Milton Gjelhaug Levine

Mucopolysaccharides in Tissue Cultures of Human and Mammalian Synovial Membrane.

Summary 1. Growth of human and animal synovial membrane in tissue cultures showed characteristic differences from control cultures of periarticular tissues. 2. Presence of hyaluronic acid in about two-thirds of supernates of tissue cultures from synovium was demonstrated by precipitation reactions with acetic acid, enzymatic reactions, viscosity determination, tests for metachromasia and electrophoretic patterns. 3. Quantitative determinations by a turbidimetric method yielded a concentration of hyaluronic acid from 4 to 15 mg %. 4. Tests were negative for sulfonated mucopolysaccharides such as chondroitin sulfuric acid or heparin. 5. Control cultures of periarticular connective tissue were negative for hyaluronic acid and sulfonated mucopolysaccharides.

Temperature Reactions in Mice Infected with Pneumococci.

Summary The body temperature of mice is markedly influenced by the surrounding temperature. Pneumococcic infections in mice are associated with a marked drop in temperature. Infected mice held at incubator temperature succumb more rapidly than those at room temperature. Incubator temperature promotes the infective processes in mice more than the protective action of sulfanilamide.

Scleroma; complement fixation test.

Conclusion A complement-fixation test for scleroma is described which offers evidence for the relationship between an organism described and the disease. The value of the test is indicated in diagnosing scleroma.

Further studies on relationship between human serum cholinesterase and serum albumin.

Conclusion The previously observed (1,2) correlation of serum albumin and serum cholinesterase values is prlobably due to the simultaneous production of these two proteins by related synthesizing mechanisms in the liver. Albumin does not act as a precursor of serum cholinesterase. Changes in the level of this enzyme reflect the changing ability of the liver to produce both serum cholinesterase and serum albumin. It is suggested that serum cholinesterase determinations are of prognostic value in liver disease and that cholinesterase determinations may be used as an indication of albumin synthesis, particularly in the presence of administered albumin.

The Nature of Rabbit Serum Cholinesterase.

Conclusions Rabbit serum, contrary to previous observations, contains only one enzyme which hydrolyzes acetylcholine, and this one has the characteristics of a true cholinesterase. In addition, there is present in rabbit serum an enzyme which hydrolyzes only benzoylcholine, and which, since it does not hydrolyze acetylcholine significantly, should not be labeled a cholinesterase. This enzyme may be similar to the “benzQylcholin-esterase” described by Sawyer 10 as being present in guinea pig liver and kidney.

Inhibitory Action of Peptone on Sulfapyridine Adsorption.

The bacteriostatic effect of sulfapyridin on the pneumococcus in vitro and the inhibitory action of peptone have been shown previously. 1 The findings compare with those reported by Lockwood, 2 who studied the effect of sulfanilamide on the streptococcus under similar conditions. The effect of peptone in preventing drug-action suggested the possibility of an interference in adsorption of the drug. To test this, a study was made of the adsorption of sulfapyridine by activated carbon particles. Varying amounts of activated carbon were added to solutions containing 10 mg of sulfapyridine and 0.85 g of sodium chloride per 100 cc. After allowing the reaction to take place for 15 minutes, the carbon was removed by filtration, and the filtrate was tested for the presence of the drug by the method described by Marshall. 3 Adsorption was found to occur, as shown in Fig. 1 (solid line). If 1% peptone (Parke-Davis) is added to the solution containing the drug, and adsorption by carbon allowed to take place, the removal of the drug is retarded. This is shown in Fig. 1 by the broken line. That this adsorption is selective is illustrated by the following experiment: 2 cc of a 1:1000 solution of sulfapyridine were added to 8 cc of saline, and adsorbed with 20 mg of carbon. After 15 minutes, the mixture was divided into 2 parts; to one, 5 cc of 0.85% saline was added, and to the other 0.85% saline containing 1% peptone. These were allowed to stand for another 15 minutes, when they were filtered and tested for sulfapyridine. The portion to which saline had been added showed adsorption of 73% of the drug, as compared with a removal of 55% in the portion to which peptone had been added. This suggests that peptone is able to displace the drug from the surface of the carbon particles.

Protective Action of Sulfapyridine in Rabbits Infected with Pneumococci.

Most of the studies on the protective action of sulfanilamide and sulfapyridine on streptococcic and pneumococcic infections have been made on white mice. In the work here reported rabbits were used as the experimental animals. Intracutaneous inoculations were made in order to permit the observation of differences in the local lesions occurring in the treated and the control groups. Rabbits weighing approximately 2 kilos were given 0.3 cc of a 1-100 dilution of an 8-hour culture of a Type II pneumococcus. The strain used had been transferred alternately through mice and veal broth for more than a year. Intraperitoneal injections of 0.2 cc into mice, in dilutions of 1:10,000,000, kills 50% of the animals. Sulfapyridine, 2-(sulfanilamide)-pyridine, was administered orally by suspending in 10% acacia and permitting the suspension to trickle down the throat from a large syringe. The treated animals were given 0.5 g one hour before inoculation, 0.5 g 3 hours after inoculation, and 0.5 g every 12 hours thereafter for a total of 5 g. In addition to the difference in mortality, there was observed a marked difference in the lesions produced in the treated and control animals. Rhoads and Goodner 1 have reported oedema and a spread of the cutaneous lesion by gravity in rabbits inoculated endermally with pneumococci. These results were evident in our control group. The treated animals showed little or no oedema, and probably because of this, showed little or no spread by gravity. That an infection was present in the skin was evidenced by the area of inflammation in the skin of treated animals. The above authors have also stated that in occasional animals they find signs of hemorrhage in the skin. In the lesions of our control series there were extensive hemorrhages, caused either by the virulence of our strain or by the age of the culture.

Active Immunity to Pneumococci in Rabbits Protected by Sulfapyridine.

The results of experiments on the protection of rabbits, infected with pneumococci, by oral administration of sulfapyridine have been reported previously. 1 The purpose of this communication is to present data obtained in the study of the immunity in animals that had recovered from the infection produced under such circumstances. Rabbits weighing approximately 2 kilos were inoculated intracutaneously with 0.3 cc of a 1/100 dilution of an 8-hour culture of a pneumococcus type II. The inoculum contained an average of 300,000 organisms as determined by plate-count. The virulence of the organism is such that an average of 1.5 organisms are necessary to cause an infection in 66% of a statistically sized group of mice weighing 20 g. This we have taken to be our minimal lethal dose. Its virulence for rabbits is such that 0.3 cc of a 1/1000 dilution of the original culture, containing 30,000 organisms, will kill all of the rabbits inoculated. The drug-treated animals were given sulfapyridine orally in a 10% acacia suspension. Each animal received a total of 5 g of the drug, 0.5 g 1 hour before infection, 0.5 g 3 hours after the initial dose, and 0.5 g every 12 hours thereafter until the full dosage was administered. Daily record was made of the temperature, and the size and nature of the skin-lesion. Rabbits recovering from the primary infection were reinoculated intracutaneously in the same general areas as previously, but not at the site of the original lesion if a scar had persisted. Only normal areas of the skin were used for reinoculation. Type-specific Immunity. Twelve rabbits reinoculated intracutaneously with 0.3 cc of the whole 8-hour culture of the homologous type II pneumococcus 30 days after the original infection, showed no rise in temperature, and no local reaction around the point of inoculation.

Protective Action of Sulfapyridine Against Type II Pneumococcal Infections in Mice.

Summary With subcutaneous inoculations of 4000 to 8000 average lethal doses of a Type II pneumococcus in mice, the survival rates at both 30 and 60 days were (1) with 0.5% sulfapyridine in the food, 44%, and (2) with 1.0% sulfapyridine in the food, 63.4%. It is believed that the slight variations in drug-intake from day to day are more than counterbalanced by a more or less continuous drug-absorption from ingested food plus drug.