINES

1065. RIBBERT, HUGO : Betrag zur Rachitis. Deutsch. med. W. 39, 8, 1913.

1066. HAQENBACH AND BTJRCKHARDT: Klinische Beohachtungen iiber die

Muskulatur der rachitischen. J. f. Kinderh. 69, 471.

1067. BING: Ueber atonische Zustande der kindlichen Muskulatur. Med.

Klin. No. 11, 1907.
1067a. BAKU, G.:L4sions rachitiques des muscles. Nourrisson. 7,229,1921.

1068. MOHR: Noorden's Handb. der Path. d. Stoffw. 2, 865.

1069. Dtr CASTBL R. : Le thymus rachitique. C. r. soc. biol. 65, 725, 1908.

1070. STOLZNER: Ueber Behandlung der Rachitis mit Nebennierensubstanz.

J. f. Kinderh. 51, 73, 199; 53, 515, 672.

1071. CATTANEO: Die Nebennieren bei Rachitis. V. Kongr. d. italien. Ges.

f. Kinderh. Rom. 1905.

1072. GASSMANN, TH. : Untersuchungen ,von gesunden und rachitischen

Knochen. H. 70, 161, 1910-11.

1073. SCHABAD: Zur Bedeutung des Kalkes in der Pathologic der Rachi-

tis. V. Der Mineralgehalt gesunder und rachitischer Knochen.
Arch. f. Kinderh. 52, 47, 1909.

1074. SIMONINI, R: Soc. med. chir. June 11, 1913.

1075. ROST, WILLIAM L. : RICKETS, N. Y. Med. J. 102, 505, 1915.

1076. ASCHENHEIM AND KAtTMHEiMER: Ueber den Aschegehalt der Musku-

latur bei Rachitischen. M. f. Kinderh. 10, 435, 1911.

1077. ASCHENHEIM, ERICH: Rachitis und Spasmophilie. I. J. f. Kinderh.

79,446.
1077a. DENIS, W., AND TALBOT, F.B.: Calcium in blood of children. Amer.

J.Dis.Childr.21,29, 1921.
1077b. HOWLAND, J., AND KRAMER, B.: Calcium and phosphorus in the

serum in relation to rickets. Am. J. Dis. Childr. 22, 105, 1921.

1078. DIBBELT: Aetiologie der Rachitis und der Kalkstoffwechsel. Deutsch.

med. W. No. 12, 555, 1913.

1079. DENTON, MINNA C. : How much food does it take to supply us with the

calcium we need. J. Home Econ. 10, 168, 1918.

1080. ARON, H. : Kalkbedarf und Kalkaufnahme beim Saugling und die

Bedeutung des Kalkes fur die Aetiologie der Rachitis. Bio. Z.
12, 28, 1908.

1081. SCHABAD: Der Kalkgehalt der Frauenmilch. J. f. Kinderh. 74, 511,

1911.

1082. ORGLER: Der Kalkstoffwechsel des gesunden und des rachitischen

Kindes. Erg. d. inn. Med. u. Kinderh. 8, 142, 1912.

1083. CRONHEIM AND MULLER, ERICH: Stoffwechselversuche an gesunden

und rachitischen Kindern mit besonderer Beriicksichtigung des
Mineralstoffwechsels. Bio. Z. 9, 76, 1908.

1084. SCHABAD: Der Kalkstoffwechsel bei Rachitis. Arch. f. Kinderh.

53, 380, 1910.

1085. SCHABAD: Zur Bedeutung des Kalkes in der Pathologic der Rachitis.

IV-Mitt.Der P-stoffwechsel bei Rachitis. Arch. f. Kinderh. 54,
83, 1910.

1086. FLAMINI: Arch, farmacol. 6, No. 12.

1087. PEISER, AMALIE: J. f. Kinderh. 81, 437, 1915.

LITERATURE TO THE TEXT 453

1088. SCHWARZ: Der Stickstoff- und Schwefelstoffwechsel in Fallen von

rachitischem Zwergwuchs. J. f. Kinderh. 72, 549, 712, 1910.

1089. SCHABAD: 2 Falle von s.g. Spatrachitis. Der Mineralstoffwechsel

in Vergleich mit kindlicher Rachitis. Mitt. a. d. Grenzg. d. Med.
u. Chir. 23, 82, 1911.

1090. DIBBELT: Die experimentelle Erforschung der Rachitis. Erg. d.

wiss. Med. 2,64,1910-11; Neue experimentelle Untersuchung iiber
die Pathogenese der Rachitis. Deutsch. med. W. 38, 316, 1912.

1091. MEYER, LTJDWIG F. : Zur Kenntnis des Mineralstoffwechsels im Saug-

lingsalter. Bio. Z. 12, 422, 1908.

1092. TAD A: Die Sauglingsnahrung "Buttermilch," eine kohlenhydrat-

reiche Magermilch. M. f. Kinderh. 4, 118, 1905.

1093. ROTHBERG, Ueber den Einfluss der organischen Komponente auf den

Kalkansatz kiinstlich genahrten Sauglinge. J. f. Kinderh. 66,
69, 1907.

1094. ORGLER: Ueber den Kalkstoffwechsel bei Rachitis. M. f. Kinderh.

10, 373, 1911.
1094a. HESS, A. F.: Newer aspects of some nutritional disorders. (Harvey

Lecture) J. A. M. A. 76, 693, 1921.
1094b. TELFER, S. V.: Influence of cod liver oil and butter fat on retention of

calcium and phosphorus. Proc. Physiol.Soc., J.Physiol.54, cv, 1921.

1095. HUTCHINSON, H. S. : Is fat starvation a causal factor in production

of rickets? Glasgow Med. J. 11, 8, 1920.

1096. HUTCHINSON, H. S. : Fat metabolism in health and disease with special

reference to infancy and childhood. Quart. J. Med. 13, 277, 1920.

1097. HOLT, L. E., COURTNEY, A. M. AND FALES, H. L. : Fat metabolism in

infants and young children. III. Fat in stools of children on mixed
diet. Amer. J. Dis. Childr. 18, 107, 1919.

1098. HOLT, L. E.: Calcium metabolism in infants and young children, and

relation of calcium to fat excretion in stodls. Ibid. 19, 97, 201, 1920.

1099. DIBBELT: Die Pathogenese der Rachitis. Arb. a. d. path. Inst. Tu-

bingen 4, 670, 1908.

1100. MASSANECK: Ueber Buttermilch. J. f. Kinderh. 60, 756, 1904.

1101. HOWLAND AND MARRIOTT: Amer. J. Obstetr. 74, 541, 1916.

1102. SCHABAD, J. A. : Die gleichzeitige Verabreichung von Phosphorleber-

tran mit einem Kalksalze bei Rachitis. J. f. Kinderh. 72, 1, 1910;
Z. f. klin. Med. 49, 435, 1910; Schabad and Sorochowetz, R. F.:
M. f. Kinderh. 10, Orig. 2, 12, 1911.

1103. SCHABAD : Dietetic Treatment of rickets. Russ. Vratch. 15, 1067, 1916.

1104. BIRK, W. : Untersuchungen iiber den einfluss des P-lebertrans auf

den Mineralstoffwechsel gesunder und rachitischer Kinder. M. f .
Kinderh. 9, 450, 1908; Birk and Orgler: Ibid. 9, 544, 1910.

1105. FRANK, L. AND SCHLOSS, E. : Zur Therapie der Rachitis. J. f. Kinder.

79, 539, 1914.

1106. SCHLOSS, ERNST: Deutsch. med. W. 39, 1505, 1913; J. f. Kinderh.

78, 694, 1914; 79, 40, 1914; Schloss and Frank: Bio. Z. 60, 378;
Schloss: Erg. d. inn. Med. u. Kinderh. 15, 1917; Die Pathogenese
und Aetiologie der Rachitis, sowie die Grundlage ihrer Therapie.

1107. GROSSER, PAUL : Stoffwechselfragen bei Rachitis. Med. Klin. 10, 577.

454 THE VITAMINES

1108. Y ELARDI, P. PEREIDA: Nutritional derangement and rachitis. Pro-

gresos de la clin. 7, 117, 1919; Medic, ibera 7, 161, 1919.

1109. MEYER, KURT: Salzstoffwechsel bei Rachitis. J. f. Kinderh. 77,

28, 1914.

1110. TOBLER: Ueber Spatrachitis. Munch, med. W. 58, 2357, 1911.

1111. RICKLIN: Calcium- und Phosphorstoffwechsel in einem Falle von

Rachitis tarda. Korr. Blatt. Schweiz. Aerzte. No. 47, 1917.

1112. PHEMISTER, D. B. : Effect of phosphorus on the growth of normal

and diseased bones. J. A. M. A. 70, 1737, 1918.
1112a. PHEMISTER, D. B., MILLER, E. M. AND BONAR, B. E.: Effect of

phosphorus in rickets. J. A.M. A. 76, 850, 1921.
1112b. SHERMAN, H. C. AND PAPPENHEIMER, A.M.: Dietetic production of

rickets and its prevention by an inorganic salt. Proc. Soc. Exp.

Biol.Med.18, 193, 1921. J. Exp. Med. 34, 189, 1921.
1112cw PAPPENHEIMER, A.M., McCANN, A. F., ZUCKER, TH. F. AND HESS,

A. F.: The effect of various modifications of a diet producing

rickets in rats. Proc. Soc. Exp. Biol. Med. 18, 267, 1921.

1113. LIENAUX AND HUYNEN: Bull. acad. roy. m£d. beige. 29, 855, 1919.

1114. HESS AND UNGER: Prophylactic therapy for rickets in a negro com-

munity. J. A. M. A. 69, 1583, 1917.

1115. FERGUSON, M. : Medical Res. Committee Rep. No 20. A study of

social and economic factors in the causation of rickets. 1918.

1115a. MACKAY, HELEN M. M.: Observations on cases of rickets in an out-
patient department. Brit. Med. J. II, 929, 1920.

1115b. ARON, HANS: Die Bedeutung von Extraktstoffen fur die Ernahrung.
II, M. f. Kinderh. (Orig.) 15, 561, 1920.

1115c. RACZYNSKI, JAN: Recherches expe'rimentales sur le manque d'action
du soleil comme cause du rachitisme. C. r. de 1'assoc. internat.
deped. ler Congrgs. Paris, 1912. p. 308.

1115d. HULDSCHINSKY, KURT: Die Behandlung der Rachitis durch Ultra-
violettbestrahlung. Dargestellt an 24 Fallen. Z. f. orth. Chirurg.
, 39, 426;Deutsch.med.W. No. 26, 1919.

1115e. MENGERT, E.: D. Med. W. 47, 675, 1921.

1115f. ERLACHER, P.: Phototherapie bei Rachitis. Wien. klin. W. 34, 241,
1921.

1115g. HESS, A. F., AND UNGER, L. J.: The cure of infantile rickets by sun-
light. J. A.M. A. 77, 39, 1921.

1115h. HESS AND UNGER: Interpretation of seasonal variation of rickets.
Proc. Soc. Exp. Biol. Med. 18, 298, 1921.; Amer. J. Dis. Childr. 22,
186, 1921.

1116. HOWLAND, JOHN AND PARK, EDWARD A.: Amer. Ped. Soc. May 31-

June 2nd, 1920; J. A. M. A. 75, 198, 1920.
1116a. DUFOUR, H.: Bull. soc. m4d. hopit. Paris. 36, 1261, 1921.

1117. v. HANSEMANN: Ueber den Einfluss der Domestikation auf die Ent-

stehung der Krankheiten. Berl. klin. W. No. 20-21, 1906.

1118. KASSOWITZ: Die Pathogenese der Rachitis. 1885.

1119. FINDLAY, R.: Rickets. Brit. Med. J. II, 13, 1908.

LITERATURE TO THE TEXT 455

1120. PATON, D. N., FINDLAY, L. AND WATSON, W. : Observations on the cause

of rickets. Brit. Med. J. II, 625, 1918.

1121. PIERSON, EDITH AND DTJTCHER, R. ADAMS: Rhubarb as an antiscor-

butic. Science, 51, 70, 1920.

1122. DICK, J. LAWSON: Defective Housing and the Growth of Children.

London, 1919; R. Soc. Med. Febr. 27, 1920. Lane. March 13th, 1920.

1123. MANN, H. COURT: 88th Meeting Brit. Med. Ass. June 30-July 2nd,

1920. Brit. Med. J. July 31st, 157, 1920.
1123a. CZERNY, ADALBERT: Rachitis. Spez. Path. Therap. inn. Krankh.

Kraus-Brugsch. IX a, 317, 1921.
1123b. SIEGERT, F.: Die Erblichkeit der Rachitis. J. f. Kinderh. 58, 929,

1903.

1124. SCHABAD: Der Kalk in der Pathologic der Rachitis. Berl. klin. w. 46,

823, 923, 1909.
1124a. HERTER, C. A.: J. Exp. Med. 3, 293, 1898.

1125. HOLT, L. E. : Diseases of Infancy and Childhood. 1907.

1126. CHEADLE, W. B.: Artificial Feeding and Food Disorders of Infants. 1906.

1127. STOLZNER. W. : J. f. Kinderh. 50, 268, 1899.

1128. ESSER: Munch, med. W. 1907.

1129. PRITCHARD, ERIC: Causation of rickets and its treatment. Ped.

Sept. 1916; N. Y. Med. J. 110, 921, 1919; Brit. Med. J. Nov. 15th,
1919.

1130. MELLANBY, EDWARD: Accessory food factors (vitamines) in the

feeding of infants. Lane. I. 856, 1920.

1131. MELLANBY, E.: R. Soc. Med. Febr. 27, 1920; Lane. March 13th, 1920.
1131a. PATON, D.NOEL AND WATSON, A.: Etiology of rickets. Brit. Med.J.

1,594, 1921. Brit. J. Exp. Path. 2, 75, 1921.

1132. HOLT, COURTNEY AND FALES: Fat metabolism of infants and young

children. IV. Amer. J. Dis. Childr. 18, 157, 1919.

1133. v. GROER, FRANZ: Bio. Z. 97,311, 1919.

1134. McCoLLUM, SIMMONDS AND PARSONS: The etiology of rickets. Proc.

Amer. Soc. Biol. Chem. J. Biol. Chem. 41, XXXI, 1920.

1135. ROBB: The influence of dry vs. fresh green plant tissue on calcium

metabolism. Science. 52, 510, 1920.

1135a. McCoLLUM, SIMMONDS, PARSONS, SHIPLEY AND PARK: Studies on
experimental rickets. I. The production of rachitis and similar
diseases in rats by deficient diets. J. Biol. Chem. 45, 333, 1921.
45, 343, 1921.

1135b. HESS, McCANN, AND PAPPENHEIMER : The failure of rats to develop
rickets on a diet deficient in vitamine A. Soc. Exp. Biol. Med.
May 18, 1921. Proc. 18, 266, 1921. J. Biol. Chem. 47, 395, 1921.

1135c. TURK,M.: Munch, med. W.45, 521, 1919.

1135d. MCCOLLUM. E. V., SIMMONDS, NINA, SHIPLEY, P. G. AND PARK, E. A.;
Studies on experimental rickets. III. Pathological condition
bearing fundamental resemblances to rickets of the human being
resulting from diets low in phosphorus and fat-soluble A: the
phosphate ion in its prevention. Johns. Hopk. Hosp. Bull. 32, 160,
1921., VIII, J. Biol. Chem. 47, 507, 1921.

456 THE VITAMINES

1136. NATHAN, M: Presse m&i. 28, 577, 1920.

1137. OGATA, M. : Rachitis und Osteomalacie in Japan. Hegar's Beitr.

z. Geburtsch. 18, Heft No. 1.

1138. LIESEGANG, R. E. : Puerperal Osteomalacia. Zentr. Gynakol. 39, No. 15.

1139. SCIPIADES, E.: Osteomalazie. Z. f. Geburth u. Gynakol. 81, 156, 1919.
1139a. KOLTONSKI, H.: Osteomalazie. M. f. Geburtsh. u. Gynakol. 52, 253,

1920.

1140. JANUSZEWSKA: Ueber Osteomalaeie mit Anhang tiber Tetanie. Wien.

klin. ther. W. No. 21, 1910.

1141. EDIT. : Brit. Med. J. Nov. 15th, 1919. Food in Europe p. 640.

1142. PAKTSCH, F.: Deutsch. med. W. 45, 1130, 1919.

1143. SATJER: Ein Fall von Osteomalazie und Tetanie in Folge von Unter-

ernahrung. Ibid. 46, 45, 1920.

1144. HAMEL, O. : Ibid. 46, 68, 1920.

1145. HEYER: Osteomalazie in Miinchen. Munch, med. W. 67, 98, 1920.

1146. KOPCHEN, A. : Z. f. inn. Med. 40, 961, 1919.

1147. CURSCHMANN, H. : Ueber Osteomalazie senilis und tarda. Med. Klin.

7, 1565, 1911.

1147a. CHELMONSKI, A.: Les maladies alimentaires des os. Presse. mecl. 29,
115,1921.

1148. CROFTAN, ALFRED C. : Osteomalacia. Amer. Med. 11,775, 1916.

1149. LOOSER, E. : Rachitis und Osteomalazia. Corr. Bl. Schweitz. Aerzte

49, 1065, 1919.

1150. ALWENS: Die Beziehungen der Unterernahrung zur Osteoporose

und Osteomalazie. Munch, rned. W. 66, 1071, 1919.

1151. BOHME, A. : Pathologische Veranderungen im Skelett durch Unter-

ernahrung bedingt. Deutsch. med. W. 45, 1160, 1919.

1152. IMHOF, A. : Psychosen un4 Osteomalazie. Z. Neurol. Psych. 14, 137.

1153. GOTJLD, ERIC P.: The bone changes occurring in v. Recklinghausen's

disease. Quart. J. Med. 11, 221, 1918.
1153a. WEISS, R. S.: v. Recklinghausen's disease in negro. Curvature of

spine in v. Recklinghausen's disease. Arch. Dermatol. Syphil. 3,

144, 1921.
1153b. COMBY, J.: Bull, de 1'assoc. med. d. hopit. 45, 382, 1921.

1154. HANAU, A. : Bericht iiber die Knochen bei Osteomalazie. Korr.

Bl. Schweitz. Aerzte. 497,1892; Fortschr. d. Med. No. 7, 1892.

1155. WILD, C. R. : Anatomische Untersuchungen uber das puerperale

Osteophyt. Diss. Lausanne, 1901.

1156. SCOTT, AGNES C. : Calcium content of the urine and blood with reference

to its variation in the condition of Osteomalacia. Ind. J. Med.
4, 169, 1916.

1157. ZUNZ, LEO: Stoffwechselversuche bei der Osteomalazie. Arch.

Gynakol. 99, 145.

1157a. ELFER, A. AND KAPPEL, J. : Z. exp. Path. Therap. 21, 104, 1920.
1157b. HUTCHINSON, H. S., AND PA TEL, P. T.: Etiology of Osteomalacia in

Bombay. Glasgow Med. J. 95, 241, 1921.

1158. LUZZATTI, F.: Spasmophilia in children. Policlin. 26,769, 1919.

LITERATURE TO THE TEXT 457

1159. STHEEHMAN, H. A. AND ARNTZENIUS, A. K. W. : Signs of calcium

deficiency. Ned. Tijdsr. v. Gen. 1, March, 27, 1030, 1920.

1160. CYBULSKI: Ueber den Ca-stoffwechsel der tetaniekranken Sauglings.

M. f. Kinderh. 5, No. 8, 1906.

1161. PEPPER, O. H. PERRY: Disorders of nutrition and metabolism, Pro-

gressive Med. June 1918.
1161a. ROHMER, P., AND VoNDERWEiDT, P.: Traitement de spasmophilie.

Nourrisson9, 87, 1921.
1161b. SCHERER, C. A.: Manifestations of the spasmophilie diathesis in

older children. Minn. State Med. Assoc. Aug. 24-26, 1921. J.

A. M. A. 77, 966, 1921.

1162. THIEMICH: Ueber die Behandlung der Krampfe im friihen Kindes-

alter. Deutsch. med. W. No. 12, 537, 1913.

1163. PETRONE, G. A. AND VITALE, C. : Infantile spasmophilia and the

parathyroid gland. La pediatria 20, 16.
1163a. PINCHERLE, M., AND MAGGEsi, B.: Riv. clin. pediatr. 18, 577, 1920.

1164. TAKASU, K. : Spasmophile Dyspepsie bei natiirlichen Ernahrung.

Sauglingskakke. J. f. Kinderh. '80, No. 5, 1914.

1165. KLOSE, E. : Altersgrenzen fur Spasmophilie. Arch. f. Kinderh.

67, 439, 1919.

1166. BLUHDORN, K. : Schicksal der Kinder mit Spasmophilie. J. f. Kinderh,

92, 294, 1920.

1167. FAAS, JAKOB. Graviditatstetanie. Diss. Erlangen 1913.

1168. GTJTHRIE: Tetany; GARROD, BATTON AND THURSFIELD: Diseases of

Children, p. 679.

1169. LUST, F. : Die Pathogenese der Tetanie im Kindesalter. Deutsch. med.

W. No. 23, 1087, 1913.

1169a. BOSSERT, O. AND GRALKA, R.: Myographie der Spasmophilie. J. f.
Kinderh. 94, 145, 1921.

1170. v. MEYSENBUG, L. : Spasmophilia and vitamines. Amer. J. Dis.

Childr., 20, 206, 1920.

1170a. JEPPSON, J., AND AF KLERCKER, K.O.: Phosphate bei der Spasmo-
philie. J. f. Kinderh. 28, 71, 1921.

1170b. IBRAHIM, J.: I), med. Woch. 46, 1359, 1920.

1171. BOSSERT, O.: Oedem bei der Spasmophilie. J. f. Kinderh 92, 121,

1920.

1172. HOWLAND, JOHN AND MARRIOTT, W. McKiM: Observations on the

calcium content of the blood in infantile tetany and the effect of
treatment with calcium. Bull. Johns Hopk. Hosp. 29, 235, 1918',
Quart. J. J. Med. 11, 289, 1918.

1172a. ELIAS, H. AND SPIEGEL, E. A.: Tetanie. Wien. Arch. inn. Med. 2, 447,
1921.

1172b. TISDALL, F. F., KRAMER, B. AND HOWLAND, J.: The concentration of
sodium and potassium as compared with that of calcium and mag-
nesium in the serum of patients with active infantile tetany. Proc.
Soc. Exp. Biol. Med. 18, 252, 1921.

1173. STHEEHMAN AND ARNTZENIUS: Calcium level in the blood. Ned.

Tijdsr. v. Gen. Apr. 3, 1168, 1920.

458 THE VITAMINES

1173a. v. MEYSENBUG, L.: Studies in spasmophilia II. Electrical reactions
of older children. Amer. J. Dis. Childr. 20, 539, 1920; III. Blood
calcium and calcium therapy in older children with Thiemich's
sign. Ibid. 21, 150, 1921.

1174. FLETCHER, ALMON: Some considerations in the study of infantile

tetany, with report of a case. Arch. Internal Med. 16, 383, 1915.

1175. LIEFMANN, ELSE: Die Azetonausscheidung im Harn von gesunden

und spasmophilen jungen Kindern. J. Kinderh. 77, 125, 1914.

1176. SHABPE, JOHN SMITH: The guanidine content of feces in idiopathic

tetany. Biochem. J. 14, 46, 1920.

1177. HOOBLEK, B. RAYMOND: Calcium-magnesium-phosphorus balance in

children subject to convulsive disorders. Amer. Fed. Soc. May
31-June 2, 1920; J. A. M. A. 75, 199, 1920.

1177a. BOLTEN, G. C.: Electrical responses with Fragilitas ossium. Ned.
Tijdar. v. Gen. 1, 952, 1921.

1178. BROWN, ALAN AND FLETCHER, ALMON: The etiology of tetany. Metab-

olic and clinical studies. Amer. J. Dis. Childr. 10, 313, 1915.

1179. STHEEHMAN, H. A. : Spasmophilia in children over three. Ned.

Tijds. v. Gen. No. 14, Apr. 1, 1145, 1916.

1180. BROWN, A., MACL.ACHLAN, J. AND SIMPSON, F. R. : Effect of intravenous

injection of Ca in tetany and influence of cod-liver oil and phos-
phorus in relation to calcium in blood. Amer. J. Dis. Childr.
19, 413, 1920.
1180a. HULDSCHINSKY, K. : Z. f. Kinderk. 26, 207, 1920.

1181. CZERNY: In Czerny-Keller. Handbuch der Kinderkrankheiten.
1181a. BLOCH, C. E.: Diseases of infants due to prolonged feeding with

excess of carbohydrates. Brit. Med. J. I, 293, 1921.

1182. RIETSCHEL: Ueber Mehle und Mehlfiitterungen bei Sauglingen und

ihre Beziehungen zum Stoffwechsel. Deutsch. Med. W. 34, 826,
1908.

1183. NOEGGERATH: Zwei Falle von Mehlnahrschaden beim Saugling. Berl.

klin. W. 44, 1423, 1907.

1184. BOGEN: Ueber Mehlnahrschaden. Deutsch. Med. W. 35,326,1909.

1185. GRUNEBERG : Nahrschaden im Kindesalter. Munch, med. W. 55, 140, 1908.

1186. LANGSTEIN, L: Jahreskurs f. arztl. Fortb. 3, 24, 1913.

1187. MEYER, L. F. : Idiopathische Oedeme im Sauglingsalter. Erg. d.

inn. Med. u. Kinderh. Bd. 17.

1188. RIETSCHEL: Mehlnahrschaden. Munch, med. W. No. 19-26, 1918.

1189. BENJAMIN: Mehlnahrschaden. Z. f. Kinderh. 10, 216, 1914.

1190. KELLER: Mehlkinder. Berl. klin. W. No. 36, 1906.

1191. STEINITZ AND WEIGERT: Ueber den Einfluss einseitiger Ernahrung

mit Kohlenhydraten auf die chemische Zusammensetzung des
Sauglingskorpers. Hofm. Beitr. 6, 206, 1905.

1192. SALGE: Ein Beitrag zur Pathologic des Mehlnahrschadens der Saug-

linge. Munch, med. W. 58, 1915, 1911.

1193. AST, D. A.: Injuries produced by starch. J. A. M. A. 61, 1275.

1194. CAUTLEY: Atrophia. Diseases of Children. Garrod, Batten & Thurs-

field. London, 1913.

LITERATURE TO THE TEXT 459

1195. FRANK, A. AND STOLTE, K. : Mehlnahrschaden. J. f. Kinderh. 78, 167, 191.

1196. HOHLFELD : Rohe Milch als Sauglingsnahrung. J. f . Kinderh. 12, H.

1, 1905.

1197. BENDIX, B. : Chronic digestive disturbances of infancy. Jacobi's

Diseases of Children. Appleton, New York, 1910.

1198. MOESE, J. L. AND TALBOT, F. B.: Diseases of Nutrition and Infant

Feeding. II. Ed. Macmillan. 1920, p. 28.

1198a. MARRIOTT, W. McK.: Pathology of nutrition in infancy. Amer. J.
Dis. Childr. 20, 461, 1920.

1199. PEHU: Alimentation des enfants malades. Paris, 1908.

1200. MORO: Karottensuppe bei Ernahrungstorungen des Sauglings. Munch.

med. W. 55, 1637, 1908.

1201. STARK, MORRIS: Some studies on the deficiency diseases of infancy

and childhood; interrelation, prophylaxis, and management.
Amer. Med. 11, 762, 1916.

1202. ARON AND SAMELSON: Karottenextrakt in Sauglingsernahrung.

Deutsch. med. W. 46, 772, 1920.
1202a. MCCLENDON, J. F. AND SEDGWICK, J. P.: Powdered spinach and sea

salts in diets of adults and infants. Proc. Soc. Biol. Chem.; J.

Biol. Chem. 46, XXVII, 1921.
1202b. ARON, HANS: Beitrage zur Frage der Wirkung und Verwertung der

Mehle bei der Ernahrung des Sauglings. J. f . Kinderh. 42, 82, 1920.

1203. KOHLBRUGGE: Ueber die Gahrungskrankheiten. Centr. f. Bakt.

I. 60, 223, 1911.

1204. HOLT, COURTNEY AND FALES: The effect of cod liver oil on a case

of "Intestinal infantilism". Amer. J. Dis. Childr. 14, 222, 1917.

1205. WEILL AND MOURIQTJAND: Lyon med. No. 6, 125, 1916.

1206. DANIELS, BYFIELD AND LOUGHLIN: The role of the antineuritic vita-

mine in the artificial feeding of infants. Amer. J. Dis. Childr.
18, 546, 1919.

1207. EDDY W. H. AND ROPER J. C. : The use of pancreatic vitamine in cases

of infant malnutrition. Proc. Soo. Exp. Biol. Med. 14, 52, 1916: Amer.
of J. Dis. Childr. 14, 189, 1917.

1208. FRIEDBERG, E. AND NOEGGERATH C. : Fettfreie Brustmilch. Arch.

f. Kinderh. 68, 195, 1920.

1209. SALGE: Ueber Sauglingsatrophie und Resorption. Munch, med. W.

54, 41, 1907.

1210. BAHRDT, H. AND EDELSTEIN, F. : Energie- und Stoffwechsel in atro-

phischen Saugling. Z. Kinderh. 12, 15, 1914.

1211. UTHEIM, K. : A study of blood and its circulation in normal infants

and in infants suffering from chronic nutritional disturbances.
Amer. J. Dis. Childr. No. 5, 366, 1920.

1212. MATTILL, P. M., MAYER, K. M. AND SAUER, L. W. : Dextrose tolerance

in atrophic infants. Ibid. 19, 42, 1920.

1213. GLADSTONE, H. B. : Fruit juices for atrophic infants. Practit. 97,

472, 1916.
1213a. MARFAN, A.B.: Nourrisson. 9, 65, 1921.

460 THE VITAMINES

1214. CZERNY, A.: M. f. Kinderh. 4, 1, 1906; 6, 1, 1908; 7, 1, 1909.

1215. SCHIPPERS, J. C. : Fat content of blood. Ned. Tijdsr. v. Gen. 2, 1081, 1920.
1215a. MARFAN, A. B.: Pathogenesis of athrepsia. Nourrisson. 9, 193, 1921.

1216. GIGON, ALFRED: Krankheiten mit gestortem Stoffwechsel. Corr.

Bl. f. Schweiz. Aerzte. 49, 1529, 1919.

1217. GRUMME, D. : Der Nahrwert der Proteine. Therapeut. Mon. Nos.

1-3, 1919.

1218. HINDHEDE, M. : Adequate protein minimum in dietaries. Ugeskr. for

Laeger, 79, Nos. 13-14-15, 1917.

1219. HINDHEDE, M. : The protein minima on a bread diet. Skand. Arch.

Physiol. 31, 259, 1914; 4th Rep. on protein minima with bread diet.
Copenhagen, Jacob Lund, 1914.

1220. HINDHEDE, M. : Ernahrungsversuche mit Gerstenwassergrutze. Skand.

Arch. Physiol. 35, 294, 1917.

1221. HINDHEDE, M. : Fett in menschlicher Ernahrung. Molk. Ztg. 28, 152,

1918; Fettminimum. Skand. Arch. Physiol. 39, 78, 1919.

1222. BANG, S. : Minimum ration of fat. Ugeskr. for Laeger. 80, 105, 1918.

1222a. McCoLLUM, E. V., SIMMONDS, NINA AND PARSONS, H. T.: Supple-
mentary protein values in foods I-V. J. Biol. Chem. 47, 111, 139,
175,207,235, 1921.

1223. NEUMANN, R. O. : Kriegsnahrung in Bonn im Winter 1916-7. Viert.

Jahrs. f. ger. Med. 57, 1, 1919.

1224. DOLLNER: Hungerodem. Munch, med. W. No. 20, 1917.

1225. KRASZEWSKI: Arch. Hyg. 86, 54, 1916.

1226. JANSEN, W. H. : Untersuchungen iiber die Stickstoffbilanz bei ka-

lorienarmen Nahrung. D. Arch. klin. Med. 124, 1, 1917.

1227. SHERMAN, H. C. : The protein requirements of maintenance in man.

Proc. Nat. Acad. Sci. 6, 38, 1920.

1228. TAYLOR, A. E. : Condition of diet and nutrition in the Internment

Camp at Ruhleben. London. Govern. Publ. 1916.

1229. BENOIT, ALB..: C. r. soc. biol. 82, 151, 1919.

1230. BOTAZZI, F. : Food requirements. Rif . med. 34, 461, 1918.

1231. KRTJSE AND HINTZE. Munch, med. W. 67, 445, 1920.

1232. PRAUSSNITZ, W. ; Minimum Nahrungsbediirfnis des Menschen. Ibid.

67, 696, 1920.

1233. STILLE: Ernahrungslehre und Kriegsernahrung. Schweitzer & Co., 1917.

1234. FUNK, LYLE AND MCCASKEY. : The nutritive value of yeast, polished

rice and white bread, as determined by experiments on man. J.
Biol. Chem. 27, 173, 1916.

1235. HAWK, P. B., SMITH, C. A. AND HOLDER, R. D. : Baker's yeast as food

for man. Amer. J. Physiol. 48, 199, 1919.

1236. RUTGERS, J. : Haben vegetabilischen Eiweisstoffe den gleichen

Nahrwert fur den Menschen wie die animalischen. Z. f. Biol.
24, 351, 1888.

1237. BORUTTAU, H. : Wie wird pflanzliches Eiweiss im Tierkorper verwer-

tet? Bio. Z. 69, 225, 1915; 82, 196, 1917.

1238. BOTAZZI, F. : Importance des proteines et des graisses d'origine animal

dans la nutrition deFhomme. Bull. soc. d'hyg. alim. 7, 179, 1919.

LITERATURE TO THE TEXT 461

1239. ROSE, C., AND BERG, RAGNAR: Abhangigkeit der Eiweissbediirfnisse

von dem Stoffwechsel. Munch, med. W. 65, 1011, 1918.

1240. HART, E. B. AND STEENBOCK, H. : At what level do the proteins of

milk become effective supplements to the proteins of cereal grain?
J. Biol. Chem. 42, 167, 1920.

1241. MAIGNON, F. : C. r. soc. biol. 82, 398, 1919.

1242. GRAFE, E. : D. Arch. klin. Med. 113, 1.

1243. EMMETT, A. D. : Polyneuritis as influenced by the amount of proteins

and carbohydrates present. Amer. Chem. Soc. Chicago, Sept.
. 6-10, 1920; Science, 52, 566, 1920.

1243a. OSBORNE AND MENDEL: Results of feeding experiments with mixtures
of foodstuffs in unusual proportions. Nat. Acad. Sci. Wash.
Apr. 26, 1921. Proc. Soc. Exp. Biol. Med. 18, 136, 1921; Growth on
diets containing more than 90 per cent protein. Ibid. 18, 167, 1921.

1244. TACHAU, PAUL: Versuche iiber einseitige Ernahrung. I. Mitt. Bio.

Z. 65, 253, 1914; II. 67, 338, 1914.

1245. HAUPT, HUGO: Chem. Ztg. 43, 134, 142, 1919.

1246. MASON, C. C. : German nutrition 1914-19. Bull. Johns Hopkins Hos-

pital, 31, 66, 1920.

1247. BORNSTEIN, K. : Berl. klin. W. 56, 968, 1919.

1248. ZUNTZ, N. AND LowT, A. : Einfluss der Kriegsnahrung auf den Stoff-

wechsel. BerL klin. W. 53, 825, 1916; Bio. Z. 90, 244, 1918.

1249. ROSENTHAL, F. : Deutsch. med. W. 45, 570, 1919.

1250. HOFFMAN, H. : Arch. Gynakol. 110, 451, 1919.

1251. JAHREISS: Munch, med. W. 66, 1421, 1919.

1252. BLOCH: Untersuchung der unterernahrten deutschen Kinder. Munch.

med. W. 67, 1062, 1920.
1252a. KAUPE, W. : Muttermilch und Kreig. M. f . Kinderh. 15, 83, 191S.

1253. ABDERHALDEN, E. : Munch, med. W. Okt. 3, 1919.

1254. PFAUNDLER, M. : Ueber Korpermasse von Mtinchener Schulkindern

wahrend des Krieges. Munch, med. W. Aug. 1, 859, 1919.

1255. BLANTON, S. : Mental and nervous changes in the school children of

Trier, Germany, caused by malnutrition. Mental. Hyg. 3, 343, 1919.

1256. DEMOOR: Bull. acad. roy. m6d. beige. 29, 37, 1919; Demoor J. and

Slosse, A. : Ibid. 30, 457, 1920.
1256a. DUTHOIT, RAOUL: Ibid. 30, 141, 1920.

1257. NOBECOURT, P. : Presse meU 27, 653, 1919.

1258. FRONCZAK, F. E. : Poland in World War from medical aspect. N. Y.

Med. J. Ill, 59, 1920.

1259. ROSENFELD, G. : Z. f . inn. Med. 41, 305, 1920.

1260. BERG, RAGNAR: Volksernahrung im Kriege. Blatt, f. biol. Med.

Sept. 1917.

1261. THOMS: Pharm. Ztg. 64, 295, 1919.

1262. OSBORNE AND MENDEL: Growth upon diets poor in true fats. Proc.

Soc. Exp. Bio. Med. Nov. 17, 1920; J. Biol. Chem. 45, 145, 1920;
45, 277, 1921.

1262a. DHTTMMOND, J. C. : Nutrition on diets practically devoid of fat.
Proc. Physiol. Soc. July 10, 1920., J. Physiol. 44, xxx, 1920.

462 THE VITAMINES

1263. HINDHEDE, M. : Nahrungsinittelverbrauch danischer Familien. Munch.

med. W. No. 28, 948, 1915; Deutsch. med. W. No. 45, 1236, 1919;
Lessons from war restrictions in food. Ugeskr. for Laeger. 81,
183, 1919.

1264. KUCZYNSKI, R. AND ZuNTZ, N. : Unsere bisherige und unsere kiinf-

tige Ernahrung im Kriege. Vieweg. 1915.

1265. HINDHEDE : The influence of the rationing on the health. Report to

the minister of the interior. Copenhagen. 1920. The effect of
restriction during the war on mortality in Copenhagen. J. A. M. A.
74, 381, 1920.

1266. NEUHAUS: Behandlung der Falle durch Kriegsnahrung entstanden.

Deutsch. med. W. No. 25, 1917.

1267. NILSSON, A. : Z. f. Gynakol. 44, 876, 1920.

1268. GESSNER, W. : Der Krieg und die puerperale Eclampsie. Ibid. 44, 570,

1920.

1269. MOMM: Munch, med. W. 67, 783, 1920.

1270. KLOTZ: Z. f. Kinderh. 26, 150, 1920.

1271. ZERNIK, F. : Kriegsernahrung und Arzneimittelwirkung. Deutsch.

med. W. Juli 31, 858, 1919.

1272. KIEFPER, O. : Einfluss des Krieges auf Lungentuberkulose. Z. f. Tuberk.

32, 65, 1920.

1273. Report of the Registrar General, for 1917. Brit. Med. J. May 10, 1919.

1274. RUMPEL: Kriegswirkung auf die Ernahrungsverhaltnisse, Morbiditat

und Mortalitat. Deutsch. med. W. Juni 19, 1919.

1275. ELIAS, H. AND SINGER, R. : Diabetes mellitus und Kriegsernahrung

Deutsch. med. W. 46, 561, 1920.

1276. SEEPELDER, R. : Einfluss des Krieges auf Augenkrankheiten. Wien.

klin. W. 32, 1245, 1919.

1277. HARRIS, SEALE. : Food conditions and nutritional disturbances in

Europe with some remarks on the etiology of pellagra. South.
Med. J. 12, 294, 1919.

1278. BOUCHARD, Ch. : Recherches nouvelles sur la pellagre. Paris. 1862.

1279. ROUSSEL, THEOPHILE: Trait6 de la pellagre et des pseudopellagres.

Paris 1866.

1280. MARIE, A. : La pellagre. Paris. Giard & Briere. 1908.

1281. ROBERTS, STEWARD R. : Pellagra. London. Henry Kimpton, 1912.

1282. SCHILLING, V. : Pellagra. Spez. Path. Ther. inn. Krankh. Kraus-Brugsch

II a, 72, 1915.

1283. NILES, G. M. : Pellagra; an American problem. 2nd ed. Saunders Co.

1917.

1284. HARRIS. H. F.: Pellagra: A Study of its Etiology, Pathology and Treat-

ment, Macmillan 1919.

1285. WEISS, E. : La pellagra nel Tirolo meridionale e 1'azione del governo

contra la stessa. Riv. pell. 13, 90, 1913; Oesterr-Sanitatsw. 26, 309,
1914.

1286. SAMBON AND CHALMERS: Pellagra in British Islands. Brit. Med. J.

p. 1093, 1913.

LITERATURE TO THE TEXT 463

1287. Box AND MOTT: Fatal pellagra in two English boys, with the result

of the pathological investigation in one case. Trans. Soc. Trop.
Med. Hyg. 6, No. 5, 1913.

1288. Low, R. C. AND YELLOWLEES, H. : Case of pellagra. Edinb. Med.

J. 25, 315, 1920.

1289. PINAULT, L. G. : Pellagra. J. Can. M. A. 6, 228, 1916.

1290. ROLPH, F. W. : Ibid. 6, 323, 1916.

1291. LAVINDER: The prevalence and geographical distribution of pella-

gra in the U. S. U. S. Publ. Health Rep. Dec. 13, 1912; 28, 1555, 1913.

1292. PETERSEN: Pellagra. J. A. M. A. 69, 2096, 1917.

1293. DEEKS, W. E: Pellagra in the Canal Zone. Med. Rec. 81, 566, 1912;

South. Med. J. 9, 123, 1916.

1294. TUTTLE, H. K. : Pellagra in Chile. J. A. M. A. 69, 2105, 1917.

1295. MCDONALD, W. M. : Pellagra in Antigua. Lane. 188, 127, 1915.

1296. NICHOLLS, L.: Tropical pellagra. J. Trop. Med. Hyg. 15, 241, 1912.

1297. DE KOCK, P. J. AND BONNE, C. : Pellagra in Surinam. Ned. Tijdsr.

v. Gen. 2, 965, 1920.

1298. ROBERTS, S. R. : Types and treatment of pellagra. J. A. M. A. 75,

21, 1920.

1299. SILER AND GARRISON: Amer. J. Med. Sci. 146, 42, 238, 1913.

1300. GRIMM: Pellagra: some facts in its epidemiology. J. A. M. A. 90,

1423, 1913.

1301. WELLMAN AND SPARKES: On "winter cases" of pellagra. J. Trop. Med.

Hyg. 15, 131, 1912.

1302. SNYDER, J. R. : Pellagra in children. Amer. J. Dis. Childr. 4, 172, 1912.

1303. WESTON, W. : Pellagra in early childhood. Ibid. 7, 124, 1914.

1304. VOEGTLIN, C. AND HARRIES, R. H. : The occurrence of pellagra in

nursing infants with observations on the chemical composition of
the human milk from pellagrous mothers. Hyg. Lab. Bull. Wash.
116, 73, 1920.

1305. BYFIELD, A. H. : Polyneuritic syndrome resembling pellagra acro-

dynia seen in young children. Amer. J. Dis. Childr. No. 5, 347, 1920.

1306. MURPHY, W. A. : Pellagra in children. Arch. Fed. 34, 254, 1917.

1307. GOLDBERGER, JOSEPH: The etiology of pellagra. The significance

of certain epidemiological observations with respect thereto. U. S.
Pub. Health Rep. 29, 1683, 1914.

1308. WOOD, EDWARD J. : The diagnosis of pellagra. Arch. Diagn. 10, 139,

1917.

1309. JOHNSON: Cited according to Roberts: Pellagra (1281).

1310. GIVENS, M. H. : Chemical analysis of the stomachal content of 100

pellagrins. Amer. J. Med. Sci. 155, 221, 1919.

1311. SILER, J. F. : Discussion on pellagra. II. Pan-Amer. Sci. Congr. Wash.

1915-16; Proc. X, 12, 18, 1917.

1312. LYNCH: Pellagra. South Med. J. 10. 286, 1917.

1313. GTJRD: Pellagra. J. Exp. Med. 13, 98, 1911.

1314. GOLDBERGER, J. : Pellagra and a method of prevention. J. A. M. A.

66, 471, 1916.

464 THE VITAMINES

1315. DEIACO, Pius: Ueber Lokalisation und Natur der pellagrosen Haut-

symptome. Wien klin. W. 20, 967, 1907.

1316. MERK, LUDWIG: Die Hauterscheinungen bei Pellagra. Innsbruck.

p. 24, Fig. 6, 1909.

1317. CROSBY, C. E. : Pellagra with erythema of scrotum as initial skin

manifestation. J. A. M. A. 68, 1403, 1917.

1318. FIND LAY, G. M.: Study of the leucocyte changes in pellagra com-

pared with those occurring in beriberi. J. Path. Bact. 23, 480, 1920.

1319. SANDY: Amer. J. Insanity. 73, 609, 1917.

1320. MILLER, R. S. AND ISMAIL, A. C. : Pellagra and pellagra psychoses.

Lane. Oct. 16, 788, 1920.

1321. CHALMERS, A. J.: Trans. Soc. Trop. Med. Hyg. 6, 163, 1913.

1322. BARDIN, J. : Note on the differential blood counts in three cases of

pellagra. Old. Dom. J. Med. & Surg. 17, July, 1913.

1323. NAGAMATSU, T. : Pellagra-like disease. Hifukwa, Hitsunyokwa Zasshi

17, 49, 1917.

1324. LORENZ, W. F. : The cerebrospinal fluid in pellagra. Publ. Health

Rep. 29, 2360, 1914.

1325. LOMBROSO: La pellagra. Torino 1892.

1326. BABES AND SIGN: Cited according to Roberts, Pellagra. (1281).

1327. CALHOUN: Ophtalm. Rec. 26, 63, 1917.

1328. BEESON, CHAS. F.: The thyroid gland in pellagra. J. A. M. A. 63,

2129, 1914.

1329. MODINOS, P. : Bull. soc. m6d. d. hop. de Paris Apr. 6, 640, 1916.

1330. WILSON : Appendix to report No 2 on a pellagra epidemic at Armenian

refugees camp. Port Said Publ. Health Dep. Egypt 1916.

1331. MORSE, PLINN F. : The general pathology of pellagra, with special

reference to findings in the thyroid and adrenals. J. Lab. Clin.
Med. 1, 217, 1916.

1332. KOCH, MATHILDE L. AND VOEGTLIN, C. : Chemical changes in the cen-

tral nervous system in pellagra. Hyg. Lab. Bull. 103, 51, 1916.

1333. RIDLON, J. R. : Pellagra. Laboratory examination in connection with

the disease. U. S. Publ. Health Serv., No. 339, 1916.

1334. MURLIN, JOHN R. : The amino acid fractions and hippuric acid in the

urines of pellagrins. Hyg. Lab. Bull. 116, 45, 1920.

1335. LEWIS, ROBERT C. : The chemical composition of the blood of pel-

lagrins. Ibid. 116, 37, 1920.

1336. JOBLING, J. W. AND MAXWELL, E. S. : Alkali reserve in the blood of

pellagrins. J. A. M. A. 69, 2026, 1917.

1337. SULLIVAN, M. X. AND STANTON, R. E. : Alkali reserve in pellagra. Arch.

Internal Med. 26, 41, 1920.

1338. SULLIVAN: Proc. Soc. Amer. Biol. Chem.; J. Biol. Chem. 41, LXX, 1920;

Sullivan and K. K. Jones. The saliva in pellagra. Ibid. 41,
LXX, 1920; Publ. Health Rep. May 16, 1919.

1339. MYERS, VICTOR C. AND FINE, M. S. : Metabolism in pellagra. Amer.

J. Med. Sci. 114, 705, 1913.

1340. NICOLAIDI: Untersuchungen liber Ernahrungsbilanz der Pellagrosen.

Riv. stiinz. med. 9, No. 6, 1913.

LITERATURE TO THE TEXT 465

1341. ALBERTONI, C. "AND TULLIO, P.: L'alimentazione maidica nel sano

e nel pellagroso. R. acad. scienze Bologna 11 Jan. 1914.

1342. HUNTER, A., GIVENS AND LEWIS: Preliminary observations of metab-

olism in pellagra. U. S. Publ. Health Serv. Hyg. Lab. Bull. 102,
39, 1916.

1343. BOYD, F. D. : Pellagra. Edinb. Med. J. 24, 366, 1920.

1343a. SULLIVAN, M. X., STANTON, R. E. AND DAWSON, P. R.: Metabo-
lism in pellagra: Study of urine. Arch. Int. Med. 27, 387, 1921.

1344. VISWALLINGAM, A.:Etiol. of pellagra. J. Trop. Med. Hyg. 23,46, 1920.

1345. SHEPPARD, S. W.: The etiol. of pellagra. Brit. Med. J. 1773, 1912.

1346. STANNUS: Pellagra in Nyasaland. Trans. Soc. Trop. Med. Hyg.

5, 112, 1912; 7, 32, 1913; Ann. Med. Rep. Nyasaland 1913; Trop.
Dis. Bull. 4, 444, 1914.

1347. SCHUFFNER AND KuENEN: Die gesundheitliche Verhaltnisse des

Arbeiterstandes des Sebembah-Maatchappy. Arch. Schiffs-u.
Tropenhyg. 16, 277, 1912.

1348. NIGHTINGALE: Zeism or pellagra? Brit. Med. J. 300, 1914.

1349. EDWARDS, C. R. : Peripheral neuritis in Jamaica. J. Trop. Dis. Hyg.

19, 53, 1916.

1350. WEISS: Oesterr. Sanitatsw. 24, 497, 1912.

1351. ALPAGO-NOVELLO : Pellagra nel provincia di Belluno. Riv. pell.

ital. 13, 67, 1913.

1352. WOOD, EDWARD J. : Vitamine solution of the pellagra problem. J.

A. M. A. 66, 1447, 1916; Soc. Trop. Med. Hyg. London. May
14, 1920; Lane. 1166, 1920.

1353. BLOSSER, R. : Sugar-cane products as a cause of pellagra in the South.

J. A. M. A. 64, 543, 1915.

1354. JOBLING, J. W. AND PETERSEN, W. : The epidemiology of pellagra

in Nashville. J. Inf. Dis. 18, 501, 1916; 21, 109, 1917.
1355 Babes: Pellagre. Bull. sect. sci. de 1'acad. roumaine. Nov. 28, 1913.

1356. CAMURRI: Einige Betrachtungen iiber die Pathogenese und die Be-

kampfung der Pellagra. Zentr. f. Bakt. I. Abt. Orig. 53, 438, 1910.

1357. DEVOTO, L. : Aetiologie und Klinik der Pell. Wien. med. W. No.

1, 1913.

1358. WILSON: Rep. of comm. appointed by the Direct. Med. Serv. Egypt.

Exp. Force, regarding the prevalence of pellagra among Turkish
prisoners of war. J. R. M. C. 33, 42t>, 508; 34, 70, 1918.

1359. BOYD, F. D. AND LELEAN P. S. : Rep. of comm. of inquiry regarding

the prevalence of pellagra among Turkish prisoners of war. Alex.
Egypt. Dec. 31, 1918.

1360. LELEAN, P. S.: Pellagra. Soc. Trop. Med. Hyg. May 14, 1920; Lane.

May 29, 1166, 1920.

1361. GOLDBERGER, J., WHEELER, G. A. AND SYDENSTRICKER E. : Study of

the diets of nonpellagrous and pellagrous households. J. A. M. A.
71, 944, 1918; U. S. Publ. Health Rep. 35, 648, 1920; Arch. Internal
Med. 25, 451, 1920.

466 THE VITAMINES

1362. SYDENSTRICKER, EDGAR: The prevalence of pellagra. Its possible

relation to the rise in the cost of food. U. S. Publ. Health Rep. No.
308, 1915.

1363. SULLIVAN AND JONES: The chemical composition of the Rankin Farm

pellagra-producing experimental diets. Hyg. Lab. Wash. Bull.
120, 117, 1920.

1364. SULLIVAN, M. X. : Ibid. p. 127, 141.

1365. GOLDBEKGER: The pell, outbreak in Egypt. Lane. II. 41, 1920.

1366. RATER: Cited by Rayer. N. Orl. Med. Surg. J. 66, 718, 1914.

1367. KLEIMINGER: Z. ges. Neur. u. Psych. 16, H. 5, 1913.

1368. LORENZ W. F. : The treatm. of pell. U. S. Publ. Hea. Rep. 29, 2357, 1914.

1369. ELEBASH, C. C. : Treating the gastro-intestinal tract in pellagra.

South. Med. J. 7, 447, 1914.

1370. ALLISON, W. L. : Texas State J. Med. 10, 123, 1914.

1371. SYLVESTER, R. E. : Pellagra. South. Med. J. 7, 449, 1914.

1372. BRAVETTA, EUGENIO : Riv. pell. ital. 15, 43, 1915.

1373. WILLETS: South. Med. J. 8, 1044, 1915.

1374. RIDLON: Pell. U. S. Publ. Health Rep. No. 353, 1916.

1375. GOLDBERGER, WARING, C. H. AND WILLETS, D. G. : The treatment and

prevention of pellagra. U. S. Publ. Health Rep. No. 228, No.
307, 1906. Goldberger. Ibid. No. 461, 1918.

1376. VOEGTLIN, NEILL, M. H. AND HUNTER, A. : The influence of vitamines

eta the course of pellagra. Hyg. Lab. Bull. No. 116, 1920; Voegtlin:
Harvey Lecture. Jan. 24, 1920; Voegtlin: Recent work on pellagra.
U. S. Publ. Health Rep. No. 597, 1920.

1377. SHERMAN: Proc. Amer. Publ. Health Ass.; Amer. J. Publ. Health,

10, 86, 1920.

1378. SILER, J. F., GARRISON, P. E. AND MACNEAL, W. J. : Further studies

of the Thompson-McFadden Comm. J. A. M. A. 63, 1090, 1914.
1378a. TANNER, W. F. AND ECHOLS, G. L.: The occurrence of pellagra in
patients apparently receiving an ample diet. J. A. M. A. 76, 1337,
1921.

1379. BOYD, F. D. : Qualitative problems of food supply. Lane. 197, 934, 1919.

1380. ROAF: Ibid.

1381. BIGLAND, A. D. : Pellagra outbreak in Egypt. I. Pellagra among

Ottoman prisoners of war. Lane. I. 947, 1920.

1382. ENRIGHT, J. I.: II. Pellagra among German prisoners of war: food

factor in disease. Ibid. 998, 1920.

1383. ECKLES, C. H., PALMER, L. S. ANDSWETT, W. O. : Factors influencing

the composition of milk. Miss. Agr. Exp. Sta. Bull. 163, 33, 1919.

1384. VAN DER SCHEER: Aphtae tropicae. Handb. d. Tropenk. von Mense

Bd. II, S. 1.

1385. THIN: Psilosis (Sprue) 2nd Ed. London 1897.

1386. BROWN, W. C. : Sprue and its Treatment. London 1908.

1387. BEGG: Sprue, its Diagnosis and Treatment. London 1912.

1388. WOOD, EDWARD J. : The recognition of tropical sprue in the United

States. Amer. J. Med. Sci. 110, 692, 1915; J. A. M. A. 73, 165, 1919.

LITERATURE TO THE TEXT 467

1389. BOTD, M. F. : Is sprue endemic in South? South. Med. J. 13, 229, 1920.

1390. STTJRTEVANT, M. : Trop. sprue in New York. N. J. Med. Soc. J. 17,

44, 1920.

1391. Hi ATT, H. B. AND ALLAN, W. : Notes on cases of sprue invalided from

the tropics; will it become endemic here? J. A. M. A. 58, 395, 1914.

1392. BAHR, P. H. : Res. on sprue. Trans. Soc. Trop. Med. Hyg. 7, 161, 1914.

1393. HALBERKANN, J. : Harn und Faces Untersuchung bei der Sprue.

Arch. Schiffs. Tropenhyg. 20, 225.

1393a. BROWN, T. B.: The absence of pancreatic secretion in sprue and the
employment of pancreatic extract in the treatment of this
disease. Amer. J. Med. Sci. 161, 501, 1921 ; Bull. John. Hopk. Hosp.
26, 289, 1916.

1394. BASSETT-SMITH, P. W. : Sprue associated with tetany. Lane. I. 178,

1919.

1395. BARACH, A. L. AND MURRAY, H. A.: Tetany in a case of sprue. J.

A. M. A. 74, 786, 1920.

1396. SCHMITTEH, F. : Sprue treated by emetine hydrochloride. Milit.

Surg. 34, 330, 1914.

1397. MUHLENS, P: Behandlung bedrohlicher Zustande bei Trope nkrank-

heiten. Deutsch. med. W. 40, 1249, 1914.

1398. SIMON, SIDNEY K. : Sprue. South. Med. Ass. Meeting Nov. 15-18,

Louisville, Ky. 1920.

1398a. CASTELLANI, A.: Treatment of sprue by massive doses of sodium
bicarbonate. Brit. Med. J. I, 338, 1921.

1399. WEGELE: Ueber die diatetische Behandlung gewisser Formen chron-

ischer Diarrhoen, speziell von "Indian Sprue." Med. Klin. 9,
866, 1913.

1400. Low, G. C. : Arthritis in sprue. J. Trop. Med. Hyg. 17, 1, 1914.

1401. CANTLIE : Some recent observations on sprue. Brit. Med. J. II, 1296,

1913.

1402. CONRAN, P. : Meat diet in sprue. Ibid. II, 206, 1920.

1403. CANTLIE: The diet in sprue. J. Trop. Med. Hyg. 9, 277, 1906.
1403a. BOVAIRD, DAVID: A study of tropical sprue, or psilosis. J. A. M. A.

77, 753, 1921.

1404. MICHAEL, CARL: A study of toxins and the serological reactions in

sprue. J. Amer. Med. Sci. 154, 171, 1917.

1405. ASHFORD: Cited according to Siler (1311).

1406. STEWART, CHARLES E. : The probable identity of pellagra and sprue.

Trans. 17 Intern. Congr. Med. London Sect. 21, Trop. Med. Part
II, 125, 1913.
1406a. HEATON, T. : Etiology of sprue. Ind. J. Med. Res. 7, 810, 1920.

1407. WERNER, H. : Skorbutsymptome bei Sprue. Arch. Schiffs. Tropenhyg.

No. 7, 1914.

1408. LEEDE: Ein Fall von Sprue durch Erdbeeren gebessert. Z. f. Hyg.

Infektionsk. 75, 578, 1913.

1409. STICKER: Erkaltungskrankheiten und Kalteschaden. Springer. Ber-

lin. 1916.

468 THE VITAMINES

1410. PRINTING: Epidemics resulting from wars. Oxford Univers. Press, 1916.

1411. MALIWA: Wien. klin. W. 30, 1477, 1917.

1412. WHEELER: War edema. Brit. Med. J. No. 2, 1902.

1413. DIGBY: The famine campaign in Southern India. 1876-77.

1413a. McLsoD, K. : Epidemic dropsy in Calcutta. Ind. Med. Gaz. 16,
148, 1881.

1414. PATTERSON, A. B. : Starvation, edema. Med. Rec. Nov. p. 715, 1899.

1415. LANDA: Deficiency edema. Gaceta med. Mexico. 11, 67, 1917.

1416. RUMPEL: Kriegsodem. Miinch. med. W. No. 30, 1915; Berl. klin.

W. 1916.

1417. JURGENS: Berl. klin. W. No. 9, 1916.

1418. BONHEIM: Miinch. med. W. No. 24, 1917.

1419. LANGE, F. : Deutsch. med. W. No 28, 876, 1917.

1420. KNACK: Wien. klin. W. No. 32, 1916.

1421. SCHIFF: Oedemkrankheit. Ibid. No. 22, 1917.

1422. JAKSCHE: Ibid. 68, 1030, 1918.

1423. VANDERVELDE, PAUL AND CANTINEAU, GASTON: Bull. acad. roy. med.

belg. 29, 129, 1919.

1424. BREUER, MARCEL: Ibid. 30, 99, 1920.

1425. BEYERMAN, W. : "Edema disease" in the Netherlands, Ned. Tijdsr.

v. Gen. 1, 2265, 1919.

1426. STRAUSS: Med. Klin. No. 39, 1915.

1427. GUILLEMIN, R. AND GuYOT, F. : Rev. me"d. de la Suisse romande. 39,

115, 1919.

1428. BUDZYNSKI B. AND CHELCHOWSKI, K. : Hunger swelling in Poland.

J. Trop. Med. Hyg. 19, June 15, 1916.

1429. WELLS, H. G. : War edema. J. A. M. A. 71, 954, 1918.

1430. TONIN, ROMANO: Gazz. degli osped. e delle cliniche. 40, 636, 1919.
1431." ENRIGHT, J. J. : War edema in Turkish prisoners of war. Lane. I,

314, 1920.

1432. MANN, W. L., HELM, J. B. AND BROWN, C. J. : An edema disease in Haiti.

J. A. M. A. 75, 1416, 1920.

1433. VACHER: La mortality a Paris en 1870. Gaz. meU de Paris p. 9, 1871.

1434. DE WOLF, H. : A report of 13 cases of edema apparently epidemic in

character. Arch. Ped. 19, 895, 1902.

1435. POTTER, P. A. : The relation of proteins to edema in marantic children.

Med. News Jan. 9, 1904; Arch. Ped. 29, 206, 1912.

1436. CHAPIN, H. D.: Cases of edema in infants. Arch. Ped. 31, 5, 1914.

1437. WATERMAN, L. : Osmosis and edema in infancy and childhood. Arch :

Ped. 31, 135, 1914.

1438. HUME, W. E. : General edema following gastro-enteritis in children.

Brit. Med. J. II, 478, 1911.

1439. ASHBY, H. T. : Practit. p. 686, 1914.

1440. KLOSE, ERICH. : J. f . Kinderh. 80, 154, 1914.

1440a. WESTON, WILLIAM.: Acrodynia. Arch. Ped. 37, 513, 1920.
1440b. CARTIN, H. J.: Acrodynia. Penn. Med. J. 24, 287, 1921.

1441. McCAY, D.: Epidemic dropsy. Scient. Mem. Gov. India. No. 37, 204,

1910.

LITERATURE TO THE TEXT 469

1442. MAVER, MARIA B. : Nutritional edema and "war dropsy." J. A. M. A.

74, 934, 1920.

1443. SCHITTENHELM, A. AND SCHLECHT H. : Die Oedemkrankheit. Springei .

' 1919. Z. exp. Med. 9, 1, 1919.

1444. SCHIFP, A. : Zur Pathologie der Oedemkrankheit. Wien. med. W.

No. 48, 1917.

1445. HULSE, WALTER: Munch, med. W. No. 28, 1917.

1446. RUMPEL AND KNACK: Berl. klin. W. No. 36, 1916. Deutsch. med. W.

No. 44, 1916.

1447. ZONDEK, HERMANN : Berl. klin. W. 55, 502, 1918.

1448. JANSEN, W. H.: Munch, med. W. No. 1, 1918.

1449. JESS : Deutsch. med. W. No. 22, 681, 1917.

1450. KNACK, A. V. AND NEUMANN, I. : Deutsch. med. W. 43, 901, 1917.

1451. FALTA: Wien. klin. W. 30, 1736, 1917.

1452. BURGER, MAX: Epidemisches Oedem und Enterokolitis. Z. f. ges.

exp. Med. 8, 309, 1919.

1453. KRAUS: Berl. klin. W. 56, 3, 1919.

1454. HULSE, W. : Virch. Arch. 225, 234, 1918.

1455. LIPPMANN: Z. f. arztl. Forth. No. 18, 1917.

1456. PALTAUF: Wien. klin. W. No. 46, 1917.

1457. JACOBSTHAL: Sitzungsb. arztl. Ver. Hamburg. 3 Sept. 1917.

1458. WOLTMANN: Wien. klin. W. 1916.

1459. MAYNARD, F. P. : Preliminary note on increased intraocular tension

met with in cases of epidemic dropsy. Ind. Med. Gaz. 44, 373, 1909.

1460. FRANKE, M. AND GOTTESMANN, A. : Wien. klin. W. 30, 1004, 1917.

1461. FEIGL, JOHANN: Bio. Z. 83, 365, 1918.

1462. ZAK: Wien. klin. W. No. 19, 1917.

1463. MAASE AND ZONDEK: Deutsch. med. W. No. 44, 45, 46, 1916; No.

16, 1917; Berl. klin. W. No. 36, 1917.

1464. REACH: Wien. klin. W. 31, 1249, 1919.

1465. V.HOSSLIN, HEINRICH: Klinische Eigenthumlichkeiten und Ernah-

rung bei schweren Inanition. Arch. Hyg. 88, 147, 1919.

1466. SCHITTENHELM, A. : Eiweisstherapie. Munch, med. W. 66, 1408, 1919.

1467. ISENSCHMID, R. : Behandlung von Oedemen. Schweiz. med. W. 50,

381, 1920.

1467a. ARON, HANS: NahrstoffmangelalsKrankheitsursache. Berl. klin. W.
No. 33, 773, 1920.

1468. PARK, F. S. : War edema. J. A. M. A. 70, 1826, 1918.

1469. RUBNER, M. : Bericht an das Reichsgesundheitsamt. Dez. 20, 1917.

1470. DETERMANN, H. : Die Bedeutung der Kriegsnahrung fur Stoffwechsel

und Gesundheit. Z. f. Physik. u. diatet. Ther. 23, 147, 1919.

1471. EPSTEIN, ALBERT A. : The nature and treatment of chronic paren-

chymatous nephritis. J. A. M. A. Aug. 11, 444, 1917. Amer. J. Med.
Sci. 154, 638, 1917.

1472. ALLBTJTT: Dropsy. Brit. Med. J. II, 395, 1918.

1473. DENTON, MINNA C. AND KOHMAN, EMMA: Feeding experiments

with raw and boiled carrots. J. Biol. Chem. 36, 249, 1918.

470 THE VITAMINES

1474. KOHMAN, EMMA A. : The experimental production of edema as related

to protein deficiency. Proc. Soc. Exp. Biol. Med. 16, 121, 1919;
Amer. J. Physiol. 51, 378, 1920.

1475. BIQLAND, A. D. : Edema as symptom in so-called food deficiency

diseases. Lane. I. 243, 1920.

1476. NIXON, J. A.: Famine dropsy as a food deficiency disease. Bristol

Med. Chirurg. J. 37, 137, 1920.

1477. LITTLE: Boston Med. Surg. J. 158, 253, 1908; 176, 642, 1917.

1478. BKTJNTZ, L. AND SPILLMANN, L. : C. r. soc. biol. 81, 1243, 1918; 82, 8, 1919;

Progres me'd. 34, 9, 1919.

1479. MERCIER, R,: 600 cas de froidures des pieds. Bull. acad. de me'd.

Paris. 82, 80, 1919.

1480. CHAUVIN, E. : Rev. de chir. 38, 793, 1919.

1481. McCARRisoN, R. : Deficiency disease: special reference to gastro-

intestinal disorders. Brit. Med. J. I, 822, 1920.

1482. LANE, SIR WILLIAM ARBUTHNOT: Chronic intestinal stasis. Brit.

Med. J. II, 795, 1919.

1483. STILL, G.: Coeliac disease. Lane. II, -163, 1918.

1484. WALLIS, R. L., MACKENZIE: 88th Ann. Meet. Brit. Med. Ass. Cam-

bridge, June 30- July 2nd 1920; Brit. Med. J. 160, 1920, July 31st.

1485. REYNOLDS, EDWARD AND MACOMBER, DONALD: Deficient diet as a

cause of sterility; a study based upon feeding experiments with
rats. J. A. M. A. 77, 169, 1921.

1486. ISHIWARA: Nature of essential hemeralopia. Nippon Gankakai Zas-

shi. June 1912; Klin. Monatsh. Augenh. 15, 569.

1487. TRICOIRE, R. : He'me'ralopie epide*mique. Paris me'd. 10, 152, 1920.
1487a. SMITH, H.: Night-blindness and the malingering of night-blindness.

J. A.M. A. 77, 1001, 1921.

1487b. APPLETON, V. B.: Observations on deficiency diseases in Labrador.
Amer. J. Public Health. 11, 617, 1921.

1488. PICK, L. : Einfluss des Krieges auf die Augenkrank. Deutsch. med.

W. 46, 44, 1920.

1489. FEILCHENFELD, W. : Augenkrankheiten im Kriege. Ibid. 46, 575, 1920.

1490. HEKTOEN, LUDWIG: The formation of antibodies in rats fed on pure

vegetable proteins (Osborne-Mendel diet). J. Inf. Dis. 15, 278, 1914.

1491. ZILVA, S. S. : The influence of deficient nutrition on the production of

agglutinins, complement and amboceptor. Biochem. J. 13, 172, 1919.

1492. KLEINSCHMIDT, H. : Ernahrung und Bildung von Antikorpern. M. f .

Kinderh. 12, 423, 1914.

1493. THOMAS, E. : Beziehung zwischen chronischer Unterernahrung zu

den klinischen Zeichen einerverminderten Immunitat. Z. f. Kinderh.
No. 4, 1914.

1494. VALAGUSSA, F. : Protein therapy of infectious diseases of children.

Policl. Med. Sektion, 27, 361, 1920.

1495. PEISER: Fett in der Kinderdiat. Berl. klin. W. 51, 1065, 1914.

LITERATURE TO THE TEXT 471

1496. Morrison, J. R.: Some of the dangers of too greatly restricted diet in

typhoid. Kentucky Med. J. 12, No. 22, 1914.

1497. BARKER, L. F. : Diet in typhoid fever. J. A. M. A. 63, 929, 1914.

1498. COLEMAN, W. : Influence of the high-caloric diet on the course of

typhoid fever. J. A. M. A. 69, 329, 1917.

1499. WALTON, ALICE C. : The new feeding in the treatment of typhoid fever.

J. Home Econ. 9, 14, 1917.

1500. CARTER, H. S., HOWE, P. E. AND MASON, H. H. : Nutrition and Clini-

cal Diabetes. Lea & Febiger. Philadelphia 1917.

1501. COMBE, A. : Comment se nourir en temp de guerre. Payot, Paris. 1917.

1502. HERDLIKA, A. : Bur. of Amer. Ethnol. Bull. 34.

1503. McCARRisoN, R. : 88th Ann. Meet. Brit. Med. Ass. Cambridge. Brit.

Med. J. II, 154, 1920.

1504. MORI, M. : J. f . Kinderh. 59, 175, 1904.

1505. CZERNY AND KELLER: Ernahrung des Kindes. p. 2, 67, 1906.

1506. KNAPP, P. : Experimenteller Beitrag zur Ernahrung von Ratten mit

klinstlicher Nahrung und zum Zusammenhang von Ernahrungs-
storungen mit Erkrankungen der Conjunktiva. Z. f. exp. Path,
u. Ther. 5, 147, 1909.

1507. FREISE, E., GOLDSCHMIDT, M. AND FRANK, A. : Experimentelle Bei-

trage zur Aetiologie der Keratomalazie. Vorl. Mitt. M. f . Kinderh.
13, 424, 1914-16.

1508. GOLDSCHMIDT, M. : Experimenteller Beitrag zur Aetiologie der Ke-

ratomalazie. Arch. f. Ophtal., 90, 354, 1915.

1509. NELSON, V. E. AND LAMB, ALVEN, R. : Further studies on the effect

of a deficiency of fat-soluble vitamine. Amer. Chem. Soc. Chi-
cago Meet., Sept. 6, 1920, Science 52, 566, 1920.

1510. GUERRERO, L. E. AND CONCEPTION, I.: Xerophtalmia in fowls fed on

polished rice and its importance. Philipp. J. Sci. 17, 99, 1920.

1511. BLOCH, C. E. : Eye disease and other disturbances in infants from

deficiency in fat in the food. Ugeskr. forLaeger 79, 309, 1917; 80,
815, 868, 1918; Ringhosp. Medd. 2, 17, 1918; 3, 57, 1918.
151 la. BLOCH, C.E.: Clinical investigation on xerophthalmia and dystrophy
in young children. J. Hyg. 19, 283, 1921.

1512. MONRAD: Ugeskr. f. Laeger. 79, 1177, 1917.

1513. R0NNE, H. : Ibid. 79, 1479, 1917.

1514. SZTARK, C. H.: Arch, de malad. d. enf. 22,23,1919.

1515; PARKER, R. H. : Xerophtalmia. Iowa State Med. J. 10, 71, 1920.

1516. MAGPIE, J. W. S. : Xerophtalmia in native of Gold Coast. Ann. Trop.

Med. Paras. 13, 343, 1920.

1517. BULLET, E. C. : Note on xerophtalmia in rats. Biochem. J. 13, 103,

1919.

1518. STEPHENSON, M- AND CLARK, A. B. : A contribution to the study of

keratomalacia among rats. Biochem. J. 14, 502, 1920.

1518a. WASON, I.M.: Ophthalmia associated with dietary deficiency in fat-
soluble vitamin A. Study of the pathology. J. A. M. A., 76, 908,
1921.

472 THE VITAMINES

1519. EMMETT, A. D. : The fat-soluble A-vitamine and xerophtalmia. - Science

52, 157, 1920.

1520. MENDEL, L. B. : N. Y. Path. Soc. Nov. 10, 1920.

1521. GUIRAL, R. : Keratomalacia and its treatment. Riv. med. y. cirurg.

Havana. 24, 157, 1919.

1522. HAMBURGER, RICHARD: Die Ernahrung der deutschen Kinder in der

Kriegszeit und der Gegenwart. Z. f. Krankenpfl. H. 5, Sept. 1919.

1523. STOLZNER, W.: Diat in Tuberkulose. Miinch. med. W. 67, 981, 1920.

1524. GEOGHEGAN, JOSEPH: Tuberculosis from a West Indian standpoint.

Lane. July 12th, 1919.

1525. RICHET, CHARLES: Traitement de la tuberculose expe'rimentale par

la viande et le serum musculaire (zomothe'rapie) Trav. du lab. de
C. Richet. 5, 1902.

1526. WOODCOCK, H. de C. AND RUSTIN, A. G. : Food values in tuberculosis.

Lane. II, 842, 1920.

1526a. GARDE Y, F.: Vitamines as important factor in treatment of pul-
monary tuberculosis. Semana me"d. 27, 759, 1920.

1527. WEIGERT: Berl. klin. W. No. 38, 1904.

1528. THOMAS, ERWIN AND HORNEMANN, O. : Beziehung zwischen der In-

fektion und Ernahrung. I. Bio. Z. 57, 456.

1529. RENON: La tuberculose et les vitamines. Bull. ge"n. de therap. 30

Juillet, 1914.

1530. MUTHU, C. : 88th Meet. Brit. Med. Ass. Brit. Med. J. II, 160, 1920.

1531. BUTTON, A. S. : Some deficiency diseases and leprosy. Med. Press

N. S. 109, 313, 1920.

1532. HUTCHINSON, SIR JONATHAN: On leprosy and fish eating. 1906.

1533. DEYCKE, GEORG. : Die Lepra. Kraus-Brugsch. Path. Ther. d. Stoffw.

II, 1, 469.

1534. UNDERBILL, F. P., HONEIJ, J. A., BOGERT, L. J. AND ALD RICH, M. L. :

Calcium and magnesium metabolism in leprosy. J. Exp. Med.
32, 41, 1920.
1534a. VOKURKA: Korrespondenz aus Budapest. J. A. M. A. 77, 215, 1921.

1535. REACH, FELIX: Studien iiber die Nebenwirkung der Nahrungsstoffe

Sitzungsb. Wien. Akad. 122, Abt. Ill, 1, 1913.

1536. SALANT, W. : The importance of diet as a factor in the production of

pathological change. J. A. M. A. 69, 603, 1917.

1537. SALANT, W. AND SWANSON, A. M. : Further observations on the influence

of diet on the toxicity of sodium tartrate. Proc. Soc. Exp. Biol.
Med. 14, 100, 1917.

1538. PEARCE, RICHARD M., AUSTIN, HAROLD, J. AND PEPPER, O. H. PERRY:

The relation of the spleen to blood destruction and regeneration
and hemolytic jaundice. XIII. The influence of diet upon the
anemia following splenectomy. J. Exp. Med. 22, 682, 1915.

1539. MADSON, E.: The vitamines. Ugeskr. f . Laeger. 80, 613. 1918.

1539a. CEILING, E. M. K. AND GREEN, H. H.: Studies in regeneration of
blood. Proc. Soc. Exp. Biol. Med. 18, 191, 1921.

LITERATURE TO THE TEXT 473

1540. DAVIS, N. C., HALL, C. C. AND WHIPPLE, G. H. : Rapid construction

of liver cells protein on a strict carbohydrate diet contrasted with
fasting. Arch. Internal Med. June 23rd, 689, 711, 1919.

1541. CAMPBELL, H. : Etiology, prevention and nonoperative treatment of

adenoids. Brit. J. Childr. Dis. 16, 140, 1919.

1541a. VAN DER BOGERT, FRANK: Diet as a factor in the etiology of adenoids.
Amer. Med. Assoc. Boston Meeting. June 2-10, 1921.

1542. HAMMER, U. : Deutsch. med. W. 46, 738, 1920.

1543. PECKAM, FRANK E. : Many orthopedic deformities due to calcium de-

ficiency, as direct result of sterilized and pasteurized food. J. A.
M. A. 75, 1317, 1920.

1544. MAGNUS-LEVY, A.: Diabetes in Kriege. Deutsch. med. W. 45, 1379,

1919.

1545. GERHARDT, D. : Diabetes in Kriegszeiten. Schweiz. med. W. 50,

141, 1920.

1546. MAGNUS-LEVY: Diabetes im Kraus-Brugsch. Spez. Path. Ther. d.

inn. Krankh. Bd. I, 1913.

1547. BORUTTAU, H. : Spezifische antidiabetische Substanz. Bio. Z. 88,

420, 1918.

1548. ROSE, CARL W. : Alkaloide der Driisen der inneren Sekretion. Berl.

klin. W. 51, 1217.

1549. JACOBSEN, AAGE: Th. B. Bio. Z. 56, 471.

1550. v. MORACZEWSKI, W. : Bio. Z. 71, 268. 1915.

1551. McCAY, D., BANERJEE, S. C., GHOSTAL, L. M., DUTTA, M. M. AND

RAY, C. : Sugar of blood and sugar in urine in varying conditions
of health in Bengali. Ind. J. Med. Res. 6, 485, 508, 1919.
1551a. WILDER, R. M. AND BEELER, C.: Plasma chlorides and edema in
diabetes. Amer. J. Physiol. 55, 287, 1921.

1552. Editorial: An unsuspected occasional menace in low protein diets.

J. A. M. A. 75, 1426, 1920.

1553. MCCARRISON, R. : Occurrence of recently developed cancer of stomach

in monkey fed on food deficient in vitamine. Ind. J. Med. Res.
7, 342, 1919.

1554. HOFFMAN, FREDERICK L. : The mortality from cancer in the Western

Hemisphere. Proc. II. Pan-Amer. Sci. Congr. Wash. X, 586, 1917.

1555. FUNK, CASIMIR : The application of chemical methods to the study of

cancer. Ibid. X, 388, 1917.

1556. EWING, J. : Pathological aspects of some problems of experimental

cancer research. J. Cane. Res. 1, 71, 1916; Proc. II. Pan-Amer.
Sci. Congr. X, 512, 1917.

1557. LOEB, LEO: General problems and tendencies in cancer research.

Ibid. p. 347.

1558. CALKINS, GARY N. : Effects of cancer tissue and of normal epithelium

on the vitality of protozoa. J. Cane. Res. 1, 205, 1916.

1559. v. GRAFF, E. : Der Einfluss der Schwangerschaft auf das Wachstum

maligner Tumoren. Wien. klin. W. 27, 7, 1914.

1560. SLYE, MAUD: J. Cane. Res. 5, 25, 1920.

474 THE VITAM1NES

1561. JANSEN: Z. f. Krebsf. 20, 682, 1909.

1562. HAALAND: Cited according to Ehrlich. Exper. Path. u. Chemotherapie..

Leipzig 1909.

1563. CRAMER, W. AND PRINGLE, HAROLD : The influence of diet on tumor

growth. Proc. R. Soc. 88 (B), 307, 315, 1910.

1564. Rous, P. : The rate of tumor growth in underfed hosts. Proc. Soc.

Exp. Biol. Med. 8, 128, 1911.

1565. v. JAWORSKI, JOSEF: Wien. klin. W. p. 1646, 1916.

1566. SWEET, CORSON-WHITE AND SAXON: The relation of diets and cas-

tration on the transmissible tumors of the rats and mice. J.Biol.
Chem. 15, 181, 1915; 21, 309, 1915. Corson-White E. P. : Penns. Med.
J. 22, 348, 1919.

1567. HOPKINS, F. G. : Discussion on defic. diseases. Proc. R. Soc. Med.

VII, 1, (Pharm. Sect. ) 1913.

1568. CENTANNI, EUGENIO: La dieta aviride per lo sviluppo dei tumori speri-

mentali. VI. Tumori II, 466, 1914.

1569. Rous, P. : The importance of diet in transplanted and spontaneous

mouse tumors. J. Exp. Med. 20, 433, 1914.

1570. DRUMMOND, J. C. : A comparative study of tumor and normal tissue

growth. Biochem. J. 11, 325, 1917.

1571. BENEDICT, S. R. AND RAHE, ALFRED H. : Studies on the influence of

various factors in nutrition upon the growth of experimental tumor.
I. J. Cane. Res. 1, 159, 1917.

1572. SUGIURA, K. AND BENEDICT, S. R. : Influence of certain diets on tumor

susceptibility and growth in albino rats. J. Cane. Res. 5, 373, 1920.

1573. VAN ALSTYNE AND BEEBE: The effect of non-carbohydrate diet upon

the growth of sarcoma in rats. J. Med. Res. 29, 219, 1913.

1574. LEVIN, I. : Proc. N. Y. Pathol. Soc. 16, 93, 1916.

1575. FRANKEL, SIGMUND AND FURER, EDINA: Wien. klin. W. 29, 483.

1576. RONDONI, P. : VII. Riun. soc. ital. patol. Pisa 1913.

1577. MURPHY: Transplantability of malignant tumors to the embryo of

foreign species. J. A. M. A. 59, 874, 1912; J. Exp. Med. 17, 482,
1913.

1578. FUNK, CASIMIR: The transplantation of tumors to foreign species.

J. Exp. Med. 21, 571, 1915.

1579. BULKLEY, L. DUNCAN: On the cure of cancer. Med. Rec. 97, 941, 1920;

The medical treatment of cancer. Davis Co. Phila. 1920.

1580. COPEMAN, S. MONCKTON: 88th Ann. Meet. Brit. Med. Ass. Brit. Med.

J. II, 159, 1920.

1581. KUNERT: Unsere heutige falsche Ernahrung, Breslau. 1914.

1582. DURAND, J. I. : Influence of diet on the development and health of

teeth. J. A. M. A. 67, 564, 1916.

1583. BLACK, G. V. AND McKAY, F. .S. : Mottled teeth. Dent. Cosmos.

58, 129, 1916.

1584. CASTILLA, C. R. : Changes in the teeth of children from nutritional

derangement. Semana med. Buenos Aires. 26, 599, 1919.

1585. MILLER, E. G. AND GIES, W. J. : Further nutritive studies on denti-

tion. J. Allied Dent. Soc. 11, 47, 69, 70, 1916.

LITERATURE TO THE TEXT 475

1586. MELLANBY, MAY: The influence of diet on teeth formation. Lane.

II, 767, 1918.

1587. ZILVA, S. S. AND WELLS, F. M. : Changes in the teeth of the guinea

pig by a scorbutic diet. Proc. R. Soc. 90 (B) 505, 1919.

1588. HOWE, PERCY R. : Effect of scorbutic diet upon the teeth. Dent.

Cosmos. 62,586, 1920; 62, 921, 1920.
1588a. ROBB, E. F., MEDES, GRACE ; MCCLENDON, J. F., GRAHAM, MARGARET,

AND MURPHY, T. J.: A study of scurvy and its bearing on the

preservation of the teeth. J. Dent. Res. 3, 38, 1921.
1588b. HOWE, P. R.: Food accessory factors in relation to teeth. J. Dental

Res. 3, 7, 1921.

1589. BALLANTYNE: Brit. Med. J. July 26th, 1919.

1590. SINCLAIR, J. F. : Influence of diet affecting second dentition. Penn.

Med. J. 12, 789, 1919.

1591. OSBORNE AND MENDEL: Incidence of phosphatic urinary calculi in

rats fed on experimental rations. J. A. M. A. 69, 32, 1917.

1592. PADUA, REGINO G. : Cystolithiasis among Filipinos in association

with dietetic deficiency. Philipp/ J. Sci. 14, 481, 1919.

1593. KIRSCHNER: Therapeut. Monatsh. 33, 300, 1919.

1594. CLEMM, W. N. : Ibid. p. 302;

1595. DRUMMOND, J. C.: Vitamines and certain aspects of the relation to

public health. Amer. J. Publ. Health. 11, 593, 1921.

INDEX TO SECTIONAL DIVISIONS OF THE LITERATURE

Historical Part Nos. 1-68

Vitamine Requirements of Plants Nos. 69-191

Vitamine Requirements of Animals Nos. 191a-462

Chemistry, Physiology and Pharmacology of the Vitamines:

Vitamine B Nos. 462a-569

Vitamine A Nos. 570-619

Vitamine C Nos. 620-672

Vitamine Content of Various Foodstuffs Nos. 673-837

Beriberi Nos. 838-913

Scurvy Nos. 914-1028

Rickets, Osteomalacia, Tetany, Spasmophilia and Carbohydrate

Dystrophy Nos. 1029-1215a

Nutrition in Man Nos. 1216-1277

Pellagra Nos. 1278-1383

Sprue Nos. 1384-1408

Hunger Edema Nos. 1409-1476

Pathological Conditions in Which the Lack of Vitamines may be

Suspected Nos. 1477-1595

476

AUTHOR INDEX

Abderhalden, 20, 25, 37,
53, 54, 80, 134, 189,
190, 198, 206, 207,
211, 255, 256, 264, 265,
266, 348

Abt, 336

Acree, 56

Adams, 34, 384

Agulhon, 61

Alb, 396

Albert, 294

Albertoni, 362

Aldrich, 385

Alincastre, 196

Allan, 369

Allbutt, 376

Allen, B., 245

Allen, F. P., 85, 86, 121

Allison, 366

Almy, 85

Alpago-Novello, 363

Alsberg, 395

van Alstyne, 390

Alting, 293

Alwens, 330

Amand, 52

Andersen, 141

Anderson, 257

Andrews, 257, 291

Anrep, 209

Antoine, 138

Appleman, 68

Appleton, 380

Arima, 293

Arneth, 299 (#954)

Arnold, 141

Arntzenius, 332, 333,
334

Aron, 114, 117, 122, 229,
317, 321, 338, 376, 397

Aschenheim, 317

Aschoff, 303, 304, 305,

308, 309
Ashby, 373
Asher, 398
Ashford, 371
Aston, 142, 147
Auer, 256, 265, 266, 267
Aulde, 397
Austin, 386
Autran, 297
Avery, 62
Ayers, 57
Azzi, 396

B

Babes, 360, 364
Bachmann, 54, 198
Bachstrom, 27, 297
Balz, 278, 283
Baglioni, 250, 251
Bahr, 369
Bahrdt, 311, 339
Bainbridge, 56
Ballantyne, 392
Bang, 342
Banner jee, 388
Banu, 316
Barach, 370
Bardin, 359
Bardouin, 316
Barger, 174
Barker, 381
Barlow, 27, 257, 302
Barnes, 259, 270
Barsickow, 186 •
Bass, 92

Bassett-Smith, 234, 370
Bauman, 133, 308
Baumbach, 299 (#954)
Baumberger, 84
Beck, 314
Beebe, 390
Beeler, 388

477

Beeson, 361
Beger, 135
Begg, 369
Belonowsky, 77
Bendix, 302, 337
Benedict, S. R., 44, 109,

125, 195, 269, 389, 390
Benjamin, 336
Benoit, 299, 308, 342
Berg, 343, 348
van den Bergh, 223
Berthenson, 298
Bertrand, 56
Beyerman, 372
Bezzola, 250
Bickel, 209
Bidault, 398
Bierman, 207
Bierry, 79, 118, 214
Biester, 263, 265, 271
Bigelow, 242
. Bigland, 367, 377
Bing, 316
Birk, 320
Birkenthal, 315

( # 1056a)
Black, 391
Elaine, 297
Bland-Sutton, 136
Blanton, 348
Blatt, 299 (#954)
Blau, 297

Bloch, C. E., 335, 382
Bloch, 348
Bloombergh, 211
Blosser, 363
Blount, 242
Bluhdorn, 333
Blunt, 397
Bogdanow, 77
Bogen, 335
Bogert, 385
van der Bogert, 386

478

AUTHOR INDEX

Bohme, 330

Bohringer & Sons, 186

Bokorny, 52

Bolten, 334

Bonar, 320

Bonheim, 372

Bonne, 352

Bonniger, 136

Bookman, 314

Borich, 299

Bornmann, 245

Bornstein, 347

Boruttau, 209, 256, 343,
387, 395

Bossert, 333

Botazzi, 342, 343, 398

Bottomley, 65, 66, 67,
68, 69, 70, 71

Bouchard, 351, 365

Boutwell, 221, 222, 223,
224, 239, 254, 258, 265,
268

Bovaird, 371

Bowditch, 312

Box, 351

Boyd, 362, 365, 367, 369

Boye, 138

Boyer, 60

Brachi, 302

Braddon, 29, 210,. 212,

279, 281, 282, 363
Brade-Birks, 314
Brandt, 308
Bravetta, 366
Breaudat, 31
Breuer, 372
Brickman, 260 (#773)
Brill, 52, 172, 196
Broomel, 248
Brown, A., 307, 334
Brown, C. J., 372
Brown, T. B., 369
Brown, W. C., 369, 371
Briining, 114, 122
Bruntz, 378
Brusa, 314
Buckner, 89
Buckuoy, 297

Budd, 297
Budzynski, 372
Buell, 221, 222
Buhrer, 396
Bulkley, 390, 391
Bull, 138, 220
Bulley, 382
Bunge, 21, 22, 26
Bunker, 60
Burckhardt, 316
Burge, 208
Burger, 374, 388
Burnet, 245
Burrows, 82
Byfield, 41, 43, 123, 157,
158, 338, 353

Cahn, 136
Cajori, 269
Caldwell, 257
Calhoun, 361
Calkins, 79. 389
Campbell, H., 386
Campbell, L. H., 133
Campbell, Mabel, 243,

266, 267
Camurri, 364
Cantineau, 372
Cantlie, 370, 371
Carr, 257, 302, 313
Carrel, 81, 82
Carter, 381
Cartin, 373
Caspari, 280
du Castel, 316
Castellani, 277, 369, 370
Cattaneo, 316
Cautley.. 312, 336
Centanni, 389
Cessna, 55
Chalmers, 277, 351, 359,

369
Chamberlain, 32, 207,

211, 268, 281
Chambers, 80
Chantemesse, 279
Chapin, 373

Chapman, 220

Charrin, 78

Chauvin, 378

Cheadle, 301, 302, 307,
324

Chelchowski, 372

Chelmonski, 330

Chevalier, 278

Chi Che Wang, 397

Chick, 131, 155, 159,
233, 236, 238, 243, 245,
246, 256, 257, 258, 259,
264, 266, 267, 269,
270, 301, 367, 398

Chittenden, 25, 67, 140,
341, 342

Christensen, 396

Chun, 283

Clark, 296, 385

Clark, A. B., 383

Clark, G. W., 50

Clark, E., 92, 93, 103,
206

Clementi, 251

Clemm, 393

Clinton, 65

Cohen, 131, 241, 265,
266, 267, 271, 272

Cohendy, 77, 78

Cole, S. W., 59, 262,

270, 271

Cole, W. C. C., 133, 135
Colebrook, 59
Coleman, 381
Collatz, 208
Combe, 381
Comby, 302, 306, 330,

397

Comrie, 300
Conception, 258, 270,

382

Conklin, 260
Conran, 370
Cook 297
Cooper, 33, 98, 167, 187,

206, 209, 210, 212, 228,

229, 237, 255, 262, 266,

271, 281

AUTHOR INDEX

479

'Copeman, 391
Coppin, 70
€oppola, 22
Corin, 278
Cornalba, 396
Corson-White, 389
Courtney, 319, 327, 338
Coutts, 260, 270
Coward, 223, 224, 225,

226, 265, 266, 269, 270,

272

Cowgill, 209
Cox, 294
Cozzolino, 307
Cramer, 229, 230, 389
Croftan, 330
Cronheim, 228, 317
Crosby, 357
Crowell, 105, 280
Curjel, 246
Curran, 262, 298
Currie, 64

Curschmann, 330, 331
Cushny, 209
Cybulski, 332
Czerny, 324, 335, 339,

381

D

Dahle, 258 (#754a)

Dalyell, 158, 159, 238,
301, 398

Damianowich, 208

Daniels, 40, 41, 43, 123,
157, 158, 240, 241, 243,
259, 265, 266, 267, 272,
338

Darling, 296

David, 60

Davis, M., 38, 39, 43,
60, 62, 92, 112, 116,
220, 221, 224, 245, 259,
264, 265, 270, 272, 344,
386

Davis, W. A., 75, 147

Dawson, 362

Dechambre, 297 (#924)

Decks, 352, 357

Deiaco, 357

Delage, 79

Delcourt, 77

Delf, 226, 239, 240, 243,
246, 266, 267, 268

Delille, 311

Delpech, 295, 297

Demoor, 348

Denis, 317

Dennett, 312, 317

Denton, 241, 376

Desgrez, 118

Determann, 376

Devloo, 52

Devoto, 365

Deycke, 385

Dezani, 126

Dibbelt, 316, 317, 319,
320

Dick, 323

Dienert, 60

Digby, 372

Disque", 299

Djenab, 209

Dollner, 342

Doryland, 56

Douglas, M., 105

Douglas, S. R., 59

Doyle, 283

Drescher, 20

van Driel, 397

Driscoll, 251

Drummond, 39, 43, 88,
89, 112, 121, 123, 174,
188, 189, 203, 207, 209,
221, 222, 223, 224, 225,
226, 265, 266, 269, 270,
271, 272, 348, 389, 390,
393

Dubin, 55, 117, 123, 124,
125, 187, 195, 199, 200,
201, 202, 204, 205, 210,
214, 215, 227, 234, 246,
248, 264, 338, 344, 346,
347, 383, 388, 398
Dubois, 278

Dufour, 323
Dufourt, 301
Durand, 391
Dtirck, 291
Durlach, 136 (#400)
Dutcher, 97, 206, 207,
208, 245, 258, 263, 268,

270, 271, 272
Duthoit, 348
Dutta, 388
Dutton, 385
Dyke, 300

E

Eberson, 59
Echols, 366
Eckles, 257, 258 (#754-

a), 368
Eddy, 55, 79, 189, 196,

271, 338, 397, 398
Edelstein, 44, 311, 339
Edie, 32, 186
Editorial: Brit. Med.

J. 298
Editorial: J. A. M. A.

388
Editorial: Trop. Dis.

Bull., 300
Edwards, 363
Ehrlich, 389, 390, 391
Eijkman, 21, 30, 31, 32,

33, 87, 92, 103, 206,

211, 212, 245, 397
Eisenhardt, 209
y Elardi, Pereida, 320
Elebash, 366
Elfer, 331
Elias, 215, 334, 350
Ellis, 30, 257, 292
Emmett, 41, 44, 85, 86,

115, 121, 194, 203, 204,
238, 271, 346, 383, 398
Engel, 314
Engstrand, 133, 135
Enright, 367, 372
Epstein, 302, 376
Erdheim, 307

480

AUTHOR INDEX

Erlacher, 322
Esser, 324
Eustis, 92, 172
Evans, 186
Evvard, 142
Ewald, 206
van Eweyk, 209
Ewing, 266, 389

Faas, 333

Faber, 234, 272

Fahrion, 224

Fales, 319, 327, 338

Falk, 242

Falta, 23, 374, 382

Feig, 299 (#954)

Feigenbaum, 309

Feigl, 375

Feilchenfeld, 380

Feuille", 316

Ferguson, 321, 323

Ferrari, 299

Ferry, 43, 60, 74, 112,

136
Findlay, 109, 140, 278,

323, 359
Fine, 251, 362
Fingerling, 20
Fingerling, A., 141
Fink, 103

Finks, 214, 252, 265, 269
Fisch, 313
Fischer, Emil, 19
Fischer, L., 395
Fish, 302, 308
Fitch, 398
Flack, 58, 92
Flamini, 318
Flather, 79, 80
Fleischmann, F., 147
Fleischmann, L., 315

(#1056a)

Fleming, 59, 296, 385
Fletcher, A., 334
Fletcher, W., 30
Fodor, 255

Folin, 181, 197

Forbes, 147, 397

Fordyce, 302

Forster, 21

Fowler, 235

Fraga, 279, 280

Franchetti, 306

Frangenheim, 314

Frank, A., 337, 382

Frank, L., 320

Frank, M., 308

Franke, 375

Frankel, E., 307, 309

Frankel, S., 133, 205,
390

Fraser, 30, 31, 93, 94,
95, 172, 196, 245, 247,
248, 278, 280, 288, 290

Freedman, 63

Freise, 125, 233, 261,
318,382

Freudenberg, 225, 233

Friedberg, 338

Friedberger, 146

Froehner, 146

Frolich, 27, 128, 129,
131, 134, 137, 144, 231,
232, 233, 235, 241, 259,
264, 295, 302

Fronczak, 348

Fuji, 109

Fujitani, 103

Fuller, 142

Fulmer, 55

Funk, 32, 33, 36, 39, 40,
41, 43, 45, 53, 55, 87,
88, 89, 96, 97, 98, 105,
109, 112, 113, 116, 117,
118, 119, 122, 123, 124,
125, 130, 133, 134,
135, 150, 167, 169, 170,
171, 174, 177, 179, 186,
187, 192, 195, 196, 197,
199, 200, 201, 202, 203,
204, 205, 206, 210, 211,
212, 214, 215, 218, 220,
227, 233, 246, 248, 252,

256, 264, 270, 324, 342,
344, 346, 347, 383, 388,
389, 390, 395, 396, 398

Fiirer, 390

Furst, 41, 246

G

Gaglio, 210
Gams, 172
Garcia, 295
Gardey, 384
Garrison, 352, 366
Gassmann, 317
Gautier, 219
Geiling, 386
Geoghegan, 384
Gerhardt, 387
Gerstenberger, 260, 270,

302, 306
Gessner, 349
Ghostal, 388
Gibson, 258, 270
Gies, 122, 392
Gigon, 341
Gingui, 299
Givens, 129, 234, 239,

240, 241, 242, 256, 263,

266, 267, 268, 271,

355, 362
Gladstone, 339
Goldberger, 45, 353, 357,

365, 366, 368
Goldfarb, 81
Golding, 226
Goldschmidt, 382
Gomez, 397
Gomi, 172
Goodby, 397
Gordon, 58
Gottesmann, 375
Gould, 330
Gouzien, 296
Goy, 205
Grafe, E., 26
Grafe, V., 72
v. Graff, 389
Graham, 392

AUTHOR INDEX

481

Gralka, 333

v. Hansemann, 323

Green, H. H., 102, 137,

Hansen, 23

141, 144, 146, 148, 150,

Harden, 40, 43, 55, 85,

151 (#447), 196, 197,

123, 129, 154, 193, 233,

210, 211, 212, 386

234, 235, 263, 270, 311,

Green, Helen S., 266

398

Greig, 246; 265, 288,

Harlan, 298

289, 296, 373

Harney, 91

Grenet, 297 (#924)

Harries, 353, 368

Grijns, 31, 87, 92, 194,

Harris, 60, 351

238

Harris, H. F., 351

Grimm, 352

Harris, S., 350

v. Groer, 327

Harrow, 398

Gross, 249, 264, 266,

Hart, C., 153, 154

267, 268

Hart, E. B., 89, 91, 142,

Grosser, 320

143, 144, 145, 150, 234,

Grumme, 341

257, 260, 270, 343

Griineberg, 335

Harvier, 299

Guareschi, 248

Haupt, 347

Guerrero, 382

Hausermann, 22, 130

Guidi, 397

Hawk, 343

Guillemin, 372

Hayem, 297 (#924)

Guillemonat, 78

Heaton, 371

Guillerd, 60

Hebrant, 138

Guiral, 383

Hedinger, 149

Gurd, 357

Heft, 55, 189

Guthrie, 333

Hehir, 279, 281, 299

Guyenot, 77, 84

Heim, 130

Guyot, 372

Heisig, 240

Hektoen, 380

H

Helm, 372

Haaland, 389

Henderson, 140

Haas, 397, 398

Henriques, 23, 141

Haberlandt, 65

Henry, 147

Hagenbach, 316

Hepburn, 278, 283, 294

Haguinea, 308

Herdlika, 381

Halberkann, 369

Herter, 324

Hall, 22, 386

Hertz, 299 (#954)

Halliburton, 272, 397,

Hess, 42, 129, 131, 132,

398

137, 158, 225, 234, 235,

Halpin, 20, 89, 91

240, 242, 243, 258, 260,

Hamburger, 384

264, 267, 268, 269, 270,

Hamel, 330, 331

271, 272, 297, 298, 301,

Hamilton, 313, 314, 384

302, 303, 307, 308, 309,

Hammer, 387

310, 311, 314, 319, 321,

Hanan, 330

322, 326, 327, 328, 336,

Hannemann,299 ( #954)

339, 387

Heubner, Otto, 315

( # 1056a)
Heubner, Wolfgang,

136, 137, 257, 302, 316
Hewlett, 155
Heyer, 330
Hiat, 369
Hift, 305
Hill, J. R., 241
Hill, L., 92, 299
Hindhede, 241, 245, 255,

264, 267, 341, 342, 343,

349,350
Hine, 58
Hintze, 342
Hirota, 257, 290
Hirsch, 297
Hoare, 141
Hocson, 33
Hogan, 43, 251
Hoffman, F. L., 388
Hoffmann, G. L., 129
Hoffmann, H., 347
Hofmeister, 175, 176,

177

Hohlfeld, 337
Hojer, 397
Holder, 343
Holm, 207
Holmes, 265
Hoist, 27, 128, 129, 131,

134, 137, 144, 231, 232,

233, 235, 241, 250, 267,

295, 302

Holt, 319, 324, 327, 338
Honeij, 385
Hoobler, 334
Hoogenhuyze, 211, 212
Hopkins, F. G., 24, 25,

26, 37, 38, 113, 115,

116, 125, 223, 224, 256,

324, 326, 389, 395, 397,

398
Hopkins, G. R., 298,

362

Hoppert, 150
Hornemann, 384

482

AUTHOR INDEX

Horschelmann, 299

v. Hosslin, 376

Houlbert, 92

Howard, 133, 154, 308

Howe, P. E., 381

Howe, P. R., 392

Rowland, 314, 320, 334

Hughes, 226, 254

Huldschinsky, 322, 334

Htilse, 373, 374, 375

Hume, 128, 131, 155,
233, 236, 238, 245, 246,
257, 258, 259, 264, 267,
269, 270, 367, 373

Humphrey, 56

Hunt, 386

Hunter, 109, 362, 366

Huntoon, 57

Hutchinson, H. S., 319,
331, 339

Hutchinson, Sir J., 385

Hutinel, 316

Hutyra, 146

Hussy, 395

Huynen, 321

Ibele, 142
Ibrahim, 333
Ide, 55

Igravidez, 279, 281
Imhoff, 330
Ingier, 130
Ingvaldsen, 154
Iscovesco, 218, 228
Isenschmid, 376, 381
Ishiwara, 379
Ismail, 358
Issoglio, 172
Iwata, 115 (#323)

Jackson, 111, 114, 129
Jacob, 23
Jacobs, 396
Jacobsen, 388
Jacobsthal, 374

Jacoby, 397

Jahreiss, 348

Jaksche, 372

v. Jaksch, 299 (#954)

Jansen, 389

Jansen, B. C. P., 103,

207, 208, 269
Jansen, W. H., 342, 373
Januszewska, 329, 333
Japha, 314
v. Jaworski, 389
Jeppson, 333
Jess, 373
Jida, 283
Jizuki, 291
Jobling, 361, 364
Johns, 214, 252, 265, 269
Johnson, 55, 189, 355
Johnston, 95
Jones, 69, 141, 261, 361,

365

Juaristi, 315
Judson, 125
Jiirgens, 372
Juritz, 253

K

Kappel, 331
Karger, 314
Karr, 133, 134, 137, 204,

270
Kassowitz, 313, 316,

320, 322, 323
Kastle, 89

Katakura, 115 (#323)
Kato, 104, 283
Kaumheimer, 317
Kaupe, 348
Kaupp, 91
Kawakami, 146
Keller, 336, 381
Kempster, 90, 91
Kennedy, 39, 222
Kent, 223, 239, 249, 258,

264

Kianizin, 78
Kieffer, 349

Kilbourne,211
Killian,308
Kimura, 103. 104
King. 135
Kirschner, 393
Kitamura, 305
Kleiminger, 366
Klein, 143
Kleinschmidt, 380
Kleissel, 396
af Klercker, 333
Kligler, 62
Klocman, 225, 308
Klose, 333, 373
Klotz, 349
Knack, 372, 373, 374,

376

Knapp, 382
Koch, M., 109, 361
Koch, W., 303, 304, 305,

308, 309
de Kock, 352
Kohlbrugge, 338
Kohler, 53, 80
Kohman, 376, 377
Kolb, 215
Koltonski, 329
Kondo, 172
Kopchen, 330, 331
deKorte, 155 (#459)
Kossel, 19
Kramer, 27, 297, 317,

334

Kraszewski, 342
Kraus, 374
Kraut, 175
Kriwuscha, 255
Kruse, 342
Kubota. 220
Kuczynski, 349
Kuenen,31,238,241l363
Kiilz 103
Kunert, 391
Kurijama, 135
Kurono, 52
Kusama, 32, 153
Kuster, 78

AUTHOR INDEX

483

Labbe, 308

Labor, 307, 308

Lagane, 291

Lake, 119, 135, 137, 245,

254

Lamb, 135, 382
La Mer, 133
Lampe, 25, 37, 134, 206,

211

Lampitt, 53
Lanceraux, 298
Landa, 372
Lander. 117
Lane, Sir Arbuthnot,

379

Lane-Claypon, 256
Lange, 372
de Langen, 293
Langstein, 44, 336
Lasegne, 297 (#924)
Lavinder, 352
Leavenworth, 188
Lecoq, 397
Leede, 371
van Leent, 29
van Leersum, 245, 398
Leggate, 279
Legroux. A., 297 (#924)
Legroux, R., 61, 62
Leitner, 308
Lelean, 365
Lemaire, 397
Leri, 314

Leslie, 260 (#773)
Lessing, 153, 154
Leven, 297 (#924)
Levin, 290
Lewi, 234. 338, 398
Lewis, 133, 134, 197,

269, 361, 362
Lichtenstein, 315
v. Liebig, 245
Liefmann, 334
Lienaux, 321
Liesegang, 329
Lind, 27, 297, 298

Linossier, 65, 397
Linton, 397
Lippmann, 374
Lipschiitz, 136 (#400)
Little, 279, 378
Lloyd, 57, 58, 59
Lobmeyer, 300
Lockeman, 56
Loeb, Jacques, 83, 84
Loeb, Leo, 389
Lombroso, 360, 364, 366
Looser, 302, 330, 331
Lorenz, 360, 366
Lotsch, 146
Loughlin, 40, 41, 43,

123, 157, 158, 259, 265,

266, 272, 338
Lovelace, 279
Low, 351, 370
Lowy, 347
Luce, 292
Luckett, 127
Lucksch, 250
Lumiere, 79, 102, 208
Lund, 79

Lunin, 21, 22, 105
Luros, 41, 44, 115, 194,

203, 238, 271
Lust, 308, 333
Lutz, 64
Luzzatti, 332
Lyle, 342
Lynch, 355

M

Maase, 375

Macalister, 70

Macallum, 39, 40, 43,
88, 111, 112, 113, 117,
118, 119, 123, 181, 197,
203 270, 344, 383

Macaulay, 296, 385

Macauley, 146

Macfie. 383

Mackay, 321

Macomber, 379

MacArthur, 127

MacDonald, 55
MacLachlan, 334
MacNab, 97
MacNeal. 366
MacPherson, 297
Madsen, 386
Magendie, 245
Maggesi, 333
Magnus-Levy, 387
Maignon, 73, 74, 214,

346

Maliwa, 372
Mangkoewinoto, 207
Mann, 324, 372
Maranon, 294
Marek, 146
Marfan, 316, 339
Marie, 351, 361, 362
Marriott, 320, 334, 337
Martinez, 279
Mason, 347, 381
Massalongo, 208, 279
Massaneck, 320
Masslow, 137
Masucci. 60
Matsunaga, 171
Matsuyama, 115 ( #323)
Mattei, 248, 270
Mattill, 122, 261, 339
Maurer, 210
Maver, 373
Maxwell, 361
Mayer, 339
Maynard, 375
McCarrison, 105 106,

107, 108, 109, 133, 153,

207, 216, 217, 227, 377,

379, 381, 388, 398
McCann, 321, 327, 328
McCaskey, 342
McCay, 373, 388
McClanahan, 314
McClendon, 133, 135,

234, 270, 338, 392, 397
McClugage, 234, 239,

240, 242, 256, 263, 267,

268, 271

484

AUTHOR INDEX

McClurg, 240, 241, 243,

265, 267
McCollum, 20, 23, 34,

38, 39, 40, 43, 44, 55,
91, 92, 110, 112, 113,
116, 117, 122, 123, 129,

130, 131, 134, 142, 143,
144, 145, 193, 195,, 202,
203.220,221,223,224,
239,245,249,251,253,
255, 257, 259, 264, 265,

266, 267, 270, 272, 327,
328, 342, 344, 380, 382,
396, 397

McCrae, 253

McDonald, 352

McGuire, 242

McKay, 392

McKim, 44

McLean, 228, 268

McLeod, 372

Mead, 258 (#754a)

Medes, 392

Medical & Surgical His-
tory of the War of the
Rebellion, 297 (#923)

Meier, 151 (#447)

Melka, 299 (#954)

Mellanby, E., 42, 138,
139, 324, 325, 326, 328,
331, 380

Mellanby, M., 380, 392

Melocchi, 395

Meltzer, 340

Mendel, 37, 38, 39, 40,
41, 43, 44, 89, 90, 113,
114, 115, 116, 117, 119,
121, 122, 123, 125, 127,

131, 195,210,218,221,
224,227,237,249,251,
252, 255, 257, 258, 262,
264, 2a5, 266, 267, 268,
269, 270, 271, 272, 346.
348, 382, 383, 393, 395

Mengert, 322
Mercier, 378
v. Mering, 137

Merk, 357
Merklen, 102
Me>y, 313
Mesnard, 62
Metchnikoff, Mme., 77
Meyer, A., 302
Meyer, Kurt, 320
Meyer, L. F.; 319, 336
v. Meysenbug, 135, 333,

334

Michael, 371
Michel, 135, 243, 271
Middlekauff, 133, 135
Miller, 258, 306
Miller, E. G., 392
Miller, E. M., 320
Miller, E. W., 238, 241
Miller, R., 28
Miller, R. S., 358
Miller, W.S., 144, 145
Mitchell, 41, 127, 150,

203
Miura, 220, 225, 235,

278, 283, 284, 285, 286,

328

Mockeridge, 70, 71
Modinos, 361
Mohr, 316
Molinari, 398
Moll, 308
Momm, 349
Monrad, 382
Montagnani, 130, 132
Moody, 129
Moore, 129, 186, 258
v. Moraczewski, 388
Morawitz, 299 (#954)
Morgen, 134
Morgulis, 85, 122
Mori, 381
Morini, 59
Moro, 77, 338
Morpurgo 125
Morrison, 147, 381
Morse, 301, 337, 361
Mosc, 396
Moszkowski, 280

Mott, 292, 358, 359

Moufang, 52

Mourges, 219

Mouriquand, 78, 92,
104, 135, 195, 212, 239,
243, 245, 246, 248, 254,
264, 271, 272, 291, 299,
338, 396, 397

Muckenfuss, 210

Mueller, 62, 344

Muhlens, 370

Muller, 223

Muller, 298

Muller, Erich, 301, 317

Muller, W., 314

Mulvany,278

Munk, 245

Murai, 172

Murlin,241,361

Murray, 370

Muthu, 385

Myers, C. N., 96, 97,
191, 196, 209, 229, 245,
246, 249, 254

Myers, Victor, 362

Murphy, T. J., 392

Murphy, W. A., 353

Murphy, 390

N

Nagamatsu, 360
Nagasawa, 271 (#836)
Naish, 260
Nambu, 297, 303
Nansen, 262
Nathan, 328
Naumwerk, 307, 387
Neill, 208, 366
Nelson, B. A., 127
Nelson, E. M., 222
Nelson, V. E., 55, 135,

257, 382
Nemoto, 283
Neppi, 398
Nepreux, 308
Netter, 301
Neuhaus, 349

AUTHOR INDEX

485

Neumann, 257, 302, 315
J#1056a), 342, 374

v. Neusser, 250

Neville, 38

Nichols, 265

Nicholls, 352

Nicolaidi, 362

Nicolet, 197

v. Niedner, 303

Nightingale, 363

Niles, 351, 353

Nilsson, 349

Nitzesco, 251

Nixon, 377

Nobecourt, 348

Nocht, 295

Noe, 153

Noeggerath, 23, 335,
337, 338, 382

Noguchi, 56

Nollau, 89

v. Noorden, 387

Norris, 63

Northrop, 83, 84

Nutall, 78

O

Ochsner, 260 (#773)

Odake, 171, 172

Ogata, 291, 329

Ohler, 92, 250

Okimoto, 115 (#323),
271 (#836)

Okuda, 115 (#323), 271
(#836)

Ono, 293

Oppermann, 308

Orenstein, 296

Orgler, 317, 319

Osborne, 37, 38, 39, 40,
41, 43, 44, 74, 89, 90,
112, 113, 114, 115, 116,
117, 119, 120, 221, 122,
123, 125, 127, 136, 188,
190, 195, 210, 218, 221,
224, 249, 251, 252, 255,

257, 258, 262, 264, 265,
266, 267, 268, 269, 270,
271, 272, 346, 348, 382,
393, 395

Oseki, 245, 264, 265, 266

O'Shea, 305

Ostheimer, 313

Otabe, 60

Ottow, 103, 196, 244

Paal, 220

Pacini, 57

Padua, 393

Paget, 311

Pagniez, 293

Paguchi, 104

Palmer, 89, 90, 91, 222,

257, 368
Paltauf, 374
Pappenheimer, 321, 327,

328
Park, E. A., 314, 323,

328

Park, F. S., 376
Parker, 279, 382
Parsons, 44, 73, 134,

255, 263, 265, 267, 271,

272, 327, 342, 380
Partsch, 330
Parturier, 313
Pasqualis, 22
Pasteur, 51
Patel, 331
Paton, 140, 323, 325
Patterson, 372
Paul, 252, 265, 269
Paus, 314
Pearce, 386
Peckam, 387
Pehu, 337
Peiser, 318, 380
Pepper, 332, 386
P6rronet, 246
Peters, 80
Petersen, 364
Peterson, 154

Petrone, 333

Pfaundler, 158, 159,348

Pfliiger, 136

Phemister, 320, 323

Pick, 380

Pickard, 269

Pickens, 298

Pieper, 56

Pierson, 263, 268, 271

Pilado, 208

Pinault, 352

Pincherle, 333

Pitz, 110, 129, 130, 221,

249, 253, 257, 262,

264, 266
Place, 148
Plimmer, 144
Pol, 32, 95, 192, 243,

245, 265, 266, 396, 398
Pommer, 316, 324
Poole, 62
Poppe, 253
Portier, 79, 134, 210,

214, 398
Potter, 373

Poynton, 301, 302, 307
Praussnitz, 342
Prescott, 241
Pringle, 389
Pringsheim, H. H., 52
Prinzing, 372
Pritchard, 260, 324
Pugliese, 245, 397

Q

Queal, 398

R

Raczynski, 322

Rae, 298

Rahe, 389

Ramoino, 207, 299, 398

Ramond, 277

Ramsden, 396, 398

Ranc, 79

Randoin, 79, 134, 210

Ranwez, 237

486

AUTHOR INDEX

Ray, 242, 266, 267, 268,

388

Rayer, 366
Reach, 376, 386
Reakes, 147
v. Recklinghausen, 315
Reed, 279
Reid, 142
Reimers, 138
R6non, 385
Report of American

Pediatric Society,

392 (#974)
Report of Eng. Med.

Res. Comm., 118, 175,

237
Report of Food Comm.

of Royal Society, 298
Report of Health of

City of Manchester,

298
Report to Local Govt.

Board, 260
Reprint U. S. Public

Health Service, 260
Retterer, 313
Rettger, 56
Reynolds, 379
Rheindorf, 303
Rhodehamel, 189
Rhodes, 267, 269
Ribbert, 316
Richardson, 266
Richter, 304
Richet, 77 (#200), 384
Ricklin, 320
Riddel, 279, 281
Ridlon, 361, 366
Rietschel, 335, 336
Rimbaud, 279
Risquez, 397
Rivers, 62
Roaf, 367
Robb, 327, 392
Roberts, 145, 351, 352,

353, 356, 360

Robertson, 50, 84, 125,

228

Robinson, H. C., 56
Robinson, R. K., 85
Robison, 234
Roger, 279, 283
Rogers, 220
Rohl, 126
Rohmann, 25, 26, 37,

125, 126, 127, 245
Rohmer, 333
Roily, 308
Roloff, 138
Rolph, 352
Rommel, 145
Rondoni, 130, 132, 133

251, 390
R0nne, 382
Roper, 338
Rose, 255, 387
Rose, 255, 343
tel Rosario, 294
Rosemann, 136
Rosenfeld, 348
Rosenheim, 68, 71, 222
Rosenthal, 347
Rosin, 307
Rossi, 239
Rost, 317
Roth, 220
Rothberg, 319, 320
Rous, 389
Rouse, 245
Roussel, 351, 352, 366
Roxas, 291
Rubner, 52, 245, 255,

341, 342, 376
Rueck, 311
Ruh, 260 (#773)
Rumpel, 350, 372, 373,

375

Rumpf , 292
Rupp, 57
Rupprecht, 318
Russel, D. W., 57
Russel, H. L., 147

Rustin, 384
Rutgers, 343

S

Saito, 52
Salant, 386
Saleeby, 172,206,294
Salge, 336, 339
Sambon, 351
Samelson, 338
Sandy, 358
v.Samson-

Himmelstiern, 30
Saneyoshi, 264
Sato, 297, 303
Satta, 125

Sauer, 314, 330, 333, 339
Saxl, 299 (#954)
Saxon, 389
Sazerac, 60
Schabad, 28, 266, 270,

314, 317, 318, 319, 320,

324

Schaefer, 258 (#754a)
Schaeffer, 78, 395, 396,

397

Schattke, 146
Schaumann, 20, 32, 36,

51, 53, 54, 109, 127,

134, 135, 137, 141, 153,

155, 170, 172, 189, 196,

198, 207, 208, 256, 270,

324, 395
Schelenz, 297
Scherer, 295. 300, 303,

305, 309, 333
Scheunert, 146
Schiff, 372, 373, 376
Schilling, 277, 279, 351,
. 369

Schippers, 339
Schittenhelm, 373, 374,

375, 376, 377, 388
Schlapfer, 26 (#30)
Schlee, 314
Schlecht. 373, 374, 375,

377,388

AUTHOR INDEX

487

Schloss, 318, 320
Schmidt, 125
Schmitter, 370
Schmorl, 137, 313, 316
Schneider, 299 (#954)
Schnyder, 104
Schodel, 307, 387
v. Schonborn, 109, 215
Schottelius, 77, 78
Schreiber, 172, 189
Schreiner, 70
Schroder, 297
Schiiffner, 31, 238, 241,

279, 363
Schulhof, 311
Schumm, 308
Schut, 293
Schwartz, E., 205
Schwartz, H., 313
Schwarz, 318
Scipiades, 329
Scott, Agnes C., 331
Scott, E. L., 215
Scott, L. C., 172
Seaman, 206
Sedgwick, 338
Seefelder, 350
Segawa, 94, 95
Seidell,40,163,188,189,

193, 196, 398
Sekine, 260, 270, 272
Sell, 221, 222
Sharp, 234
Sharpe, 140, 334
Shearer, 58
van der Sheer, 369
Sheppard, 362
Sherman, 245, 264, 321,

327, 342, 366, 398
Sherwood, 55
Shiga, 32, 153
Shimamura, 171, 172
Shipley, 328
Shizume, 104
Shorten, 242, 266, 267,

268
Sicard, 279, 283

Siegert, 324

Siler, 352, 355, 366

Simon, 370

Simmonds, 44, 110, 113,
130, 193, 195, 221, 223,
239, 249, 251, 253, 257,
264, 266, 267, 327, 328,
342, 380

Simonini, 317

Simpson, 32, 186, 293

Simpson, F. R., 334

Simpson, K., 278

Sinclair, 313, 392

Singer, 350

Sion, 360

Sittler, 302

Skelton, 243, 257, 258,
259, 267, 269

Skinner, 70

Slonacker, 114

Slye, 389

Van Slyke, 186

Smith, Alice, H., 233

Smith, C. A., 343

Smith, C. H., 279, 281

Smith, D. W., 260, 270

Smith, H., 379

Smith, M. L., 133

Smith, W. J., 309

Smith, Theobald, 128

Snyder, 353

Socin, 22, 125

Sommer, 234

Sorochowetz, 266, 270

de Souza, 202, 203, 344

Sparkes, 353

Speroni, 398

Sperry, 56

Speyer, 300, 303

Spiegel, 334

Spillman, 378

Spitzer, 257

Sprawson, 279

Spriggs, 73

Spruell, 150

Stammers, 225

Stannus, 362

Stanton, H. T., 30, 31,
93, 94, 95, 172, 196,
245, 247, 248, 280

Stanton, R. E., 361, 362

Stark, 338

Stead, 69, 150, 151

Steenbock, 89, 91, 143,
150, 193, 221, 222, 223,
224, 239, 249, 254, 257,
260, 264, 265, 266, 267,
268, 270, 271, 343, 396

StSfansson, 262

Steinitz, 336, 337

Stephenson, 222, 383

Stepp, 20, 21, 36, 96,
115, 117, 125, 126, 229,
396

Sternberg, 395

Stetter, 314

Stevenson, 301

Stevenson, H. C., 55,
189, 198

Stewart 146

Stewart, C. A., 114

Stewart, C. E., 371

Stheehman, 332, 333,
334

Sticker, 372

Stiles, 395

Still, 154, 307

Stille, 342

Stilling, 137

Stockholm, 204

Stocklasa, 245

Stolte, 337

Stolzner, 316, 324, 384

Strauss, 372

Strong, 280

Strongman, 312

Strudwick, 239

Stuart, 189

Stuessy, 259

Sturtevant, 85, 369

Suarez, 251

Sugiura, 44, 125; 190,
195, 267, 269, 389

488

AUTHOR INDEX

Sullivan, 192, 229, 254,
264, 265, 271, 273, 361,
362, 365 '

Supplee, 40, 258

Sure, 115

Sutton, 314

Suzuki, T., 293, 317

Suzuki, IL, 115, 171,
172, 220, 264, 271, 272

Swanson, 383

Sweet, 389

Swett, 257, 368

Swoboda, 271

Sydenstricker, 365

Sylvester, 366

Szalagyi, 251

Sztark, 382

Tachau, 346, 376
Tada, 319, 320
Takaki, 29
Takasu, 333
Talbot, 154, 317, 337
Tanaka, 175
Tanner, 366
Tasawa, 95
Taussig, 298, 303
Taylor, 342
Telfer, 319
Teruuchi, 32
Theiler, 102, 137, 141,

144, 146, 148, 150, 151,

210, 211

Thiemich, 333, 334
Thierfelder, 78
Thin, 369
Thjotta, 62
Thomas, E., 380, 384
Thomas, K., 20, 343
Thompson, 32, 293
Thompson, Helen B.,

125

Thorns, 348
Thurlow, 52
Tisdall, 334
Tixier, 316

Tobler, 301, 320
Todd, 154

duToit, 151 (#447)
Tonin, 372
Torrey, 129
Towles, 192,271
Toyama, 103
Tozer, 128, 136, 239
Trappe, 136
Travis, 279
Tricoire, 379
Trop. Dis. Bull., 299

(#941)

Tschirch, 398
Tsujimoto, 220
Tsuli, 122
Tsuzuki, 33
Tuchler, 299 (#954)
Tullio, 362
Turk, 328
Turner, 299
Tuttle, 352
Turban, 26 (#30)

U

Uhlmann, 209

Underbill, 25, 140, 385

Unger, 131, 132, 225,
234, 235, 240, 242, 243,
258, 260, 267, 268, 269,
270, 271, 272, 302, 307,
310, 311, 314, 321, 322,
326,328

Urbeanu, 251

Urizio, 305

Utheim, 339

Vacher, 373
Valagussa, 380
Vallery-Radot, 293
Vandervelde, 372
Vansteenberge, 52
Vanutelli, 299
Vedder, 32, 56, 92, 93,
103, 145, 172, 206, 207,

212, 213, 234, 268, 277,
293, 295, 396

Viljoen, 102, 137, 141,
144, 146, 148, 150, 151
(#447), 210, 211

Viswallingam, 362

Vitale, 333

Vlahuta, 52

Voegtlin, 96, 97, 109,
119, 135, 137, 189, 191,
192, 193, 196, 209, 229,
245, 246, 249, 254, 264,
265, 271, 272, 353, 361,
366, 368, 396

Voit, 341, 342

Vokurka, 385

Volkmann, 315 ( # 1056-
a)

Vonderweidt, 333

Vordermann, 29

W

Wakeman, 39, 43, 74,
112, 136, 190

Walker, 150

Wallis, 57, 265, 379

Walshe, 211, 291

Walton, 82, 381

Ward, 109

Waring, 366

Washburn, 261

Wason, 383

Wassermann, 303, 308

Waterman, 373

Watson, 23, 225, 323,
325

Watson-Wemyss, 395

Webster, 186

Wechuizen, 293

Wegele, 370

Weigert, 336, 337, 384

Weill, 78, 92, 104, 135,
195, 212, 239, 244, 245,
246, 248, 254, 264, 271,
272, 291, 301, 338, 396,
397

Weiss, 314, 330, 351, 363

AUTHOR INDEX

489

Wellman, 92, 172, 353
Wells, 266, 372, 392
Werner, 371
Wernich, 29
Weston, 353, 373
Wheeler, 127, 265, 271,

365,372

Whipple.238,241,386
White, 189, 193
Wieland, 109, 313
van der Wielen, 270
Wilcock, 24
Wild, 330
Wilder, 388
Wildiers, 51, 52
Willaman, 64, 65
Willard, 314
Willcox, 262, 271, 279,

294, 299
Willets, 366 •
Williams, R. J., 53, 198,

199, 201, 202

Williams, R. R., 95, 105,

Y

172, 188, 193, 194, 206,

Yamada, 283

207, 268, 293, 294

Yano, 283

Wilson, 361, 365, 367

Yellowlees, 351

Wiltshire, 266, 303, 311

Yoshikawa, 283

Winfield, 260, 270

Winton, 245

Z

Wintz, 26 (#30)

Zak, 305, 375

Wise, 248

Zeller, 356

Wolcott, 279

Zernik, 350

Wolf, 60

Zilva, 40, 43, 55, 85, 123,

de Wolf, 373

129, 146, 154, 193, 195,

Wolff, 397

220, 221, 224, 225, 226,

Wollmann, 77
Woltmann, 375

233, 234, 235, 263, 267,
270, 307, 311, 328, 380,
392

Wood, 353, 357, 363, 369

Zlocisti, 303

Woodcock, 384

Zondek, 373, 375

Woods, C. D., 252

Zucker, 321

Woods, E., 245

Zuntz, 347, 349

Wrampelmeyer, 40

Zunz, 331

SUBJECT INDEX

Adenine, attempt to synthesize

vitamine B from, 193
Adenoids, possible dietetic origin of,

386
Adrenaline, in beriberi, increase of,

133

in scurvy, decrease of, 133
Adrenals, in scurvy, enlargement of,

133
Adsorption of vitamine B from yeast

with colloidal arsenic sulfide,

189
of vitamine B from yeast with

fullers earth, 189
of vitamine B from yeast with

mastic, 189
of vitamine B from yeast with

norit, 62, 63

Allantoine, in rice polishings, 170
Aminoids (casein preparation),

growth substance in, 63
Amphibia, vitamine requirements of,

85
Anemia, influence of vitamines on,

386
Animal experiments under sterile

conditions, 76, 77, 78, 79
Antiberiberi vitamine (see vitamine

B)
Antirachitic vitamine (see Vitamine

A)
Antiscorbutic vitamine (see Vitamine

C)
Arthritis deformans, occurrence of,

392

Artificial diet, inadequacy of, 75
"Aschamine," from yeast, 190
Ash-free diet, experiments with,

21, 22

Atrophy, symptoms of, 336, 337, 339
therapy of, 337, 338

"Auximones," similarity of, to

vitamines, 67, 68
Avitaminoses, general discussion and

classification of, 15, 16, 17, 18
Avitaminosis, in cattle, 75, 76
in horses, 75, 76

B

B. coli, growth of, 60

B. diphtheriae, growth of, 60, 61, 62

B. histolyticus, growth of, 60

B. influenzae (B. Pfeiffer), growth

, of, 60, 61, 62
B. pertussis, growth of, 62
B. proteus, growth of, 60
B. sporogenes, growth of, 60
B. welchii, growth of, 60
B. xylinum, growth of, on yeast

extract, 56
Bacteria, certain types of, vitamine-

containing medium for, 57
probable function of, in intestinal

canal, 76, 78

synthesis of vitamine by, 57
vitamines in, demonstration of,

57
Bacterized peat, effect of, on growth

of nitrifying bacteria of the soil

(B. radicicola and Azotobacter

chroococcum) 70

effect on growth of plants of

extract of, 65, 66, 67
Banana fly, growth of, under sterile

conditions, 83

Barley, nutritive value of, 248, 249
Barlow's disease, 309 (see Infantile

scurvy)
Beriberi, effect of pilocarpine on,

206

effect of pituitary extract on, 206
effect of purine derivatives on,

206

491

492

SUBJECT INDEX

Beriberi, effect of pyrimidine deriv-
atives on, 206
effect of quinine on, 206
effect of strychnine on, 206
effect of thyroid extract on, 206
first clear conception as to cause of

development of, 31
human, acute pernicious or cardio-
vascular form of, 289
human, chemical pathology of,

293

human, dry atrophic form of, 287
human, general pathology of, 291,

292

human, general therapy of, 293
human, infantile, 290
human, infantile, therapy of, 294
human, mode of occurrence of and

diets leading to, 280, 281, 282
human, mortality in, 289
human, prevalence of, 278, 279
human, relationship of, to rice con-
sumption, 29, 30, 31, 32, 33
human, relationship of, to scurvy,

295

human, sensory-motor form of, 283
human, symptomatology of, 228,

283

in birds, blood sugar in, 109
in birds, changes in the gastro-
intestinal tract, 107, 108
in birds, chemical pathology of,

109
in birds, glycogen content of liver

in, 109
in birds, increase of adrenaline in,

133

in cats, 135
in chickens, 92, 93, 94
in dogs, 137
in pigeons, 97, 98
in pigeons, endocrine glands in,

105, 106, 215

in pigeons, glucose in, 215
in pigeons, glucosuria in, 215, 217
in pigeons, heart in, 104
in pigeons, muscles in, 104

in pigeons, nervous system in, 103,

104
in pigeons, pathological anatomy

of, 103

in pigeons, phlorizin in, 215
in pigeons, time of development

of, 99
in pigeons, types of, 99, 100, 101,

102
mention of. in Neiching (oldest

medical book 2697 B. C.) 278
nature of substance curative for,

31, 32, 33
original conception of cause and

prevention of, 30, 31
starvation, 211

"Bios," characteristics of, 51, 52
effect of, on growth of yeast, 55
nature of, controversy as to, 52
similarity of, to vitamine B, 51,

52
Bow legs, possible dietetic origin of,

387

BreadC "Soldiers' bread"), 245
Butter, centrifugation of, 38

C
Calculi, formation of, influence of

vitamines on, 393
Cancer, influence of vitamines on,

388, 389, 390, 391
possibility of chemical substance

as etiological factor in, 388
Carbohydrate dystrophy, atrophic-

hydremic form of, 335
etiology of, 336
hypertonic form, of, 335
pure atrophic form of, 335
occurrence of, in artificially-fed

babies, 335

Carcinomas, supposed acceleration of
growth of, by underfeeding,
389

Carrel's antiseptic method of wound
treatment, possible use of vita-
mines as nutrient in, 82
tissue cultures in vitro, 81, 82

SUBJECT INDEX

493

Carrotinoids, effect of absence of,

90, 91, 222, 223

supposed relationship with vita-
mine A, 90
Casein, in hunger edema, supposed

curative action of, 376, 377
nutritive value of, 43, 44
nutritive value of, on autoclaving,

344

Cattle, "bush-sickness" in, 147
diseases in, theory of poverty of

soil as cause of, 147
"impaction paralysis" in, 147
"lamziekte" in, 146, 147, 148, 149,

150, 151

"stallmangel" in, 146
"stijfziekte" in, 146, 147, 148
Cats, beriberi in, 135

experimental rickets in, inability

to produce, 136
Cereals, baking of, 246
milling of, 245

nutritive value and vitamine con-
tent of, under various methods
of preparation, 244
Chicken beriberi, 92
sarcoma, effect of vitamines on,

390
Chickens, growth of, importance of

vitamines A and B for, 92
"leg-weakness" in, 89
normal nutrition of, 87, 88, 89,

90, 91, 92
ophthalmia in, 87
polyneuritis in, 87
"Cholesterol gland" (see "Lipoid

gland")

Choline in rice-polishings, 170
Cod liver oil, chemistry of, 218
fractionation of, 219, 220
reduction of, 220
specificity of, for rickets, 328
"spinacen" from, 220
"squalen" from, 220
use of for chemical isolation of
vitamine A, 218

Colpidium colpoda, growth of, in-
fluence of vitamine extracts on,
80
Cooking water, vitamine content of,

241
Corn, as cause of "emmaisadura,"

250
consumption of, in relation to

pellagra, 250, 251
dried, absence of vitamine C in,

254
feeding of, as cause of scurvy and

beriberi, 250, 251
nutritive value of, 250
vitamine content of, as affected by

milling, 252, 253
Coxa vara, in scurvy, 387

D

Desmotopa, growth of, 84
Diabetes, influence of vitamines on,

387

limitation of diet in, possible pro-
duction of avitaminosis by,
388

Diet, artificial, inadequacy of, 75
ash-free, experiments with, 21, 22
influence of, on growth of neo-
plasms, 389
weekly, in Denmark, during the

War, 349
weekly, in Dresden, during the

War, 348

Dietary composition in Italian pella-
gra districts, 364
constituents, necessity for proper

relationship between, 340
Dogs, beriberi in, 137
osteoporosis in, 140
pellagra-like condition in, 140
rickets in, 137, 138
scurvy in, 137

scurvy in, symptoms of, 138, 139, 140
Dropsy, (see Epidemic dropsy)
Drosophila, growth of, on solution
of cane sugar and salts, 83, 84

494

SUBJECT INDEX

Drosophila, ampelophila, growth of,

84
melanogaster, growth of, 84

E
Egg production, factors influencing,

91
"Emmaisadura," rotted corn as

cause of, 250
Epidemic dropsy, characteristics of,

288, (see 373)
Essential food factors, opposition to,

existence of, 25
Exophthalmic goitre, as suspected

avitaminosis, 380
Exudative diathesis, 337, 339

Ferments, comparison between

vitamines and, 163
Fish, growth of, 85
Flat feet, possible dietetic origin of,

387
Flies, development of, part played by

vitamine B in, 84
Fly, (see Drosophila, Musca domes-

tica, Desmotopa, Sciaria, Ty-

roglypha)

Foodstuffs, non-protective and pro-
tective against rickets, 325
table showing vitamine content of,

264

Fragilitas ossium, in rickets, 313
Frogs, growth of, influence of vita-
mines on, 85, 86, 87
Fruit juices, antiscorbutic properties

of, erroneously ascribed to

laxative action, 129
Fullers earth, use of, in adsorption

of vitamine B from autolyzed

yeast, 188
Fungi, vitamine requirements of,

64, 65

Aspergillus niger, 64
Oidium lactis, 65
Sclerotinia cinera [(Bon) Schroter],

64

G

Gal — lamziekte (See Lamziekte)
Goats, vitamine requirements of,

141

stijfziekte and lamziekte in, 148
Gonococcus, growth of, 59
"Gossypol," toxicity of, in pigs,

145
Graves disease (see Exophthalmic

goitre)
Guinea pigs, vitamine requirements

of, 127, 128
use of, in study of scurvy, 127

H

"Helles Mark," appearance of, in

scurvy, 132, 309

Hermeralopia, as suspected avi-
taminosis, 379

Hemophilic bacteria, factors neces-
sary for growth of, similarity to
those required for growth of rats,
60

Higher plants, growth of, extract
of bacterized peat in, 67, 68, 69
"Hikan" (see Ophthalmia)
Horses, edema in, 146

"sukumi" or "gokosukumi" in,

146
Hunger edema, casein as supposed

curative for, in rats, 376, 377
differentiation of, from beriberi

and scurvy, 377
development of, r61e of adrenaline

in, 377

etiology of, 376
in rats, production of, 376
metabolism in, 375
mode of development of, 374
pathology of, 374
prevalence of, 372
relationship of, to epidemic dropsy,

373

symptomatology of, 373
therapy of, 375

SUBJECT INDEX

495

Insects, 83

Intestinal stasis, as suspected avi-
taminosis 379

K

"Kaffir," as protective against

scurvy 300
Kallak, as suspected avitaminosis,

378
Keratomalacia (see Ophthalmia)

Lactalbumin, nutritive value of,

43, 44, 115

Lactic acid bacteria, growth of, in-
fluence of autolyzed yeast on,
52, 53,

growth of, inhibition of, 53
Lactose, supposed antiscorbutic ac-
tion, of 129

•Lamziekte, etiology of, 150, 151
in cattle and goats, 146, 147, 148,

149

pathology of, 149
six phases of, 151
symptoms of, 149
vitamine B not a causative factor

in, 150, 151

Laxatives, supposed antiscorbutic ac-
tion of, 129
Lecithin, action of, due to vitamine

impurity, 81

influence of, on growth, 81
in nutrition, supposed influence of,

24
supposed curative action of, on

beriberi, 52

"Lecksucht," in reindeer, 142
Lemna minor, growth of, influence
of crude nucleic acid deriva-
tives on, 69
Leprosy, as suspected avitaminosis,

385
"Leptom," function of, in plant

nutrition, 65
Life without bacteria, 76, 77, 78, 79

Lions, rickets in, 136

"Lipoid gland," in development of

avitaminoses, relation of, 229,

230
Lipoid group, classification of those

substances essential to life in, 21
Lipoids, in the body, synthesis of,

20

M

Maize (see Corn)

Man, vitamine requirements of, 155,

156, 157
"Maniok," consumption of, as cause

of beriberi, 278
Mammals, importance of vitamines

for, 109

Marasmus, occurrence of, 337
"Marmite" (yeast extract) 154, 294
Meat, significance of, as dietary com-
ponent, 262

"Mehlnahrschaden," 122, 335 (see
Carbohydrate dystrophy and
Atrophy)
Meningococcus, conditions for

growth of, 57, 58, 59
probability of necessity of two

vitamines for growth of, 58
Metazoa, growth of, 81
Mice, vitamine requirements of, 125,

126, 127
Milk, in nutrition, importance of,

256

nutritive value of, 261, 262
vitamine content of, 258
vitamine content of, influence of

drying on, 260
vitamine content of, influence of

evaporation on, 260
vitamine content of, influence of

heat on, 259

vitamine content and composition
of, influence of vitamine content
of fodder on, 256, 257, 258
vitamines in, demonstration of,
256, 257

496

SUBJECT INDEX

Milk, synthetic, 260
Milk dystrophy, 337
Milk powder, ability of mice to thrive

on, 22

Moller-Barlow's disease, 22 (see In-
fantile scurvy)
Monkeys, beriberi in, 153

pellagra-like disease in, 155

routine feeding of, 155

suitability of, for study of scurvy,

153, 154

vitamine requirements of, 153,

154, 155

N

"Neiching" (oldest medical book,
2697 B. C.), mention of beriberi
in, 278
Neoplasms, growth of, influence of

diet on, 389, 390, 391
effect of pregnancy on, 389
Nicotinic acid, attempted synthesis

of vitamine B from, 193
in rice polishings, 171
Nucleic acid derivatives, supposed
influence on growth of plants, 70,
71
Nucleins, in the body, synthesis of,

20
Nutrition in Europe during the war,

347, 348, 349
in man, 340

on high fat diet, 338, 339, 346, 347
on high protein diet, 346, 347
on high starch diet, 346, 347
on high sugar diet, 346, 347
Nutritional requirements, 73
Bunge's statement, 22
Hopkins' prophetic view, 24

O

Oats, nutritive value of, 249
"Oocytin," growth substance from

ox-blood, 50
Ophthalmia, absence of in Germany

despite lack of vitamine A, 349
as suspected avitaminosis, 381, 382

cure of, by yeast, 117

cure of, by zinc sulfate and boric
acid wash, 117

in chickens, 87

in rats, 117

on diet containing butter, 117
"Opsin," in growth of microorgan-
isms, 56

'Orypan," in rice polishings, 172
"Oryzanin," in rice polishings 173
Osteogenesis imperfecta, relation to

congenital rickets, 313
Osteomalacia, etiology of, 331

metabolism in, 331

pathology of, 330

prevalence of, 329

symptoms of, 330

therapy of, 331
Osteoporosis, in dogs, 140

relation to rickets, 313, 315, 329

Parabiosis, in mice, 125, 126
Paramecium aurelia, growth of, effect
of vitamine extract (pancreas) on,
79

Pellagra, absence of, in Central
Europe despite small amount of
animal protein available, 350

association of, with the diet, 363,
365, 366, 367

chemical pathology of, 361

condition in dogs similar to, 140

etiological factors in, 368

etiology of, 366

etiology of, in relation to corn con-
sumption, 250

experimental, production of, 365

infantile, 368

in monkey, supposed production
of, 367

lack of protein as cause of, unlike-
lihood of, 347

metabolism in, 362

mode of development of, 363

occurrence of, ratio in men and
women, 352

SUBJECT INDEX

497

Pellagra, prevalence of, 351, 352
prognosis, 362
progress of, 352, 353
relationship of, to beriberi and

scurvy, 362, 363
study of, importance of pre-pella-

grous diet in, 365'
symptomatology and pathology of,
355

bones, 360

circulatory system, 359, 360
endocrine glands, 361
eyes, 361

gastro-intestinal tract, 355
nervous system, 357
"pellagra glove," 357
"pellagra shoe," 357
sexual organs, 360
skin, 357
therapy of, 366
types of, 353
acute, malignant, 354
chronic, 354
light, sub-chronic, 354
severe, cachectic, sub-chronic,

354

Phosphorus insufficiency, theory of,
as explanation of beriberi and
similar diseases, 20
"Phyllanthus emblica," as cure of

scurvy, 297
Phytin, in beriberi, supposed value

of, 33

Pica, cure of, 152
etiology of, 152

in cattle and goats, 147, 148, 151
in ostriches and poultry, 151
Pigs, growth of, influence of alfalfa

grass on, 145

growth of, influence of corn on, 143
growth of, influence of wheat on,

143
nutritive requirements of, 142, 143,

144, 145

susceptibility of, to development of
experimental rickets, 146

vitamine requirements of, uncer-
tainty as to, 146
Pigeons, beriberi in, 97, 98
beriberi in, time of development of,

99
beriberi in, types of, 99, 100, 101,

102
nutritional requirements of, 95,

96, 97
Plant extracts, growth promoting

effect of, 65

Pneumonia, influence of diet on, 385
Poisons, toxicity of, influence of

vitamines on, 386
"Polyneuritis gallinarum," 21, 30,

87
substitution of by "experimental

beriberi of animals,'' 92
Potatoes, nutritive value and vita-
mine content of, 255, 256
"Protein-free milk," as an unknown
factor in nutrition, 115
inadequacy of, 116, 127
Protein minimum, 341, 342, 343, 350
Proteins, biological value of, 343
of various origin, table showing

nutritive value of, 264
vegetable and animal, 74
vegetable and animal, comparison
between nutritive value of, 343
vegetable and animal, possible ex-
planation of difference in food
value of, 43
vitamine sparing action of, 214,

325

Protozoa, growth of, 79, 80
Pyorrhoea alveolaris, in scorbutic

guinea pigs, 392
Pyridine derivatives, in vitamine

fractions, presence of, 165, 166
attempts to synthesize vitamine
B from, 193

R

Rabbits, unsuitability of, for study

of scurvy, 135 «
ophthalmia in, 135

498

SUBJECT INDEX

Rats, ability of, to choose between
two diets one that is adequate,
114

ability of, to resume growth after
period of repression, 114

food intake of, influence of vita-
mine B on, 119, 120

food intake of, necessity of measur-
ing, 113, 114

growth of, effect of lack of vitamine
A on, 118, 119, 120, 121, 122

growth of, effect of lack of vita-
mine B on, 118, 119, 120, 121

growth of, effect of small quantities
of milk on, 116

normal growth curves of, compari-
son of, 122, 123, 124

preparation of diets for,' 112, 113

types of cages for, 111

vitamine requirements of, 110, 114

vitamine requirements of, on diets
rich in protein, fat starch and
sugar, 345, 346, 347
Reindeer, suspected avitaminosis in,

142

Rice, changes in, as result of milling,
248

polished, ability of pigeons to live
on, 248

phosphorus content as possible in-
dicator of nutritive value of,
31, 32

removal from, on polishing, of sub-
stance protective against beri-
beri, 30

Rice-polishings, active phosphorus-
free solution from, 168

attempts to isolate vitamine B
from, 172, 173, 174, 175, 176

"beriberi vitamine" from, 169

chemical investigation of, 167

fractionation of (diagram), 169

nutritive value of, 248

substances isolated from, analyses

of, 170, 171

Rickets, association of, with lack of
vitamine A, 323,

association of, with lack of phos-
phorus, 323

association of, with lack of sun-
light, 323

association of, with a particular
dietary, 28

chemical pathology of, 315, 316

cod liver oil in, specificity of, 328,
329

congenital, 313

Czerny-Keller diet in, 320

diagnosis of, 315

dietetic theories with regard to,
324

effects of, in later life, 314, 315

etiology of, 323

foodstuffs not protective against,
325

foodstuffs protective against, 325

in dogs, 137, 138

in dogs, symptoms of, 138, 139,
140

in breast-fed children, 314

in lions, 136

metabolism in, 317, 318, 319

minimal quantitative factors in,
24

occurrence of, 313

pathological anatomy of, 315, 316

prevalence of, 312, 313

symptomatology of, 315

therapy of, and its therapeutic
effect on metabolism, 320, 321,
322

vitamine etiology of, 324, 325, 326,
327, 328

X-ray in study of therapy of,
322, 323

S
Salt mixture, of McCollum, 113

Osborne-Mendel, 113
Schlatter's disease (late rickets), 314
Schwann's sheath, in beriberi, 292
Sciaria, vitamines in growth of, 84
Scoliosis, possible dietetic origin of,
381

SUBJECT INDEX

499

Scurvy, association of definite mode
of nutrition with etiology of,
24, 27

experimental, conditions for dem-
onstration of, 131, 132

experimental, decrease of ad-
renaline in, 133

experimental, metabolism in, 133,
134

experimental, symptoms of, 131,
132

in guinea pigs, discovery of, 27,
128, 129

in guinea pigs, identity of, with
scurvy in man, 130

infantile, 27, 301, 302, 306, 307

infantile, latent form of, 306

in man, diagnosis of, "white line"
characteristic in, 307

in man, differential diagnosis of,
by means of administration of
vitamine C, 307

in man, hematology in, 308

in man, metabolism experiments
in, 308

in man, mode of development of,
300, 301

in man, pathology of, 308, 309,
310

in man, prevalence of, 297, 298, 299,
300

in man, relationship between beri-
beri and, 295, 296

in man, symptomatology and
progress of, 303, 304, 305

in man, therapy of, 311
Secretin, non-identity of, with vita-
mine B, 208

Sheep, vitamine requirements of,
141

"staggers" in, 141, 142
Ship beriberi, occurrence of, 294,

295

Soil conditions and vitamines (dia-
gram), 76
Spasmophilia (see Tetany)

"Spindling sprout disease," of pota-
toes, 69
Spirochetes, testicular substance in

cultivation of, 56
Sprue, as suspected avitaminosis,

371

pathology of, 370
prevalence of, 369
symptomatology of, 369
therapy of, 370

Sterility, as influenced by diet, 379
Stijfziekte, in cattle and goats, 146,

147, 148

cure of, by change of diet, 148, 149
Streptococci, growth of, influence
of tryptophane fraction of some
proteins on, 344
explanation of, 344

Streptococcus hemolyticus, con-
ditions necessary for growth of,
62, 63

failure to grow on beef-heart
extract decolorized with norit,
62, 63
growth of, on beef-heart extracts,

62

Substances essential to life, failure
to recognize the existence of, 25,
26

Synthetic diets, deficiency of, ex-
planation of, 23

Teeth, development of, influence of

vitamines on, 391, 392
Testicular substance, in cultivation

of spirochetes, 56
Tetany, nature of, 332
etiology of, 332
symptoms of, 332, 333
therapy of, 334
Thyroid, influence of food poor in

vitamine on, 105
Tike-tiki (rice polishings extract),

294

Trench sickness, as suspected avita-
minosis, 378

500

SUBJECT INDEX

Tubercle bacilli, vitamines in growth

of, 60
Tuberculosis, as influenced by diet,

384
Tumors, growth of, inhibition by

underfeeding, 389
Tyroglypha, vitamines in growth of,

84

Vegetable soups, as therapeutic
measure in nutritive disturb-
ances of children, 337, 338
Vibrion septicus, growth of, 60
Vitamine, antipellagra, possible oc-
currence in animal protein of,
45

choice of the name, 18, 36

cycle, theory of, 71, 72

content, of bacterial intestinal
flora, 210
cooking water, 241

most frequently used foodstuffs,
table showing, 264

various food-stuffs, influence of
ageing on, 243
baking on, 246
canning on, 242
cooking on, 238
drying on, 241
heating on, 238
storing on, 243

natural difficulties involved in
determination of, 236, 237

relative effect of temperature and
time of heating in presence
and absence of air on, 239,
240

hypothesis of Portier, 79

-like action of certain substances,
206

preparations on the market, in-
efficiency of, 393, 394

requirements, effect of dietary com-
position on, 210

dependence of, on amount of food
assimilated, 213

-sparing action of protein, 214, 262,

263, 346

synthesis of, by bacteria, 57
theory, importance of, effect of

clinical data on, 26
types, 35, 36

Vitamines, classification of, 36
comparison of, with ferments, 163
conception of, research leading to,

19, 20
demonstration of existence of A,

B and C, 40, 41
definition of, by Willaman, 65
gradual recognition of existence of,

37, 38, 39

inability of animal body to synthe-
size, 49
influence of, on anemia, 386

calculi, formation of, 393

cancer, 388, 389, 390

carcinomas, 389

chicken sarcoma, 390

development of teeth, 391, 392

diabetes, 387

neoplasms, 388, 389, 390

toxicity of poisons, 386
in animal kingdom, r61e of, 73
in bacteria, demonstration of, 57
in grain kernels, localization of,

245, 246
in higher plants, localization of,

49
in infections, significance of, 380,

381
in nutrition, relative importance

of, 73

in pure state, possible action of, 394
in tissues, 210
in typhoid bacilli, 57
in vegetable kingdom, r61e of, 49,

50, 51

nomenclature, 39
practical aspect of, 393, 394
relation of, to condition of soil, 75
synthesis of, by some plants, 49,

50
synthesis of, by yeast, 51

SUBJECT INDEX

501

Vitamine A, chemistry of, 218

demonstration of, use of animal

tests in, 225
effect of heat on, 224
effect of lack of, on growth of

rats, 118, 119

effect of, oxidation on, 224
effect of, reduction on, 224
effect of, ultra-violet light on, 224
in butter, 220

in cod liver oil, 218, 219, 220
in fat-free milk, 223
in grasses, 221
in green leaves, probable formation

of, 72, 226

in physiology and pathology, sig-
nificance of, 223
in synthesis of neutral fats, role of,

226, 227

in vegetables, 221
nature of, 222
possible differentiation -of, from

antirachitic vitamine, 42
properties of, 223
relation of, to carrotinoids, 222,

223
relation of, to edema, prophylaxis

of, 227

relation of, to lipoids, 228, 229
relation of, to ophthalmia, 229
relation of, to urinary calculi,

227

solubility of, in water, 223
stability of, 224
Vitamine B, adsorption of, from auto-

lyzed yeast, with colloidal ar-
senic sulfide, 189
with fuller's earth, 188
with mastic, 189
adsorptive capacity of proteins for,

43
attempted isolation of, from rice

polishings, 172, 173, 174, 175, 176
attempted isolation of, from yeast,

177

attempted synthesis of, from
adenine, 193

attempted synthesis of, from nico-
tinic acid, 193

attempted synthesis of, from o-
oxypyridine, 193

attempted synthesis of, from p-
oxynicotine, 193

as possible mother-substance of
product of endocrine glands, 208

chemistry, physiology and pharma-
cology of, 163, 207

demonstration and estimation of,

195

animal tests, 195, 197, 198
phenol reaction, 197
phosphorus content, 196
nitrogen content, 196
uric acid reaction, 197
yeast fermentation test, 198
yeast growth test, 198, 199, 200,
201

suitability of for water-soluble
vitamines but not for vita-
mine B, 203

effect of, on scurvy, 41

effect of lack of, ori growth of rats,
118, 119

fractionation of, difficulties in-
volved in, 164, 165

in carbohydrate metabolism, role
of, 212

influence of, on food intake of rat,
119

in yeast, theory of occurrence of,
as nucleoproteid, 189, 190

its possible differentiation from the
growth-promoting substance, 41,
42, 44, 53, 203, 204

nature of, means for studying,
165, 166, 167

non-identity of, with secretin, 209

preparation of, from yeast, as ac-
tive crystalline substance of con-
stant melting point, 165 178, 179,
180, 181
analyses of, 182, 183, 184

502

SUBJECT INDEX

Vitamine B, animal tests with, 185
solubility of, 194

in olive oil, 191
sources of supply of, 192
stability of, 167
stability of, against heat, and

chemical and physical agents,

194, 195
Vitamine C, chemistry and nature of,

231

effect of, on beriberi, 41
effect of, on growth, 43
formation of, on germination of

grains, 41, 49, 246
in differential diagnosis of scurvy,

307
influence of, on growth and weight

of infants, 158, 159
stability of, 235
various means of fractionation and

concentration of, 232, 233, 234,

235
Vitamine D, in growth of certain

bacteria, 205
in growth of rats, importance of,

205
in yeast, demonstration of, 204,

205
separation of, from vitamine B,

204, 205

W
War edema, unlikelihood of lack of

protein as cause of, 347
Wheat, vitamines in, localization of,

249

X

X-ray, in diagnosis of scurvy, 307
in study of therapy of rickets, 322,
323

Yeast, active crystalline substance
obtained from, 178, 179, 180, 181
analysis of, 182, 183, 184
animal tests with, 185

as rich source of vitamine B, 51
attempts to isolate vitamine B

from, 177

chemical investigation of, 177
curative influence of, on beriberi,

51

dialysis of, 190
fermentation of, as affected by

certain organic substances, 51,

52, 53
growth of, influence of autolyzed

yeast on, 52, 53
growth of, influence of size of

inoculation, 51
growth of, inhibition of, 53
growth of, in presence of as-

paragine, 52
growth of, in presence of peptone,

52
growth of, in presence of vitamine

extracts, 52, 54
growth of, method of Bachmann,

54, 198

growth of method, of Eddy, 198
growth of, method of Funk-Dubin,

200, 201, 202

growth of, method of R. J. Wil-
liams, 53, 54, 199
growth of, not a measure of vitamine

B, 53, 203

growth of, unaffected by vita-
mine C, 54
in growth of bacteria, influence of,

56

in nutrition of flies, role of, 84
methods for fractionation of, 186,

187, 188, 189, 190, 191
substance isolated from, 184, 185
substances isolated from, animal

tests with, 185

synthesis of vitamine by, 51, 55
treatment of, with colloidal arsenic

sulfide, 189
treatment of, with fuller's earth,

188, 189

treatment of, with mastic, 189
treatment of, with norit, 62, 63