Legionella pneumophila serogroup (SG) 1 has been known as a most frequent and important causative bacteria of Legionella pneumonia. It was reported that antigenicity of the serogroup was composed by numerous antigenic factors. To study antigenic factor formula of Japanese isolates from both clinical and environmental sources together with Philadelphia 1 strain, we examined slide latex agglutination system with monoclonal antibodies for subsergorouping. Philadelphia 1 and GIFU10102 strain (environmental isolate) were used as immune strains based on a cross absorption test result by rabbit antisera. Five anti L. pneumophia SG1 monoclonal antibody-producing clones were established. All monoclonal antibodies were SG1 specific but showed different reactivities with strain to strain. We prepared monoclonal antibody sensitized latex and used for subserogrouping of isolated strains. Twenty-two clinical isolates from Legionella pneumonia patients and 26 environmental isolates were examined. Each strain was reacted with at least one of the 5 sensitized latex and its antigenic formula was designed by sensitized latex reaction pattern. As a total of 6 patterns demonstrated in clinical isolates and 7 patterns in environmental isolates, 48 L. pneumophila SG1 islates were divided into 11 subserogroup patterns. The antigenic formula of Philadelphia 1 was same as one of clinical isolate patterns. It was considered that this subserogrouping system would be a useful tool for epidemiologic study.
Ecological behaviors of uropathogenic bacteria in the infected urine were characterized, and they were compared to those in vitro cultivation. The organisms directly collected from the infected urine revealed no piliation, but when they were isolated and cultured in heart infusion broth (HIB), the pili appeared. The pili were identified as Type I pili as examined by immunogold electron microscopy. Although the organisms directly collected from the infected urine did not have pili and did not agglutinate against anti-Type I pili anti-serum, the whole cell lysate analysis by western blotting revealed that the organisms express Type I pili antigen intracellularly. The organisms in some cases revealed a capsule formation in the urine, but it disappeared when cultured in vitro. Hemolytic Escherichia coli were isolated from 9 cases out of 20 with acute cystitis, however, hemolysin was not detected from the infected urine and the organisms. When they were cultured in HIB, all strains produced the hemolysin. The composition of outer membrane proteins (OMPs) of the organisms in the infected urine was clearly different from those in vitro cultivation. As the most prominent finding, a 70 kDa OMP of the organisms in the infected urine disappeared in vitro. Antibacterial activities of several drugs examined in healthy human urine and in HIB were variable from drug to drug. The mean value of minimum inhibitory concentration (MIC) of ofloxasin was 25 times higher in the urine than in HIB, whereas the MIC of erythromycin was almost equal in both conditions.
To assess the effectiveness and side effects of sequential interferon (IFN)-α and β treatment for patients with chronic hepatitis C, 25 patients were enrolled in a trial of this regimen. The patients were given 6 million units (MU) of natural human INF-β daily for 2 weeks followed by 6 MU of natural human IFN-α three times a week for 10 to 22 weeks. Serum alanine aminotransferase (ALT) levels normalized for at least 24 weeks in 10 patients (40%), of whom 4 (40%) had no detectable serum hepatitis C virus (HCV) RNA. Three variables were significant in predicting a sustained response: a low serum HCV RNA level, a low Knodell's fibrosis score, and a low indocyanine green retention rate at 15 minutes. Elevated serum ALT and proteinurea were observed with IFN-β treatment but these side effects were mild and disappeared when INF-β treatment ended. While all patients completed the entire regimen, we concluded that sequential IFN-α and β treatment provides no additional antiviral effects in chronic hepatitis C.
Recently, low dose and long term use of Macrolides (Mls) has been reported to be effective in treatment of chronic lower respiratory tract infections, however its mechanism is still obscure. We evaluated the effect of Mls (EM, AZM, RKM) on cytokine mRNA expressions. We preincubated the whole blood with several concentrations of Mls and removed the Mls and then stimulated human whole blood with LPS as an experimental vivo model. In order to examine cytokine mRNA expressions, we used the RT-PCR method. Cytokine mRNA expressions were suppressed significantly (p<0.05) by pretreatment with EM, AZM; moreover, the suppression was peaked at low concentrations (0.04-0.2μg/ml). Although, Cytokine mRNA expressions were not suppresed by pretreatment with RKM. These results suggest that EM, AZM have suppression on Cytokine mRNA expressions, and consequently, this suppression has a reasonable effect for DPB patients.
Since 1989, outbreaks of Salmonella ser. Enteritidis food poisoning has dramatically increased in Tokyo, and a total of 95 outbreaks has been reported between 1989 and 1993. As the results of examination of phage type by the method of Ward, et al., strains isolated from 31 (32.6%) of 95 outbreaks were phage type 34 which is a very rare type in Europe and America. To characterize these unique strains of phage type (PT) 34, 302 isolates obtained from patients and vehicle foods of the outbreaks, eggs and environments in Tokyo were examined for plasmid DNA plofiles and antimicrobial resistance patterns. Plasmid DNA was extracted by the Kado's method, and analysed by agalose gel electrophoresis. Antimicrobial susceptibility was tested for CP, TC, SM, KM, ABPC, NA, SXT, FOM and NFLX by the K-B disc method. The strains of PT 34 were further subdivided to 4 types by plasmid profile and antibiogram. The prevalent pattern of PT 34 strains was type plasmid profile which carried 2 different plasmids (> 250 and 60 kb) and was streptomysin-resistant. A total of 28 (90.3 %) of 31 outbreaks were found to be caused by this specific type of Salmonella ser. Enteritidis. Several kinds of egg-related foods were suspected for the vehicles of transmission among 12 outbreaks. Especially, in 5 outbreaks, this specific type (type B plasmid profile and streptomysin resistant) of Salmonella ser. Enteritidis strains were isolated both from patients and incriminated food which were cooked with eggs. The phase type of Salmonella ser. Enteritidis strains isolated from raw egg and poultry or environmental specimens from poultry farm showed that the prevailing phage types were almost the same as those of foods poisoning outbreaks. This strongly suggests that these foods may be considered as a source of infection in Salmonella ser. Enteritidis outbreaks.
A rapid and sensitive PCR-based microwell plate assay (PCR-MWP) system to detect the 16 S ribosome RNA gene of Helicobacter pylori was developed. Analytical sensitivity, evaluated with purified recombinant plasmid DNA and genomic DNA of H. pylori, was one copy of DNA per PCR. Specificity was validated with a panel of DNA from 75 kinds of microorganisms including Helicobacter showed weak positives, when 1 pg of DNA was input. Other microorganisms gave negative signals even when 100 pg of DNA was used for PCR. When compared with a Nested-PCR system to detect the urease A-subunit gene performed by a commercial reference laboratory, the results obtained (sensitivity 93.3% and specificity 73.3%) was almost equivalent. The PCR-MWP was rapid and easy for the detection of H. pylori DNA in gastric juice specimen.
The acquired immunodeficiency of the host plays an essential role in the occurrence of infections even with low pathogenic bacteria. The increase of cases with MRSA and/or pseudomonas infection is one of the serious problems in hospital management in Japan for the elderly as well as pediatric patients. In the present study, mitomycin C (MMC)-treated hosts were prepared in young, adult and old mice to test the immunopotentiating action of the promising Chinese herbal medicine, Tohki-Rikuoh-Toh (TRT), Hotyu-Ekki-Toh (HET) and Juzen-Taiho-Toh (JTT). The effect of these herbal medicines on organ structure and its function in the MMC-treated hosts is clarified and discussed for medical use. 4-5, 8-10 and over 50 week old male C57BL/6 mice (Clea Japan Inc.) were injected with MMC at a dosage of 3 to 5 mg/kg to inhibit the bone marrow, thus creating a mouse model with reduced immunopotential. A powder extract of TRT, HET and JTT was administrated orally at a dosage of 500 mg/kg/day for seven consecutive days. The white cell number and the subset analysis were carried out by the FACS method. The bactericidal effect of the host was monitored by NBT reduction test. Peritoneal macrophages were prepared by the adherence technique. The macrophage phagocytic activity was examined by an ACAS system. After the administration of TRT, HET and JTT, the body weights recovered as much as 90%, especially in young animals which had been reduced to 75% of their normal values. After MMC-treatment, with the herbal medicines, HET was good for young mice while JTT was effective for the old ones. As for the effect on B cells, the plaque-forming cells (PFC) of spleen cells were compared among the groups. As a result, PFC in the HET group was 184% and the other two were 80'-95% as compared to 76% in the MMC-treated ones. The number of white blood cells in the MMC-treated mice returned to 80% of their normal value. In addition, the phagocytic activity of macrophages increased to 50% although that of the non-treated group was only 20%. The phagocytic activity also recovered in the JTT and TRT of 131% to 95%, respectively compared to 11% in the MMC-treated control. When TRT, HET and JTT were administered orally to mouse models whose immunopotential had been inhibited, the herbal medicines activated both quantitatively and qualitatively, showing themselves to be effective interstitial medicines. In addition, the data from the animal models showed no side effects, comfirming the complete efficacy of the drug. Moreover, there was no direct anti-bactericidal effect from these medicines, suggesting that the immunomodulating action of this medicine is host-mediated. It is interesting that quantitative and qualitative recovery were seen when HET was administered to MMC-treated young hosts while JTT was good for the old. With this investigation, the effective components are still unknown for different generations, and we need to clarify this aspect for better understanding of the efficacy of herbal medicines.
Prulifloxacin (PUFX), a new quinolone antimicrobial agent, was administered to a total of 122 patients and carriers to investigate its clinical efficacy, safety and usefulness in infectious enteritis (bacillary dysentery, enteritis caused by Salmonella spp. and enteropathogenic E. coli, cholera and so on). In addition, the minimum inhibitory concentration (MIC) of UFX (active compound) was determined against each clinical isolate, and compared with that of ciprofloxacin (CPFX), ofloxacin (OFLX), tosufloxacin (TFLX) and nalidixic acid (NA). The correlation between the concentration of UFX in feces and the change of the fecal microflora were also investigated when PUFX was administered to the patients with acute infectious enteritis. A daily dose of 400 mg of PUFX was administered orally in two divided doses (morning and evening) for 5 days, with the exception of 7 days administration against salmonella enteritis and 3 days administration against cholera. 84 cases were adapted for evaluating the usefulness. The clinical efficacy was 100% in all the enteritis except salmonella enteritis, in which it was 88.9%(8/9 cases). On the bacteriological efficacy, the elimination rate was 100% in all isolates except Salmonella spp., in which it was 75.0%(12/16 cases). As for the adverse effect, uriticaria in moderate degree was observed in 1 (0.9%) of 109 cases. Abnormal changes in laboratory findings were seen in 3 (3.0%) of 100 cases, consisting of 1 with eosinophilia and 2 with elevated S-GPT, although they were all slight in degree. The usefulness rate was 65.5%(55/84 cases) for “very useful” and 95.2%(80/84 cases) for “very usuful” and “usuful”. MIC90 of UFX against Shigella spp., Salmonella spp., E. coli and V. cholerae, was 0.025, 0.05, 0.025 and 0.05μg/ml, espectively. These values were the same as those of CPFX and TFLX, and superior to OFLX and NA. UFX concentrations in feces followed by administration of PUFX in 3 cases with acute infectious enteritis were higher than that of MIC90 of UFX against Shigella spp., Salmonella spp., E. coli and V. cholerae. The changes of the fecal microflora, which influence the efficacy and safety of PUFX, were not observed.
We reported two cases of pulmonary dirofilariasis in Nagasaki Prefecture. Each case was admitted to our hospitals for evaluation of an abnormal shadow on chest X-ray film taken for examination. We strongly suspected lung cancer before operation. It was resected in order to confirm the pathological diagnosis. There was a Dirofilaria immitis in the vessels surrounded by the necrotic tissues. The case of pulmonary dirofilariasis tends to increase in Japan, but the positive rate of dogs with Dirofilaria immitis tends to decrease, because of decreasing breeding places of mosquitoes by the spread of the public sewage system in Nagasaki City.
A 38-year-old hemophiliac, who had been infected with HIV by the administration of blood products and had been diagnosed as AIDS by the onset of Pneumocystis carinii pneumonia, was admitted to our hospital with the complaints of headache and vomiting. After he was diagnosed as cryptococcal menigtis using the microsopy, cryptococcal antigen detection and culture of cerebrospinal fluid, treatment with amphotericin-B and fluconazole was started. As there was no clinical improvement, spinal drainage was performed and acetazolamide administered in order to reduce the intracaranial pressure. Treatment was changed from AMPH-B and FLCZ to a combined therapy of AMPH-B and itraconazole. As his clinical features showed improvement, he was discharged home on a maintenance dose of ITCZ and acetazolamide after having been hospitalized for three months. This case-report may be of use in the management of cryptococcal menignitis in patients with AIDS.
A case of unilateral hilar lymphadenopathy due to mycoplasmal pneumonia in an adult patient is presented. A 54-year-old female was admitted to our hospital because of high grade fever and abnormal shadows on chest roentgenograms. She did not have any respiratory symptoms before admission. Chest roentgenograms on admission revealed a tumor-like shadow in the right hilus resembling lung cancer. On the seventh day after admission, abnormal shadows on chest roentgenograms spontaneously improved without therapy. The patient was diagnosed as having mycoplasmal infection based on the serological tests.