Penicillin as a paradigm for

medical research
ex
THEORIES
AND
EXPLANATIONS:
Do theories explain anything
in clinical medicine?
NDCL
S
St John’s College Oxford
Formal Hall and
scruffy students
St John’s College
Oxford
Research Centre
But to return to
the theme
on “Theories
and
explanations
in various
disciplines-
 But first…

What is a theory?
 The noun
"theory"
has 3 senses
in WordNet.
(web dictionary for those too lazy to
visit the library!)
1. theory -- (a well-substantiated
explanation of some aspect of the
natural world; an organized system of
accepted knowledge that applies in a
variety of circumstances to explain a
specific set of phenomena; "theories
can incorporate facts and laws and
tested hypotheses"; "true in fact and
theory")
2. theory -- hypothesis, possibility, (a
tentative theory about the natural
world; a concept that is not yet verified
but that if true would explain certain
facts or phenomena; "a scientific
hypothesis that survives experimental
testing becomes a scientific theory";
"he proposed a fresh theory of alkalis
that later was accepted in chemical
practices")
3. theory -- (a belief that can guide
behavior; "the architect has a theory
that more is less"; "they killed him
on the theory that dead men tell no
tales")
 In summary

•An explanation
•A possibility
or
•a belief
"When I use a word," Humpty-Dumpty said, in
rather a scornful tone, "it means just what I
choose it to mean -- neither more nor less."

So what have others

considered
theory to be
 Theory becomes a
spiritual force when it is
gripped by the masses

Karl Marx “ Critique of Hegel’s Philosophy of Right

"A hypothesis or theory is clear,
decisive, and positive, but it is
believed by no one but the man who
created it. Experimental findings, on
the other hand, are messy, inexact
things, which are believed by
everyone except the man who did
that work."    — Harlow Shapley
(1885-1972).
 The great tragedy of Science
—the slaying of a beautiful
hypothesis by an ugly fact.

Thomas Huxley (1825-1895)

"The most exciting phrase to
hear in science, the one that
heralds new discoveries, is
not 'Eureka!' ('I found it!') but
rather 'hmm....that's
funny...'"    — Isaac Asimov.
 And finally…
Why think?  
Why not try the
experiment?

John Hunter (letter to Edward Jenner)

Some major achievements in modern
medicine
•Asepsis
•Anaesthesia
•Pasteur’s “germ theory”
•X rays
•Aspirin
•Penicillin
•CT/MRI
•Monoclonal Antibodies
•Human genome project
Let’s take Penicillin as a paradigm for
looking at the mechanics of medical
research

Norman Heatley 1911-2004

A look at the story of Penicillin in a little
more detail.

Today, the use of penicillin and other antibiotics

is common place. It is something that we all take
for granted, today. However, many diseases and
simple wounds that are so easily treated today
because of the availability of antibiotics has not
always been available. Antibiotics are a relatively
recent discovery and the first practical one,
penicillin, was not available until the early 1940s.
Is the concept (theory) of using fungal
products new?
c1000BC- Chinese put mouldy
soybean curd on boils.
Other cultures have used warm earth,
which contains moulds and other
fungi, as first aid in injuries. So,
although the concept of antibiotics is
relatively recent, its use has been
around for some time.
Theory Experiment
The discovery of
penicillin has often
been described as
a miracle…
The usual story is that penicillin was
discovered accidentally, at St. Mary's
Hospital, in London, by Dr.Alexander
Fleming who noticed that the growth
ofStaphylococcus aureus,
contaminated by a
species of
Penicillium was
inhibited. Fleming
wrote a paper on
his findings in
1929 and the rest
is history.
was it that simple?
did Fleming discover
penicillin by chance?
are any others deserving
of credit?
Luck was involved at several
points -Alexander Fleming did
not intend to be a doctor.
1900 Boer War (LSR-shooting,
swimming and water polo).
1901- Uncle John died and left
him and his brothers with a
sizeable inheritance (£250
each).
His older brother, Tom, advised him
to invest his money on his career
and suggested that he attend
medical school. Fleming selected St.
Mary's Hospital, in London, only
because he had once played water
polo against them. After graduation,
he had the option of leaving St.
Mary's to be a surgeon or he could
join the Inoculation Service and stay
at St. Mary's. The major influence
was St. Mary's rifle club.
St Mary’s c1900
St Mary’s c1900
That Alexander Fleming discovered
penicillin by chance is a myth. Before
Fleming, there were a series of
observations that influenced his research
and allowed him to come to the correct
conclusion when a chance contamination
in his bacterial culture was observed.
This same opportunity came to others as
well but their only response to the
contamination was that it had ruined their
experiment and they had discarded the
cultures and thought nothing more of it.
Discoveries in science are rarely
made by chance.

"stood on the shoulders of giants.”

Einstein on Newton

1500 B.C. moulds and fermented

materials in the treatment of
diseases.
C19. Pasteur Germ Theory of Disease,
i.e., diseases were caused by
microorganisms.
This led to the search for "The Magic
Bullet".

The most common problem was the

contamination of bacterial cultures
by other microorganisms especially
fungi.

These contaminations led to a

number of observations in the late
1800s:
1871 Joseph Lister -urine samples
contaminated with mould did not
allow the growth of bacteria.

1874 William Roberts -bacterial

contamination is generally absent in
cultures of the mould Penicillium
glaucum.

1877 Louis Pasteur & Jules Francois

Joubert anthrax contaminated with
unidentified moulds was inhibited.
1897 Ernest Duchesne M.D. observed
that E.Coli was eliminated
by penicillin when both
were grown on the same
culture and in inoculation
of laboratory animals
prevented the animal from
contracting typhoid.
Although, he urged that further research be
carried out, he was unable to do so. Upon
completing his degree, he went into the army
and while serving, died of tuberculosis.

1874-1912
1920's Andre Gratia and Sara
Dath (Brussels)
fungal contamination in one of
their Staphylococcus aureus
cultures was inhibiting the growth
of the bacterium.

They identified this as a species

of Penicillium and presented their
observations as a paper.
GRATIA, A. and S. DATH, 1925  Propriétés
bacteriolytiques de certaines moisissures. C. R. Soc. Biol.
91:1442-1443. 92: 461–462 .
There was little attention paid to this paper and the two scientists did
not pursue this line of research further. missed being a major part of
the penicillin story in not pursuing this research further.
 Fleming and WW1
Lister's concept of antiseptic
surgery. Fleming argued that
antiseptics had an adverse effect by
destroying leukocytes more rapidly
than the invading bacteria. They
recommended the use of a saline
solution to cleanse wounds.
(ignored till WW2)
1922 Fleming discovered lysozyme,
("little brother" of penicillin).
Lysozymes are enzymes present in
biological substances as varied as
egg whites, tears and mucus that
causes bacteria to lyse or burst. The
first biological substance that he
tested was mucus from his nose (he
had a cold at the time).
This would later be a major discovery, but at the time,
though interesting there was no obvious clinical
application.
1928 staphylococcal research
frequent contamination with airborne
moulds.
‘sloppy scientist’ lab in disarray.
month long vacation threw the plates
in a tray of lysol.
Retrieved for a visitor.
It was only then that he
noticed the unusual
inhibition zone around
the fungus.
Isolated and named it penicillin
Published 1929 but no interest

Failed to purify and concentrate

penicillin

unable to prove that it had any

therapeutic value and doubted it
himself at this time.
the verge of a great discovery.

"yellow magic"

P. rubra incorrect but penicillin.

Charles Thomas Penicillium notatum.

penicillin diluted 1,000 times effective
in killing bacteria

?translated to practical use

Fleming only worked on and off with

with penicillin between 1928-1931
quantity and quality problems

colleagues bored xc Howard Florey

Cecil Paine (Sheffield)
“the forgotten man” in the
development of penicillin.

Inspired by paper not

Fleming

"a shocking lecturer, the worst you could

possibly imagine-
not because of inherent shyness, but
because he never appeared enthusiastic
about his subject”
sycosis barbae (staphyloccus)

soaking gauze with the crude penicillin

extract- failure

miner’s eye Pneumococcus

baby’s eye congenital gonorrhea

irrigated the eye with the crude

penicillin extract-success
Paine never published his results
concerning these patients nor did he
ever present an oral paper on these
patients

Crude extract + new job

Fleming was still floundering

required the expertise of a chemist
Sheffield University 1932
Paine discussed his penicillin work
with a newly arrived Professor of
Pathology. The professor, said Paine
"took not the slightest interest at that
time."
Yet six years later, he was to
begin a program of research
that would lead to the mass
production of penicillin.
The Professor's name was
Howard Florey
mid 1930's
Sulfa drugs Germany
M and B May and Baker

Fleming began working with both

these drugs and had stopped
working with penicillin by 1934
 1928-1938 Howard W. Florey lysozyme
1938 penicillin
Team: Ernst B. Chain,
Leslie Falk, Norman
Heatley and twenty
other scientists and
technicians.

Where Fleming had a

poorly equipped lab
with no staff support,
Florey's lab was well
staffed and equipped.
Florey and Chain
Chain began the work with sub-
cultures from Fleming's original isolate
of P. notatum.
Florey in charge of the research
program

-on the editorial board

-knew Fleming quite well
-Paine in 1932 (Florey forgotten)
former students remembered quite distinctly that Florey
mentioned Paine's work in a lecture, at Oxford, in 1936.
Florey and Chain and scientific interest

“I became interested - immediately - in Fleming's

paper, not because I hoped to discover a
miraculous drug for the treatment of bacterial
infection which for some reason had been
overlooked, but because I thought it had great
scientific interest. In fact, if I had been working at
that time in aim-directed scientific surroundings,
say in the laboratory of a pharmaceutical firm, it is
my belief that I would never have obtained the
agreement of my bosses to proceed with my project
to work with penicillin”
(Chain)
Chain purified penicillin for two mice
injected by J.M. Barnes not Florey
The last time he requested his assistance,
Florey turned to Mrs. Margaret Jennings,
who was in the lab, and said, pointing to
Chain, "in one of my weak moments I
promised this man to test his fractions
and here he comes pestering me again."
After what Chain described as "these
humiliating remarks in front of others not
involved in the project, it became clear to
me that Florey was not really interested in
penicillin…"
fifty white mice Streptococcus
25 penicillin alive 25 no penicillin died

ready for human subjects

in 1940, on human subjects, it had

required two professors, five
graduates and ten assistants working
almost every day of the week for
several months to produce enough
penicillin to treat six patients.
Albert Alexander, a 48 year old
London policeman had nicked
himself while shaving.
sulfa drugs failed.
"purified" penicillin administered
By 5 days the patient was
recovering. Urine recovery but no
more penicillin
relapsed and died
"the treatment was a success, but the patient
died."
15 year old boy hip -success

48 year old laborer carbuncle -success

4 year old boy infected eye ?failure

following measles. Sulpha drugs were used
unsuccessfully. semi-comatose meningitis
third day of penicillin treatment swelling
down and emerging from the coma.
sudden convulsion and he died four days
later. ( burst blood vessel)
Fleming obtained Penicillin from
Florey for old friend Harry Lambert

Fleming injected the penicillin into his

friend's spine and the treatment
proved completely successful.

Florey injected penicillin into the

spines of artificially infected animals,
with the result that they had all died!
Although the relationship between Florey and Fleming
seemed to be an amiable one. It would later turn sour.
 Florey's "famous pinched
smile”always gave him a
look of caustic severity but it
allegedly hid tooth erosion
caused by his drinking dilute
hydrochloric acid prescribed
for achlorhydria

An alternative suggestion concerns

a nervous dentist pulling out the
wrong front tooth
1941: Florey&
Heatley to USA
Rockefeller
Foundation
mouldy cantaloupe
in a market in
Peoria, Illinois
(Penicillium
chrysogeum)
bed pans to 25,000
gallon metal tanks
Fleming the discoverer of penicillin,
was almost forgotten by the time
penicillin was being mass produced.

However, he would be rescued from

oblivion, in 1943, when he was
knighted along with Howard Florey,
and two years later was awarded the
Nobel Prize in Physiology and
Medicine along with Florey and Chain.
Fleming predicted that the use of
penicillin would, in time, be of limited
value because bacteria would
eventually recombine genetically to
resist the effects of penicillin. By as
early as 1952, as much as three-fifths
of all staph infections were penicillin
resistant.
So does any theory explain any of the
penicillin story possibly the greatest
medical advance in the 20c?

or was it a case of
experimental preparation
careful observation
hard work
and luck?
 Maureen and Bernie's Wine Travel Guides

"Penicillin cures, but wine makes people happy.”

-- Sir Alexander Fleming