A review was made on studies of tuberculosis using germfree animals appeared in the literature including the present authors'. Miyakawa was the first who studied experimental tuberculosis in germfree animals, and described a lesser degree of abscess formation following subcutaneous infection, and smaller number of granulomas following intravenous infection in germfree guinea pigs in comparison with those in conventional ones. On the contrary, Suter and Kirsanow, and Huempfner and Deuschle reported no marked difference between germfree and conventional *mice on the viable bacterial unit in organs or lesions produced following infection with BCG or laboratory maintained H₃₇Rv strain. Hobby et al reported that germfree mice showed more uniform sensitivity than conventional mice to infection with virulent tubercle bacilli, and further that the immunizing effect of BCG was revealed more efficiently in germfree than conventional mice.
We compared the fate of organisms in organs and lesions produced in germfree (GF, axenic CD-1 and ICR-GF) and conventional mice (SPF, reared in SPF barrier or in vinyl isolators) following intravenous. infection with virulent bovine type tubercle bacilli strain Ravenel. The results revealed a remarkable difference between GF and SPF mice in later stages of infection, i.e., at 3 weeks in the large infection dose (10⁶ viable unit level) and at 4 weeks and later in the small infecting dose (10⁴ viable unit level). A greater propagation of organisms was found in the organs of GE mice than those of SPF mice with both large and small infection doses. A lesion produced in GF mice with the large dose was characterized by exudative and necrotic changes in the lung, kidney and heart, and lesser developed granulomas in the liver and spleen, while productive changes appeared in all the organs of SPF mice. A lesion in GF mice following small infection dose showed more progressive and extensive proliferation than that in SPF mice, , and was characterized by many histiocytes engorged with acid fast bacilli alike in Yersin type tuberculosis. By summing up the findings, it was considered that GF mice responded to phagocytosed bacteria with a reaction of the reticuloendothelial system and formed granulomas as seen in SPF mice, however, an inhibitory activity of phagocytic mononuclear cells in GF mice to the multiplication of intracellular mycobacteria seemed to be less comparing with that of SPF mice. Consequently, GF mice showed a higher bacterial unit in the organs and more extensive lesions than SPF mice later than 4 weeks, after infection.
The above described characteristics of tuberculous infection of germfree mice led the authors to the studies on delayed type hypersensitivity (DTH) reaction in germfree mice. DTH represented by footpad reaction with PPD on infected GF mice showed very poor reaction 2e-d3 weeks after infection. During the same period, SPF mice responded markedly. Immunization of mice with a mycobacterial component (phenol extract residue) suspended in paraffin also showed an impaired DTH reaction in GF mice while a typical DTH reaction appeared in SPE mice. Depressed DTH reaction in GF animals was also reported by Lerner, and Lev and Battisto in guinea pigs. This impaired DTH reaction in GF mice seems to be related to, if not all the cause of, the low resistance of germfree mice to tuberculous infection.

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Phage typing of M. tuberculosis isolated from the patients was performed to identify the source of tuberculous infection in families.
1) The phages used were derived from the National Institute of Health in Japan. One RTD was tested by spotting 0.01ml of ten fold dilutions starting from 1×10⁸/ml onto RVA plates overlaid with semisolid RVA containing 0.5% agar and host bacilli. Complete lysis was observed by 100 particles per 0.01ml or more for all phages.
Thus it was defined that one RTD in our laboratory was 10⁴ particles per milliliter for each phages.
In 1954, Dr. S. Froman originally reported the phage D34. However the D34 which was given to me by Dr. Murohashi did not lyse H₃₇Rv. Consequently this particular phage has been termed D34'.
2) Using one RTD, each phage was spotted on the 160 strains isolated from pulmonary tuberculosis patients. As for susceptibility, the common receptors were observed among DS6A, Y10, C3, B1, GS4E and BK₁ phages each other. D34' phage showed the most specific lysis. Accordingly it was suggested that strains of M. tuberculosis were divided into two lytic patterns. In one pattern, the strains show susceptibility to some of the DS6A, Y10, C3, B1, GS4E and BK₁ phages. In the other pattern, only one strain was susceptible to D34'. Since the phage types, to which the least number of strains show susceptible, seemed very useful for epidemiological studies, the BK1 and D34' were selected for the nexexperiments.
3) For the reason just stated, BK₁ and D34' were selected and applied to the typing of isolated strains, and further phage typing were carried out by the following procedure. About 0.5mg of semi-dried isolated strains in 0.5ml of saline and 3ml of RVA were poured over a plate of 20ml of solidified RVA. After incubation at 37°C for three days, 0.01ml of phage suspensions containing 1×10⁴ and 1×10³/ml were spotted. After additional incubation at 37°C for another 11 days, the degree of lysis was recorded.
Final reading were recorded by classifying the strains “highly susceptible”, “intermediately susceptible” and “less susceptible” to each phage. That is, two concentrations of one RTD and 1/10 RTD were spotted on each bacterial lawn of the strains to be tested. When 10 phages (1/10 RTD) spotted produced plaques, such a strain was regarded as “highly susceptible” to the phage; and if 100 phage particles (1 RTD) produced no plaques, such a strain was regarded as less susceptible to the phage; the other strains were regarded as having intermediate susceptibility to the phage.

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In order to clarify the mechanism of fatty liver formation by ethionamide, the effects of ethionamide on the fatty acid synthesis in rat liver were investigate.
Male Donryu rats weighing 100 to 130g were used. Pure ethionamide (2-ethyl-thioisonicotinamide) suspended in distilled water was given orally through stomach tube. Rats were sacrificed within 48 hours after single administration of ethionamide, livers removed and the specific activities of NADPH₂ generating enzymes in the soluble fraction of liver were measured spectrophotometrically.
Another experiments were performed with regard to the incorporation of [2-¹⁴C] acetate into the liver lipids of ethionamide-administered rats. Each rat received intraperitoneally 10μCi of sodium acetate-2-¹⁴C (3.75mCiim mole) at some time after the administration of ethionamide and was sacrificed 3 hours later. Liver total lipids were extracted according to the procedure of Folch et al. Total lipids evaporated to dryness under nitrogen were dissolved in a small amount of chloroform, treated with acetone and then separated into two portions; acetone soluble and insoluble fractions. The former neutral fat fractions were evaporated to dryness and dissolved in 10ml of chloroform. The latter phospholipid fractions were dissolved in 3ml of chloroform. After evaporation of aliquots of the lipid extracts to dryness, total lipid, neutral fat and phospholipid were measured gravimetrically. Another aliquots of the lipid extracts were added 15ml of scintillation fluid and the radioactivities were assayed with a liquid scintillation spectrometer (Packard Model 3003). The results obtained were as follows.
There were no remarkable changes in the NADPH₂ generating enzyme activities of both 200mg and 400mg per kg body weight of ethionamide-administered rat liver.
Incorporation of ¹⁴C-acetate into the total liver lipids of ethionamide-administered rats increased remarkably, and showed approximately three times the specific radioactivity of the control rat liver 3 hours after the administration of ethionamide. Furthermore, it was found that the incorporation of ¹⁴C-acetate increased particularly in the neutral fat fraction.
It has been demonstrated that incorporation of ¹⁴C-palmitate into the liver lipids increased in carbon tetrachloride-administered rats, and that incorporation of ¹⁴C-acetate into the liver triglyceride increased remarkably also in orotic acid-administered rats. Other investigators suggested that the stimulation of hepatic fat synthesis might be a significant factor contributing to the development of fatty liver. Another report from this laboratory and the results presented above suggest that the utilization of acetate in the liver of ethionamide-administered rats increases and, as a result of the stimulation of fatty acid synthesis, the activity of glucose-6-phosphate dehydrogenase rises in accordance with increased requirement of NADPH2. As described in another report, however, an increased import of lipid into the liver from adipose tissue was observed in the case of a high dose administration of ethionamide. It is necessary, therefore, to investigate more minutely the relation between these factors contributing to the development of fatty liver.

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多剤耐性有空洞肺結核41例を対象とし, この研究開始直前に用いられていた経口投与の抗結核剤に新抗結核剤Tuberactinomycin-N (TUM-N) を加えて6カ月間治療し, その治療効果ならびに副作用について検討した。TUM-Nは原則として1日1回19を初めの3カ月間は毎日, その後は週2日筋肉内に注射した。
1. 併用薬剤に感受性が認められずTUM-N単独治療とみなしうる症例14例における培養陰性化率は1カ月42.9%, 6カ月28.6%であり, TUM-Nと感受性の薬剤とを併用した16例では1カ月62.5%, 6カ月72.7%であつた。
2. TUM-N治療中にaudiometryにより8, 000c/sで20db以上の低下の認められたものは40例中1例 (2.5%) のみであつた。検尿成績については, 1例に一過性の軽度の蛋白尿を認めたにすぎなかつた。そのほかの副作用としては, 耳鳴りと発疹が各1例に認められた。
以上のごとく, TUM-Nは副作用が少なく, 聴力低下の出現頻度も低く, 連日使用の可能なことは抗結核剤として本剤に有利な点であると考えられる

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It is wellknown that the incidences of post-operative complications is high among the surgically treated cases with drug resistant pulmonary tuberculosis. To improve the results, a new anti-tuberculous drug, Rifampicin, was used before and after surgical treatment at 7 National Sanatoria in Japan.
Pulmonary resection cases were selected based on certain criteria from SM, INH, KM resistant cases, and the results were compared with matched pairs selected from SM, INH resistant cases who were administered Kanamycin prior to, during and after surgical operation. RFP 0.45g daily was administered once a day for 4 months prior to and after operation.
According to the present study, only one case of bronchial fistula was found at 40 days after operation out of 21 cases of the RFP group, while five post-operative complications, such as bronchial fistula, empyema thoracis, relapse of remaining lesions, were observed among 21 cases in the control group.
There was no remarkable side-effects or toxicity due to RFP.
It is concluded that RFP is as effective as or more effective than Kanamycin in preventing the incidence of post-operative complications for drug resistant cases.

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Although the lung surgery has been safely performed recently, the radical operation for empyema thoracis is still difficult and its results are not always satisfactorily. In these circumstances, even in surgeons, the radical operation does not urge for the patients with quiescent empyema which has not any, symptoms and the patients without symptoms are kept under observation.
Since 1967, at the Chest Disease Research Institute Hospital of Kyoto University, one hundred and forty cases of empyema were treated. In this series it is included 8 cases with acute empyema, 57 cases with post-operative empyema and 75 with chronic one which had so-called silent phase of the empyema, namely some quiescent periods over three months. Seventy-three out of 75 cases with silent phase were, operated and reviewed as the object of this series.
CAUSES of EMPYEMA; In sixty-eight patients out of 75, an underlying lessions can be estimated; 34 cases (50%) are pleurisy. 18 cases (26%) following artificial pneumothorax. Three quarters of the chronic empyema are presumed to be tuberculous.
INTERVALS of SILENT PHASE; More than half of these cases have been 4, uiescent during ten years or more and one of them has been asymptomatic for 40 years.
SYMPTOMS; Except 27 cases without symptoms, symptoms such as hemoptysis (15 cases), formation of bronchopleural or cutaneous fistulae (9), swelling of the chest wail (4), etc., are developed in 48 cases. Only three cases develope some signs of acute inflammation. Empyema with fistulae is found frequently in patients with pleural calcification
OPERATIVE METHODS and their RESULTS; Seventy three patients are operated. Resection which included decortication combined with lobectorny is performed on 26 cases. Decortication is done on 27 cases and empyema-space reducing procedures such as thoracoplasty is employed on 5 cases. Another 15 cases are treated with preliminary open treatment and thereafter radical operation is done. In the cases without symptoms (so called silent empyema), the excellent operative results are obtained. Postoperative death among these 27 cases is only one case who died because of pneumonia due to agranulocytosis at 18 months later. Death rate in cases with symptoms is as high as in the postoperative empyema. (Operative mortality rate is 12% and 14%, respectively.)
HISTOLOGICAL FINDINGS; On the microscopic examination of the pleura, interstitial tissue layer of the visceral pleura is well preserved in the cases in which decortication is easily performed; while in those patients in whom such procedure is difficult, usually all vestiges of the mesotherial layer are lost, and the fibrotic process extends into the substance of the pleura and even into the lung. Even in the clinically quiescent cases, infiltration with leucocytes and giant cells is observed in the lung parenchyma.

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