This paper is concerned with the antituberculous activity of ethambutol combined with isoniazid against the original susceptible strain and isoniazid resistant strains by the in vitro, and in vivo experiments.
1) The H37Rv strain of human type tubercle bacilli was inoculated in Kirchner's semi-solid agar media containing ethambutol and isoniazid in graded concentrations and inhibition of the growth was determined after incubation of three weeks. No enhancement of antituberculous activity of the former drug was observed on addition of the latter.
The isoniazid-resistant variant of H37Rv strain and three strains freshly isolated from sputa of tuberculosis patients were tested in Dubos' liquid media for the influence of isoniazid on the in vitro growth inhibiting effect of ethambutol. The results were the same as in the experiment above mentioned.
2) Albino dd/y male mice weighing approximately 20 mg were used in the in vivo experiments. The animals were infected intravenously with H37Rv strain, and divided into four groups. Three groups were subcutaneously injected 2 mg of ethambutol, 0.02mg of isoniazid and 1 mg of ethambutol plus 0.01mg of isoniazid respectively, and the last group, the control, was untreated. The treatment were started from the next day of the iefection and continued for twenty-one days. All animals were sacrificed two day after the last treatment, and the therapeutic effects were evaluated by comparison of the viable units of tubercle bacilli cultivated from the lungs and spleens of the animals.
The enhancement of antituberculous activity was demonstrated in the combined use of the drugs, where tubercle bacilli were cultivated to the least extent from the animals of the group of this treatment. The experiment with a single or combined use of 1 mg of ethambutol and 0.01mg of isoniazid was carried out, showing the superiority of the combined treatment as. in the former experiment.
Subsequently, the groups of mice were infected with the isoniazid-resistant variant of H37Rv and treated with ethambutol and isoniazid as in the experiment with the sensitive strain. No effect of isoniazid therapy was found in those animals, but the addition of isoniazid to ethambutol did suppress more significantly the cultur able units of tubercle bacilli in organs of the animals than in those treated with ethambutol alone. The experiment was further conducted with the mice infected with two other isoniazid-resistant strains freshly isolated from tuberculosis patients yielding the same results.

抄録全体を表示

Studies were carried out on the acid production patterns from carbohydrates by the sapro phytic, rapid growing Mycobacteria and the availability of the patterns in the classification of them was discussed.
Media used were the agar slants of the following basal media composition: NaCl 0.5g, (NH₄) ₂SO₄ 0.5g, MgSO₄ 0.5mg, KH₂PO₄ 40mg, K₂HPO₄ 160mg, bacto-agar 1.5g and 1.5ml of a 0.04% BCP (Brom Cresol Purple) solution. Compounds added as the substrates were: glucose, mannose, fructose, galactose, arabinose, xylose, rhamnose, sucrose, trehalose, raffinose, dulcitol, mannitol and sorbitol. All these compounds solution were sterilized by filtration through porcelain filter and added to the basal media in a final concentration of 1%. Onto the agar slants of the above mentioned compositions was inoculated a certain amount of the bacterial suspension prepared from the 2 to 3 day old culture on Ogawa's egg slant, and the slants were kept in an incubator at 37°C for 4 weeks. Color change of the media due to. the acid production from carbohydrates was recorded as follows:
+complete color change into yellow within 7 days.
+complete color change into between 8 days to 3 weeks.
(+): complete color change intolater than 3 weeks.
-: no color change even after 4 weeks.
In the first place, acid production patterns were investigated both on the well known strains such as M. phlei, M. smegmatis, M. fortuitum, M. pisicium, M. marinum, M. platypoecilus and on the rather recently classified strains such as M. balnei, M. salmonifilum. As shown in the Table 1 conspicuous differences were observed in the acid production patterns from carbohyd rates among M. phlei, M. smegmatis, M. fortuitum and Mycobacteria isolated from fishes, and they were grouped into 4 different types, namely, M. phlei type (Group 1), M. smegmatis type (Group 2), M. fortuitum type (Group 3), and M. salmonifilum type (Group 4). M. balnei and Mycobacteria isolated from fishes were rather weak in the production ability and no significant difference was found amoung them suggesting their closed relationship with each other. Next, 38 strains of saprophytic Mycobacteria with a slightly yellowish colonial pigmentation kept in our laboratory were examined on their acid production patterns. As shown in the Tables 2a and 2b they were grouped into 4, namely, 2 strains to group 1 (M. phlei type), 18 to group 2 (M. smegmatis type), 14 to group 3 (M. fortuitum type) and remained 4 to group 4 (M. salmo nifilum type). Between the ranges of the growth supporting temperature and the acid production patterns fair agreement was observed in the foregoing 3 groups, while no agreement was de tected among 4 strains which belonged to the group 4 (M. salmonifilum type).
It was concluded that the method to investigate acid production patterns from carbohydrates seems to be available in the classification or the identification of the non-deeply pigmented saprophytic Mycobacteria to a considerable extent.

抄録全体を表示

Microscopic observations were made on tuberculous lesions in the lymphnodes obtained from 65 autopsy cases of tuberculosis and in the surgically removed ones of 36 patients, giving priority to the histological behavior of argyrophil fibers in the caseous lesion. Besides, there was an opportunity of reexamining the primarily infected lymphnodes of tuberculosis that had previously been studied by Dr. Tamura. The latter materials were obtained from a group of persons inoculated with typhoid vaccine which was incidentally contaminated with tuberculous bacilli of the human strain.
Results obtained were as follows:
1) The formation of tuberculous lesion in the lymphnodes began with features of sinus catarrh and, occasionally, of a nodular form. Then, there was a slight infiltration of leucocytes, a micro-abscess being formed in the central part of the lesion. The central micro-abscess came to being surrounded by a zonal infiltration of epithelioid cells transformed from reticulum cells which manifested sinus catarrh. As epithelioid transformation was advanced, argyrophil fibers disappeared for the time being from granulation tissue of epithelioid cells. Soon, the abscess fell into necrosis and successively became homogenous. At the same time, argyrophil fibers began to extend again into the external layer of the epithelioid cell zone from its outside, especially from the circumference of the blood vessels, and an argyrophil network was then formed. In the central necrotic area, however, no argyrophil fiber was found.
In case a living body was in good condition of the host-parasite relationship, collageniza tion of these argyrophil fibers took place from the site of stem-fibers, finally extending to their slender branches. At this moment, lymphoid cells containing microhistiocytes appeared and moved from the outside of the lesion into the central necrotic part through the epithelioid cell zone. These cells took part in scavengery of the central necrotic substances. Thus, the lesion as a whole looks like a doughnutshaped collagenous ring. When the lesion was still smaller, it was replaced by scar tissue.
2) While the processes above-mentioned were fundamental in the formation of the granuloma and in its healing in the lymphnode, findings might be modified according to an immunological status of an individual.
The lesion in the lymphnode attached to the primary complex and of the early generali zation showed a severe infiltration of leucocytes, being accompanied by large mononuclear cells, and formed a large caseous tubercle which occupied a greater part of the lymphnode. Under the circumstances, argyrophil fibers disappeared extensively in the caseous lesion.
On the contrary, caseous lesions in the lymphnodes of the late generalization and of tuber culosis of the third stage consisted of numerous tubercles which fell into caseation and were fused one another. On this occasion, no leucocytic infiltration was observed and, consequently, a plenty of nodular argyrophil network was found in the lesion.
As such, the histological behavior of argyrophil fibers in the caseous lesion appeared to represent well the immunological status of the human subject against the tubercle bacillus.

抄録全体を表示

In recent years, the causes of death in pulmonary tuberculous patients have been changing in accordance with the use of anti-tuberculous chemotherapeutics and the marked developement in tuberculosis control measures. Useful informations on the mothod of treatment for far advanced pulmonary tuberculous patients will be obtained by analysing the causes of death in pulmonary tuberculous patients, and in order to investigate this problem, the authors made, clinical and pathological studies on 45 cases of pulmonary tuberculosis died during the period from January 1957 to December 1962. The results obtained were the following:
1. The average age of death in pulmonary tuberculous patients has become higher in recent years.
2. Extent of pulmonary lesions in died cases was far advanced in 82%, and among them 69% were bacilli positive.
3. In 76% of far advanced cases, death was caused by tuberculous origin, such as chronic respiratory insufficiency, pulmonary edema, hemoptysis and pneumothorax. In the remaining 24% of far advanced cases, death was caused by non-tuberculous diseases developed during the course of treatment, and among them, malignant tumor, apoplexy, pneumonia and infection of urinary tract were frequently observed.
4. Mixed infection of lung, loading on right ventricle and complications in gastrointestinal systme and urinary tract were frequently observed by pathological-anatomical examination.
The results mentioned above indicate that the aggravation of pulmonary tuberculosis was still the major cause of death in pulmonary tuberculous patients. But, in accordance with the increase of average age of death in pulmonary tuberculous patients, death caused by nontuberculous complications has been playing more and more important role in the causes of death in pulmonary tuberculous patients in recent years.

抄録全体を表示

We have repeatedly reported that in our clinical experience combined use of 1314Th and INH for patients having INH-resistant tubercle bacilli is more effective than the use of 1314Th alone.
In the present study, experiments were conducted in order to confirm the combined effect of these drugs in vitro to INH-resitant tubercle bacilli.
1) By using Dubos' liquid medium, a hundred combinations of 1314Th and INH were prepared, each containing different concentrations of these drugs. In these media, H Rv strain resistant to 50γ/ml/ of INH was cultured for two weeks. Consequently, when 2. 5γ/ml and 1γ/ml/ of 1314Th were contained in the medium, the favourable effect of the addition of INH was observed (Table 1).
2) A similar effect was observed when Kirchner's medium was employed in place of Dubos' medium (Table 2).
3) Combination effect of 1314Th and INH was examined by using Ogawa's vertical diffusion method. When INH was used in combination with 1314Th in such high concentrations as 25γ/ml/ and 50γ/ml, the resulting inhibitory zones were much higher than those observed when 1314 Th was used alone (Fig. 1, Table 3).

抄録全体を表示

Using CF #1 male mice infected with 50γ/ml-INH-resistant H₃₇Rv, the author examined the effect of combined use of 1314 Th and INH to INH-resistant tubercle bacilli.
Mice were divided in the following five groups.
Group 1: No treatment.
Group 2: Treated with 500γ of INH every day.
Group 3: Treated with 500γ of 1314Th every day.
Group 4: Treated with 500γ of 1314Th and 100γ of INH every day.
Group 5: Treated with 500γ of 1314Th and 500γ of INH every day.
Consequently, the tuberculous involvement of the fifth group was the slightest both in the number of viable units recovered from the lung and in the histological findings. There was observed no significant difference between the third and the fourth group.
These results in vivo are quite in agreement with those in vitro described in the author's first report. Thus the effect of 1314Th is enhanced by the combination with INH, even in the case of INH-resistant tubercle bacilli.

抄録全体を表示