The authors have confirmed the multiplication of M. leprae in the foot-pads of mice, hamsters and rats. Subsequent to the progressive infection of M. lepraemurium in the foot-pads of mice, hamsters, rats and chipmunks, it seemed to be reasonable to examine whether M leprae would grown in the foot-pads of chipmunks.
The experimental methods of foot-pad inoculation and assessment of bacterial growth were similar to those already published by ourselves. The chipmunks used were of ages from 5 to 9 weeks. Three isolates of M. leprae were referred to: one was a homogenate of leproma tissue, and the others were mouse passage material. Portions of the same inoculum suspensions were also inoculated into the foot-pads of mice. A part of the suspension used was heated to 100°C for 30 minutes to kill the bacilli. In the chipmunks receiving this heated bacilli, there was no evidence of growth in the foot-pads. In Table I, the results of two mouse passages of M. leprae indicates that these two inoculates of different sizes progressed. Both strains rose to almost the same "ceiling", and the fresh strain rose to a plateau level of 1.0×10⁵ at 25 weeks after the inoculation. However, the bacillary counts of the chipmunks were less than those of the mice. In addition, in the chipmunks receiving M. lepraemurium, nodular swelling was observed on the footpads.
It may be concluded from the results that a successful transmission of M. leprae was apparent.

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By means of an apparatus specially devised by Dr. Mita, Professor of Physiology of the Iwate Medical University School of Medicine, which is operated with three cps sinusoidal alternating current, the impedance of the following skin of leprosy patients has been investigated: the skin considered to be normal judging from its outer appearance, and obtained from 20 different sites of the whole body in each subject. A total of 173 leprosy patients consisting of 91 males (74 of the lepromatous type and 17 of the tuberculoid type) and 82 females (58 of the lepromatous type and 24 of the tuberculoid type) have been studied. They were all patients hospitalized in the Tohoku Shinsei-en, a national leprosarium located in the Tohoku district of Japan, and the investiga-tion was made in August 1965 under a definite room condition: room temperature 27-29C, and room humidity 74-75%. The sites of the skin examined are as follows being divided largely into two groups and studied statistically because of the special status of the so-called normal skin of the leprosy patients: the first group consisting of the anterior breast, the upper abdomen, the upper part of the back, the lumbar region, and the nape of the neck; the second group consisting of the face (frontal region, cheek, and jaw), the upperarm and forearm (the extensor as well as flexor aspect), the palm and back of the hand, the thigh (the inner as well as outer aspect), the leg (the extensor as well as flexor aspect), and the sole and back of the foot. Comparing the impedance values obtained in the investigations with that of the normal skin of normal individuals examined with the same apparatus in the same season under the same room condition and reported by Dr. Takasu, the following results have been obtained.
I. Results obtained in the first group (the skin of the trunk and the nape of the neck).
(1) The impedance values of the so-called normal skin of the trunk and the nape of the neck of the leprosy patients were both found, as a rule, to be significantly lower than that of the normal skin of normal individuals.
(2) The impedance values of the so-called normal skin of the leprosy patients of the first group were noted not to have any significant age-, sex-, and site-differences.
II. The results obtained in the second group (the face and the extremities).
(1) The impedance values of the face and the upperarm were noted not to be significantly different between the leprosy patients and the normal individuals.
(2) The impedance of the skin of the extremities were noted to have a trend to become higher in values toward the peripheries.
(3) Among the sites of the extremities, the impedance values of the forearm and the hand as well as of the leg and foot were noted to be significantly higher in the leprosy patients than in normal individuals.
(4) As the so-called normal skin of the leprosy patients is occasionally associated with sensible disturbances, the impedance values were also compared between the socalled normal skin with sensible disturbances and the same with no sensible disturbances in the leprosy patients. In the study, the skin of the upperarm and the thigh were investigated, and it was noted in almost all age groups and sites that the impedance values were significantly higher in the skin with sensible disturbances.
Taking the hitherto mentioned results into consideration, the physiological meaning of the impedance of the so-called normal skin of the leprosy patients judged from its outer appearance in summer were surmised as follows.
(1) The impedance value is assumed to be closely related to the sweat function, and also has connection with insensible perspiration as well as blood circulation of the skin.

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Demasking in N-conjugates of sulfone drugs and sulfonamides by the incubation with micro-organisms was examined. Twenty-one strains of mycobacteria together with four of non-mycobacteria) strains were employed in the experiment.
Results found were as follows:
1) The deacetylation of mono-Acetyl DDS (MADDS) was generally more active in mycobacteria than in the non-mycobacterial strains, especially it was active in such strains as the rapid growing mycobacteria except M. balnei, M. tuberculosis H37 Rv, NQ bacilli, and Nonphotochromogens. As for the unclassified mycobacteria, the action in the Nonphotochromogens was more marked than that in the Photo- and Scotochromogens employed in the experiment.
2) In spite of the active deacetylation in the mycobacterial strains, the action could not be found so markedly or was nearly negative in whole the N₄-acetylates of the sulfonamides as well as in the non-mycobacterial strains.
3) Employing NQ bacilli, further studies were carried out to examine the difference between the demasking in several heterogeneous radicals and in A radical, together with the difference between the demasking in N₄-A and N₁-A of the sulfonamides. Tested compounds were: Enantoyl MADDS (MEnADDS), mono-Enantoyl DDS (MEnDDS), monoDiethylamonacetyl MADDS (MEADDS), mono-Morpholinoacetyl MADDS (MMADDS), N-acetylates of Sulfamethoxypyridazine (respectively abbreviated as N₁ASMP, N₄ASMP, and DASMP), and those of Sulfaisoxazole (N₁ASIX, N₄ASIX, and DASIX). The details are shown in Chart 1. Generally, it was found that this strain was apt to demask A more rapidly than En, E, and M from DDS, though the demasking of the latter two was comparatively lower than the former two.
The inactivity in the demasking of N4-A was again noticed in the case of the sulfonamides similarly to (2), and N₁-A suffered severe deacetylation even in the control samples. Therefore, DASMP and DASIX were detected as the corresponding N₄-A even in the controls, as well as the N₁-A were respectively detected as SMP and SIX.
4) Next, the demasking of MADDS by a crude cell-free extract of M phlei (Penso) was examined, and it was found that within 3 hours under shaking-incubation, a gradual increase of freed DDS could be detected with the lapse of the incubation periods. However, the condensation of the activity was failed, resulted in the disappearance of the activity during the fractionation.
5) Employing ³H-glucuronate (³H-GNa) and DDSG-³H, which was synthesized from DDS and ³H-GNa, the influence of E, culi K-12 on the N-G was examined, under the correction of pH during the incubation. Though the intake and the catabolic variation of 3H-GNa freed from DDSG-3H could be noticed, the demasking of N-G by the bacilli could not be elucidated.

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In the precedent paper, the possible relation between thyroid function and the healing mechanism of leprosy was discussed. Now, if the thyroid hormones are subjected to free from human serum under the interference of antileprous drug in vivo, it may intensify the affinity of the hormones to the tissue components of periphery lesions, even though a difference in the peripheral role of the hormones between the leprous lesion and the healthy skin can be considered.
Authors examined the interference of several antileprous drugs in the binding of ¹³¹I-thyroxine (¹³¹I-T₄) to human serum by the use of an in vitro experimental method, radiostereoassay. The drugs employed were di-N-acetyl DDS (DADDS), Ciba 1906, Sulfamethoxypyridazine (all of them are representative drugs adsorbed on albumin, A-type as previously reported), 1314 TH (G-type), and DDS.
The ratio of the bound thyroxine in A per G gradually increased with the concentration of 1314 TH added to the serum, which was supposed to be due to the removal of bound ¹³¹I-T₄ from G to A by the direct interfeeence in the binding of thyroxine to TBG. While, in the cases of the A-type drugs, the rate was contrary diminuted with the concentration of the drugs, or in some case, once diminuted and then increased to form a valley.
When the concentration of thyroxine increased, the valley was apt to remove to the point where the concentration of the drugs was lower than before. This phenomenon can be explained by the following concept that the every binding site of drugs or even hormones is not restricted by a single component, but by an adsorption order on the several components peculiar to a drug or a hormone. Depending upon the concept, the following explanation can be possible. During the diminution of the ratio in A per G, the saturation of TBG by both of thyroxine and one of such drugs may not be reached and the interference by the A-type adsorption of the drug to the slight binding of thyroxine to A is merely emphasized.
While, when TBG is saturated, the non-specific binding of thyroxine to A necessarily increases with the concentration of the drug, thus the valley is formed. In addition, when the concentration of thyroxine or the drug is increased, the mutual interference in TBG may considerably increase, when it is compared with that in A, because the saturation in TBG can be reached more easily than in A and the binding of thyroxine to TBG is well-known to be more stable than to A. Therefore, when the concentration of thyroxine is increased, TBG must be saturated by a lower concentration of the drug, thus the valley removes to the lower concentration of the drug.
Although not so obviously noticed, a similar result in the ratio of prealbumin (PA) per G was detected, as well as in the ratios of A per total and PA per total, in which the total means the radioactivity throughout PA, A, and G.
A diuretic Saltron (4-Chloro-1, 3-disulfonamide) also showed a shallow valley. In the case of DDS, this phenomenon could not be openly apparent, perhaps be due to the comparatively labile binding to human serum, or to the binding type as previously reported.
In conclusion, it can be said that a mutual interference or even a competative relation between thyroxine and several antileperous drugs in the binding to the protein components of human serum is positive, even though it is merely based on an in vitro experimental method.

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Elongation phenomena of M. lepraemurium in vitro were observed using silicon -coated slide glasses compared to common slides. The results obtained indicate that M. lepraemurium promptly shortened after being smeared on the silicon slide glasses, and that remarkable elongation of M. lepraemurium on the silicon slide was observed only in the medium which was the most appropriate for elongation of the bacilli studied by using common slides. It could be noted from the results mentioned above that these findings might demonstrate that the bacilli fixed more faster on the silicon slide than on the common slide. On the other hand, it could be said that subtle morphological changes in the bacilli during cultivation might be missed if the silicon slide was used for observation of the bacilli.

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A relationship between morphological index of live and dead M. lepraemurium was studied by using light and electron microscopes. First of all, it was clarified that the most important factor for staining conditions influencing morphological index of acid fast morphological index of M lepraemurium obtained by a staining method at room temperature was significantly changed by the method of staining with heat: by the latter, most of nonsolidM. lepraemuriumwere changed to solid acid-fast bacilli. Next, a comparison was made between morphological index of live M lepraemurium and dead ones that were killed by artificial treatments.M. lepraemurium was inactivated by treatments with ultraviolet-irradiation, heating at 60°C for 30min, chloroform, ether, toluene, and n-buthyl-alchol, whereas it was not affected by treatments with desoxycholate and petroleum ether. There were no differences between staining qualities of live and dead M. lepraemurium. However, the cell wall destructions were illustrated in dead bacilli thus inactivated by artificial treatments, when observed by an electron microscope.

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The adsorption of DDS and the relating drugs on NQ bacilli and E. coli K-12 (CS-2) was examined by Batch Method.
The drugs employed were DDS³⁵, DADDS³⁵, DDS•-G, and four of non-labeled sulfaagents. These were separately dissolved into 5ml of calf serum or physiological saline in a concentration of 10μg/ml. The lyophilized bacilli of 15, 30, 60, 120mg each were suspended and centrifuged, after the vigorous agitation followed by 1 or 4 hours' shaking-incubation.
As the result, it was found that the adsorption of a drug on bacteria was parallel to a certain extent with the adsorptivity to serum and was also inhibited by the adsorptivity of the calf serum when it was put to use as a basal medium.
No influence could be found by the pre-treatment of NQ bacilli with 5% TCA or with ethanol-ether. Therefore, this adsorption is supposed to be merely non-specific one.
The adsorption of DDS³⁵ on the coli bacilli was higher than that on NQ bacilli, because the coli bacilli adsorbed DDS35 even in the calf serum. It may perhaps be due to the difference in the form of the bacterial clumps between the two strains.
The adsorption on an activated charcoal was superior to that on the two strains.
The adsorbed DDS³⁵ per 100mg of NQ bacilli was increased with the weight of NQ bacilli suspended into the saline. While, it was contrary decreased in the case of DADDS³⁵, suggesting the saturation in the earlier stage.
When it was used non-labeled Sulfaisoxazole, -dimethoxine, -methoxypyridazine, Sulfonamide, and DDS, a lowering in the optical density could be detected, though not so clearly noticed.
Based on this interactive adsorption and on a presumption concerning the more predominant out-flow of labile conjugates from capillary vessels than that of the stable conjugates, the drug action mechanism in the peripheral lesions was mentioned.

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Bacillary suspension of M. lepraemurium was filtered through millipore filter type HA (pore size 0.45μ±0.02μ), type PH (pore size 0.30μ±0.02μ), type GS (pore size 0.22μ±0.02μ) and type VC (pore size 0.10μ±0.008μ) respectively under a pressure of 20mm Hg. Acid-fast bacilli in filtrates were examined microscopically after ultracentrif ugation (20, 000rpm, 45min.), and infectivity of filtrates was also examined by subcutaneous inoculation to mice.
Acid-fast bacilli were observed only in the filtrate through HA filter, and only this filtrate showed infectivity to mice. The other filtrates which were not observed acid-fast bacilli in them developed no murine leproma in mice.
Bacillary concentration of filtrate through HA filter was calculated as about 10⁴/ml by means of bacterial count and infectivity test by 10-fold dilution method.
It was concluded that it is needless to suppose the existance of special filtrable form of M. lepraemurium.

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