January 1, 1949

THEI ROCKEZELflER FOZfMIBTION

CONEIDEUTXAL
MONTHLY REPORT

For the Information of
   The Trustees

Edited by George W. Gray


CONTENTS

Chemistry Joins Borces with Biology . o o o 1

Four Books in the Natural Sciences . . . . 16

Radioactivity in Public Hoalth Research o 19


Grants from the Foundation:

$1,240,865 to Oalifornia Institute of Technology

tJH3MtSTR.Y JOINS FORCES Wi?H BIOLOGY

    More then four centuries have passed since Parase1sus of Hohenheim
gave scientific medicine its charter in his celebrated bypsthesis:
       The human body is a aonglameration of chymieal matters;
    when these are deranged illness results, and naught but
     chymical medicines may cure the same*

It has taken man a long time to learn even a small part csf these schymical
matters,w for as recently as 1849 the molecular weight af water was so
uncertainly knawn that this principal ingredient of the body's conglomeration
was still being written as HO by many chemists. Indf3ea, the idea that wlch

atom has a definite combining power was yet to be accepted. Today, what a

changei Biochemistry is now the principal battleground of science's attack

on di seas8 .  The wealth of physiologically useful chemicals whose idsntifi-

cation has come out of these studies - such compounds as the vitamins, the

hormones, and the antibiotics, to name but three groups - provides powerful

evidence in support of the Paracolsian doctrine and has spurred research in

hundreds of universities, medical schools, and institutes.

Notable among the centers of biochemical interest is the Californta

Institute of Technology at Pasadena.  Here there is no conscious seeking for

new vitamins, new hormones, new antibiotics, or any other specific nutritional

or therapeutic agent.  Put there is being conducted a systematic search far
the weys in which the body1 s molecules behave.  And becauoe the living process

is always associated with huge molecules comprisiag hundreds, thousands, aad
even tens of thousands of atoms in a single &ucture, the pregram at the


2

California Institute is being fooussed primarily on these giant molecules.

Their attractions aa repulsions, their combinations and modifications, their

breakdown into smaller units and the joining of these into new patterns of

molecular architecture - it is such goings-on that the Pasadena scientists are

prying into with all the techniques that chemistry can bring to reinforce

those of biology.  Their inquiry is directed at the most fundamental of all

biological processes: reproduction, nutrition, growth, and disease, each

studied at the molecular level.  If man can learn how these processes operate

at the molecular level, perhaps he can better control or alleviate their

disorders at the body level.  Such, at least, is one visitorls understanding

of the purpose of the program, after spending a week in the laboratories,

talking with experimenters, watching the experiments, and breathing the

atmosphere of California Institute research.

The Attack from Two Sides

    Biochemistry has two avenues of approach. 6ne may enter it from

either the biological side or the chemical, and usually the main strength

of a research program comes from one or the other,of these two directions,

seldom from both. A remarkable aspect of the dual project at Pasadena is

its balance.  This is not a case of a biological laboratory adding a chemical

department to its facilities, nor yet that of a chemical laboratory taking an

interest in biological problems.  It is, rather, a joining of forces between

two coordinate divisions, each of which is a leader in its field.

     The Division of Chemistry at the California Institute was founded by

Arthur A. Noyes.  He had previously served as acting president of the Massa-

chusetts Institute of Technology and was the director of its Research


3

Laboratory of Physical Chemistry when he came to Pasadena in 1913. At first

Dr. Noyes served the California Institute on a half-time basis, devoting the

other half of the year to the Massachusetts Institute. But in I919 he decided

to give himself wholly to the work in California and soon made it foremost

in teaohlng and research.  He was a physical chemists his emphasis was on the

inorgtic aspects of the science ) end aspiring chemists from all over America
came to Pasadena to study the fundamentals under the master;

     Among these students was Linus Pauling, a recent graduate of the

Oregon State Agricultural College.  Perhaps Noyes saw in him the man he wanted

to train as his sucoessor* At all events, the young Oregonian became a

favorite pupil, spent three years of advanced study under Royes, and was so

imbued with the physical aspects of chemistry that`he seriously considered

specializing in atomic physics.  A National Research Pellowehip enabled

Pauling to spend a year in Munich with one of the world's leading theoretical

physicists, Arnold Sommerfeld, and these studies were continued the following

year with Niels Bohr at Copenhagen and Erwin Schroedinger at Zurich. But the

problems that made the strongest appeal to him were in chemistry; so Pauling

remained a chemist, meanwhile continuing his investigation of the forces

which operate between atoms and molecules , a study which resulted in his great

book, sThe Nature of the Chemical Bond."  California Institute made him a full

professor in 1931, when he was only 30 years of age, and following Dr. Woyes

death in 1936 Pauling was appointed to succeed him as chairman of the division

and director of the chemical laboratories.

     "I was a physical chemist," ezqlained Drr Pauling, "with this

dominating interest in the forces which cause atoms to join into molecules and

molecules to react with one another.  The forces are electrical, of course,


4

and depend on the number of protons and electrons present and the order of

their arrangement in the structures.  This is essentially a physical subject:

or, rather, it belongs to that borderland where chemistry and physics merge*

In these studies I naturally selected the simpler molecular structures to

work with, such as the metals and inorganic compounds; but in the course of

the investigation I also tested en organic substance whose molecule is large

and complicated:  the hemoglobin which gives the blood cells their red colorr

I found that in arterial blood the hemoglobin was repelled by a megnet, but in

venous blood it was attracted.  This led to a study of the chemical bond

between the hemoglobin and the owgen which it picks up in the lungs. I wanted

to consult someone who had specialized on hemoglobin and found the authority

in A. I, Mirsky of the Rockefeller Institute for Medical Research. Dr. Mfrsky

came to the California Institute for a year, and we collaborated on a study

which resulted in a joint paperes

     This paper attracted the attention of Icarl Landsteiner, the dig--

coverer of blood types, and Dr. Landsteiner asked Dr. Pauling if his theory of

the chemical bond could throw light on a certain antibody reaction.

Landsteiner*s request introduced Pauling to the highly complicated specialty

of immunology; the two men became close friends and frequent conferees on the

subject, with Landsteiner supplying information and hypotheses from the bio-

logical angle and Paul& from the chemical, "From that time on I gave a

great deal of thought to the chemical aspects of immunol~gy,~ said Dr.

hlmg, strying to understand, in terms of the chemical bond, how an antibody

neutralizes a virus or other antigen."  By 1939 he had arrived at a chemical

picture of the reaction and reported his results to the Amerfcan Chemical

Society as *A Theory of the Structure and Process of Formation of Antibodies.tf


5

     Thus, under Dr. Pauling, there was a transition* In the course of a

few years the division added to its program the investigation of hemoglobin,

antibodies, and other molecular giants which originate only in living systems,

while still continuing the basic work in the chemistry of inorganic and simpler

organic substances.  The Rockefeller Founda         tributed to this devel-

opment through financial grants, beginning          th additions in 1933 and

1934, it had made appropriations to the Ca       stituts for "research in
                                                   .                                  , ..u..t "
chemistry under Dr. Linus Pauling." The shift in program was reflected i@Gx8,4,Y
                                                                           %". ".,&SZ

when the first of five annual appropriations was voted "for the development

of chemistry in relation to biological problems."  Other grants were earmarked

for "researches on the structure of antibodies and the nature of immunological

aotion.z

     Meanwhile, a transition was also taking place in the Institute's

Division of Biology.  This division had been organized in 1928 by Thomas Hunt

Morgan, who had left the chair of experimental zoology at Columbia University

to pioneer this new planting in California. Like Dr. Noyes in physical

ohemistry, Do. Morgan was already world-famous in genetics; and his coming to

Pasadena brought several strong additions to the faoulty, most of them geneti-

aists, and attracted from all parts of the country students who wished to

speaialize in this science.  The Foundation made two grants to support Dr.

Morgan*s researches.

     Genetics lends itself to mathematization more easily than most bio-

logical saiences , and perhaps it is rightly called the most "physicals of the

branahes of biology.  Certafnly Morgan had a strong urge toward collaboration

between biology, on the one hand, and chemistry, physics, and mathematics, on

the other.  In the first announcement of this division he said: trA closer


6

relation of these sciences with biology is imperative,H and during his regime

interrelations with the physical sciences were cultivated. Dut the main

effort of his division was in genetics, and particularly in classical genetics,

a field in which Morgan was the acknowledged master*  Classical genetics has

to do with the morphology 06 the meahanism of heredity, the position and order

of the genes in the chromosomes, and their identification as dominant or

recessive in the control of inherited characteristics.

     After Drc Morgan's retirement in 1941, the division was administered

for several years by a staff committee headed by Alfred H. Sturfevant. The

serological aspects of genetics interested Dr. Sturtevant, and during this

period there was increasing collaboration between the two divisions. The

Foundation made several grants for joint researches by Drs. Sturtevant and

Pding.  But direction of the division by a committee was only a temporary

qedient while the Institute was seeking a successor to Dr. Morgan, and

toward the end of 1945 he was found in the person of Stanford University*s

professor of genetics, George We Beadle.

     Beadle's history closely parallels that of Pauling. Both men were

8atlonaI Research Bellows; and as Pauling came to the California Institute

to study under Noyes, so Beadle came to study under Morgan.  It is also

significant that at the time when Pauling was turning his attention more end

more to the biological molecules, Beadle was becoming enamoured of chemistry

aa the handmaiden of genetics.  During his ten years at Stanford he devoted

laosf of his research effort to experiments with the bread mold, Neurospora,

rad was able to demonstrate in this lowly fungus that the processes of
isutrffion (which are purely chemical) are directed by the genes, Perhaps it

is not undue praise to say that Dr. Beadle's work with the mold did more


7

than any other research to establish the chemical nature of genie action.

Classical genetics now has a lively daughter known as chemical genetics, a

prolific contributor of new findings to science*

    This, then, was the situation in 19466 With chemical research in

charge of a biologically-minded chemist, and with this chemically-minded

geneticist placed in charge of biological research, the California Institute

offered an unusual opportunity.  The Divisions of Biology and Chemistry imme-

diately prepared a prospectus outlining Ita joint program of research on the
fundamental problems of biology and medicine."  The program would occupy
fifteen years and require R budget of $400,000 a year; and application was

made to foundations for support.  It was estimated that about five years would

be required to bring the program to its full operating capacity, and as
interim grants The Rockefeller Foundation appropriated $50,000 in 1946 and an

equal amount in 1947, followed this year by a long-term appropriation of
$700,008 to be paid in annual installments of $100,000, Thus, $800,000 has

been committed within the last three years*  The earlier contributions, begin-

ning with the first grant on behalf of Dr. Pauling's work in 1932, have
totaled $440,865, making a grand total of $1,240,865 given to &.lifornia

Institute for research in chemistry and biology.

     In addition, the joint program of research on the fundamentalprob-
lems of biology and medicine has attracted support from other funds8 $60,000

a year from the National Foundation for Infantile Paralysis, and lesser grants

from the Nutrit$on Foundation, the Hermann Frasch Foundation, and others. As

rapidly as competent scientists can be found and trained for the new posts

oalled for by the program, the work is being expanded to the full scope envi-

sioned by the prospectus.


Research Manpower

     Of the two essentials to successful research - manpower and equip-

ment - the human element of course is the more important. Wickliffe Rose used

to say that his main function as an officer of the Rockefeller Boards was to

discover men of superior brains and then back them up through a period suf-

ficient to demonstrate their abilities.  What makes the situation at the Cali-

fornia Institute so challenging is the presence there of the two staffs of

scientists with their already integrated teamwork of biology and. chemistry.

In 1946, when the joint program was projected, the staff in biology, including

all workers from professors to research fellows and assistants, comprised 32

persons; and the corresponding groups in chemistry totaled 86. At present

biology is employing the services of 79, and chemistry 97, a grand total of

176 for the two divisions, or an increase of 49 per cent over the status of

three years ago,  Rot all of these staff additions were financed from the

funds earmarked for the joint program; some of them represent new fellowships

that have become available in the last year or two or chairs recently estab-

lished by the Institute.  But all are engaged in research, and the joint

program is benefiting  enormously from this increase.

     hong the recent staff additions are John G. Kirkwood in chemistry

md Max Delldak in biology.  Dr. Kirkwood is in the distinguished line of

physiaal chemists and had held research and teaching positions at Massachu-

86th Institute of Technology, University of Chicago, and Cornell. He was

%dd professor af chemistry at Cornell in 1948 when called to the newly-

ortablished Arthur A. Noyes professorship of physical chemistry at the

4ulifornia Institute.  Like Dr. Pauling, he has a predilection for the giant

JyIlties, and recently developed a new type of electrophoresis apparatus


9

with which to study their properties.  Tests made at Pasadena within the last

few months show that the Kirkwood apparatus will separate the proteins of

blood plasma to a finer degree than any other device heretofore used.

    Dr. Delbr&ck is a physicist turned biologist. His primary training

was in Germany in theoretical physics, but he became interested in bacteri-

ology, and came to the United States as a Rockefeller Fellow in biologyr He

has made many contributions to our knowledge of bacteriophages, the invisible

Piruses which prey upon bacteria.  Although these viruses multiply with great

rapidity when they penetrate a bacterium, and in some of the experiments seem

to do this by a kind of sexual reproduction, in other respects they appear as

inert as any chemical.  Indeed, each virus seems to be made up of but a few

giant molecules.  The viruses thus occupy a borderland between the living and

the non-living, between biology and chemistry, and study of them constitutes

an Important part of the joint program.  Dr. Delbr%ck joined the California

Institute faculty in 1947, coming from Vanderbilt University.

     Other members of the combined staffs could be cited with equally

interesting detail.  There is Laszlo Zechmeister, formerly of the University

of Peas, Hungary, who came to the California Institute as professor of organic

ahsmlstry in 1940.  Dr. Zechmeister is an authority in chromatography, an

a#z~zing technique for separating carotenoids and other organic pigments out

of afxtures - and his specialty is contributing directly to the joint research

program, Another worker is Dan H. Campbell, an immunochemist, brought here

in 1942 with the assistance of a grant from the Foundation. Dr. Campbell

kas been collaborating with Dr. Pauling in an effort to synthesize sntibodies

b direct chemical means - a daring project which if successful may revolu-

Moaaice the control of infectious disease.


10

Research Equipment

    The laboratories and other campus buildings of the California Insti-

tute are in the Mediterranmnslyle af architecture, low structures arranged in

pleasing quadrangles, with connecting colonnades. Chemistry is housed in the

Gates end Crellin Laboratories, which in effect are one large building in the

form of a TJ.  Biology has its headquarters in an adjoining edifice 'known as

the William 8. Kerckhoff Laboratories.  The juxtaposition of the two buildings

has proved to be of great convenience to the growing collaboration between

the chemists end biologists, end symbolizes the partnership in research.

     In addition to its main facilities in KercMloff, biology has several

smaller establishments off the campus.  Across the street are three labor-

atories of plant physiology, one of them just being completed at a cost of
$440,000, a gift from the Earhart Foundation of Detroit. Half a mile away

is another center of botanical research, the Orlando Road Greenhouses; and

five miles distant is Arcadia Farm, ten acres devoted to the study of genetics

in uorn* Fifty miles southeast, at Corona de1 Uar on the Pacific shore, is

the marine laboratory.  This is both a center for research and a means of

supplying marine animals for experiments in biology and chemistry on the

campus Last summer still another unit was completed, a large animal house
&lch adjoins a section of Kerckhoff on the north. 'This animal house was
built entirely undergrwnd, cost $150,000, and embodies all current conven-
iences for breeding, housing, and caring for experimental animals. It is

Uspensable to the joint biology-chemistry program.

    With this combination of laboratories and auxiliary facilities, the
division of biology is really superbly equipped. There is plenty of working


11

BpaC8, but of COW88 as the research program 8XpandS to its full SCOp8 this

will b8COm8 Crowded and additional quarters will be needed* The laboratories

of the division of ch8mistrg, with its larger staff, are already serfous~

crowded; and plans haV8 been drawn for a new btilding to be erected on the

campus in the vacant area north of Kerckhoff and west of Crellin and Gates.

'phis new laboratory, d8Sign8d in the same architectural style as the older

buildings, will be US8d for the joint chemistry-biology program* It is

planned to connect with both Xercl&off and Crellin and will increase the

research quarters of the two divisions by 75 per cent, The estimated Cost Of
the proposed building is $Z,OOO,OOO, and there are good prospects that a

sufficiently large fraction of the necessary funds will be obtained to permit

aonsfxuction to begin this coming year.

Research Objectives

    SW8 are seeking to uncover basic phenomena and learn the principles

Mrich govern them, rather than to solve specific practical problems," explain-

ed Dr. Beadle.  "The action of genes in controlling inheritance, the action

ef antibodies in neutralizing the destructive effects of bacteria, viruses,

aad Other antigens, the action of hormones in promoting or retardfng growth -

these are examples of basic ph8nOm8Ila, processes that are fundamental to life.

ti W8 Understood the basic processes, if we knew precisely what is happening

&l 8aCh in terms of molecular action, we believe the solution of the prac-

&iaal problems would follow almost inevitably."

And so the biology-chemistry team at the klifornia Instftute is

%!&erested in genes, antibodies, viruses, hormones, biological pigments, and

  tea strUctur0s.  HOW does each behave biologically, and how can this

Wmior be aCCOUZIt8d for chemically?


12

     Chemical behatior is related directly to the mol8cUlar St3XCtUP3

of the reacting substances; therefore one Of the principal objectives Of the

joint research program is chemical analysis. What are the buflding blocks
that enter into the construction of genes and the other entities? How are

these building blocks put tOg8th8r, in what order of &rrangement, and what

ar8 the resulting Size and shape Of the StrUCtUre?

     "&i8nC8 is Still far from CO2I$8t8ly analyzing these biological

agents," said Dr. Beadle, "but the investigations tend to show that the mole-

cular form lolown as protein is the key structure. Apparently most of the

bodies that we are studying in our program are either simple proteins or

conjugated proteins,"

    Simple proteins are "simple" only by contrast with the vaster

arohit8CtUr8 of the conjugated molec~8s.  Actually, a simple protein will

consist of hundreds, sometimes thousands, of atoms. When placed beside

f8milia.r inorganic molecules, such as thOS8 of water, sulfuric acid, ammonia,

and table salt, even the smallest protein molecule is like a Whale among

WItlOWS*  But a protein is simple in this respect: when it is broken down
it dO8S not separate into its hundreds or thousands of individual atoms, but

divides into characteristic groups of atoms which the chemists know as amino

acids.  It is as though when a house was demolished, it broke up into base-

ment, roomsi and attic, rather than into individual bricks and boards.

'Pwenty-three different amino acids have been found in proteins, and the

pOss%ble combinations that may be formed from these twenty-three building

blocks run into cauatless millions.  It is no wonder that proteins occur in

the wide variety which makes one man's meat another man's poison. But a
number of the most familiar and wholesome substances of the body?, equipment


13

am3 simple proteinst pepsin and many of the other dfgestive enzymes, insulin
and meny of the hormones, albuznin, fibrinogen, and many other components of

the blood plasma.

    The conjugated proteins represent a further step in structure.
After a simple protein molecule has been built by the joining together of
molecules of different amino acids, it may hook on to a pigment and form a
conjugated protein such as the hemoglobin of the blood. Or, it may attach
itself to a complicated chain of sugar molecules known as a polysaccharide
and form a conjugated protein of another type, such as the mucin of saliva.
Another possibility is the joining of a protein with a vitamin - the enzyme

carboxylase is of thfs type.  Finally, proteins may be linked with nucleic

acids to form nucleoproteins - and here we reach the ultimate of giantism

ambng molecules.  For if a simple protein fs pictured as a whale among the
minnows, a nucleoprotein may be likened to a leviathan with form so tre-

mendous that it might swallow the whale.  Nucleic acid alone is a large

structure - some of its molecules contain 160,000 atoms - and when units of
this size combine with units the size of proteins, the combination is truly

enormous.  Some of the viruses which Wendell M. Stanley isolated in his

studies at the Rockefeller Institute were identified as nucleoproteins and

weighed up to 8,000,OOO times the weight of hydrogen.  Such structures

comprise nearly a million atoms.

     It is believed that both viruses and genes are nucleoproteins,
while the antibodies are thought to be simple proteins consisting of chains
of amino-acid residues folded together fn a certain way. According to Dr.
Pauling*s theory, these folded chains of interlinked amino-acid residues
(polypeptides, they are aalled) are afloat in the bloodstream: end whenever


14

they encounter the bacterfum, virus, or other odd body against which they

serve to protect the organfsm, the mutual attractions between the two cause

the chain to approach and attach itself to the intruder. The action of the

chemical bond thus brings the antibody to overlay an area of the surface of

the foreign body with a shield or encrustation which blocks the latter*s

activity*  This explanation is necessarily brief and oversimplified, but

perhaps it suggests the aation by which an antibody neutralizes a microbe.

    w The genes, we believe, exercise an overruling control on all these

activities ,It  said Dr. Beadle.  "They do this) we think, by serving as the

master patterns for the many proteins which function in the processes of

life. Thus, there is probably a gen(, which serves as the template for the
I&s manufacture of insulin, another which provides the mold for pepsin,

and so for albumin, fibrinogen, the polypeptide chain that forms antibodies,

rrnd all the rest.  There are several thousand genes distributed among the 48

chromosomes of the human body cell, a number sufficient to provide templates

for the thousands of big molecules requfred for health. Diabetes, on this

theory, is a consequence of a missing or defective gene, leaving its victim

unable to manufacture insulin.  Similarly, the bleeders or hemophiliacs lack

bhe nozmal gene for manufacturing a gamma globulin which is an essential com-

ponent of the blood-clotting equipment.  Our experiments with the bread mold,

hrospora, have demonstrated this genie control of the biochemical processes

In numerous instances. We found, for example, that after exposure to ultra-

violet radiation, the Neurospora lost its ability to make certain vitamins.

like genes which controlled this manufacture had been destroyed, and there-

ufter the Neurospora languished unless these vi tamins were supplied in its

foode Similarly, Sterling Emerson of our laboratory found that a minute


15

change in tts genes caused the Neurospora to accept as food a compound that
before the change had acted as a poison. Indeed, after mutaffon, the
Surospora would not grow unless fed a sulfonamide which previously had
blocked growth and caused death.*

    The strongest impression that one brings back from a visit to the

Oaltech team is the magnitude of the task of analyzing these invisible

molecules .  As a step toward understanding the proteins, the group fs working

first on the amino acids, trying to map precisely the structure of these

protein building-blocks.  Robert B. Corey of the chemistry staff spent a year

and a half analyzing the configuration of glycine, the simplest of the amino

acids.  He bombarded it with x-rays, and measured the angles at which the rays

bounded off the molecule.  In this way he not only determined the position of

each carbon at on, each oxygen, and each hydrogen in the glycine, but actually

ttsasured the distances between the atoms, After completing this job, Dr.

&my went on to alanine, which is larger and more complicated. The experi-

(mae he had gained on glycine stood him in good stead, and he requfred only

a year to work out the exact pattern of alanine.  He has now taken up a still

)#)re complicated amino acid, threonfne , and this winter is deep in a study of

flk In this way, the group plans to move from the amino acids to more compli-

safe& structures, with the hope that eventually they may be able to dissect

801~ of the proteins, perhaps even nucleoprotefns, into their integral parts.


16

POUR BOOKS IN THE NATURAL SCIENCES

Among the books in the natural sciences which were published

during 1948, four may be listed as of special interest because of the

relationship of the authors and their subjects to projects fostered by

the Foundation.

     Mathematical Biophysics, by Nicholas Bashevse.
            The University
of Chicago Press, Chicago, 1948, pp. 669, $6.

       This is a revision and extension of a briefer book pub-

lished in 1938 with the assistance of a grant from the Founda-
tion.  Dr. Bashevsky is professor of mathematical biophysics
in the University of Chicago.  His special field of research
has been the attempt to interpret biological processes such
as cell division, growth, nutrition, and communication in terms
of physical phenomena.  His treatment includes a physico-
mathematical theory of the operation of the central nervous
system.

    Cybernetics, by Norbert Wiener. The Technology Press, John Wf1-e~
and Sons, New York, 1948, pp. 194, $3.

  Another book which undertakes to interpret biological
proaesses in terms of physical phenomena is this new work by
Dr. Wiener, professor of mathematics in Massachusetts Institute
of Technology.  Professor Wiener dedicates the book to Arturo
Posenblueth, physiologist of the Mexican National Institute
of Cardiology, and there is a strict appropriateness in this
dedication, for ,Cybernetics treats of a problem on which the
mathematician and the physiologist have been collaborating for
a number of years, and since early 1948 with the help of a
grant from the Foundation.  The problem is that of nervous
control, how the brain communicates with the muscles and directs
their action. As the book discloses, the brain does this in ways
quite parallel to those used by dial telephones, thermostat
regulators, ,electronic calculators, and other automatic machines.
"In their more elaborate forms," says Professor Wiener, smodern
aosrputing machines are capable of memory, association, choice,
and many other brain functions.  Indeed, the experts have gone
so far in the elaboration of such machines that we can say the
human brain behaves very march like the machines. The construction
of more complex mechanisms actually is bringing us closer to an
understanding of how the brain itself operates." The word


17

cybernetics was coined from the Greek "kybernetes," meaning
steersman, and Wiener uses it as the name for the new science
ncommunfcation and control in the animal and the machine."  His
book is heavily loaded with mathematical equations end other
technicalities, and fs hardly a manual for the laymen, but a very
readable interpretation was written by Wiener for the "Scientific
American" and appeared in its November number.

     Submicroscopic Morphology of Protoulasm and Its Derivatives, by
Albert Frey-Wyssling.  Elsevier Publishing Company, Amsterdam, 2nd edition,
19443, ppm 26_, $6.

  Professor Frey-Wyssling was a fellow of the International
Education Board in 1925-26, and in recent years his work at the
Eidgen0ssische Technische Hochschule in Zurich has received sup-
port from the Foundation.  He and a small group of associates
in Zurich are specialists in the study of structures beyond the
reach of the optical microscope - such as the tin&e morphology
of chromosomes, slime molds, cellulose, and silk. The present
work is a new edition, in an English translation, of a work that
has long been standard in this difficult field.

     Iladioactive Indicators. Their Applications in Biochemistrv, Animal
Efursiob~. and Patholom
London, 1948, ppo 556, $16. by George Heresy.  Interscience Publishers, Inc.,

  Professor Hevesy was among the first, perhaps the very first,
to use radioactive elements as tracers in biochemical research.
As early as 1923 he published an article under the title, "The
Absorption and Translocation of Lead by Plants: A Contribution to
the Application of the Method of Radioactive Indicators in the
Investigation of the Change of Substance in Plants.n The experi-
ments which produced this paper were pioneer studies, made at a
time when the only available radioactive elements were those of
n&ural occurrence.  Hevesy used thorium B, an isotope of lead
with a radioactive half-life of 10.6 hours, to show that lead was
heavily concentrated in the root tips of the bean plant and that
it was eventually transported to other locations throughout the
tfssues of the plant.  In the ensuing quarter century Hevesy's
research has reached into magy areas of the rapidly growing science
of radiobiology, enabling him to write this authoritative text-book.
With the discovery (by the Joliot-Curies in 1934) that ordinary
elements like carbon, iodine, iron, and phosphorus can be made
radioactive by bombardment, the usefulness of the tracer tech-
nique was greatly enhanced.  The method is now in wide use in many
fields of biology, including the investigation of fundamental life
processes, such as photosynthesfs in plants and nutrition in
animals, as well as in the study of pathological conditions, drug
action, end other medical problems.  Someone has said that its


38

introduction constitutes as significant an adv-anci9 in the develop-
ment of biology as the introduction of the microscope more than
two and a half centuries ago.  Beginning in 1930 when he was
at the University of Freiburg, continuing at Copenhagen where he
was a refugee from Hitler in 1934, and later at Stockholm where
he went when the Nazis overran Denmark, Professor Hevesyls
studies have received the support of grants from the Foundation.
In 1946 he spent six months in the United States as a visiting
professor on a Foundation grant.


19

R4DIOACTIVITY IN PUBLIC HEALTH XESXARCH

     The tracer technique which Heresy discusses in his book on

Radioactive Indicators is being applied by the International Eealth Division

of the Foundation in the study of three kinds of disease-transmitting agents:
(1) viruses, (2) malaria parasites, and (3) mosquitoes.

In Virus Research

     The incubating hen* s egg with its growing embryo has become a

universal medium for the cultivation of viruses.  It takes about 21 days for

an egg to hatch, and in the course of this process various systems develop

to serve the growing embryo.  The most conspicuous are the amniotic sac with

its fluid in which the embryo lies, the larger allantoic sac which overhangs

and partly envelopes the amniotic, the yolk sac with its rich store of

nutrients, and the interconnecting system of blood vessels.  Virus injected

into any of these systems eventually reaches the embryo, but the selection

of the route and technique of inoculation depends on the nature of the virus

that is to be cultirated.  Usually the virus is introduced into one of the

extra-embryonic sacs by means of a needle inserted through a window cut in

the shell of the egg.  The window is then closed, the egg returned to incuba-

tion, and in two to three days the embryo is teaming with the virus.

      Most but not all of the viruses which are known to infect animals

have been cultivated in egg embryos.  Their growth may be recognized in any

one of several ways:  by the death of the embryo; by the production of

characteristic pathological changes in the embryo; by the agglutination of

red blood cells, a phenomenon limited to influenza and a few other viruses:


20

and by the inoculation of material from the embryo into a susceptible animal.

If, however, a virus does not kill or produce recognizable changes in the

embryo, ar fails to cause agglutfnation of red blood cells, or if no suscep-

tible animal is readily available, the growth of the virus within the egg

may escape notice.  The problem is to find other means of identifying the

presence of unknown viruses - aa it WEM with this in mind that Riohard M.

Taylor and his associates of the International Eealth Division Laboratories

decided to experiment with radioactive phosphorus.

     Phosphates are among the mineral compounds necessary to metabolism,

and therefore it was felt that the introduction of such substances into the

developing egg would have no adverse effect on the embryo. By using a radio-

active isotope of phosphorus, ~32, which has a half-life of 14.2 days, the

compouzd was provided with a tag which betrayed its presence,  The idea was

to inject the phosphate into various Darts of the developing egg and see if

its transport to other parts was affected by the presence of virus.

     For the first series of experiments only healthy eggs were used,

the object being to get the normal result unaffected by the virus. Dr. Taylor

and his oo-workers injeoted the radioactive phosphate into the allantoio sac,

and then at intervals of two, three, and four days took sampling of the

allantoic fluid, amniotic fluid, yolk-sac fluid, blood, and embryonic tissue,

and tested each for radioactivity to see where and to what degree the phos-

phorus had dispersed.  They found that most of it remained within the allan-

toic sac, though small quantities were detected fn the blood stream and in

the embryonic tissue.  Taking other healthy eggs, they next thrust the needle

into the amniotic sac of each end deposited the phosphate there, Tests for

radioactivity showed that the phosphorus rapidly passed to the other systems,


21

and by the third day precious little remained in the amniotic fluid. Similar

experiments tested the effect of introducing the phosphate into the yolk sac -
it'passed into the embryo, the blood, to some degree into the allantoic fluid,

but none went into the amniotic sac.  Similarly, when the compound was

injected directly into a blood vessel, the larger amount remained in the

blood, some entered the allantoic sac - but again, none passed into the

amniotic.  The experiments thus indicated that the allantoic sac tends to

retain the phosphorus and will also receive it when introduced by other

rout es.  Contrarily, the amniotic sac neither retains nor receives the

element.

     The question was whether or not the presence of virus would make
any difference in this pattern of phosphorus transport. To test it, the

experiments which had been made with healthy eggs were repeated with virus-

infected eggs.  It was found that if the egg was infected with influenza

virus, and the phosphate was introduced into the allantoic sac, the radio-

activity no longer remained concentrated there, but rapidly diffused to other

parts, particularly into the blood stream.

     The testing technique is as follows. Samples are taken of the
various fluids, a standard quantity of each is placed in proximity to the
tube of a Geiger-M&er counter, and the number of radiations given off is

counted.  Since the radioactive phosphorus atoms discharge beta particles at

a rate which is statistically constant, the number of radiations per minute
is a direct index to the number of phosphorus atoms present.  In one series

of experiments, in which the phosphate was injected into the allantoic sac

of a healthy egg, specimens were taken of the allantoic fluid and of the

blood at intervals of 48, 72, and 96 hours, and equal quantities were tested

for radioactivity, with the following results;


22

                  Ailanmc fluid    Blood

At the end of 48 hours         1483
At the end of 72 hours          1332           99
At the end of 96 hours          1308         134

The concentration of the redioactive phosphorus did not change

much, and the amount that had passed into the blood stream at the end of the

fourth day was about a tenth of that retained in the allantoic sacs But

when this eJrperiment was repeated with the virus of influenza injected into

the allantoic sac along with the radioactive phosphorus, the results as

picked up by the Geiger-Nller counter were strikingly different.

                   Allantofc f hid   ii!a!A

At the end of 48 hours          2039
At the end of 72 hours           831         307
At the end of 96 hours          300          460

Obviously the presence of the virus had a profound effect on the

permeability of the membrane enclosing the sac; for four days after its

receipt of the injections the allantoic sac has lost more phosphorus to the

blood stream than it had been able to retain, and corresponding or even

larger amounts had passed into other fluids and into the embryo itself.

As far as the experiments have gone, influenza has a more potent

effect on the dispersion of the phosphorus than any other virus~  Mumps v-irus

shows an influence, but to a lesser degree , whereas yellow-fever virus has

no apparent effect at all.  Dr. Taylor and his associates are working at

present with throat washings from various undetermined respiratory infections.

St would be a great boon if this tracer technique should point the way to a

simple test for the identification of specific viruses, or to a sure means of

separating viruses out of a mixture.


23

In Malaria Research

     The plasmodium of malaria is a parasite of the red blood cell. It
enters a red cell and, after a period of time which varies with the parasite

type, the cell bursts and out come a number of new plasmodia. Immediately

each of the new brood attacks a fresh red cell, bores into it, and begins a

new cycle of growth and multiplication.  Without red cells the plasmodium

dies, and therefore it is dependent on the cell for certain metabolic func-

tions.  But presumably there are other functions which ft performs itself,

and one of the key problems is to identify the two sets.  If we hew what the

malaria parasite does for itself and what it must depend on the red cell to do

for it, the biochemists and pharmacologists would be better prepared for their

effort to develop new ant imalarial drugs.  To be effective, the drug must

block some essential step in the parasite's metabolism - and a preliminary

task is to identify that vulnerable function.

     Several approaches to the enigma of the plasmodium are being

explored in the program of malaria research conducted by Max Theiler in the

International Health Division Laboratories.  One is of special interest in

thfs oonnection because it is using radioactive phosphorus. Delphine H.

Clarke was intrusted with this research, and before beginning it she spent

several weeks at the Radiation Laboratory  o f the University of California

studying tracer technique.

     Most investigations of plasmodia have been in animals that were ill

of malaria.  But for study of its function Dr. Clarke wanted to isolate the

parasite as much as possible.  She made a rich broth of nutrients, including

an extract of ruptured red cells and a phosphate containing radioactive phos-

phorus.  When this nutrient solution was blended in a test tube, she placed


24

fn it a number of living red cells which were infested with the parasites.

And then for 24 hours she watched what happened. At the end of that time,

the red cells were removed, and the plasmodia measured under the microscope.

They were larger:  there was no doubt they had grown. Then Dr. Clarke f rac-

tionated the plasmodia, grinding them up, and by means of chemical agents

separated out the fatty compounds, the nucleo-proteins, and other components,

3ach of these fractions in turn was tested under the Geiger-Miller counter,

and the lively responses of the indicator needle showed radioactivity from

each.  It was clear that the tagged phosphorus had passed from the solution

into the parasite and had entered into the construction of nucleoprotein, fat,

and other compounds.  But which was responsible for the synthesis - the plas-

medium or the cell in which it lived?

     Nucleoproteins, those supergigantic molecules which Drs. Beadle and

Pauling are studying in the joint biology-chemistry program at Caltech, are

believed to be the fraction which indicates growth. So Dr. Clarke directed

a series of experiments at the investigation of nucleoprotein synthesis.  She

worked with animals first, selecting chickens because they are susceptible to

malaria,  She injected phosphates containing p32 into the blood of healthy

chickens, and after 24 hours took blood specimens, fractionated the red cells,

and by counts of the radioactivity determined the amount of nucleoprotein

that had been synthesized.  Then she took malarjous chickens, injected them

with the phosphate, and after the usual period of time fractionated red cells

taken from the infected birds.  These cells contained ten times as much

nucleoprotein as the healthy cells.  The studies were repeated in the test

tube, again with healthy red cells and then with malaria-infected cells, end

the result was the same: the cells containing the parasites synthesized ten


25

times as much new nucleoprotein as the healthy cells produced. From these

results Dr. Clarke concluded that the synthesis of nucleoprotein can

assuredly be used as a measure of the rate of parasite growth.

     The experiments indicate that the synthesis of new nucleoproteins
(and thus the growth of the parasite) is dependent upon the presence of en

abundance of normal red cells or of an extract of such cells.  Therefore,

althou& it seems that the formation of nucleoproteins is a function of the

parasite itself, nevertheless the normal cells must supply certain materials

which enable this synthesis to take place. It is these as yet unknown

materials which are now being studied and whose identification would seem to

answer the question why the parasite must live within a cell.  This in turn

may show what kind of drugs we must devise to stop the growth of the parasite

ma lead to its complete destruction.

In Mosquito Research

     Because they serve as vectors of malaria and yellow fever, the

study of mosquitoes is as pertinent to the control of these diseases as the

study of the plasmodium and virus.  For many years the International Health

Division has been interested in the flying abilities of the insects.  It was

important in the establishment of control measures to know how far from an

infected village or plantation they could travel. To determine this, mos-

quitoes were marked w dusting with bronze powder, fluorescent material, or

spraying with dyes, and the marked mosquitoes were then released from a

central station,  When recaptured, it was possible to identify them and thus

determine their flight range and how long they had endured the buffeting of

imclement weather,


26

     But each of these methods of marking subjected the mosquitoes to

handling.  Many were injured in the process , and one could not be sure how

the survivors were affected physiologically, perhaps weakened in endurance.

There was need of a less-violent marking method, and it was found and effec-

tively demonstrated last year in the use of the radioactive tracer teChniqUea

      The demonstration was made at the West African Yellow Fever Labora-

tory near Lagos, Nigeria, by John C. Bugher and associates of the International

Health Division.  Dr. Bugher explored the possibilities of the method first

in 1946, when he was in the United States, working with Dr. Whitman. They

showed at that time that if a phosphate containing radioactive phosphorus

were dissolved in the water in which larvae were hatching, the phosphorus

would be assimilated by the insect and could be identified in the mature

mosquito,  When Dr. Bugher returned to Africa he took with him a Geiger-

Miiller counter and other needed apparatus, and soon had en experiment going

with radioactive marking.  In addit ion to the phosphorus, he used a radio- *

active isotope of strontium, Sr 89
                                     .  The method is essentially the same with

both, but the strontium has a half-life of 55 days, therefore lasting longer

as a marker.

     "What we did in Lagos ,s said Dr. Bugher, VW&S to put mosquito eggs

in a tray of water, end after the eggs had hatched we added the radioactive

material - either strontium chloride or disodium phosphate. We did not

handle the adults; simply let nature take its course. In one experiment we

marked 5,000 mosquitoes in this way, in two others 10,000 each, and in the

final test 250,000.  Our release point was in the compound of the Yellow

Fever Laboratory, and catching stations were set 9 at intervals of a few

hundred feet in all directions.  Each catching station consisted of a man,


27

hired for the purpose, who took off his shirt and sat or stood in the

appointed place.  Whenever a mosquito landed on him, he captured it in a

tube and waited for the next arrival.  At the end of the night's vigil each

man brought his catch to the laboratory, where the mosquitoes were identified

and subjected to the counter for recognition of the radioactive ones. We

found that some, flying in the direction of the prevailing wind, traveled

as far as 4,000 feet. Against the wind few made any great headway. Some of

the mosquitoes were picked up 30 days after their release, but these were

rare; most of those recovered were found within the first fifteen daysIn

     The familiar yellow-fever vector Aedes aemti-, which is easy to

colonize in the laboratory, was used in the flight-range studies. Dr. Bugher

thought it would be helpful to see if the tracer technique could be applied

to the forest mosquitoes which breed in the tree holes, such as Aedes

africanus, the vector of jungle yellow fever in Central Africa. Accordingly,

the strontium compound was added to the water of the tree hole, and later the

adult mosquitoes resulting from larvae in this water were easily recognized

by the Geiger-M'&ler counter.  Bg treating a considerable number of tree

holes in a restricted area, flight-range and longevity studies may be made

of all the tree-hole breeders, with the added advantage that the observations

are made under natural conditions.

     In the course of identifying the mosquitoes, it was noticed that

the females were twice as radioactive as the males. This led Dr. Bugher to

test various parts of the insect's body, to see if the radioactive elements

were more concentrated in some organs than in others. He found that the

larger size of the females accounted in part for their greater absoqtion of

phosphorus, but even more important was their larger mass of malpighfantubes,


28

This is an abdominal organ which functions somewhat as kidney, and it has a

high capacity for absorbing the radioactive material.

     When the dissections of different species were compared, some

striking contrasts showed up.  *In Aedes aemti we found that about 25 per

cent of the radioactive strontium went into the legs and about 30 per cent

into the malpighian tubes, whereas in the malaria mosquito Anonheles gambia

only 4 per cent was found in the legs and 70 per cent in the malpighian

tubes.  These contrasts can only mean that there are mar'ked physiological

differences between the two species,s said Dr. Bugher.

     `IIt is also significant ,fl he continued, "that the newly-emerged

mosquito, just changed from pupa to winged insect, has very little of the

strontium in the legs, but 24 hours later the legs have acquired their full

quota.  This points to the fact that the series of profound changes which go

on in the pupa do not produce a complete adult.  Internal changes are still

happening during the `drying outt process."

     Thus, the application of the radioactive tracer technique to mos-

quito marking, for the purpose of determining flight range, has opened the

door to a new way of investigating mosquito physiology - a subject that

Dr. Bugher hopes to pursue more fully.