In order to investigate the distribution of Bordetella pertussis in Nagoya City and estimate efficacy of the vaccine, we tried to isolate B. pertussis from patients with Pertussis-like symptoms who went to the department of pediatric general hospitals in Nagoya City from 1989 to 1992.
B. pertussis were isolated from 43 patients among 164 patients with Pertussis-like symptoms All of these isolates were classified into 1, 3, 6 serotype. It was impossible to isolate any B. pertussis in 1992 because of yearly variations. B. pertussis was only isolated from patients who were not vaccinated.
Among the 28 siblings of the patients, 18 were vaccinated and 2 fell ill, but the latter had only mild symptoms. Three did not fall ill although they had an incomplete vaccination. According to these results we suggest that this component vaccine is effective.

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A total of 16 strains of Enterohemorrhagic Escherichia coli (EHEC) isolated from diarrea patients in Saitama from 1990 to 1992 were tested for their serotype, verotoxin production, biochemical characteristics, antibiotics sensitivity and plasmid profiles.
By serotype analysis, 14 strains from two outbreaks and 12 sporadic cases were classified as type 0157: H7, one as O111: H-(not motility) and one as 0128: H2. Typing of verotoxin by gene analysis using Polymerase Chain Reaction (PCR) showed that 9 of 0157: H7 strains including two cases from outbreaks and O128: H2 have VT1 and VT2 genes, other 0157: H7 have the VT2 gene and 0111: H-has only the VT1 gene. Biochemical characteristic anlaysis indicated twostrains of 0157: H7 type from outbreaks were biotype II and the rest of 0157: H7 wre biotype I.
One of the 0157: H7 strain from a sporadic case showed positive for urease production.
According to sensitivity tests against antibiotics, out of the 0157: H7 group, one strain was resistant against ABPC, one against SM and two strains resistant to SM-TC.
For plasmid profiles, all strains had 94 Kb plasmids and several smaller sizes of plasmids. But 5 strains of 0157: H7 had 94 Kb plasmid only.

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To determine the incidence of enteric adenovirus, 447 stool specimens were collected between January 1991 to October 1992 from children with gastroenteritis at a clinic in Fujieda, Japan. Each specimen was tested for adenovirus group antigen and for specific enteric adenovirus types 40 and 41 (Ad40/41) by using monoclonal antibody enzyme immunoassays. Adenovirus was found in 10.7%(48 of 447) cases and 77.1%(37 of 48) of them were Ad40/41. There were two peaks of adenovirus infeciton May and Novenber 1991. The virus was isolated with 293 cells from 48 of adenovirus positive specimens, and then was typed according to the SmaI cleavage pattern of the viral DNA and by neutralization with specific antisera. Ad41 was predominantly isolated in 64.6%(31 of 48), and Ad40 was in 10.4%(5 of 48). The remaining virus isolates were typed as Adl, 2, 5, and 6. Ad41 was founded in 89.3%(25 of 28) from October 1991 to March 1992. Though Srna I and Hind III cleavage identified six Ad41 variants in the period, the variants identified from October 1991 to March 1992 were only two. The results indicated two variants of Ad41 strains caused an outbreak during the end of 1991 to early 1992 in Fujieda.

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A nested PCR assay was developed for the diagnosis of Candida albicans infection. A segment of the C. albicans β-tubulin gene was amplified in this assay that was sensitive with the detection limit of 10 fg DNA of C. albicans. This assay was found to be specific by obtaining the negative results with 13 organisms (5 other Candida, 2 Cryptococcus, 3 Aspergillus, Escherichia coli and others). Cerebrospinal fluid of a patient with C. albicans meningitis was analyzed and was found to be positive. These results suggest that this assay can be clinically useful because of its high sensitivity, specificity and rapidity.

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We performed a clinical study of 20 cases (33 episodes) of respiratory infections due to mucoid Pseudomonas aeruginosa by transtracheal aspiration (TTA) in the recent 10 years.
There was only one pneumonia without underlying chronic lower respiratory infection (CLRTI) case positive for mucoid P. aeruginosa and others were all CLRTI among 33 TTA trials. In contrast, nonmucoid P. aeruginosa was recovered from 9 cases of respiratory infections without underlying CLRTI among 46 TTA trials. Monomicrobial infection of mucoid P. aeruginosa was 69.7%, and polymicrobial infection containing mucoid P. aeruginosa was 30.3%, and Haemophilus influenzae was the most frequent microorganism recovered with mucoid P. aeruginosa. The recovery rate of mucoid P. aeruginosa among P. aeruginosa-colonized cases was 56.3% in diffuse panbronchiolitis, and that was 42.9% and 40.0% in bronchiectasis and chronic bronchitis, respectively. Mortality due to pneumonia with nonmucoid P. aeruginosa was 46.1%, but there was no fatal pneumonia case with mucoid P. aeruginosa. In CLRTI, laboratory data were not remarkably different between mucoid and non-mocoid P. aeruginosa-colonized cases.
Thus, these results suggest that mucoid P. aeruginosa is a more important organism in persistent infections in the lower respiratory tract compared with nonmucoid P. aeruginosa, and further investigations is required on the mechanism and clinical role of this infection.

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We isolated the virus from the respiratory specimens of 16 patients (7.9%) when we investigated the bronchopulmonary infections of 203 adult patients by the viral isolation method for the diagnosis of the viral infection and transtracheal aspiration (TTA) for taking directly the specimen from the lower respiratory tract.The bronchopulmonary infections, isolated the virus, consisted of acute bronchitis (3 cases), pneumonia (8 cases) and the acute exacerbation of chronic lower respiratory infection (5 cases).The positive viral isolation consisted of Rhinovirus (11strains), Herpes simplex virus I (3 strains), Parainfluenza virus III (1 strain) and Respiratory syncytial virus (1 stain).There were 9 transtracheal aspirates, 8 pharyngeal swabs and 5 nasal washings in the positive viral specimens.We experinced 5 cases who had the same virus isolated from both the upper respiratory tract and the lower respiratory tract.H.influenzawe, P. aeruginosa and other bacteria were found in 14 of the 16 TTA cultures.
These results suggest that the role of the viral infection is important either independently or relating the bacterial infection in the adult bronchopulmonary infections.

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The prevalence of influenza in Japan was studied, dividing the whole country into six sections such as Tohoku, Hokkaido and Kanto, to determine regional differences from the nationwide viewpoint. This study was based on data obtained from a total of 720 prefectures where three of the first index (the time the epidemic outbreak developed), second index (the time the virus was isolated) and third index (the time the prevalence reached its peak) were determined for the 16 prevalence periods of influenza between 1976 and 1993. The mean value (standard deviation) of each index was obtained for each year of prevalence and for each area. The differences obtained were analyzed statistically, and the following results were obtained:
1. There was a marked regional difference in the development of epidemic outbreak of cold; it was earliest in the Kanto and Kinki areas (late November to early December) and latest in the Kyushu area (mid January).
2. Isolation of the virus was earliest in the Kanto and Kinki area (mid January) and latest in the Chugoku and Shikoku areas (late January to early February), showing a regional difference.
3. The peak of prevalence was earliest in the Kanto area (early February) and latest in the Tohoku, Hokkaido, Chugoku and Shinkoku areas (mid February), showing a regional difference.
4. The difference between the earliest and latest areas was 4.2 weeks regarding the first index, while it was decreased to 1.8 weeks for the second index and to 0.8 weeks for the third index.
5. Analysis of the difference between the first and second indices in each area revealed that the earlier the development of epidemic outbreak of colds, the earlier the isolation of the virus, although the interval between these indices was relatively long.
In contrast, it was apparent that the later the development of such outbreak, the later the viral isolation, with a relatively short interval between the indices.

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The seroprevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) in 303 serum samples collected from that apparently healthy population inhabitating different areas in eastern Nepal was studied. Samples were collected at Dharan Municipality, Sunsari85, Pancha Kanya Village Development Committee, Ilam86, Dhankuta Hile, Dhankuta82 and Basantapur Village Development Committee, Tehrathm50. HBsAg and anti-HBsAg antibody was screened by reverse passive haemagglutination (RPHA) and passive haemagglutination (PHA) respectively and positivity was confirmed by enzyme linked immunosorbent assay (ELISA). Anti-HCV antibody was detected by ELISA. None of the samples were positive for HBsAg. Anti-H BsAg antibody was positive in 1.9%(6/303). The positive rate increased with age reaching 25% positivity among the elderly. The anti-HBsAg antibody positivity was 2.35, 2.32, 1.22 and 2.00 in Dharan, Ilam, Dhankuta and Tehrathum respectively. Anti-HCV antibody was detected only in one sample (15-year-old boy) collected in Dharan. These findings indicate that the HBV and HCV infections are not active in eastern Nepal.

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We studied the γ/δ T cells, which are thought to be one of the local immune-defense system, in an experimental ascending urinary tract infection model in mouse.
The experimental infection was induced in the BALB/c mouse by transurethral instillation of Escherichia coli.
γ/δ T cells were stained immunohistochemically with ABC method and the localization in the uninfected and infected murine urinary tract was determined. Then, to determine the CD4 or CD8 phenotype of γ/δ T cells in the infected site, a two-color immunohistological study was done.
In the uninfected state, γ/δ T cells were observed in the bladder epithelium and submucosa, not in the kidney. It is thought that the difference of γ/δ T cell distribution between kidney and bladder is affected by the bacteriological milieu, because the chance for foreign antigen exposure of bladder may be greater than that of the kidney.
Six hours after inoculation of E. coli, the γ/delta; T cells infiltrated in to the bladder and kidney, following a gradual increase in number. Especially at 105 days after inoculation, severe accumulation of y/6 T cells in the renal scarring lesion was observed.
Two-color immunohitological study revealed that the phenotypes of γ/delta; T cells in the renal scarring lesion were both CD4^-CD8^- double negative and CD4^-CD8⁺ single positive. The CD4^- CD8⁺ γ/delta; T cells were probably activated by the bacterial antigen.

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Previous epidemiological studies of Cryptococcus neoformans infection in Japan showed that only C. neoformans var. neoformans is present and serotype A is the most common with frequencies in excess of 95%. A DNA fingerprinting method, using a genomic DNA probe (UT-4p), has become available recently which discriminates between individual isolates in a population that are morphologically and serologically indistinguishable. Fifty-two serotype A isolates of C. neoformans were obtained from three different institutions (in Nagasaki, Chiba, and Tokyo) in Japan. Only two of these strains were isolated from AIDS patients and one from pigeon excreta. Of the nine reported fingerprinting patterns in serotype A, only three types (IV, V and VII) were observed in Japanese isolates. Pattern IV was almost exclusively observed in Nagasaki isolates (21/22) with only one of pattern VII. In Chiba, however, patterns VII and IV appeared to be equally distributed. In Tokyo, patterns IV and V (which included two isolates from AIDS patients) were observed at similar frequencies.
Restriction fragment length polymorphism analysis of four isolates of serotype AD showed a typcal serotype A pattern which also contained a serotype D-specific band. This finding suggests the independence of serotype AD. These data could enhance the survey of the epidemiology of cryptococ-cosis.

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The antibacterial activity of epigallocatechin gallate (EGCg), a catechin, against 53 clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) was evaluated and expressed as minimal inhibitory concentration (MIC). The MIC₅₀ and MIC₉₀ of the strains were found be 64 and 126 μE g/ml, respectively by the microdilution method. A time-kill study using an isolate showed that EGCg appeared to be bacteriostatic at 1-2 × MIC and bactericidal at 6 × MIC against MRSA. In addition, the activity of EGCg was stable to various physical conditions including boiling or freezing. These findings suggest that EGCg could be a useful agent for treating MRSA infection.

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A sixteen year old female was feverish from June 12, 1993. Methicillin-resistant Staphylococcus aureus was isolated from the blood, the diagnosis of MRSA sepsis was established. Vancomycin (2g/day) was administered for eighteen days, but MRSA was not eradicated in the blood culture. Then she was administered a combination therapy of arbekacin (200 mg/day) and imipenem/cilastain (1g/day) for seven days, but MRSA in the blood was cultured continuously. The sequential combination therapy of netilmycin (200 mg/day) and minocycline (200 mg/day) was started, MRSA was eradicated from the blood culture after four days. The sequential combination therapy netilmycin and minocycline was seemed to be effective for MRSA infection.

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We report a case of Corynebacterium jeikeium septicemia associated with malignant lymphoma. The patient is a 58-year-old male who was diagnosed as malignant lymphoma on August 1992. May 15, 1993, he was admitted to our hospital because of oliguria, abdominal flatulence and vomiting which developed a few days before admission. Anticancer regimen were started.
In the middle of July, white blood cell (WBC) count dropped to 100/mm³ and body temperature rose to 39°C. He was been treated with Ceftazidime and Piperacillin.C. jeikeium was recovered from blood culture. Antibiotics were switched to minocycline and vancomycin. He died of septic shock and pneumonia. Autopsy revealed the presence of the colonies of Rods. Which were morphologically compatible with C. jeikeium were observed in lung tissue and in the small pulmonary vessels.

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Since penicillin-resistant Streptococcus pneumoniae first recognized in 1967, the rate of penicillin-resistant strains has been increasing worldwide. There have been up to 50% from pediatric specimens in Japan.
We reported three pediatric cases with penicillin G resistant Streptococcus pneumoniae infection to show some important clue from these cases for clinical practice against resistant pneumococcal infection. The first case was a typical acute mastoiditis, although we have expierienced only masked mastoiditis recently. The second case was meningitis with septicemia, which did not show any abnormality in the first obtained cerebrospinal fluid. The third case was recurrent bronchitis in a child with cerebral palsy. The minimum inhibition concentrations of these isolated strains were 0.25μg/ml in the second case an 2.0μg/ml in the first and third cases.

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A 48-year-old female was seen at our hospital after having a severe fever of nearly 40°C, for a period of 9 days. She complained of pain in the left side of her chest. An X-ray examination revealed a slight infiltration of the upper and middle lung fields. At this time, it was learned that the women's pet bird had recently died.
This case was diagnosed as acute pneumonia due to psittacosis. Therefore the administration of Roxithromycin was started. After a few day her condition improved.
During the course of treatment, serum was taken and a throat swab was done_ A mirroimmunofluorescence (MIF) test asperformed to check the serum antibody levels against Chlarnydia psittaci. The serum titer rose from 1: 8 to 1: 256 in 15 days after admission. The final diagnosis was made after positive isolation of C. psittaci by means of the cell culture method.

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A case of uncommon iritis due to Chlarnydia pneumoniae (C. pneumoniae) is reported. The patient was a 9-year-old boy who had suffered from cough, pharyngeal pain, and low grade fever. The symptoms persisted for more than 1 month in spite of an oral cephem antibiotic. Ophthalmalgia, congestion around the iris and cough had lasted with alleviation and exacerbation. A diagnosis of C. pneumoniae infection was made by specific polymerase chain reaction (PCR) method and microimmunofluorescence test (MIF). The symptoms subsided with administration of clarithromycin (CAM: 300 mg/day) for 2 weeks. Because of the simultaneous alleviation of iritis, C. pneumoniae infection was considered to introduce the iritis.
Much remains to be clarified about this pathogenesis of iritis and more detailed evaluations are required.

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