In the previous paper, Ishikawa reported that putrescine is always observed in the blood of lepromatous leprosy patient with M. leprae in his skin (LA LEPRO 36, 238 (1967)). It is well known that putrescine is one of the ptomains and causes the ptomaine poisoning. So the Ishikawa's report presents a serious problem confronting the lepromatous leorosy patients. In this paper we described the method of determination of putrescine and cadaverine in serum and then the result of measuring these ptomaines in the serum from lepromatous leprosy patients.
The determination of putrescine and cadaverine is as follows;
Serum was extracted with n-butanol by McIntire's method. Butanol layer was acidified with 1N-HCl and then butanol was removed by lyophilization. Lyophilized residue was applied to a small column of Amberlite CG-50 according to the method of D. R. Morris with some modification (cf. Fig. 1).
By the use of Amberlite CG-50 putrescine and cadaverine could not mutually be separated, but could be separated from lysine, histidine, arginine, istamine etc. Standard curve of ninhydrin reaction showed that the concentration of putrescine or cadaverine was in proportion to the rate of ninhydrin reaction. The recovery of putrescine in normal human serum after butanol extraction and column chromatography was about 90%.
About 2ml (varying from 1.6 to 4.5ml) of serum from lepromatous leprosy patient with bacilli in his skin was tested for detection of putrescine and/or cadaverine, but in no case out of seven we did find more than 0.05μ moles of putrescine and/or cadaverine in the serum.
Ishikawa obtained the serum from 5ml of blood, so about 2 to 2.5ml of serum was used for detection of putrescine. The quantity of serum used for determination of putrescine by Ishikawa is almost the same as used by us. We do not believe that the Ishikawa's method is far better than that used by us.
Putrescine and cadaverine are known to be formed by decarboxylation from ornithine and lysine respectively in living organisms. From our result we can not decide whether M. leprae has decarboxylase activity or not, because the products of this bacillus may be metabolized to non-toxic materials by human nzyme system to maintaine the homeostasis. For example, even if the putrescine is produced, it will perhaps be oxidized by diamine oxidase in human liver. It seems to be necessary to measure directly decarboxylase activity in the bacillus.

抄録全体を表示

Boiden meshcement coated slide glasses (BM slides) were used for smears of M. lepraemurium which were cultivated in the medium for elongation in vitro. It was obvious that the BM slides had the following advantages for the experiments of elongation of M. lepraemurium; (1) the smear spread easily on the BM slides, because of non-hydrophobic, (2) the slides fasten the materials, even if they contained a lot of tissue protein, nevertheless, (3) slight elongations of M. lepraemurium were able to occur on the BM slides, even when they were not seen on the conventional slides. Therefore, the BM slide is recommended as a tool for experiment of elongation of M. lepraemurium in vitro .

抄録全体を表示