As reported previously, ⁵⁾ Promin injected intraveneously in the aural vein of the rabbit was recognized to present in the urine and plasma of the animal as N-glucosides of DDS. Similar investigations were made on the urine of both normal subject and Promin resistant patients.
Likewise in rabbits, N-glucosides were detected as main metabolites without any appreciable difference on the urine of resistant patients. Further reports discuss the mode of the acquisition of Promin resistance.

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The selection of laproma for test material of sufficiently active bacilli and a revision of Ito and Sonoda' s collection method for a purer form of the bacillary emulsion were studied for the experiments of the metabolic investigation of murine leprosy bacillus.
The results are summarized below.
1) Leproma in the rat produced 2-3 months after the intratesticular inoculation is most suitable for collecting fresh, active and viable bacilli for metabolic experiments on the marine leprosy bacillus.
2) For testing the purity of the bacillary collection, tissue components without affinity for methylene blue should be detected by dyeing with Nile blue, while some tissue components not stained with Nile blue should be demonstrated by determining of adenosine monophosphatase activity.
3) The Nile blue staining tissue debris in the bacillary fraction can be removed by repeated washing and low speed centrifugation, while the minute particles which show adenosine monophosphatase activity can be taken away be treating with 1/8 N NaOH.

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A number of screening tests concerning various drugs for murine leprosy have been performed by the authors. The present report deal with several drugs since 1959 up to this day.
1) Ciba 1906 was unable to suppress the onset of the disease.
2) Griseofulvin (7-chloro-4. 6. 2'-trimethoxy-6', -methyl-gris-2, -en-3. 4'-dione) indicated no suppressing nor healing effect.
3) Zygomycin A showed no effect by oral administration, but brought up a sign of slight suppression by subcutaneous injection.
4) P-Ethoxyphenyl-methyl-dithiocarbamate (No.255 TAKEDA) and P-Ethoxyphenyl-benzyl-dithioc-arbamate (No.258 TAKEDA) were ineffective, either administered separately or combined with DDS.

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To establish the best method of the preparation, standardization and preservation of lepromin, comparative studies on the skin reactions in leprosy patients were commenced using the following lepromin samples:
1) In the first experiment, two lepromins mady by different methods (Mw, by Wade in 1956 and M8, by the Mitsuda-Hayashi's method in 1958) were compared with each other. With respect to the early reaction, the two lepromins gave no significant difference. The Mw, however, elicited weaker late reactions than the M8. The results of bacillary counting of the Mw and M8 were 150 and 220 millions per ml, respectively.
2) To ascertain the effect of heat on the potency of lepromin, fresh nodules from leprosy patients were homogenized and divided into two parts, the one was autoclaved to make the Wade's lepromin (Ma) and the other was boiled with saline to make the Mitsuda-Hayashi's (Mb). The bacillary counting of the two lepromins gave nearly equal results, 276 millions per ml in Ma and 289 in Mb. The Ma antigen, however, elicited stronger late reactions than the Mb, while no difference was found with respect to the early reactions. From these results and from practical considerations, lepromin used actually in Japan are always made by the Wade's method.
3) An attempt was made to preserve the standard lepromin by freezdrying in vaccuo. After the storage in ice-box during one year, the dried lepromin was resuspended in 0.5% carbol water and its potency was compared to the standard lepromin stored in ice-box during one year. No significant difference was observed between the two lepromins with respect to the early as well as late reactions

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Comparison of the potencies between Mitsuda's s antigen presented by Wade or that presented by Mabalay and Mitsuda' s antigen (Lot 13) prepared in National Institute for Leprosy Research, (Japan) were made by means of the bacillary counting and in animal test and human test. As the results obtained, no differences were found in these tests, in spite of these antigens which were made from different leproma, in different laboratory and at different calendar year. Therefore, these three Mitsuda' s antigens were not suitable as materials used for the comparison of potency tests.

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The healed patients discharged from leprosaria are increasing graduall in Japan by the remarkable developement of antileprosy drugs in this country, where leprosy patients have been isolated in leprosaria by the law. The number of discharged patients at present, however, is not sufficient because about one half of all patients in leprosaria are bacilli-negative ; for example about 70% of patients iu National Leprosarium, Kuryu-Rakusen-en. are bacilli-negarive.
Why the percentage of discharged cases in all patients is low at precent in spite of the remarkable incsease of the bacillinegative patients? This question is for the management of leprosaria in future. As a result, nine factors which influenced upon the rehabilitation of leprosy patients were obtained, and a kind of the tetative plan of forecast methods were made for rehabilitation. This plan will be investigated in other leprosaria for the practical use.

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Murine leprosy bacilli were collected with the Nakamura-Hanks' method from the subcutaneous leproma of rats on the three months after inoculation, and lyophilized.
The authors' studies were examined on the ribonucleic acid and amino acid of bacilli. The findings to be described were as follows :
1) The ribonucleic acid was extracted with the phenol method and fractionated. The r-RNA, represented for the constituent to alternate rapidly, was for the most part and the p-A-RNA (s-RNA ?), persumably taking part to protein synthesis, was for scanty part : thus, the p-A : r value was such a low value as 1: 5. From these findings it may be suggested the murine leprosy bacilli were standing in rather resting phase concerning to the cell growth.
2) The total free amino acids of leprosy bacilli, the source of protein biosynthesis, was such a small quantity as to be less than two times of the total protein-amino acids, and in the former the amino acid such as glycine, arginine, leucine, methionine and cystine were less than in the latter.
The authors' idea from the evidence at hands seems to suggest the slinghtness of the ability of protein biosynthesis of murine leprosy bacilli, namely, it may be called almost impossible of rapid multiplication of murine leprosy bacilli.

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The C-reactive protein (CRP) test was applied to the sera of 291 leprosy patients, among which. 250 were of the lepromatous and 41 of the tuberculoid type. CRP tests in the 84 lepromatous cases which were in the state of Erythema Nodosum Leprosum (ENL), showed 72 cases (85.7%) to be positive and 12 (14.3%) negative. On the contrary, no CRP was found in the sera of 27 cases which were in the state of lepromatous exacervation and of 4 cases of "Epithelioid Reaction" (Hirako), i. e. the other type of lepra reactions including "Pseudoexacervation" (Souza-Lima) and "Acute Infiltration" (Tajiri). Only two cases (1.5%) were positive in residual 135 leproma tous patients who were not in the reactional state at the test, though many of the cases hadd experienced ENL. Of the 41 tuberculoid cases, only one (2.4%) was positive and 40 (97.6%) were negative.
CRP was abscent in sera of ENL patients whose symptoms were relatively light' while always present in the severe cases. Repeated CRP tests in certain ENL patients showed that CRP appeared in serum within a few days after the onset of ENL and disappeared at the arrested stage. These findings suggest that the CRP test will be useful for the differentiation of ENL from the other lepra reactions and as an indication for cortisone treatment of ENL.

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Regarding the effectiveness of the treatment of ENL (Erythema nodosum leprosum) with Vitamin C (1-ascorbic acid) injection, one of the authors had presented a paper at the 14th Japanese Leprosy Conference in 1940.
The present authors studied again on the same subject with a large dose of the preparation which was administered intravenously with considerable good effects. The results are summarized as follows :
1. The number of leprosy cases with ENL under this treatment amounted to 11 (male 5, female 6).
2. The preparation of Vitamin C employed was Vitacimin (TAKEDA): 1cc. (100mg) and 2cc. (500mg) of 1-ascorbic acid solution in each ampule for intravenous use.
3. The administration of the drug was repeated every day as a daily dose of 100mg, 200mg or 500mg.
4. The total effectivity rate was 73% (8/11).
5. Discolouration of the erythema occurred in 2 to 6 days (4 days on an average), the total amount of ascorbic acid administered being 200 to 3000mg (1328mg on an average).
6. Complete disappearance of the erythema was seen in 4 to 15 days (10 days on an average), the amount of ascorbic acid injected being 660 to 7500mg (3120mg on an average).
7. No side effects were encountered. Furthermore, even in a case with ENL, which brought about mental disorder following cortisone treatment, the Vitamin C therapy was well tolerated and gave a remarkably favorable effect in the treatment of ENL.

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