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Gamma Globulin

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Gamma Globulin MAY 26, 1945 BRITISH 73 TRINITROTOLUENE MEICALMEDICAL J OURNAL 739 presence in the urine of animals fed on a-T.N.T. of throat. The symptoms came on after an initial period 2: 6 dinitro-4-hydroxylaminotoluene, 2 6 dinitro-4-amino- when there was an increased sense of well-being accom- toluene, and 2: 4 dinitro-6-aminotoluene (which may be panied by a voracious appetite. There was evidence of estimated accurately by spectrophotometric methods7) haemolysis in over 85 % of the students, with a fall in points to some reduction mechanism. T.N.T. may also haemoglobin, red cell, and haematocrit readings. There be oxidized: experiments with trinitrobenzyl alcohol in were also an increase in reticulocytes, bilirubinaemia with attempts to establish the origin of the red pigment indicate urobilinuria, and a marked erythroblastic response in the the probable existence of such a mechanism, since only marrow. As Noro found, the reticulocyte response was a-T.N.T. and 2 : 4: 6 : trinitrobenzyl alcohol would cause not maximal until a few days after exposure had ceased. excretion of red pigment. Present knowledge of detoxica- This suggests that T.N.T., in addition to a destructive action tion mechanisms suggests that the glucuronides of T.N.T. on the circulating red cells, also affects the bone marrow. urine must arise from T.N.T. oxidation products: they are probably trinitrobenzyl glucuronide,.or dinitroaminobenzyl glucuronide, or a mixture of both. The hydroxylamine GAMMA GLOBULIN derivative provides an example of the making of a more toxic product in a detoxication mechanism, and it is a The story of how human blood has been harnessed to powerful former of methaemoglobin in vitro. Since the war effort will prove to be among the most fascinating the first stage in the biological reduction of T.N.T. is the tales of our generation. In the early days of the war formation of dinitrohydroxylaminotoluene, there is pro- blood banks and improved methods for the preserva- duced a very reactive compound which may be harmful to tion of whole blood were successfully established. Then, the blood and may poison some of the enzyme systems. with the emphasis on shock, harvesting of the plasma The hydroxylamine can be detoxicated by conversion proteins became important, and large-scale methods of into the relatively non-toxic dinitroaminotoluenes, and its freezing and drying plasma and serum were evolved. poisonous effects will depend on the rate at which this Meanwhile in America before the war, but accelerated conversion occurs. Although the conversion takes place by the war, methods for the fractionation of the plasma in vivo it is by no means complete, for the hydroxylam.no proteins were being steadily developed in the department and the amino compounds are excreted in the urine in of physical chemistry at Harvard under the direction of roughly similar amounts. The formation of dinitro- Prof. E. J. Cohn. The methods themselves were revo- hydroxylaminotoluene may therefore partly explain the lutionary in their utilization of new physical and chemical toxic action of T.N.T. Doses of T.N.T. up to 150 mg./kg. processes: high-speed centrifugation, electrophoresis, diffu- are eliminated in 24 hours; larger doses require up to 48 sion, viscosity, and precipitation at iso-electric charges have hours; 47% of, the T.N.T. administered is excreted as all been used for the separation of different protein frac- glucuronides and 30% as aromatic amino compounds. The tions with the least possible damage to the constituents. aromatic amino compounds include not only dinitroamino- The story has lately been summarized by Cohn' himself, toluenes but possibly dinitroaminobenzyl alcohol conjugated while various chemical, immunological, and clinical with glucuronic acid. Trinitrobenzylglucuronide is also a aspects are described in a special symposium2 on the likely constituent. The red 2:2':6:6'-tetranitro-4':4-azoxy- subject. toluene isolated by previous workers is not a metabolic pro- Plasma has been conveniently divided into five fractions: duct; it is absent from freshly voided urine and is formed I is largely fibrinogen; II and III contain gamma globu- from dinitrohydroxylaminotoluene during the extraction lin, the iso-haemagglutinins and Rh antibodies, and pro- procedures. Rimington and Goldblattl have shown that thrombin; IV is mostly alpha and beta globulin; and workers handling aromatic nitro and amido compounds have V is albumin, which constitutes nearly 60% of the total the a greatly increased urinary coproporphyrin excretion,. and proteins. Indeed, fractionation of plasma on a large this change is also found in T.N.T. workers. scale was first undertaken in order to obtain the albumin Little clinical work has been possible on T.N.T. poison- in concentrated form for the treatment of shock. Having ing during the present War, but during the Russo-Finnish a small, fairly regular molecule, albumin contributes 80% war Noro12 observed that in Finnish T.N.T. workers of the osmotic pressure of plasma proteins and can be pre- as an 25 reticulocytosis was not very great during the period of pared iso-viscous % solution which osmotically four actual exposure, but that it became more evident when the is times as concentrated as plasma. Besides its value 4vorker left contact with T.N.T.; this suggests that reticulo- in shock, concentrated albumin may be used in the treat- ment of and its cytosis was a reparative phenomenon which is to some hypoproteinaemia, oedema, (because of low salt as a diuretic. Of the other extent inhibited by T.N.T. In order to establish the early content) proteins in a of effects of exposure to T.N.T. Higgins, O'Brien, Stewart, thrombin supplied matrix fibrin, called fibrin foam, used as a haemostatic in and Witts13 examined a number of Oxford students has been neurosurgery and in from surfaces and in who* volunteered to work in a filling factory during their controlling bleeding oozing haemo- of human vacation. The chief subjective complaints were nausea philia. Being composed entirely proteins, fibrin the minimum of with variable loss of appetite, diffuse abdominal pain, foam is absorbed with reaction. Again, worked like a to vomiting and diarrhoea, fatigue, and irritatiqo of nose and fibrinogen has been up plastic prepare fibrin- films which-, With the properties of a rubbery sheet, lLancet, 1940, 1, 73. 12Acta med. scand., 1941, Suppl. No. 120. lScience, 1945, 101, 51. 13 Brit. J. industr. Med., 1945, 2 (in the press). 2 J. clin. Invest., 1944, 23, 417 et seq. 740 MAY 26, 1945 GAMMA GLOBULIN BRITISH ' ~~~~~~~~~~~~~~~~~~~~~~~~~MEDICALJOURNAL have been used as dural substitutes and in the prevention FOOD CONTROL of meningo-cerebral adhesions. The long controversy over preservatives in food ended with Probably the most important derivative of plasma will the passing of the Public Health (Preservatives, etc., in be the gamma globulin, which contains a large part of the Food) Regulations of 1925. Many experts had advocated natural antibodies of the blood. Gamma globulin con- that the use of particular preservatives should be forbidden, stitutes about 11% of the plasma proteins, and the greater but the Ministry of Health adopted in these regulations part of this globulin has now been fractionated in practi- the much sounder plan that all preservatives (except the common types like sugar or vinegar) should be prohibited cally pure form from the combined 11+Ill fraction. The except those specifically permitted for indicated foods and concentration of antibody as judged by the estimation of only in amounts which must not be exceeded. It was never the diphtheria and streptococcus antitoxin and influenza suggested that the last word had been said, but the prin- virus A antibody is 15 to 20 times that of the natural ciple was established that nothing might be added unless pooled plasma, and its content of certain antil,odies equals specifically authorized and under defined conditions as to or exceeds that of convalescent serum. There has been the preservative to use, its quantity, and the food to be of different pools preserved. Under war conditions the conserving of food remarkable uniformity in the potency supplies became urgent, and the central authorities con- from various regions of America. Gamma globulin has cerned-i.e., the Ministries of Health and of Food-had already been used on a large scale for the prevention and to face the question of relaxing certain requirements if it attenuation of measles in familial contacts and in institu- could be shown that it favoured the conserving of more tions; analyses indicate that a dose of 2.0 to 2.5 c.cm. will food and was not harmful to the consumer. Obviously protect 80% of young, susceptible, and intimately exposed such decisions are not easy, involve the balancing of children, with marked attenuation in the remainder. Where various opposing factors, and demand an intimate know- ledge of the conditions of food preparation and preserva- attenuation only is desired the recommended dosage is tion. This need accounts for the rather bewildering around 0.5 c.cm. In other words, results equal to those number of Orders that have been issued-some allowing obtainable with convalescent measles serum have been certain relaxations, others cancelling relaxations previously achieved with one-quarter to one-half the dosage of that permitted. One of the latest is Regulation 60 CAA of rather rare commodity. Although the gamma globulin the Defence (General) Regulations, 1939, and is a good now issued cannot be used intravenously, progress has illustration of the way in which preservatives are being con- made with the removal of the depressor substance trolled. It allows oranges to be imported in wrappers been treated with diphenyl; meat and dehydrated vegetables which precludes intravenous therapy, and it seems likely may contain sulphur dioxide, and margarine and bacon that such a preparation will be useful for the treatment boraX; jam may contain more sulphur dioxide than in of early cases of measles and other infections.
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