(Vol.74 No.8 August 1999) <1> Kekkaku Vol.74,No.8:585-598,1999 INVESTIGATION OF PULMONARY HEMODYNAMICS AND CHEST X-RAY FINDINGS IN HYPERCAPNIC PATIENTS WITH PULMONARY TUBERCULOSIS SEQUELAE (1)*Jun-ichi YASUDA, (1)Osamu OKADA, (1)Takayuki KURIYAMA, (2)Keiichi NAGAO, (3)Fumio YAMAGISHI, (4)Ikko HASHIZUME, and (5)Akira SUZUKI (1)*Department of chest Medicine, Institute of Pulmonary Cancer Research, School of Medicin, Chiba University,(2)Health Sciences Center, Chiba University, (3)Division of Thoracic Disease, National Chiba-Higashi Hospital, (4)Department of Chest Medicine, Hamamatsu Medical Center, (5)Department of Respiratory Diseases, Tokyo Metropolitan Fuchu Hospital We investigated pulmonary hemodynamics and chest X-ray findings to explore pathophysiological significance of chronic hypercapnia in patients with pulmonary tuberculosis sequelae. One hundred and seven patients underwent examinations of blood gases and right cardiac catheterization. The patients were divided into two groups, according to arterial carbon dioxide tension under room air breathing (Paco(2)). Group I (n=35) was defined as 45 Torr or lower of Paco(2), and Group II (n=72) was the hypercapnic group whose Paco(2) was over 45 Torr. In addition, spirometry was done in 34 patients of Group I and 68 of Group II. First, the values of blood gases, spirometry and pulmonary hemodynamics were compared between the two groups. Secondly, between 22 of Group I and 50 of Group II, the values of pulmonary arteriolar resistance (PAR) before and after 100% oxygen breathing for 10 minutes were compared. These comparisons were made by exploratory data analysis. Lastly, we described in all cases with five items of chest X-ray findings and the extent of each finding we had defined. The items were emphysematous change;fibrosis, bronchiectasis, and/or cavity (hereafter abbreviated as "fibrosis"); lung resection and/or atelectasis;pleural thickening;and thoracoplasty. We explored the items of X-ray findings which may relate to hypercapnia by ridit(abbreviation for "relative to an identified distribution") analysis. The results were as follows. (1) Hypercapnic patients tended to have severer restrictive ventilatory impairment and hypoxemia. Under an even level of arterial oxygen tension (Paco(2)), tissue oxygenation was not poorer in Group II than in Group I. (2)Hypercapnic patients tended to have more unfavorable pulmonary hemodynamics. More than half of them had pulmonary hypertension defined as 20 mmHg or higher of pulmonary artery mean pressure (PAm). Under an even level of Paco(2), PAm was higher in Group II. Although 34 patients of Group II showed Paco(2) over 60 Torr, 23 of them had pulmonray hypertension. (3)PAR after oxygen breathing was more likely to decrease in Group II than in Group I. (4)As any mean ridit was standardized and adjusted to 0.5 in Group I, the maximum was the mean ridit of "pleural thickening" (=0.67), nextgfibrosish(=0.65) in Group II. The above two items of X-ray findings, in which each mean ridit was higher than in any other item, were more influential on hypercapnia. We conclude as follows. (1)Pulmonary hypertension is severer in hypercapnic patients with pulmonary tuberculosis sequelae;it may be mainly attributable to hypoxic pulmonary vasoconstriction. (2)An important cause of chronic hypercapnia may be pathological changes such as gpleu- ral thickeninghand gfibrosishseen on the radiogram. Key words:Pulmonary tuberculosis sequelae, Hypercapnia, Pulmonary hypertension, Hypoxic pulmonary vasoconstriction, Tissue oxygenation, Chest X-ray findings *1-8-8 Inohana, Chuo-ku, Chiba 260-8670 Japan. (Received 20 Nov. 1998/ Accepted 12 Apr. 1999) <2> Kekkaku Vol.74,No.8:599-609,1999 DEVELOPMENT OF SLIDE-METHOD TO DISTINGUISH ALIVE AND DEAD MYCOBACTERIA BY FLUORESCENT STAINING -A Trial for Solving the Biohazard Problem in TB Laboratories- *Masamichi KINOMOTO *Department of Bacterial and Blood Products, National Institute of Infectious Diseases An acid-fast staining can detect mycobacteria in clinical specimens rapidly and specifi- cally. It equally stains living and dead bacteria. It would be of more clinical use if the viability of mycobacteria in a sample was deter- mined by the staining. In the present paper, the problems of FDA/EB staining, which de- tects live or dead bacteria, were solved by establishing a new technique, a slide-method. An air-dried smear of Mycobacterium bovis BCG (Tokyo 172) on a glass slide was cov- ered by a filter paper fully soaked in the staining solution (500 ƒÊg FDA and 40 ƒÊg EB per ml PBS). This was kept in an incubator at 37Ž for 20 min. The filter paper was re- moved after incubation and the slide was examined using a fluorescent microscope with a blue filter. Live bacteria were stained greenish yellow taking the FDA stain in while dead bacteria were stained red with EB. This new slide technique eliminated the problems associated with FDA/EB staining. Moreover, stained smears appeared to be more stable compared with the conventional tube method. To overcome the biohazard problems in smear examination of tubercle bacilli, heating of the slides on a heat block at 100Ž for 20 min or passing air dried smears in a flame 5 to 30 times was tried to kill the bacteria. The heat-treated slides were stained with FDA /EB and the number of green and red bacteria were counted. Samples of the smeared bac- teria were taken after heating and cultured on a solid medium to determine the presence of any colony-forming unit. It was found that no CFU was observed after heating and the morphology of the stained sample was the same to that before heating. These facts suggest that the above mentioned method is a simple, safe yet inexpensive diagnostic tool for mycobacterial clinical specimens. Key words:Mycobacteria, Rapid diagnosis, Fluorescein staining, Esterase activity, Slide method *Gakuen 4-7-1, Musashimurayama-shi, Tokyo 208-0011 Japan. (Received 27 Jan. 1999/ Accepted 18 May 1999) <3> Kekkaku Vol.74,No.8:611-616,1999 THE CLINICAL EVALUATION OF MTD FALSE-POSITIVE & FALSE-NEGATIVE RESULTS (1,2)*Junko MIYAMOTO, (1)Atsuro HASHIMOTO, (1)Ryusuke MIZUKANE, (1)Toyohiro SASAKI, (1)Takakazu KIYA, (1)Masao NAKATOMI, and (3)Shigeru KOHNO (1)Department of Medicine, Nagasaki National Hospital, (2)*Department of Medicine, Mitsugi Public General Hospital, (3)Second Department of Internal Medicine, Nagasaki University School of Medicine The usefulness of MTD (Amplified Mycobacterium Tuberculosis Direct Test) for a rapid diagnosis of tuberculosis was evaluated. A total of 400 clinical samples obtained from July, 1995 to June, 1997 were tested by MTD, direct microscopy and culture, The results of MTD and smear/culture were coincident in 387 out of 400 samples. Eight samples (2%) were MTD false-positive (i. e. they were MTD positive but smear and cluture negative), and 5 (1.25%) were MTD false-negative (i. e. MTD negative but smear and/or culture positive). Despite a careful review of the clinical data of those patients whose samples showed discrepant results, the reasons of discrepancy were not clear in 2 (0.5%) of the 8 false positives and 3 (0.75%) of the 5 false negatives. In the other cases, the MTD false positives may be accounted for the presence of previous M.tuberculosis infection, the influence of anti-tubercxulous medication and so on, and the MTD false negatives are most likely due to the presence of inhibitors (blood, for example) or to the small number of organisms in the specimens. It can be concluded that adequate samples should be obtained, and that MTD should be repeated in case of discrepant results. Key word:MTD, Clinical specimens, False positive, False negative *124 Ichi, Mitsugi-cho, Mitugi-gun, Hiroshima 722-0393 Japan. (Received 8 Feb.1999/Accepted 26 May 1999) <4> Kekkaku Vol.74,No.8:617-621,1999 THERAPEUTIC EFFECTS OF BENZOXAZINORIFAMYCIN KRM-1648 ADMINISTERED ALONE OR IN COMBINATION WITH GLYCYRRHIZIN AGAINST MYCOBACTERIUM AVIUM COMPLEX INFECTION IN MICE (1)Toshiaki SHIMIZU, (1)*Haruaki TOMIOKA, (1)Katsumasa SATO, (2)Tatsuya AKAKI, (3)Keiko OGASAWARA, and (4)Shin KAWAHARA (1)*Department of Microbiology and Immunology, (2)Department of Dermatology, (3)Department of Otorhinolaryngology, Shimane Medical University, and Department of Internal Medicine, (4)National sanatorium, Minami-Okayama Hospital We previously examined the effects of a Chinese medicine gMao-Bushi-Saishin-Toh (MBST) which has anti-inflammatory activity on the therapeutic efficacies of a benzoxazinorifamycin, KRM-1648 (KRM), against Mycobacterium avium complex (MAC) infection induced in mice. MBST potentiated the therapeutic activity of KRM aginst MAC infection. In the present study, we examined the effects of another anti-inflamma- tory drug Glycyrrhizin, which is effective for chronic hepatitis, on the therapeutic efficacy of KRM aginst MAC infection induced in mice. First, KRM significantly inhibited the bacterial growth in the lungs and apleen of MAC-infected mice. Glycyrrhizin exhib- ited no therapeutic activity against MAC infection and did not affect the expression of the therapeutic efficacy of KRM. Secondly, treatment of murine peritoneal macrophages (MƒÓs) with Glycyrrhizin caused no significant changes in the MƒÓ anti-MAC activity. Key words :Mycobacterium avium complex, KRM-1648, Glycyrrhizin *89-1 Enya-cho, Izumo-shi, Shimane 693-8501 Japan. (Received 10 Mar. 1999/ Accepted 26 May 1999)