The isolation frequency of cultivated acid-fast organisms from nasal washings of leprosy patients was 90.0% by the method that has been reportedl1).
The present communication describes the experience whether the organisms produce virulence in experimental animals or not.
The cultivable organisms of Mycobacterium sp. I strain isolated from nasal washings of the patient with lepromatous leprosy were suspended in strile saline solution on the living condition. Then, the suspension was inoculated subcutaneously, intraperitoneally and into foot-pads of rabbits, guinea-pigs and mice.
The organisms did not produce progressive lesions in any partes of used experimental animals macro-and micro-scopically.
However, in case of inoculation into mice foot-pads, the acid-fast organisms similar to leprosy bacilli were observed in the inoculated sites, and these organisms seemed to multiply in numbers.
The organisms harvested from infected foot-pads of mice were tried to be cultivated onto the egg-yolk media, but there was no growth of acid-fast organisms.

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The observations reported here are concerned with the cyto-morphological studies on the cellular organelle of Mycobacterium lepraemurium (Hawaiian strain), and the capsular structure around the bacilli in the host cells.
As the peripheral parts of the bacilli, the capsular structure, the cell wall and the cytoplasmic membrane were clearly observed.
In the cytoplasm of the bacilli, two kinds of typical organelle, that is, the intracellular membranous organelle and the electron dense or homogenous granules were generally found.
The large electron dense and homogenous bodies were found in murine-leproma cells, and there were the small electron dense granules in these bodies.
In the cell division process, the division site on the surface of cell wall became weakly, and then the cytoplasmic membrane, adjacent to this wall site, distorted towards the center of cytoplasm. Thus, the septum formation occurred and some of intracellular membranous organelle arranged parallel to the edge of septum. These membranous organelle, at this time, seemed to play a role in the cell division process.
After the construction of cross wall (septum), the old wall around two new halves of the new cell was separated and released from new cell wall.
These disrupted old walls seemed to remain meanwhile as a thin layers, and then to become amorphous substances.
From the facts mentioned above, it could be explained that the capsular structure around M. lepraenzurium was mostly originated from bacterial side.

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The name Lingular Disease, 10) 19) regardless of etiology, altogether with the middle lobe syndrome11)12)13)14)15)16)17)18) is suggested to be included in the middle lobe disease20)21) and is for all cases of atelectasis and pneumonitis which are not caused by active tuberculosis or by neoplasm. While conceivably some cases might have been caused originally by tuberculous lymphadenitis in the earliest time, the sequential pneumonitis later in life is non-specific and not distinguished from pneumonitis brought about by nontuberculous lymph nodes or by mucus plugs and poor drainage. Lingular Disease10)19) can thus be defined as characterized clinically by hemoptysis, blood-tinged sputum and recurrent episodes of pulmonry infection and pathologically by atelectasis, fibrosis and bronchiectasis and is occasionally made an erroneous diagnosis as pulmonary tuberculosis, chronic bronichitis, bronchial asthma, cardiopathy and neuropathy, 16, 18) in spite of its characteristic and peculiar roentgenological appearance.
For aught we know, this disease has not been previously described in the Japanese literatures, except one introduction of its conception, 16) though this disease is never rare. The primary aim of the repart is not to impart knowleoge but to increase understanding concerning the dignity of human life and reverence to more advanced medicine.

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Since 1912, when Murata first described this syndrome called Erythema Nodosum Leprosum (ENL) as a clinical entity, we have had so many papers on ENL. The characteristics of ENL, which were clinically observed on 177 patients with lepromatous leprosy, could be arranged as follows;
1. ENL is essentially the manifestation of antigen-antibody reactions and occurs in the pure and near lepromatous patients. But some of lepromatous cases (49.4%) escape ENL in their course.
2. This condition usually starts several months to 24 months after treatment begins.
3. ENL can be graded as slight, moderate or severe in degree. This condition is definitely more frequent and more severe in clinically more advanced, more mature cases than in those with slight lesions.
4. Although ENL has become much more common since the initation of sulfone therapy, it can be caused by other antileprosy drugs including chaulmoogra oil. 5. As far as bacterial negativity is concerned, prognosis of patients with severe ENL is not always worse than that of the patients without ENL.
6. We have tried to notice previously the occurrence of ENL in the lepromatous patients. Twenty three cases of 34 new cases (68%), who took the serologic test for syphilis (STS), showed positive STS at the start of chemotherapy. These biological false positive cases of STS were confirmed by RPCF and TPHA. There was only one case who showed latent syphilis. The correlation between the occurrence of ENL and the positive STS in the lepromatous cases was statisically significant.
7. The Leproagglutination (Ogata)6), which has antigen made from cardiolipin and lecitin (1: 1) was shown in this paper as a more useful test for ENL. The lepromatous cases with positive Leproagglutination test in serum dilutions of 1: 64 or greater before chemotherapy were apt to have episodes of ENL in their clinical course (p=0.01).

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Suspensions of Mycobacterium leprae were incubated at 4°C or 30°C for 60min with rifampicin at a concentration of 2mg/ml. Before inoculating of mice, haves of the suspensions were repeatedly washed with a balanced salt solution. The unwashed bacilli did not multiply in mouse foot pads regardless of the exposure temperatures to the drug. The washed ones pretreated at 4°C multiplied normally. The organisms treated with the same procedure but at 30°C resulted in a significant growth delay.

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Eight of 5 weeks old nude mice (BALB/c-nu/nu) were infected with Mycobacterium leprae obtained from a case of the relapse lepromatous leprosy patient, and they were kept under SPF (specific pathogen free) condition in Vinylplastic-isolator to prevent wasting disease and other microbial infection. All of 3 mice survived for more than 17 months after the infection showed swelling of foot pad at the inoculation site. These 3 mice were sacrificed at 17th, 19th and 22nd month after inoculation respectively, and histopathological and bacteriological examinations were carried out. Histopathologically, lepromatoid lesions were observed not only in the foot pad of the infected site but also in low temperature parts of the body such as eyelid, earlobe, tail and nose. Invasion of acid-fast bacilli into peripheral nerves was remarkable. The acid-fast bacilli increased in lepromatoid lesions were identified as M. leprae by following identification tests: 1) Failure to growth on artificial media, 2) No granuloma formation in normal mouse, 3) Loss of acid-fastness by pyridine extraction, 4) Positive D-dopa oxidase activity, and 5) Results of lepromin reaction with patients.

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