Cathepsin B activity has a crucial role in the developmental competence of bovine cumulus-oocyte complexes exposed to heat shock during in vitro maturation.

Cathepsin B was found to be correlated inversely with the quality of bovine oocytes and embryos. The aims of this study were to evaluate i) the relationship between heat shock during in vitro maturation (IVM) of bovine cumulus-oocyte complexes (COCs) and cathepsin B activity in relation to apoptosis and ii) the effect of supplementation of cathepsin B inhibitor (E-64) during IVM of heat-shocked COCs on embryonic development. After IVM at 38.5 °C for 22 h (control group) or at 38.5 °C for 5 h followed by 41 °C for 17 h (heat shock group) either with or without 1 µM E-64, activities and protein expression of cathepsin B and caspase 3 were evaluated as well as TUNEL staining. After IVF, developmental rate, total cell number, and the percentage of apoptotic cells in blastocysts were evaluated on day 8 (day 0, IVF day). Heat-shocked IVM COCs showed significantly high activities and expressions of both cathepsin B, and caspase 3 accompanied by a significant increase in number of TUNEL-positive cells. Addition of E-64 significantly decreased the activities of cathepsin B and caspase 3, and TUNEL-positive cells in heat-shocked IVM COCs. Moreover, addition of 1 µM E-64 during IVM under heat shock conditions significantly improved both developmental competence and quality of the produced embryos. These results indicate that heat shock induction of cathepsin B is associated with apoptosis of COCs, and inhibition of cathepsin B activity can improve the developmental competence of heat-shocked COCs during IVM.

Efficacy and safety of voriconazole in the treatment of chronic pulmonary aspergillosis: experience in Japan.

BACKGROUND: Though various clinical conditions of aspergillosis can occur, depending essentially on the host's immunological status, the focus of research in North American and European countries has mainly been on invasive pulmonary aspergillosis in immunocompromised patients. There are, however, also many problems to overcome in chronic forms of aspergillosis. One of those problems is that there are no codified treatment guidelines for chronic pulmonary aspergillosis (CPA). Especially in Japan, this issue is more serious, because there are more cases with CPA due to the many aged people with past history of tuberculosis. Several clinical cases and case series have reported the usefulness of the various antifungal agents that are available. The new triazole, voriconazole, in particular, seems to be effective in the treatment of CPA. The aim of the present study is to evaluate the efficacy and safety of voriconazole in the treatment of CPA in non-immunocompromised patients. PATIENTS AND METHODS: We conducted a prospective, open-label, non-comparative, multicenter study over a 2-year period. For inclusion in the study, patients with confirmed or probable CPA were recruited in 11 hospitals of the National Hospital Organization in Japan. Clinical, radiological, serological, and mycological data were collected at baseline and 12 weeks after treatment or at the end of treatment. RESULTS: Among 77 patients enrolled in the study, 71 patients (mean age 65.9 years, 56 males and 15 females) were eligible for the study. All of the eligible patients presented with underlying lung diseases, including sequelae of tuberculosis (n = 35), non-tuberculous mycobacterial lung disease (n = 8), chronic obstructive pulmonary disease (COPD) (n = 8), interstitial pneumonia (n = 7), cystic lung disease (n = 4), pneumothorax (n = 3), bronchial cancer (n = 1), and others (n = 5). Voriconazole was indicated in 48 cases (68 %) as the first-line treatment for CPA and 23 patients previously received other antifungal therapies. Based on a composite of clinical, radiologic, serological, and mycologic criteria, good response was seen in 43 patients (60.6 %), no response was observed in 19 patients (26.8 %), and 4 cases (5.6 %) got worse. Five patients (7.0 %) were unassessable for efficacy. The common adverse events were visual disturbances (17 patients, 23.9 %), abnormal liver function test results (12 patients, 16.9 %), adverse psychological effects (3 patients, 4.2 %), and others (10 patients, 14.0 %). Treatment with voriconazole had to be stopped in 2 cases (2.8 %) because of serious adverse events (abnormal liver function test results). There was no association between adverse effects and trough voriconazole levels in serum. CONCLUSIONS: In Japan, voriconazole provides effective therapy of CPA in non-immunocompromised patients with an acceptable level of toxicity.

Consequences of physiological heat shock beginning at the zygote stage on embryonic development and expression of stress response genes in cattle.

The goal was to understand the role of heat shock at the zygote stage in causing infertility. Culture at 40°C reduced the percentage of inseminated oocytes that became a morula or blastocyst by d 6 or that were a blastocyst at d 8. An additional experiment was done to test whether effects of heat shock occur early in development or at the time of morula formation. Exposure to 40°C for 24 h decreased development to the blastocyst stage if exposure was at the zygote stage [8 to 32 h postinsemination (hpi)] but not if exposure occurred at the morula stage (116 to 140 hpi). To test effect of oxygen concentration, inseminated oocytes were cultured at 40°C for 12 or 24 h in either air (20.95% O2; high oxygen) or a 5% (vol/vol) O2 environment (low oxygen) that approximates the partial oxygen pressure of the reproductive tract. Blastocyst development was reduced by 40°C for 12 or 24 h under both atmospheres and was higher for embryos cultured in low oxygen than for embryos cultured in high oxygen. Examination of cell numbers at 72 hpi indicated that heat shock reduced developmental potential of embryos by reducing competence to complete cleavage divisions after first cleavage. Changes in expression of genes involved in heat shock and oxidative stress were measured to determine whether zygotes are more susceptible to heat shock because of reduced capacity for transcription. Heat shock was performed for 24 h at the 1-cell stage (expression examined in 2-cell embryos) or at d 5 (examined in morulae). Heat shock increased amounts of steady-state mRNA for HSPA1A but not for HSP90AA, SOD1, or CAT. We observed a tendency for a stage × temperature interaction for HSPA1A because the difference in expression between 38.5 and 40°C was greater for morulae than for 2-cell embryos. The amount of HSPA1A mRNA was less for morulae that were heat shocked than for 2-cell embryos cultured at 38.5°C. Heat shock at a temperature and oxygen tension similar to those seen in vivo can disrupt developmental competence of bovine zygotes. Increased susceptibility of the early embryo compared with the morula to heat shock was not due to reduced HSPA1A mRNA because amounts were higher for 2-cell embryos than for morulae.

Cathepsin B activity is related to the quality of bovine cumulus oocyte complexes and its inhibition can improve their developmental competence.

Recently, the quantity of cathepsin transcripts in cumulus cells was found to be associated with low-developmental competence of bovine oocytes. In the present study, we investigated (1) the relation between cathepsin B activity and the quality of in vitro-matured cumulus-oocyte complexes (IVM COCs) and denuded oocytes and (2) the effect of a cathepsin B inhibitor (E-64) on embryo development and quality. The activity of cathepsin B was evaluated in IVM COCs and denuded oocytes. After maturation of COCs with or without E-64, followed by in vitro fertilization, zygotes were cultured for 8 days. Cleavage and blastocyst rates were evaluated on days 2 and 8, respectively. Quality of embryos was evaluated by differential staining of day 8 blastocysts. TUNEL staining was conducted on IVM COCs and blastocysts. Cathepsin B activity was clearly detected in the low-quality oocytes, and in the cumulus cells of both high- and low-quality oocytes. This latter activity was diminished by addition of E-64. The presence of E-64 during IVM also significantly increased both the blastocyst rate and the total cell number, and improved blastocyst quality associated with a significant increase of trophoectoderm cells. TUNEL staining revealed that inhibition of cathepsin B significantly decreased the number of apoptotic nuclei in both the cumulus cell layer of matured oocytes and blastocysts. These results indicate that cathepsin B activity can be a useful marker of oocyte quality. Furthermore, inhibition of cathepsin B greatly improves the developmental competence of bovine oocytes and increases the number of high-quality embryos.

Intracellular cathepsin B activity is inversely correlated with the quality and developmental competence of bovine preimplantation embryos.

Recently, the activity of cathepsins B was found to be correlated inversely with the developmental competence of bovine oocytes. In this study, we investigated (1) the role of intracellular cathepsin B expression and developmental competence as well as the quality of bovine preimplantation embryos, and (2) the effect of cathepsin B inhibitor (E-64) during in vitro culture (IVC) on the development and quality of bovine embryos. After in vitro fertilization (IVF) followed by IVC for 7 days, good and poor quality embryos classified by morphology and developmental rate on days 2, 4, and 7 were assessed for cathepsin B expression and activity. To investigate the effect of cathepsin B inhibition on embryonic development, putative zygotes were cultured with or without E-64, followed by evaluation of cleavage and blastocyst rates on days 2 and 7, respectively. Embryonic quality was evaluated by both TUNEL staining and total cell number in day-7 blastocysts. In each developmental stage, cathepsin B expression and activity were significantly higher in poor quality embryos than good quality ones. Moreover, addition of E-64 during IVC significantly increased both the blastocyst rate and the total cell number. TUNEL staining revealed that inhibition of cathepsin B significantly decreased the number of apoptotic nuclei in day-7 blastocysts. These results indicate that cathepsin B activity can be useful as a marker for inferior quality embryos. Moreover, inhibition of cathepsin B greatly improves the developmental competence of preimplantation embryos and increases the number of good quality embryos.

Eosinophilia in bronchoalveolar lavage fluid and architectural destruction are features of desquamative interstitial pneumonia.

AIMS: Desquamative interstitial pneumonia (DIP) is a rare pattern of diffuse parenchymal lung disease known to overlap with respiratory bronchiolitis-interstitial lung disease (RB-ILD). The aim was to review biopsy-proven cases of DIP to investigate further the clinical, imaging and histological features of this disease. METHODS AND RESULTS: Twenty patients fulfilled the pathological criteria: 19 men and one woman with a mean age of 54 years. Clinical features, bronchoalveolar lavage (BAL) data, radiological findings, pathological findings other than criteria, effect of therapy and outcome were examined. The BAL data for 17 cases revealed marked eosinophilia (mean 18%) and moderate neutrophilia (mean 11%). Computed tomography in 17 patients showed peripheral involvement in all cases with a clear margin in 64% and thin-walled cysts in 35% of cases. Additional pathological features were a distinct lobular distribution (70%) and architectural destruction (70%) with cyst formation (55%). Eighteen of the 19 patients (95%) improved under steroid pulse and/or oral therapy. Sixteen subjects (80%) are alive, three died of other diseases and one died of DIP 74 months after the diagnosis. Percent vital capacity increased significantly and new thin-walled cysts appeared in one case. CONCLUSIONS: BAL eosinophilia, lobular distribution and architectural destruction with cyst formation are characteristic features of DIP.

Differential induction of squamous cell carcinomas and adenocarcinomas in mouse lung by intratracheal instillation of benzo(a)pyrene and charcoal powder.

Differential induction of squamous cell carcinomas, adenomas, and adenocarcinomas was observed in the lungs of male C57BL/6 and C3H/He mice after repeated intratracheal instillation of benzo(a)pyrene (BP) and charcoal powder suspended in 0.9% NaCl solution. when a high dose of BP (1.0 mg BP and 0.5 mg charcoal powder) was instilled intratracheally once a week for 8 weeks or when a low dose of BP (0.5 mg BP and 0.5 mg charcoal powder) was instilled once a week for 16 weeks, squamous cell carcinomas were induced in high incidence (77 to 87%) in the early period of observation, whereas pulmonary adenomas and adenocarcinomas were induced in low incidence (0. to 48%) in the late period of observation in both strains of mice. On the other hand, when a low dose of BP was instilled intratracheally once a week for 8 weeks, pulmonary adenomas and adenocarcinomas were induced in high incidence (76 to 91%), but squamous cell carcinomas were induced in low incidence (9 to 26%). These results show that a larger quantity of BP instilled intratracheally was needed for induction of squamous cell carcinomas than for induction of adenomas and adenocarcinomas in the lung of mice. Thus, when the carcinogen is administered to a single organ of a single mouse strain by the same route, different amounts of carcinogen have different effects on the incidences of various histological types of tumors.

Randomized controlled study of adjuvant immunotherapy with Nocardia rubra cell wall skeleton for inoperable lung cancer.

In order to evaluate the clinical benefit of Nocardia rubra cell wall skeleton (N-CWS), a randomized controlled study was performed with inoperable lung cancer patients entered in 5 institutions from October 1978 to June 1981. Patients without pleural effusions were treated initially with conventional therapies such as chemotherapy and/or radiotherapy, according to common protocol, and then patients in performance statuses 0 to 3 were randomized into control and N-CWS groups with stratification into 16 categories according to 4 histological types and 4 clinical stages In the N-CWS group, 400 micrograms N-CWS were initially injected once or twice into the bronchial tumor using a fiberoptic bronchoscope, and subsequently 200 micrograms of N-CWS were injected at monthly intervals into the skin from the shoulders to upper arms. Of 309 patients, 118 patients in the N-CWS group and 108 patients in the control group were eligible for statistical analysis. There was statistically no significant difference in survival rate between the control and the N-CWS groups. According to histological type, significant prolongation of the survival period was observed in patients with small-cell carcinoma. The 97 patients with pleural effusions were initially randomized into control and N-CWS groups. In the control group, local chemotherapy with Adriamycin was performed and, in the N-CWS group, local administrations and monthly intracutaneous injections of N-CWS were given. Tube thoracostomy was performed in both groups. The local response rate was statistically greater in the N-CWS group than in the control group, and survival period was also prolonged significantly in the N-CWS group. The main adverse reactions to N-CWS were skin lesions in the injected sites and fever, but these were temporary and not serious.

Risk of developing active tuberculosis in rheumatoid arthritis patients on adalimumab in Japan.

SETTING: Nationwide study in Japan. OBJECTIVE: To identify the clinical characteristics of and risk factors for tuberculosis (TB) in rheumatoid arthritis (RA) patients receiving adalimumab. DESIGN: A post hoc case-control study involving 7755 RA patients evaluated in an all-case post-marketing surveillance of adalimumab from 18 June 2008 to 20 January 2011 and spontaneous report of TB during adalimumab treatment. RESULTS: The analysis included 22 TB cases and 7733 non-TB controls. Despite the high frequency of extra-pulmonary TB (17/22, 77.3%), all 22 TB cases had pulmonary signs or symptoms. Of patients enrolled in all-case post-marketing surveillance, there were relatively high percentages of patients who had latent tuberculous infection (LTBI) at baseline. The percentage of patients who received prophylaxis was about six times higher in patients with LTBI than those without. Of those patients who received prophylaxis, none developed TB after treatment with adalimumab. Multivariate (non-matched and age-matched) and propensity score analyses identified age ⩾65 years (OR 4.59, 95%CI 1.52-13.89, P = 0.0070), moderate to severe RA (OR 4.61, 95%CI 1.07-20.00, P = 0.0408], past or current renal dysfunction (OR 4.65, 95%CI 1.47-14.71, P = 0.0089) and diabetes mellitus (OR 3.30, 95%CI 1.31-8.26, P = 0.0110) as potential risk factors for TB. CONCLUSION: Screening, prophylaxis and monitoring for TB are essential to ensure the safety of adalimumab treatment.

Pulmonary hyalinizing granuloma with Castleman's disease.

A case of pulmonary hyalinizing granuloma (PHG) with Castleman's disease in a 43-year-old man is presented. He was asymptomatic, but the disease was recognized due to a routine chest roentgenographic study. Anemia, multiple lymphadenopathy, hypoalbuminemia and polyclonal hypergamma-globulinemia were observed. Histological examination of cervical lymph nodes revealed the plasma cell type of Castleman's disease. The diagnosis of PHG was confirmed by video-assisted thoracoscopical lung biopsy, and the immuno-histochemical staining of lamellar fibrosis for types I and III collagen was positive.

KL-6 as a serologic indicator of Pneumocystis carinii pneumonia in immunocompromised hosts.

KL-6, a serum marker for interstitial pneumonitis, was evaluated in patients with Pneumocystis carinii pneumonia (PCP). Patient 1 was a 56-year-old woman with rheumatoid arthritis treated with immunosuppressive drugs and corticosteroids. Patient 2 was a 59-year-old man with a glioblastoma who received anti-cancer drugs and corticosteroids. In both patients, serum KL-6 showed an abnormally high level due to the complication of PCP, and it decreased following successful treatment. These results indicate that PCP is one of the diseases in which serum KL-6 increases.

Paragonimiasis miyazakii associated with bilateral pseudochylothorax.

A 37-year-old man who suffered from bilateral pleural effusions, subcutaneous abdominal induration and blood eosinophilia, was admitted to our hospital. He had ingested raw crabs at a pub-restaurant before the onset of his symptoms. His pleural effusions were chyliform containing cholesterol crystals, and a high level of immunoglobulin E (36,580 IU/ml) and anti-Paragonimus miyazakii antibody were detected. He was effectively treated with praziquantel. This case suggests that paragonimiasis should be strongly suspected if blood eosinophilia, pseudochylothorax, and a high level of immunoglobulin E in pleural effusion are detected.

Asbestos-related lung cancer and mesothelioma in Japan.

In Japan, crocidolite had been used for asbestos cement pipe and spraying, and amosite had been used for building board and spraying. These two types of asbestos had stopped to use in Japan in the late 1970s. An extreme increase in imported asbestos (all 3 commercial types) was observed between 1960 and 1974. In 1960, 77,000 tons of asbestos were imported, and reached the peak as 352,316 tons in 1974. This extreme rise of asbestos imports corresponds with the recent rapid increase in mortality of malignant pleural mesothelioma. Between 1995 and 1999, an estimated mean annual death from pleural mesothelioma was about 500. The annual number of compensated occupational respiratory cancers due to asbestos exposure has also been increasing. Up to the end of March 2000, 162 cases with malignant mesothelioma and 197 cases with lung cancer were compensated. As for lung cancer, epidemiological studies are scanty in Japan. Limited environmental data of the working places in asbestos textile factories suggests that heavy asbestos exposure in the past made deaths from respiratory diseases. Less asbestos exposure will enable exposed workers to survive enough to reach cancer age. Even now smoking rate among males in Japan are over 50%. So lung cancer deaths caused by the interaction between smoking and asbestos exposure will be continuing.

[Optimization of arbitrarily primed polymerase chain reaction (AP-PCR)-based DNA fingerprinting of Mycobacterium tuberculosis and its clinical application].

Since the number of outbreaks of pulmonary tuberculosis is increasing in Japan, epidemiological analysis is important to prevent the disease. Since Mycobacterium tuberculosis lacks the variety of biotypes among strains, genetical analysis is considered to be a promising measure to differentiate various of this pathogen. We applied arbitrarily primed polymerase chain reaction (AP-PCR)-based DNA fingerprinting to clinically isolated strains of M. tuberculosis. Although genetic analyses of M. tuberculosis by AP-PCR were reported by several investigators, reproducibilities of their results were not sufficient to be widely accepted as a reliable epidemiological tool. To attain high reprodicibulity, we attempted to optimize AP-PCR conditions including primers and annealing temperature, and to purity of DNA preparations. In this study, high reproducibility was attained by using the mixed primers of 1309F and 92R, and DNA preparations with an absorbance ratio (A260/A280) of higher than 1.50. Twenty two clinical isolates, including strains isolated from one incidence of nosocomial infection and from that of intrafamilial infection were analyzed by the optimized method; consequently they were grouped into 16 types. This AP-PCR method requires only one week subculture of M. tuberculosis and less than 24 hours for analysis. This AP-PCR method allowed us to obtain the highly reproducible results within a considerably short term, which would be applicable to clinical epidemiological investigation.

[The epidemiology of pulmonary disease caused by Mycobacterium avium complex in Japan].

The number of the patients of active lung tuberculosis and nontuberculous lung mycobacteriosis (NTM) admitted to the 97 sanatoriums in Japan were studied. The number and the prevalence rate of tuberculosis did not decrease during the years from 1985 to 1990, indicating the prevalence rate of 43.1 in 1990 per 100,000 population. Nevertheless, the number of NTM has gradually increased in these 6 years. The prevalence rate of NTM and pulmonary M. avium complex disease (MAC) calculated were 4.18 and 2.46 in 1985, increasing to 6.90 and 4.53 in 1990. Consequently, annual number of the patients newly infected with MAC was suspected to be approximately 5500 in 1990 in Japan. These infections occurred principally in South-West of Japan in the previous years, and were not so common in northern part of Japan. However, a remarkable increase in the number and the prevalence rate of MAC infection has also been noted in northern part of Japan in these 6 years. The number of MAC patients distributed equally in both sexes and the average age was 67 years. The 60% of the patients had some underlying diseases.

[Nontuberculous mycobacteriosis (NTM) in Japan--epidemiologic and clinical study].

The epidemiologic study for NTM was conducted among 211 national, provincial and private sanatoriums in Japan. The case cards of patients with NTM disease from 97 hospitals were collected by questionary method. The total number of NTM patients newly admitted in these hospitals were 2,873 in 7 years from 1985 to 1991, and the culture positive tuberculosis patients were 22,836 cases in the same period. The number of NTM patients with Mycobacterium avium complex (MAC) and with Mycobacterium kansasii were 1,675 and 240 respectively. The NTM patients were increasing year by year and the prevalence rate was estimated at 2.45 per 10(5) population in 1991, while on the other hand the rate of tuberculosis announced officially by the Ministry of Public Welfare was 15.0 in 1991 (rates were almost the same in these 7 years). The almost 3,000 patients were supposed to be affected by NTM in 1991, and one out of 7 patients infected by acid fast bacilli may be NTM case. The 3 out of 4 NTM patients are MAC cases and another one is M. kansasii case. In MAC cases M. avium are predominant in the northern half (from Kinki to Hokkaido) and M. intracellulare are predominant in the southern half (from Tyugoku to Kyushu) of Japan. For MAC cases, the number of patients were almost the same in male and female, the average age was 66 and 67 years respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

[Nontuberculous mycobacteriosis; the present status of epidemiology and clinical studies].

In Japan, The Mycobacteriosis Research Group at the Japanese National Chest Hospitals has continuously made the clinico-epidemiological study of nontuberculous mycobacteriosis (NTM) since early 1970s. The prevalence rate was determined as 0.82, 0.91, 1.22, 1.74 and 2.43 per 100,000 population per year in 1971, 1975, 1980, 1985 and in 1990 respectively. The newest datum in 1997 was 3.52. These data indicates the prevalence rate has continuously increased and became 3.8 times than 25 years ago. While on the other hand, the prevalence rate of lung tuberculosis has decreased from 133.1 to 15.2, becoming one nines in the same period. The numbers of newly detected patients of lung mycobacteriosis in 1996 were also studied at 12 hospitals in Kinki district. The rate of NTM was 16.6% in 4 sanatorium hospitals, being about the same to the datum of The Mycobacteriosis Research Group. The rate of NTM in 8 general hospitals was surprisingly high, 40.0%. The 70% of NTM patients were infected with Mycobacterium avium complex (MAC). The 24% were with M. kansasii, and the only 6% were with other miscellaneous species. That is, the about one thirds or more of total NTM patients were female MAC desease patients, another one thirds or less were male MAC patients, and the more than 90% of M. kansasii patients (about one fourth of total patients) were male. These 3 groups took the most part of NTM patients. The rate of female MAC patients with small non-cavitary lesion without underlying diseases showed a tendency to increase, and the rate of male MAC patients with cavitary lesions with underlying lung or systemic diseases decreased. In 1997, American Thoracic Society (ATS) published the official statement about the diagnosis and treatment of NTM disease. The table-1 in that statement showed the new criteria for diagnosis of NTM pulmonary disease. It is useful for precise diagnosis of lung NTM disease, and the old criteria made by The Mycobacteriosis Research Group of the Japanese National Chest Hospitals is also useful for rough diagnosis. In the ATS statement, for adult HIV-negative MAC patients, minimum three drug regimen of clarithromycin (or azithromycin), rifabutin (or rifampin) and ethambutol, with intermittent streptomycin which is option for extensive disease, is recommended. This regimen is the same that most of the Japanese specialists for NTM disease recommended. The follow-up study of 47 Japanese MAC patients treated by the regimen contained clarithromycin with other anti-tuberculous drugs revealed that 80% of cases converted into bacilli negative and that the regimen had durable effect for at least 24 months. The resectional surgery may be considered for localized disease, and supportive nutritional treatment must also be considered for the MAC patients to whom the drug therapy was not effective, as if for the tuberculosis patients of multi-drug resistant.

[Multidrug-resistant tuberculosis. 4. Treatment and prognosis of multidrug-resistant tuberculosis].

We studied the clinical characteristics, treatment and prognosis of multidrug-resistant pulmonary tuberculosis patients retrospectively. In this study, multidrug-resistant is defined as both resistant to 0.1 microgram/ml of INH and 50 micrograms/ml of RFP at least. From 1990 to 1997, out of 1841 culture positive pulmonary tuberculosis patients, 76 patients (4%) proved to be multidrug-resistant (53 males, 23 females, age 18-84, 40 originally treated cases and 36 relapse cases). Most of cases revealed resistance to other drugs in addition to INH and RFP. The combination of anti-tuberculous drugs were complicated and changed repeatedly. The incidences of administration of drugs were as follows; TH 62%, EB 58%, PZA 58%, KM 33%, PAS 33%, SM 29%, CS 20%, EVM 14%, CPM 3%. New quinolones, for example OFLX/LVFX, CPFX and SPFX, were also used frequently (62%). Eight percent of patients were operated. Bacteriologically effective drugs that meant culture negative were TH (14%), PZA (12%), KM (12%), EB (12%), SM (5%), new quinolones (16%). 67% of originally treated cases and 43% of relapse cases became culture negative. Many cases were treated for a long period. 19% of originally treated cases and 33% of relapse cases were treated more than three years. 11% of patients were died of tuberculosis. Major prognostic factors were diabetes mellitus (17%), malignancies (10%), non-adherence (9%) and other complications. Because of no absolutely effective treatment, we have to choose a treatment according to each patient. Development of new treatment is crucial.

[Epidemiologic study of the actual investigation of chronic excretors of Mycobacterium tuberculosis bacilli].

Of 1,295 patients with tuberculosis who were admitted in 33 Japanese national sanatoriums from September to October, 1994, actual investigation of patients with persistent expectorate tubercle bacilli continuously for more than 12 months was carried out by questionnaire. From the result, numbers of patients who had persistently expectorated bacteria were 85 (6.6% of inpatient). The rate of male and female was 3:1, and mean age was 63.5 years old. Many slim type patients (less than 90% of the standard body weight) were observed at the rate of 55.8%. The rate of the inpatient whose admission period was for more than 10 years were found in 22%. The past history of the tuberculosis and surgical therapy were existent at the rate of 83.3% and 18.8% respectively. The chief complications such as diabetes mellitus (12.4%), pyothorax (10.1%), alcoholism (10.1%) and hepatic dysfunction (6.7%) were observed at the respective rate. As chest X-ray findings on admission, severe cases with cavity of I or II 3 by the classification of the Japanese society for tuberculosis were found in 32.2%, and many cases with much amount persistently excreted bacteria such as more than number 7 of Gaffky scale (24.7%) or 3+ by culture (55.8%) were detected by sputum-test on admission. From the result drug-resistance, resistance of the main drugs early found from initial medication, and high degree resistance of isonicotinic acid hydrazide (INH; 80%), rifampicin (RFP; 94.4%), ethambutol (EB; 85.6%) and streptomycin (SM; 76.4%) was found. As intractable causes which were indicated by doctor in charge, drug-resistance, drug allergy, disorder of life, unfavorable medication-compliance and unsuitable treatment were considered.

[II. multidisciplinary analysis of chronic excretors of Mycobacterium tuberculosis bacilli].

For the most patients with pulmonary tuberculosis, treatment are successful and the chemotherapy can be finished within a year. Few patients, though receiving standard anti-tuberculous chemotherapy, expectorate tubercle bacilli continuously for more than a year. They are named "chronic excretors of mycobacterium tuberculosis bacilli" or "intractable tuberculosis patients". Being suspected that this phenomenon is produced by many factors, it is necessary to correspond suitably to these patients for eradication of tuberculosis. This symposium was organized to search for some factors on the bacterium and on the host, which caused patients to be the chronic excretors or intractable patients. We hope that our discussion may contribute to take the appropriate measures for such hard diseases. The main results are summarized as follows. 1) Dr. TSUCHIYA conducted a epidemiologic study by questionaire method to 33 Japanese national sanatoriums. The total number of tuberculosis patients admitted to these hospitals on a optimal day from September to October 1994 were 1,295. Numbers of patients who had expectorated bacilli continuously for more than 12 months were 85 (6.6%). The male/female ratio was about 3, their average age was 64. They had a tendency of thin-physique and most of them had past histories of tuberculosis, being the refractory cases. The average clinico-epidemiologic conditions of the patients came before our eyes. 2) Dr. OGATA analysed the results of drug sensitivity tests of mycobacterium tuberculosis bacilli from 2,630 pulmonary tuberculosis patients which contained 179 chronic excretors. The only 2.2% of bacilli from the chronic excretors were sensitive to RFP (50 gamma). The 99.4% of bacilli showed to be resistant to INH (0.1 gamma). The 39.1% of bacilli were sensitive to streptomycin (20 gamma). To EB (2.5 gamma) 34.1% of bacilli were sensitive. However, the ratio of chronic excretors in tuberculosis patients was not increased in these 17 years, and for many cases of these chronic excretors, they are all inexperienced in PZA, multi-sensitive-drug chemotherapy including PZA and OFLX were useful. Some of them underwent lobectomy combined for effective chemotherapy. The chronic excretors are likely to be affected multidrug-resistant bacilli, and it is important for such cases to treat intensively with several kinds of sensitive drugs. 3) Dr. FUJIWARA studied Interleukin-10 (IL-10) producing function of monocytes stimulated by Mycobacterium tuberculosis (M-TB). The IL-10 usually down-regulates a number of different macrophage functions, including microbicidal activity against intracellular bacteria such as Mycobacterium species. They found that the mean IL-10 production by peripheral blood mononuclear cells (PBMC) obtained from a healthy tuberculin-reactor stimulated with multidrug-resistant M-TB was greater than that with drug-sensitive M-TB. The IL-10 producing cells were monocytes, not T-lymphocytes. They moreover found that PBMC from intractable tuberculosis patients secreted greater amounts of IL-10 than those of healthy subjects. These data may give a hint for development of new immunotherapeutic methods for multi-drug-resistant tuberculosis patients. 4) Dr. HARA analysed clinical courses of 580 consecutive cases of bacilli positive pulmonary tuberculosis admitted in his hospital from a point of underlying diseases. All of dead 24 cases had some underlying diseases; diabetes mellitus 3, chronic liver diseases 4, others 17 (malignant neoplasms, central nerve vascular diseases, malnutritional condition, etc). (ABSTRACT TRUNCATED)