We mesured neutrophil's CL (CL-index) in 12 patients of the bacterialpneumonia three times per each case: before, during and after chemotherapy. Before the initiation of chemotherapy. CL-index in the six patients remained higher than that in healthy controls, while theremaining six showed lower levels of CL-index compared to the controls. In the 11 cases, their CL-indexes fell to levels lower than those obtained before treatment.
Additionally, in the 11 cases their CL-indexes increased after the termination of chemotherapy. Furthermore, in the nine cases the product of the neutrophil number and CL-index was decreased by chemotherapy, and the decrease in the product correlated with improvementin their clinical conditions.

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Trichosporon beigelii (formerly called Trichosporon cutaneum) is an emerging pathogen of disseminated trichosporonosis in immunocompromised patients. We conducted postmortem microbiological examinations of lung aspirates and blood in the heart. T. beigelii was isolated from the samples of 7 (2.24%) of 313 patients. Five of 7 patients isolated were diagnosed as having trichosporonosis based on histochemical and clinical findings. The other two were considered as colonization. Immunohistochemical study using Cryptococcus neoformans-absorbed antiserum to T. beigelii was carried out on the sections of autopsied tissues with deep mycoses. The sections from seven patients were positively stained, which were formerly diagnosed as candidiasis and/or aspergillosis. Consequently, 9 patients had disseminated trichosporonosis caused by T. beigelii. In Japan, 43 patients including the present 9, 31 males and 12 females, aged 51 (range, 2-84), were reported in the literature. Thirty-seven (86%) patients had hematologic malignancy, and the majority of them revealed profound neutropenia due to cytotoxic chemotherapy. Thirty-eight (88%) patients died despite the anti-fungal chemotherapy including amphotericin B. New strategies for refractory disseminated trichosporonosis in immunocompromised patients is needed.

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Certain bacteria produce some carcinogens such as N-nitro compounds, n-butyric acid and n-valeric acid.
From this point of view, the examination of intrauterine bacterial flora in patients with uterine endometrial cancer may provide important information.
Twenty patients with the diagnosis of uterine endometrial cancer and 20 patients without complications other than myoma uteri were enrolled in the study. Enterobacteriaceae, Streptococcus agalactiae and anaerobic bacteria were mainly detected.
The products of these bacteria might be considered to contribute to the initiation of endometrial carcinogenesis.
Mixed abnormal flora between aerobic and anaerobic bacteria were detected in all patients with uterine endometrial cancer. It is suggested that uterine endometrial cancer provides favorable conditions for bacterial growth. Mixed abnormal bacterial flora also might influence the onset and growth of uterine endometrial cancer.

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A recent nationwide increase in β-lactams-resistant Streptococcus pneumoniae has attracted a great deal of attention. We studied the drug sensitivity of S. pneumoniae isolated from various clinical specimens in Saga Medical School Hospital between April 1988 and December 1991.
To determine the drug sensitivity of the strains, we used a micro-dilution method and detemined the MIC. Drug resistance was evaluated using MIC of ampicillin (ABPC) as a reference MIC, and the results were roughly classified into the following three groups: sensitive (≤0.1μg/ml), moderately resistant (0.2-3.13μg/ml) and highly resistant (≥6.25 μg/ml). The isolation frequency was calculated on the basis of one strain from one patient. No strain of S. pneumoniae with high resistance against ABPC was found in 1988 (94 strains of S. pneumoniae were isolated) and 1990 (115 strains isolated), but one such strain (0.8%) was found among 129 strains isolated in 1989, and 2 such strains (2.4%) among 84 strains isolated in 1991. Moderately resistant strains were isolated at the frequencies of 12.8%, 15.5%, 22.6%, and 21.4% respectively, in 1988, 1989, 1990, and 1991. A sum of the frequencies of “moderately resistant” and “highly resistant”(2.4%) strains was 23.8% in 1991. The frequency of resistant strains is increasing and the intensity of resistance is also being elevated.

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Polymerase chain reaction was applied to identify enterotoxin-producing Clostridium perfringens by amplifying a segment of the C. perfringens enterotoxin gene. All of the four enterotoxin-positive reference strains tested were PCR positive while an enterotoxin-negative strain was PCR positive. All 17 clostridial strains (16 species) other than C. perfringens were PCR negative. With clinical strains isolated from various clinical specimens in Japan, Korea, and Thailand, all three enterotoxin-positive isolates were PCR positive and all 82 enterotoxin-negative isolates were PCR negative. PCR results for amplifying a region containing the initiation codon of the C. perfringens enterotixin gene also demonstrated complete agreement with enterotoxin producibility. These results suggested that the PCR assay is a rapid and simple test for identifying the enterotoxin-producing C. perfringens without using any cultures and spore treatments.

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Anti varicella-zoster virus (VZV) antibodies were detected by an immune adherence hemagglutination (IAHA) test, and were compared with the CF test, IFA test and ELISA test, respectively. Type O, Rh-positive RBC for IAHA was obtained from five healthy volunteers. All five RBCs had sufficient sensitivity as the indicator cell. Optimum incubation temperature was 37°C in the serum and in the complement. The complement was obtained from guinea pig sera, and the most suitable concentration was 1;100. The convalescent VZV antibody titers were similar to the values obtained from any of the methods previously mentioned. However, mean titers measured by CF were about twoto fourfold lower than in the values of IAHA. Seroconversion rates of the live VZV vaccine, as detected by CF and IFA were relatively low (CF; 76%, IFA; 56%). In contrast, those obtained from ELISA and IAHA showed perfectly (100%). These results indicate that IAHA has sufficient sensitivity and specificity in the detection of VZV antibodies. In addition, the IAHA test is thought to be a rapid and easy test to perform in ordinary laboratories.

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The erythrocyte membrane damaged by streptolysin-O (SLO) was observed in negative staining electron microscopy. It was confirmed that rings took arc (c-ring), sigmoidal (s-ring) or circular (o-ring) structures, and had electron-dense centers of a diameter of 24 nm and 4.9 nm width.
We found a crown structure on top of the ring in view of side projection. The ring structure was constructed by three layers of the electron lucent top which was the crown, the second dark layer, and the third, base part which embedded in the erythrocyte membrane, and the hights were 3.2, 1.6, 5.0 nm, respectively.
When the ghost membrane of erythrocyte was treated with SLO, the double of the inner and outer layers of a ring were observed by the negative-staining images. The figures of rings taken by under focus showed that one ring might be constituted between the 22 and 24 pair of inner and outer molecules. Totally 44 or 48 toxin molecules might be required for one O-ring

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We investigated the action of the ammonia produced by Helicobacter pylori urease on the cultured cells. The urease was purified from supernatant fluid of sonicated cell of H. pylori cultured on blood agar for 2 days at 37°C under microaerophilic condition. Purification was carried out by DEAE Sepharose chromatography, Phenyl-Sepharose chromatography, Sephacryl S-200 SF chromatography and fast protein liquid chromatography on Mono-Q. Vero, HeLa and Intestin 407 cells with or without the addition of 30 mM urea were exposed to the purified urease. Those cells showed cytotoxic effects within 80 minutes after addition of purified urease in the presence of urea. The ammonia production was observed on tissue culture medium within 10 minutes, and the ammonia concentration ranged from 5.56 mg/ml to 7.3 mg/ml and pH in the medium was over pH 9.0. No such effect was observed on the cells exposed to urease without urea. Ammonia water added to Vero cells showed the same cytotoxic effect within 70 minutes on the production of ammonia and raised the pH. However, when the cells were exposed to the ammonia water pre-neutralized to a given pH 7-8 using 1 NHC1 cytotoxic effect ect was not observed.
It was concluded that the cytotoxic effect ect of H. pylori urease was dependent on ammonia generated by hydrolysis of urea.

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The air-pouch model of inflammation in rats is excellent in that it allows quantitative evaluation of inflammation, and it is used for analysis of inflammatory mediators and as an evaluation system for anti-inflammatory drugs. We investigated the possibility of using this system as an experimental infection system. As a result, inflammation was found to be caused by injection of a constant amount of Staphylococcus aureus solution (10⁴-10⁸). The amount of infiltration and the number of infiltrating cells varied with quantity of bacteria. The infiltrating cells consisted mainly of neutrophils. In this experimental model of infectious disease, the severity of inflammation could be quantitively evaluated as a function of time in terms of bacterial proliferation and the body's response to bacterial proliferation based on the amount of fluid in the air pouch and the number of infiltrating cells, suggesting that the model is useful. In this eperimental system, there were no differences between the number of live bacteria, the number of infiltrating cells or the amount of infiltration when S. aureus Smith strain and clinically-isolated methicillin-resistant S. aureus (MRSA) were used, suggesting that there is no difference between the inflammation-induced activity of MRSA and MSSA.

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We tried to detect enteroadherent Escherichia coli exhibiting a diffuse pattern of adherence to HeLa cells from stock strains derived from patients with or without diarrhea in Brazil, Myanmar and Japan (Osaka). Enteroadherent E. coli was found from 16 (23 strains) out of 126 (384 strains) in diarrheal infant cases (12.7%), 26 (29 strains) out of 126 (348 strains) in healthy control cases (20.6%) in Brazil, from 15 (18 strains) out of 221 (542 strains) in diarrheal infant cases (6.8%), 4 (4 strains) out of 87 (212 strains) in healthy control cases (4.6%) in Myanmar. In Japan (Osaka), enteroadherent E. coli was detected from 7 (7 strains) out of 123 (198 strains) in diarrheal cases (5.7%). These results show that enteroadherent E. coli (diffuse) may not be associated with diarrhea. Forty-four (62.9%) out of 70 strains belonged to 19 O serogroups or 21 O: H serotypes. All strains were H antigen serotypable, and 26 strains were found to be nonmotile. The predominant O: H serotypes were O11: H15, O11: H-, O 20: H34, O21: H5, O89: H-, O99: H33 and O154: H45. Only one strain belonged to enteropathogenic E. coli serogroup O127.

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Treponema pallidum subsp. pallidum Nichols (Tp) antigens were purified by centrifugation in the presence of Hypaque (sodium diatrizoate) or MgCl₂. When the TPHA (Treponema pallidum hemagglutination) tests with crude and purified antigens were carried out in the sera of patients with untreated primary syphilis, the purification by MgCl₂ resulted in enhanced sensitivity to Tp-specific IgM as the same as the purification by Hypaque. In addition, the SPHA (solid phase hemadsorption) test with the Hypaque-purified antigen to and-Tp-IgM antibodies was carried out using the sera of patients with primary and secondary syphilis. Fifty-nine percent (27/46) of them showed positive for the crude antigen and 100%(46/46) for the purified antigen, indicating that sensitivity to and Tp-IgM was clearly enhanced with the antigen purification. The captured Tp-IgM-RIA test with the purified antigen was also undertaken in 16 patients with primary syphilis, 31 patients with secondary syphilis and 73 patients with latent syphilis. All the patients in the former two groups were positive, while all of the patients in the latter group were negative.
To elucidate the mechanism whereby the purification improves the sensitivity to and-Tp-IgM antibodies, host-derived antibodies associated with crude and purified Tp cells were measured by a fluorescent antibody technique. As the results, a great amount of rabbit anti-Tp-IgM antibodies were found on the surface of the crude antigen, whereas only a trace amount was detected on the purified antigens.
On the basis of these findings, it was considered that the purified antigen exhibits a strong reactivity to human anti-Tp-IgM antibodies due to the removal of rabbit anti-Tp-IgM antibodies from the surface by the purification procedure.

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A 24-year-old male with chronic renal failure on Continuous Ambulatory Peritoneal Dialysis (CAPD) complained of cough and dyspnea. Chest X-ray film showed a pneumonia shadow and MRSA and Candida krusei were detected in the sputum. Pneumonia improved with vancomycin and fluconazole. Treatment with methylpredonisolon was needed for retinodialysis.After this treatment, pneumonia deteriortated. Pneumonia did not improve with vancomycin and anti-fungal agents. This severe pneumonia was improved with a combination therapy of vancomycin, miconazole and G-CSF. A combination therapy of antibiotics and G-CSF is considered to be effective for severe pneumonia.

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