(Vol.74 No.9 September 1999) <1> Kekkaku Vol.74,No.9:655-660,1999 INTERNALIZATION AND REPLICATION OF MYCOBACTERIUM TUBERCULOSIS AND M.AVIUM COMPLEX WITHIN TYPE U ALVEOLAR EPITHELIAL CELL LINE Katsumasa SATO, Keiko OGASAWARA, Tatsuya AKAKI, and *Haruaki TOMIOKA *Department of Microbiology and Immunology, Shimane Medical University Profiles of internalization and replication of Mycobacterium tuberculosis (MTB) or M. avium complex (MAC) within A-549 human type U alveolar epithelial cell line (A-549 cells) were studied and the results were compared with the mode of internalizing and proliferative behaviors of the organisms within murine peritoneal macrophages (Ms) and various M-like cell lines (murine J774A. 1, human THP-1, and human MONO-MAC -6). First, MTB and MAC internalized not only in peritoneal Ms and M-like cell lines but also in A-549 cells. Secondly, MTB and MAC replicated within A-549 cells and these organisms displayed much more vigorous intracellular multiplication in A-549 cells than in murine peritoneal Ms and J774A. 1 M-like cell line. Human M-like cell lines (THP -1, MONO-MAC-6) allowed the growth of MTB and MAC equally or occasionally more rapidly, as compared to the case of A-549 cells. Key words:Mycobacterium tuberculosis, Mycobacterium avium complex, Type II alveolar epithelial cells, Macrophages *89-1 Enya-cho, Izumo-shi, Shimane 693-8501 Japan. (Received 23 Mar. 1999/Accepted 3 Jun. 1999) <2> Kekkaku Vol.74,No.9:661-666,1999 EFFECTS OF THE CHINESE TRADITIONAL MEDICINES "MAO-BUSHI-SAISHIN-TO" AND "YOKUININ"ON THE ANTIMYCOBACTERIAL ACTIVITY OF MURINE MACROPHAGES AGAINST MYCOBACTERIUM AVIUM COMPLEX INFECTION Toshiaki SHIMIZU, Chiaki SANO, Tatsuya AKAKI, Keiko OGASAWARA, Katsumasa SATO, and *Haruaki TOMIOKA *Department of Microbiology and Immunology, Shimane Medical University We previously examined the effects of two Chinese traditional medicines "Mao-Bushi- Saishin-To" (MBST) and "Yokuinin", on the therapeutic efficacies of a benzoxazinorifa- mycin, KRM-1648, against Mycobacterium avium complex (MAC) infection induced in mice. MBST but not Yokuinin potentiated the therapeutic activity of KRM-1648 against MAC infection. In the present study, we examined the effects of these traditional medi- cines on some M cell functions. First, MBST significantly potentiated M anti-MAC anti- microbial activity, while Yokuinin did so to a much lesser extent. Secondly, MBST and Yokuinin each strongly inhibited production of nitric oxide (NO) in MAC-infected Ms. Thirdly, treatment of Ms with MBST or Yokuinin caused reductions in the accumula- tion of IL-10 in culture fluids by MAC-infected Ms during the first 2-days cultivation. On the other hand, in the separate experiment, treatment of Ms with these drugs caused no significant change in the accumulation of TGF- by MAC-infected Ms at day 7. These findings suggest that these Chinese traditional medicines, particularly MBST, potentiate M anti-MAC antimicrobial activity, however, NO do not appear to be cru- cial effectors in the anti-MAC activity of MBST-or Yokuinin-treated Ms. Moreover, MBST-and Yokuinin-mediated down-regulation of the production of IL-10 in MAC-in- fected Ms may be related to their potentiating effects on M anti-MAC activity. Key words:Mycobacterium avium, Mao- Bushi-Saishin-To, Yokuinin, Nitric oxide, IL-10, TGF- *89-1 Enya-cho, Izumo-shi, Shimane 693-8501 Japan. (Received 26 Feb. 1999/Accepted 9 June. 1999) <3> Kekkaku Vol.74,No.9:667-675,1999 CURRENT EPIDEMIOLOGICAL TREND OF TUBERCULOSIS AMONG FOREIGNERS IN JAPAN *Takashi YOSHIYAMA, Nobukatsu ISHIKAWA, Hitoshi HOSHINO, Akihiro OHKADO *Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association In Japan, the proportion of foreign patients among the total tuberculosis patients is still very small, but their problems in tuberculosis case finding and treatment require in- tensive control activities as in other low prevalence countries with higher proportion of foreignborn cases. The latest national survey for the foreign tuberculosis patients con- ducted in 1996 shows the epidemiological status between 1990 and 1993. The number of foreign tuberculosis patients in 1993 was 484, consisting of 1.0% of the total new pa- tients in Japan. The new case rate among foreigners was estimated to be 53 per 100,000 against 38 for whole Japan in 1993. Compared with the figure in the 1993 survey, the nimber of foreign patients declined from 585 in 1992 to 484 in 1993. However, the num- ber of bacillary positive tuberculosis patients in 1992 was 230 and almost the same as in 1992. The decline or stagnation of total number of tuberculosis patients can be due either to the decrease in the foreign population inflow into Japan (real decline), or partially to the reduction of overdiagnosis in X ray examination and the possible loss of some cases in the 1996 survey method. A manual sorting method form the registration cards was used at each public health center, since there is no item ot country of origin in the routine tuberculosis surveillance system, and some cards might have already been displaced by the time of the survey for patients who were excluded from the registry, either cured died or defaulted. The average treatment completion rate (119911-93) among foreign patients was 51%, which was much lower than the national figure of 81% for the same years. Moreo- ver the rate showed deteriorating trend by year. For more accurate information, the for- eigner's data must be taken in the national tuberculosis surveillance system and control activities for foreigners need to be strengthened. Key words:Foreigners, Tuberculosis, Treatment result, Epidemiology *3-1-24, Matsuyama, Kiyose-shi, Tokyo 204-8533 Japan. (Received 13 May 1999/Accepted 23 Jun. 1999) <4> Kekkaku Vol.74,No.9:677-681,1999 CHEMOPROPHYLAXIS AGAINST MYCOBACTERIUM AVIUM COMPLEX INFECTION INDUCED IN MICE 1*Hajime SAITO, 1Kazuyasu MURAKAMI, 2Kazuo KOBAYASHI, 2Masaichi GIDOH, 3Takayoshi HIDAKA, 4Hyuk Han KWON 1*Hiroshima Environment and Health Association, 2Leprosy Research Center, National Institute of Infectious Diseases, 3Takasago Research Laboratories, Kaneka Corporation, 4Seoul Health College, Korea M. avium complex (MAC) is one of the important causative agents of opportunistic in- fections among AIDS patients. Recent evidence showed that the entry of infection is through the gastrointestinal tract. In the present study, we compared the prophylactic ef- fect of some antimicrobials against MAC infection induced in mice. Different groups of beige mice were fed with pellets containing 0.0067% (10mg/kg) of KRM-1648, rifabutin (RFB) and clarithromycin (CAM). Seven days after the administration of drugs, the mice were infected with M.intracellulare N-241 (5x10(8) CFU) orally, five times, the every other day and killed one and 126 days after the 1ast infection. The effect of drug was evaluated using the frequency and severity of gross lung lesions in the mice and by the total CFU recovered from the lungs and spleen. MAC infection was not likely to be established since there was no macroscopic evidence of lesion in organs and the recovery of cultures from lungs and spleen tested was negative, in 3 of 10 mice in the control group, 2 of 9 in the CAM group, 4 of 9 in the RFB group and 4 of 10 in the KRM group. These mice were excluded from the analysis of the study results. Thus, we examined 7 mice in the control group, 7 in the CAM group, 5 in the RFB group, and 6 in the KRM group. Tubercle-like lesions were observed in the lungs of all 7 mice in the control group (se- verity:3+in 5 mice;4+in 2 mice), in 5 of 7 mice (71%) in the CAM group (severity :2+in 1 mouse;3+in 4 mice), and in 4 of 5 mice (80%) in the RFB group (severity :1+in 1 mouse:2+in 1 mouse; 4+in 2 mice), while only slight lesions (severity:1+) were observed in 4 of 6 mice (67%) in the KRM group. There was no macroscopic evidence of lesion in spleen, liver and kindneys. The log CFU was determined at the next day of the completion of the last infection. The log CFU of the lungs was 2.49 and 2.28 in the control group and the CAM group, respec- tively. The bacteria were not recovered either from the lungs in the RFB and KRM groups, nor from the spleen in all the groups. The order of efficacy of the drugs on the basis of the CFUs recovered from the lungs and spleen in each gruop determined 126 days after the completion of the last infection was as follows;KRM>CAM>RFB in the lungs and KRM>CAMRFB in the spleen, although there was no significant difference among the three drugs (p<0.05). However, the significantly preferable effect was ob- tained in the three drug groups as compared with the control group. Key words:Mycobacterium avium complex infection, Chemoprophylaxis, Clarithromycin, Rifabutin, KRM-1648 *9-1 Hirosekita-machi, Naka-ku, Hiroshima 730-8631 Japan. (Received 19 Apr. 1999/Accepted 16 Jun. 1999) @ <5> Kekkaku Vol.74,No.9:683-691,1999 The 74th Annual Meeting Special Lecture U.PRESENT SITUATION AND PROBLEMS OF TUBERCULOSIS IN JAPAN -Its Prevention, Diagnosis and Treatment *Masakazu AOKI *Japan Anti-Tuberculosis Association 1. Outline of tuberculosis epidemiology after the World War II Tuberculosis was surprisingly highly prevalent during the World War 2nd in Japan, and it was reported that the incidence of TB was as high as 698.4 in 1951. Strong TB control program has been carried out throughout the country after the new Anti-TB Law had been enacted in 1951. TB incidence decreased with the annual reduction rate of 11% up to 1976, it was one of the highest speed of TB decrease in the world. The decrease of TB is stagnating since 1977, and the incidence is still 33.9 in 1997. 2. Causes of stagnation of TB in Japan It is estimated that the annual risk of TB infection was so high as 4% in 1945 that many of the children had been infected with TB bacilli in those days. As a result, the majority of the people aged 60 years old or more at present are already infected with TB bacilli, and more than half of the new cases is being occurred among them at present in Japan. Moreover, the life span of those already onfected people is extending so remarkably that the percentage of the aged is increasing and TB incidence is stagnating in Japan. The causes of the stagnation are not the spread of HIV infection or the increase of TB among the immigrants or refugees, because both of them are not so big problems in Japan. However, more severe problems of TB epidemiology in Japan are that @the incidence of smear positive cases didn't decrease more than these ten years, Athe most marked stagna- tion of the incidence of TB is being observed maong the young aged 20 to 39, Bthe incidence of TB in big cities doesn't decrease recently and Cgroup infection and nosocomial TB infection is increasing recently. The author has analyzed the factors of the stagnation of TB in Japan, and come to the conclusions that it is caused by @the increase of the population by 3.0%, Athe aging of the population by 41.0%, Bthe increase of compromised host (diabetes mellitus and so on) by 19.8%, and Cother factors by 36.3%. It was considered that the main other factor is the marked stagnation of TB infection risk mainly by socioeconomic changes such as progressed urbanization and the spread of the modern housing with aluminum sash to increase the chances of TB infection in office buildings and/or private house. 3. Present problems of TB prevention, diagnosis and treatment in Japan As BCG vaccination is os widely and so repeatedly being carried out for children that the diagnosis of TB infection is almost impossible now in Japan. Accordingly the fre- quency and/or risk factors of TB infection in the community are not so clear. Preventive chemotherapy is given for high risk groups in Japan, but its diagnosis may become often unreliable. New techniques such as PCR, MTD, AccuProbe, RFLP and so are used fre- quently in Japan, but the consideration on the false negative results of the examinations are lacking on occasion. The results of the cohort analysis of TB treatment among all the registered cases are rather satisfactory, but it was concluded that the shortening of the duration of the hospitalization and chemotherapy is advised if compared with those of Japan and those of the other developed countries. Key words:Tuberculosis in Japan, Stagnation of TB decrease, TB infection *1-3-12, Misaki-cho, Chiyoda-ku, Tokyo 101-0061 Japan. (Received 9 Jul. 1999) <6> Kekkaku Vol.74,No.9:693-697,1999 The 74th Annual Meeting Presidentl Lecture PATHOGENESIS AND THERAPY OF ACUTE LUNG INJURY *Satoshi KITAMURA *School of Health and Social Services Department of Occupational Therapy, Saitama Prefectural University. Almost all of respiratory diseases except benign lung tumors and lung dysplasia entail acute lung injury (ALI). The many clinical conditions associated with acute lung injury include aspiration pneumonia, bacterial pneumonia nad sepsis. Acute lung injury is the end results of common pathways initiated by a variety of local or systemic insults lead- ing to diffuse damage to the pulmonary parenchyma. Despite the accumulation of abun- dant information regarding the physiological and cellular basis of lung injury and increasing sophiscated intensive care, an inprovement in prognosis has lagged behind. It has become clear that there is not one mediator responsible for ALI, but rather a com- plex interplay exists between diverse proinflammatory (eg, lipopolysaccharide, comple- ment products, cytocains, chemocains, reactive oxygen species and arachidonic acid products) and anti-inflammatory (IL-10, IL-1-RA, PG12) mediators. Early in the course of ALI, large numbers of neutrophils are sequestered in and emigrate from the pulmonary capillaries. The fundamental cause of ALI is pulmonary vascular hyperpermeability caused by the activated neutrophils which release oxygen radicals and proteases. In these processes several adhesion molecules play very important roles. Neutrophil elastase in- hibitors become very useful therapeutic agents against acute exacerbation of idiopathic interstitial pneumonia (IIP), because this pathological conditions is a type of ALI. Simi- larly, N-acetyl cystein could also become a useful therapeutic agent against IIP, because it is a precursor of glutathione, which is the major antioxidant in the fluid lining of the bronchial epithelium. Key words:Acute lung injury, Oxygen radicals, Adhesion molecules, Idiopathic interstitial pneumonia, Neutrophil elastase *820 Sannomiya, Koshigaya City, Saitama 343-8540 Japan. (Received 5 Jul. 1999)