The reliability of bacillary counting for the standardization of lepromin was examined using the undiluted standard lepromin and its 1:2, 1:4, and 1:8 dilutions. A linear relationship was demonstrated between the dilution degree and numbers of leprosy bacilli contained there. Size of the skin reactions was read in leprosy patients comparing between those elicited by the undiluted standard and by diluted antigens. The result indicated that the reaction size becomes smaller with the dilution degree of the antigen. Therefore, a parallelism between the potency of lepromin and the number of bacilli in the antigen was made clear. It is possible to settle the criterion of lepromin potency in the term of bacillary number. It was found that when lepromin is prepared by the same procedure using a pool of many leprous nodules, it contains leprosy bacilli in number from 150 million to 160 million per ml.

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The influence of tissue component upon the potency of lepromins defined by the bacillary numbers was investigated. Three samples of lepromin were prepared with newly formed leprous nodules obtained from the patients untreated or under treatment, and also with those of relapse nature removed from the patients under treatment. These lepromins were adjusted by dilution or concentration to contain the standard number of leprosy bacilli (160 millions per ml), and then the potency test was conducted comparing them on leprosy patients. No statistically significant difference of the potency was revealed among the three lepromins as far as the early reaction (Fernandez) is concerned in 157 cases of the lepromatous type and 34 cases of the non-Lepromatous type. In the late (Mitsuda) reaction, however, the lepromin prepared from newly appeared nodules in untreated patients was somewhat lower in potency than the other two lepromins. Particulary, the reactions with slight induration of less than 4mm in size were most rare among the lepromatous cases injected with this lepromin. This observation was reasonably explained by the fact that the nodules from untreated patients had a larger amount of leprosy bacilli than those of the patients under treatment, and that after adjustment to the same bacillary number the lepromin from the formers had the less amount of tissue component than those from the latters, which might be responsible for nonspecific reaction. This observation suggests that a consideration is needed to uniform the amount of tissue component in case the potency of lepromin is defined by the number of leprosy bacilli contained in it.

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When the standard lepromin was treated with the sonic vibration (9.5 kilocycle per sec.), the corse tissue fragments were dispersed in fine particles, and at the same time the number of leprosy bacilli diminished exponentially. Under the conditions of the experiment, about half of the bacilli was destructed within 5 minutes. The complement-fixing ability of the lepromin with leprous serum was, however, increased by the sonic vibration showing that the active substance was not destroyed by this treatment. Comparison of potency tests on leprosy patients gave the results that the lepromin sonic-vibrated for 30 minutes elicited weaker early reactions than the untreated standard lepromin, while there was no significant difference between them with respect to the late reactions. Consequently, it is considered that the late reaction may be caused not only by the intact leprosy bacilli but also by the mechanically destructed bacilli.

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Leprosy has many peculiarities which distinguish it from all other diseases. It is a chronic disease, the histopathological features showing inflamatory granuloma which is composed of histiocyts or macrophages and lymphocytes.
The purpose of this report is to summarize the metabolic alteration of such peculiarities of leprosy and to make them for diagnosis, and for judgments on the effectiveness of treatments of the patients.
These problems have been studied by the measurement on the following enzymes and products of the metabolism, -the glucose-6-phosphatase, phosphohexose-isomerase, aldolase, lactic dehydrogenase, lactic acid and pyruvic acid in the leprosy patients and normal healthy individuals. The following results were obtained.
1) Gross histopathologenic pictures of leprosy resemble to that of malignant tumor, but the aspect of the enzymes in the serum of leprosy is different from that of malignant tumor with disseminated metastasis and metastasis to the bones.
2) Leprosy is a disease with muscular atrophy, it is not the myogenic muscular atrophy but the neurogenic muscular atrophy from the enzymological point of view.
3) It was found that leprosy had a fairly abnormal metabolic response for glycolysis. It is believed that the condition in leprosy is due to disturbance of oxidation, caused by deficiency in oxygen of the tissues.

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The growth and distribution of Myc, lepraemurium in mice transplanted with Ehrlich ascites tumor cells and the survival of tumor transplanted mice influenced by Myc. lepraemurium inoculation were investigated. Survival time of mice transplanted with Ehrlich ascites tumor cells was prolonged by previous inoculations of Myc, lepraemurium. Slight enhancements of the growth and distribution of Myc. lepraemurium by simultaneous transplantation of Ehrlich ascites tumor cells were demonstrated.

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Myc. lepraemurium suspension was inoculated into mouse or rat testicles which were removed from the bodies. These testicles were incubated in the various kinds of medium at 37°C. At an appropriate incubation time, the testicles were periodically took out, and were homogenized with sterile distilled water. The number of the bacilli of Myc. lepraemurium which were contained in each testicle was counted on the stained smear. Among the media tested, it is seemed that the medium which contained pyruvate in Hanks solution (minus glucose) is most suitable one for the increase of the bacilli in the testicle. The maximum peak of the increase of the bacilli was demonstrated on 7th day of incubation at 37°C. However, it is not yet decided whether the increase of the bacilli is due to true multiplication of the bacilli or not.

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