Barbara J. Hawgood

Doctor Albert Calmette 1863-1933: founder of antivenomous serotherapy and of antituberculous BCG vaccination.

In 1891 in Saigon (now Ho Chi Minh City), Dr. Albert Calmette established the first daughter Pasteur Institute for the protection of the local population against rabies and smallpox. Inspired by the discovery of diphtheria antitoxin by Behring, Calmette studied ways of raising serum against cobra venom. In 1895, now in Lille at the second daughter institute that he established, Calmette produced anticobra serum for therapeutic use that was to revolutionize the treatment of snakebite worldwide. The incidence of tuberculosis in the working class of the industrial north shocked Calmette. In response, firstly he organized an antituberculous dispensary to provide assistance to the sick and help limit the spread of the disease by improving social hygiene and secondly he devoted himself, with the assistance of Camille Guérin, to obtaining an attenuated live strain of tubercle bacilli with fixed biological characteristics for use as a vaccine. Such a strain developed during repeated passage of a culture of Mycobacterium bovis grown on a bile potato medium. In 1919, Dr. Albert Calmette took up the appointment of Sub-Director of the Pasteur Institute of Paris. Prolonged trials of BCG (Bacille Calmette-Guérin) vaccine showed it to be safe and vaccination of very young infants born of tuberculous mothers commenced in 1921. The use of BCG vaccine as a prophylactic against tuberculosis spread world-wide and has remained important in combatting this scourge.

Pioneers of anti-venomous serotherapy : Dr Vital Brazil (1865-1950)

Dr Vital Brazil was a great humanitarian and pioneer of medical science. His main work arose from his concern with poisonous snakebite accidents to labourers working the land. Vital Brazil estimated that, at the beginning of this century, deaths due to crotaline snakebites in the State of São Paulo, Brazil, were nearly 3000 per year, representing a mortality rate of about 25%, the majority being due to bothropic envenomation. After reading a report of Calmettes anti-Naja serum, Vital Brazil raised monovalent serum against the venom of Bothrops jararaca and the venom of Crotalus durissus terrificus. In 1989 this led to the first demonstration of the specificity of anti-venomous serum and later, the first production of polyvalent serum for therapeutic use. As Director of the newly founded Institute Butantan in São Paulo, Vital Brazil was actively engaged in every aspect of serotherapeutic treatment. This included organizing a unique system of exchanging anti-ophidic serum for snakes as well as a wide-ranging teaching programme. His many outstanding contributions to the fields of immunology, public health, toxinology and herpetology required not only a very high level of observational, deductive and practical ability but also an unswerving vision and sense of duty; this was allied to great administrative skill and exceptional energy.

Assessment of the antiarrhythmic activity of nicorandil during myocardial ischemia and reperfusion

The potential pro- and antiarrhythmic effects of nicorandil (1-100 microM) were assessed in isolated rat hearts subjected to coronary artery ligation and reperfusion under conditions of normal (5.9 mM) and lowered (3.2 mM) perfusate K+. Nicorandil dose dependently increased coronary flow, induced a moderate negative inotropic effect but had no chronotropic effects. During ligation (15 min), only high concentrations of nicorandil (50 and 100 microM) significantly reduced the incidence of ventricular premature beats and ventricular tachycardia in normal perfusate, but ventricular fibrillation was observed in 2/9 hearts. No antiarrhythmic effects were observed with hypokalemic conditions. During reperfusion, nicorandil was associated with a more rapid degeneration into ventricular fibrillation in normal perfusate while the incidence of ventricular fibrillation was only reduced by 100 microM nicorandil. No antiarrhythmic effects were observed during reperfusion with lowered K+ and all drug-treated hearts demonstrated irreversible ventricular fibrillation. Nicorandil perfusion (50 microM; 5.9 mM K+) did not affect the depression of ATP or elevation of lactate within the ischemic tissue during coronary artery ligation. These data do not support an effect of nicorandil against ischemia- or reperfusion-induced arrhythmias in the intact heart in vitro and may suggest a proarrhythmic effect particularly at lowered K+ concentrations.

Abbé Felice Fontana (1730-1805): founder of modern toxinology.

One of the great experimentalists in toxinology, Felice Fontana, worked in Tuscany on the venom of the European viper; his classic text Traité sur le Vénin de la Vipère was published in 1781. Fontana identified fundamental questions which he proceeded to test experimentally using as quantitative an approach as possible within the technical confines of the mid to late eighteenth century. Amongst his findings are the discoveries that the constituents of viper venom can be precipitated by alcohol, that the action of the venom is myotoxic and that it produces not only coagulation of blood, but paradoxically, fluidity as well. After more than 200 years, it is time to honour his name and recall his work once more.

Alexandre Yersin (1863–1943): discoverer of the plague bacillus, explorer and agronomist

Alexandre Yersin was born in French Switzerland and later took French nationality. While a medical student he worked in Paris with Emile Roux to discover the exotoxin produced by the diphtheria bacillus. Two years after graduation, he left Paris for French Indochina where he was the first European to explore and map the central highlands of Vietnam. As a member of the French Colonial Health Service he was sent to Hong Kong in 1894 to investigate the outbreak of bubonic plague. He isolated from buboes the causative bacillus that later was named Yersinia pestis in his honour. In Vietnam, Yersin established a Pasteur Institute at the coastal village of Nha Trang where he lived for the rest of his life. He developed vaccines and antisera for both men and animals and, as an agronomist, he introduced the Brazilian rubber tree and Peruvian cinchona tree (for quinine) into the country. In Vietnam, as in France, the name of Yersin continues to be venerated.

Albert Calmette (1863–1933) and Camille Guérin (1872–1961): the C and G of BCG vaccine

Early in his medical career, Albert Calmette showed a remarkable aptitude for bacteriology and, in 1891, he opened the first daughter Pasteur Institute in Saigon, French Indochina at the request of Louis Pasteur. In 1894, at the Pasteur Institute in Paris, Calmette succeeded in developing an antiserum to cobra venom and so initiated antivenomous serotherapy. In 1895 Calmette was asked to found a second daughter, Pasteur Institute in Lille. Soon he was joined by the young veterinarian, Camille Guérin, and so began a unique partnership, the two men striving to free people from the dreadful scourge of tuberculosis. Investigating the intestinal route of tuberculous infection, Calmette and Guérin began to grow Mycobacterium tuberculosis bovis in a beef bile-glycerine medium. With continuous replanting of the culture in this medium (from 70 to 235 times), an attenuated bacillus of fixed properties was discovered; this Calmette called Bacille-Calmette-Guérin (BCG). Exhaustive testing of BCG showed its safety and effectiveness in protecting young animals against tuberculosis. In 1924 vaccination of newborn infants with BCG began in France and spread worldwide.

Early induction by crotoxin of biphasic frequency changes and giant miniature endplate potentials in frog muscle.

1 Following the addition of crotoxin (250 nm) at the frog neuromuscular junction, there was an initial fall in frequency of miniature endplate potentials (m.e.p.ps), followed by a secondary rise which was characterized by the appearance of large spontaneous potentials (giants, g.m.e.p.ps) and an occasional large potential of the burst type. 2 In the presence of 2‐(4‐phenylpiperidino)cyclohexanol (AH5183, vesicamol), an inhibitor of vesicular acetylcholine uptake, the frequency of g.m.e.p.ps induced by crotoxin was reduced. 3 The characteristic changes in m.e.p.p. frequency and amplitude distribution were absent with crotoxin in Sr‐EGTA Ringer. In the presence of high concentrations of Mn (3.6 or 5.4 mM with 0.9 mM Ca), the crotoxin‐induced initial fall and the onset of the secondary rise in m.e.p.p. and g.m.e.p.p. frequencies were slower. The timing of these phases was unaffected by Ca concentrations ranging from 6.3 to 0.9 mM. 4 High concentrations of Mn ions partially inhibited the phospholipase A2 activity of crotoxin on artificial phospholipid membranes. This also supports the involvement of the Ca‐dependent phospholipase A2 subunit in both phases of the physiological action of the toxin. 5 G.m.e.p.ps were associated with a moderate increase in m.e.p.p. frequency (2–3 s−1) and were of a time‐course similar to that of m.e.p.ps. They persisted after washing with medium lacking Ca ions and in the presence of Ca‐Mn Ringer that blocked evoked responses. 6 It is concluded that crotoxin, acting through its phospholipase A2 subunit, produces specific disturbances of synaptic exocytosis and vesicle formation in the axolemma of the motor nerve terminal which lead to biphasic changes in m.e.p.p. frequency and the onset of large spontaneous potentials.

Maurício Rocha E Silva MD: Snake venom, bradykinin and the rise of autopharmacology

Maurício Rocha e Silva is well known as the discoverer of bradykinin, the powerful hypotensive and smooth muscle stimulating polypeptide which was first detected in plasma following the addition of Bothrops jararaca venom. The discovery in São Paulo, Brazil, in 1948 was the outcome of studies on proteolytic enzymes that Rocha e Silva started in 1939 at a time when circulatory shock was considered to be mediated by histamine. This line of research was prompted by the publications of Feldberg and Kellaway which identified the release of histamine and a slow-reacting substance (SRC-C) from isolated lungs perfused with Naja venom. Rocha e Silva was interested in determining whether trypsin-like enzymes, as shown for phospholipase, had a role in the release of histamine in shock. Instead, he and his co-workers demonstrated that such an enzyme released a new autopharmacological principle, bradykinin, from a plasma globulin precursor. Studies by Ferreira and Rocha e Silva on ways of blocking plasma kinin-destroying activity led Ferreira to isolate bradykinin-potentiating peptides from B. jararaca venom. These peptides were later shown to block angiotensin-I converting enzyme and so have an effect on hypertension. The discovery of bradykinin has led to a new understanding of many physiological and pathological phenomena including circulatory shock induced by venoms and toxins.

Sir Charles James Martin MB FRS: Australian serpents and Indian plague, one-hundred years ago

In 1891 as Demonstrator in Physiology at the University of Sydney, Charles Martin began the first systematic study of the chemical and physiological properties of the venoms of the Australian elapid species, Pseudechis porphyriacus and Notechis scutatus. Two major constituents were detected: a large coagulable protein which was associated with intravascular clotting, and a small proteinaceous molecule, an albumose, associated with neurotoxicity. Martin designed and constructed a high-pressure gelatin membrane ultrafilter for fractionation of venom. His studies indicated that certain physiological actions and clinical symptoms were related to the faster rate of diffusion within the tissue space of a neurotoxic constituent relative to a clotting constituent. Extending this work to toxin-antitoxin relationships, Martin provided evidence that antitoxin was a large molecule with slow diffusibility in tissue and advised the administration of curative serum (including diphtheria antitoxin) by intravenous injection. In 1903, Martin returned to London as Director of the Lister Institute of Preventive Medicine. He was soon involved in the planning of scientific work to be undertaken by the Commission for Investigation of Plague in India as the disease continued to ravage the subcontinent. Detailed epidemiological studies of possible factors involved in the spread of Pasteurella pestis showed, unequivocally, that infected rat fleas were the vector of transmission from rats to humans.

Karl Heinrich Slotta (1895-1987) biochemist : snakes, pregnancy and coffee

In 1923 Karl H. Slotta obtained his PhD in chemistry from the University of Breslau, Germany, where he continued to work. At the instigation of the gynaecologist Ludwig Fraenkel, Slotta made the first isolation of progesterone in 1933. In 1934 he proposed the correct structural formula. Slotta was appointed professor of chemistry in 1935, but with the oppression of the Nazi regime mounting, he soon left Germany with his family to take a post at the Instituto Butantan, Brazil. Initially he worked on the chemistry of coffee. In 1938 Slotta and his brother-in-law Heinz Fraenkel-Conrat isolated crotoxin from Crotalus durissus terrificus venom, the first snake toxin to be obtained in crystalline form. They had evidence to suggest that the toxicity of crotoxin was due to a phospholipolytic action on nerve lipids. In 1938 Slottas department was closed; he subsequently cofounded a biopharmaceutical company. In 1956 Slotta was appointed research professor of biochemistry at the University of Miami, USA. Slotta purified the most basic polypeptide from Naja naja venom, known as direct lytic factor, and with James Vick identified this as cardiotoxin. Karl H. Slotta will be remembered not only for his skill as a biochemist but also for his indomitable and cheerful spirit.