The concentrations of c type cytochrome and the proportions of free amino acid in various organs of mice, intraperitoneally inoculated with murine leprosy bacillus were determined along the periods of time after inoculation.
1) The c type cytochrome concentrations in spleen and kidney of the infected mice were found to be higher than that of normal mice, on the date from 4th day until 5th week after inoculation.
On the 15th and 30th week after inoculation, however, the c type cytocrome concentrations in liver, spleen and indey of the infected mice, the most of which were suffered from several leproma grown in their abdomen, were found to be reduced less than 65-78% of the normal levels.
2) The following changes in the proportions of free amino acid in liver, spleen and kidney of the infected mice were observed:
On the 3rd week after inoculation, spleens of the almost inoculated mice were enlarged extremely, and their contents of Glu., Thr. and Cys. increased, while Gly., Ala. and Leudecreased.
The content of Cys. in liver of the infected mice on the 5-15th week after inoculation were found to decrease. On the 15th and 30th week, the content of Glu. in the infected liver were found to increase, while the content of Ala. decreased.
According to the results as mentioned above, especially, the remarkable changes in free amino acid proportions of Glu., Ala., Gly. and Ser. in the organs of infected mice, the occurence of disturbances concerning lipid and carbohydrate metabolism as well as protein metabolism could be assumed in the mouse body infected with murine leprosy bacillus.

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The study was carried out in order to observe if M. lepraemurium can grow extracellularly in mouse peritoneal cavity and if this microorganism can grow in the peritoneal macrophage of mouse and guinea pig, and to know the growth pattern of non-pathogenic mycobacteria in vivo.
The bacillary suspension of M lepraemurium Hawaii strain, M. smegmatis ATCC No.14468, Mycobacterium species Jucho strain, M. species Dencho strain, and M. species Takeo strain was prepared and mixed respectively with equivalent volume of 4% Special Agar Noble solution. Small agar block containing mycobacteria was prepared and inserted into peritoneal cavity of mouse and guinea pig.
Growth of M. lepraemurium was checked by stamp-smear of agar block, and non-pathogenic mycobacteria was obseved by stamp-smear and viable unit counting on Ogawa's medium.
M. lepraemurium showed intracellular multiplication in mouse peritoneal cavity at 2 months after the insertion, and at 3 months after the insertion, numerous bacilli were observed intracellularly and extracellularly. However, no multiplication was observed when the agar blocks were inserted into guinea pig's peritoneal cavity.
Non-pathogenic mycobacteria showed multiplication in mouse peritoneal cavity in a limited period after the insertion, and M. smegmatis showed logarithmic growth until 2 days after the insertion.

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Nakamura has analyzed the cellular components of M. lepraemurium and reported that a large quantity of lipids but only a small amount of nucleic acid were present. Since an Ascaris lives in the intestines where oxgen pressure is very low, an Ascaris stores special kinds of fatty acids as the result of incomplete combustion of its nutrients to produce the energy. It has been also shown that M. lepraemurium has an irregular TCA cycle for carbohydrate metabolism and proliferates in the subcutaneous node under low oxygen pressure, lipids may be stored in the cell as the result of incomplete combustion of its nutrients to produce the energy. A comparative study of the lipid content of M. lepraemurium and other acid-fast organisms which can be cultured, together with their DNA and RNA contents were carried out.
The lipid contents of acid-fast organisms fluctuate widely according to the compositions of the culture media and conditions of cultivation so that it is somewhat irrational to compare lipid contents of acid-fast organisms grown in a culture medium and M. lepraemurium which has proliferated in a living host. It was found, nevetheless, that the lipid content of the M. lepraemurium was higher than of the cultured acid-fast organisms regardless of the media used for cultivation. The DNA content of M. lepraemurium is almost equal to the other bacteria, but the RNA content of M. lepraemurium is relatively lower. This may be related to a low metabolic activity of M. lpraemurium.
The fatty acid of the chilled acetone-soluble neutral fat fraction of M. lepraemurium was examined by gas chromatography. It was found that there was little difference in the fatty acid compositions and no fatty acid of a special structure in M. lepraemurium, particularly palmitic acid was present in the highest proportion followed by tuberculostearic acid (10-methyl stearic acid).
The high lipid content of acid-fast organisms may be due to their special carbohydrate metabolism but the biochemical background for the storage of lipids in the cells must be studied further.

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Immunofluorescent staining was performed on cryostat sections of 3 skin eruptions of ENL and 3 skin lepromas of lepromatous leprosy without ENL. In the indirect incubation method, the antibody labelled with fluorescenn isothiocyanate to the pooled serum immunoglo-bulins of the lepromatous leprosy with ENL was prepared in rabbits. The final fluorescein-protein ratio of the conjugate was to 1.3. The sera of 10 lepromatous leprosy with ENL, 15 lepromatous leprosy without ENL and of 4 tuberculoid leprosy were examined. Sections were treated with the patient's own serum, at a 1: 30 dilution, and subsequently with fluorescent antibody at a 1:100 dilution. In the sera of ENL, the prominent fluorescence indicated that immunoglobins were presented within the cytoplasma of the plasma cells, small lymphocytes and of histiocytes at the site of eruptions of ENL. The cell nuclei of the round or oval histio-cytes were also often fluorescent. These reactions were not observed, however, in the sera of lepromatous leprosy without ENL and of tuberculoid leprosy.
The results of the previous reports suggest that there are immunological tissue damage at the stage of ENL. It is probable that tissue cell degeneration in case of persistent lepromatous inflammatory changes is responsible for the production of antibodies. In advanced lepromatous leprosy, especially in those of the systemic or diffuse forms in which the episode of ENL are frequently observed, the presence of delayed hypersensitivity due to degenerated leprosy bacilli or altered self cell components may rather be important in causing the production of many humoral self factors (antibodies). One of the interesting facts is that phospholipids may be the intrinsic factors for the occurrence of ENL at the later stage of lepromatous leprosy. M. leprae has phospholipid antigen similar to that of human heart, blood vessels, nerve tissues and joints. Needless to say, antigens of the bacillary components presumably become cross antigens to those tissues, and M. leprae causes autoantibody formations against heart, vessels, nerve and/or joints. It is conceivable that the autoantibodies found in the sera of ENL may arise through this mechanism.

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Leprosy patients have been treated in the department of dermatology of Kyoto University since its start in 1903. In 1923 the special laboratory for leprosy research was built and attached to the department of dermatology. In 1938 present building of the Leprosy Research Laboratory was built together with a ward for leprosy patients. Since that time all the in-and out-patients of leprosy have been treated in this institute.
The author summerized the statistical data of the new cases of leprosy and in- and out-patients in our institute, treated in the past twenty years, which roughly corresponds to the sulphone period of leprosy treatment in Japan, and further compared the clinical features of leprosy in these 20 years to those in the 1903 to 1916 in our institute. Another intention of the author was to try to interprete the clinical data in our institute in the hope that it might throw some light on the prospect of leprosy in Japan.
1) The number of new cases of leprosy diagnosed in our institute has remarkably decreased in the past 20 years. In 1906 new cases in our institute were 154, whereas in 1967 the number decreased to only 8. 362 new cases in 1948-1967 were compared with 1852 new cases in 1903-1916.
The following statistical features were found. The ratio of the tuberculoid cases to the lepromatous ones has decreased; 4.18: 1 in 1903-1916 and 1.02: 1 in 1948-1967. The female cases have relatively increased; the ratios of male cases to female ones are 3.85:1 in 1903-1916 and 2.12:1 in the last 20 years. There were decreased young new patients, but relatively increased aged ones in these 20 years. Especially new cases in advanced age with tuber-culoid type of leprosy have curiously increased in number.
Most of new cases lived in Kyoto prefecture or neighbouring prefectures and visited our clinic within 5 years after onset of the disease. About 90 per cent of lepromatous cases were admitted in the leprosaria or in our institute. Most of the tuberculoid cases were treated in our ward and/or in our out-patient clinic.
There were 39 Koreans, about one third of new cases, who lived in Kyoto prefecture. It was discussed why Korean patients were so many in Kyoto prefecture.
2) The number of out-patients in our institute has increased little by little. But the positivity of bacilli in skin of these patients has decreased due to the effect of the anti-leprosy drugs.
3) The total number of in-patients in these 20 years was 299. 70 per cent of these patients were lepromatous leprosy. After leaving our ward, 43.4 per cent of all in-patients were admitted in the leprosaria and 51.9 per cent cases were treated in our out-patient clinic.
4) The importance of the out-patient clinic and the ward for leprosy patients outside the leprosaria was discussed for the present situation of the leprosy control in Japan.

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It is well known from old times that the destructive and absorptive changes of fingers are seen in the lepromatous type of leprosy. In one of these changes there are bamboo-shoot like changes of fingers which are elucidated in the handbook of Klingmiiller as Spina ventosa leprosa and which are also reported as bamboo-shoot finger by Inaba. The true aetiological and pathological factors of Spina ventosa leprosa are not defined. Spina ventosa leprosa is not seen in nonlepromatous leprosy, but seen only in lepromatous leprosy following on repeated severe Erythema nodosum leprosum reaction. Bamboo-shoot like bone changes in leprosy are supposed to be closely related to immune cross-reaction.
It has been very difficult to get high titered anti-M. leprae rabbit sera against crude extract of M. leprae but high titered anti-M. leprae rabbit sera are obtained by immunizing rabbit with fresh unheated human leproma and polysaccharide fraction from Klebsiella pneumoniae type 2 in incomplete Freund's adjuvant. This paper describes the cause of Spina ventosa leprosa from the immunological point of view.
The results are shown in the tables and figures. As may be seen in Fig. 1. 2, 3 in agar gel double diffusion technique the anti-M. leprae rabbit sera reacted among the extract antigens of M. leprae, BCG, M. lepraemurium and human phalanx, each forming a precipitation band which fused. On the other hand, in Fig. 4 anti-BCG rabbit sera also gave a fused precipitation band between the extracts of M. leprae and human phalanx. These results clearly indicate the presence of a common antigen among M. leprae, BCG, M. lepraemurium and human humap phalanx. The common antigen is supposed to be a polysaccharide nature judging from boil stability.
As can be seen in Fig. 5, 6, 7, 8, 9 the sections of human bones were stained with direct immunofluorescent method. The anti-M. leprae rabbit sera and anti-BCG rabbit sera contained a gamma globulin factor binding with human bones. The specificity of the reactions could be confirmed by inhibition test and absorption test.
These findings suggest that immune mechanisms might ber responsible for the occurrence of Spina ventosa leprosa.

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