Chlamydia trachomatis was detected from first-voided urine sediments of 97 male patients with urethritis by polymerase chain reaction (PCR). Since urine and urinary sediments only treated with proteinase K inhibited DNA amplification by PCR, DNA was further purified by phenol extraction and concentrated. Two oligonucleotides based on sequences within the major outer membrane gene from C. trachomatis serovar L₂ were used as primers. A DNA fragment of 242 bp specific for C. trachomatis was amplified by PCR and detected by agarose gel electrophoresis.
The DNA fragment was amplified by PCR in all specimens of urine sediments from 50 patients with Chlamydiazyme-positive urethral swab. In 38 specimens of urine sediments from 47 patients with Chlamydiazyme-negative urethral swab, PCR was negative. The overall coincidence rate between the PCR for detecting C. trachomatis in first-voided urine sediments and Clamydiazyme in urethral swab was 90.7%(88/97). Detection of C. trachomatis from first-voided urine sediments by PCR was considered to be noninvasive and useful for the diagnosis of male urethritis due to C. trachomatis.

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In order to determine the optimal dose of cefepime (CFPM) for respiratory tract infections, a dose finding study was conducted in patients with chronic respiratory tract infections, and the clinical properties of the drugs were compared with those of ceftazidime (CAZ).
Each drug was administered by intravenous drip infusion at the dose of 2 g/day of CFPM, 4g/day of CFPM or 2g/day of CAZ for 14 days.
1. The overall efficacy rates evaluated by the committee were 82.6%(19/23) for the CFPM 2g/day group, 85.0%(17/20) for the 4g/day group and 79.3%(23/29) for the CAZ 2g/day group, with no statistically significant difference among the three groups.
2. The overall efficacy rates evaluated by the attending physicians were 91.3%(21/23) for the CFPM 2g/day group, 78.9%(15/19) for the CFPM 4g/day group and 75.9%(22/29) for the CAZ 2g/day group, with no significantdifferenceerence among the three groups.
3. Bacteriological eradication rates were 88.2%(15/17) for the CFPM 2g/day group, 68.8%(11/16) for the CFPM 4g/day group and 63.2%(12/19) for the CAZ 2g/day group, with no statistically significant difference among the three groups.
4. The incidences of adverse reactions were 3.8%(1/26) for the CFPM 2g/day group, 9.1%(2/22) for the CFPM 4g/day group and 3.4%(1/29) for the CAZ 2g/day group, with no significant difference among the three groups. The incidences of abnormal laboratory findings were 37.5%(9/24) for the CFPM 2g/day group, 15.0%(3/20) for the CFPM 4g/day group and 3.4%(1/29) for the CAZ 2g/day group. There was a significant difference erence among the three groups.
5. Utility rates assessed by the committee were 81.8%(18/22) for the CFPM 2g/day group, 76.2%(16/21) for the CFPM 4g/day group and 75.9%(22/29) for the CAZ 2g/day group. Utility rates assessed by attending physicians were 90.9%(20/22), 78.9%(15/19) and 72.4%(21/29), respectively. There was no significant difference among the three groups.
From the above results, it is concluded that the optimal dosage of CFPM is 2g/day for chronic respiratory tract infections.

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The efficacy, safety and usefulness of cefepime (CFPM), a new cephem antibiotic, in bacterial pneumonia, were evaluated in a comparative study against ceftazidime (CAZ). Each drug was administered by intravenous drip infusion at a dose of 1.0 g (nominal potency) twice daily for 14 days, and the following results were obtained.
1. A total of 183 cases were enrolled in this study. Efficacy rates ( “good” or better responses) as evaluated by the subcommittee were 90.3%(65/72) in the CFPM group and 94.0%(63/67) in the CAZ group, with no significant difference between the 2 groups.
2. Efficacy rates ( “good” or better responses), as evaluated by attending physicians, (in the same bacterial pneumonia cases which were subjected to evaluation by the subcommittee) were 87.5%(63/72) in the CFPM group and 89.6%(60/67) in the CAZ group, with no significant difference between the 2 groups.
3. Bacteriologically, eradication rates were 96.9%(31/32) in the CFPM group and 96.7%(29/30) in the CAZ group with no significant difference between the 2 groups.
4. The incidence of side effects was 5.9%(5/85) in the CFPM group and 4.8%(4/84) in the CAZ group, with no significant difference between the 2 groups. No significant difference was also found between the 2 groups in the incidence of abnormal laboratory findings; 28.4%(23/81) of the case in the CFPM group and 34.1%(28/82) in the CAZ group.
5. As for overall usefulness of the drug in bacterial pneumonia cases, utility rates ( “useful” or better evaluations) as evaluated by the subcommittee were 88.9%(64/72) in the CFPM group and 92.5%(62/67) in the CAZ group. The rates as evaluated by investigators (in casesjudged as evaluable by the subcommittee) were 87.5%(63/72) and 85.1%(57/67), respectively. There were no significant differences between the 2 groups.
These results indicated that CFPM is very useful for the treatment of bacterial pneumonia.

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The clinical efficacy, safety and usefulness of Cefepime (CFPM), a new cephem antibiotics, in chronic respiratory infections were evaluated in a comparative study against Ceftazidime (CAZ). Each drug was administered by intravenous drip infusion at a dose of 1.0 g (nominal potency), twice daily for 14 days, and the following results were obtained:
1. A total of 170 cases were enrolled in this study. Efficacy rates ( “good” or better responses) as evaluated by the subcommittee were 86.2%(56/65) in the CFPM group and 84.5%(60/71) in the CAZ group, with no significant difference erence between the two groups.
2. Efficacy rates ( “good” or better responses) as evaluated by attending physicians were 83.3%(55/66) in the CFPM group and 84.5%(60/71) in the CAZ group with no significant difference between the two groups.
3. Bacteriologically, eradication rates were 83.3%(40/48) in the CFPM group and 88.2%(45/51) in the CAZ group, with no significant difference between the two groups.
4. Side effects occurred in none of the patients in the CAZ group and in 4 of the 66 patients in the CFPM group. There was a significant difference between the two groups (Fisher's test p=0.0489). The incidence of abnormal laboratory findings were 17.6%(12/68) in the CFPM group and 21.1%(16/76) in the CAZ group. There was no significant difference between the two groups.
5. The utility rates evaluated by the subcommittee were 81.8%(54/66) in the CFPM group and 84.5%(60/71) in the CAZ group with no significant difference between the two groups.
Only in the incidence of side effects, there was a significant difference between the two groups (Fisher's test p=0.0489), but there was no significant difference in other items of efficacy, safety and usefulness between the two groups. These results indicate that CFPM is useful for the treatment ofchronic respiratory tract infections.

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Anti staphylococcal activity by clinical isolates of Pseudomonas aeruginosa was tested by the reversed agar plate and the filter paper stamp methods.
Almost 40% of Pseudomonas aeruginosa inhibited the growth of both Methicillin resistant Staphylococcus aureus (MRSA) and Methicillin sensitive Staphylococcus aureus (MSSA).
Green pigment (Pyocianin) produced strains showed a strong inhibitory effect against MRSA and MSSA respectedly. But some other pigment (Yellow, Red) strains also showed anti staphylococcal activity. These data suggest the colonization of Pseudomonas aeruginosa with anti staphylococcal activity may not be eradicated by the anti pseudomonic antibiotics.

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The bactericidal effect of gentiana violet against MRSA isolated from clinical specimens was studied both in vitro and in vivo. The results obtained are as follows:
1) Minimum bactericidal concentration (MBC) of gentiana violet to MRSA was between 0.0025% and 0.08% and the MBC was not influenced even if 25% human whole serum exists in the medium.
2) The number of cells were 2.1×10⁷ CFU/ml in medium which decreased to 5.4×10⁴ CFU/ml within 5 min by existing 0.1% gentiana violet in the medium as the final concentration, and also decreased under 103 CFU/ml 15 min later.
3) Minimum inhibitory concentration (MIC) of gentiana violet to MRSA was between 0.00015% and 0.00063%, and the inhibitory activity was not influenced even if gentiana violet was incubated with the bacteria in the medium for 72 hr at 37°C.
4) By using an ointment containing 0.1% gentiana violet to 12 cases of patients with the MRSA infected skin lesions, MRSA was eliminated completely from the infected areas of the skin within 4 weeks.
5) The side effects of gentiana violet were not observed in all cases during the use of the ointment containing gentiana violet.
It is suggested that gentiana violet may be one of the useful drugs for the treatment of the skin lesions infected with MRSA.

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In recent years, new quinolones such as ofloxacin (OFLX) and tosufloxacin (TFLX) have been frequently used in the treatment of bacterial enteritis caused by unknown organisms. The agent of first choice for the treatment of Campylobacter enteritis is one of the macrolides, but new quinolones are often administered accidentally to adult patients with Campylobacter enteritis. We have detected quinolone-resistant strains of Campylobacter jejuni (C. jejuni) after the treatment of some patients with new quinolones, and accordingly we reviewed the treatment of Campylobacter enteritis.
We experienced 178 adult patients with Campylobacter enteritis from January 1989 to November 1991 at our hospital. From them, we selected 52 patients (32 males and 20 females) in whom stool culture were performed both before and after treatment. The initially administered antimicrobial agent was a macrolide (rokitamycin, RKM) in 6 cases, a new quinolone in 22 cases, and kanamycin (KM) in 24 cases. The new quinolone used was OFLX in 17 cases, TFLX in 3 cases, and norf loxacin (NFLX) in 2 cases. Fifty-one of the 52 C. jejuni strains isolated before treatment were susceptible to OFLX and erythromycin (EM) according to antimicrobial disc susceptibility tests.
C. jejuni was eradicated in all patients treated with RKM or KM. However, treatment failed to achieve bacteriological cure in 8/22 (36.4%) patients given new quinolones. In these patients, the strains of C. jejuni isolated before treatment were susceptible to OFLX, but the strains isolated after treatment were all resistant to OFLX according to disc susceptibility tests. The minimum inhibitory concentrations (MIC) of OFLX for C. jejuni strains isolated after treatment was 64-fold higher than the MIC for the strains isolated before treatment. Cross-resistance was recognized between OFLX, TFLX, and NFLX, but the quinolone-resistant strains of C. jejuni were still susceptible to macrolides.
Therefore, we recommend macrolide therapy when C. jejuni is not eradicated by treatment using new quinolones, so as to prevent quinolone-resistant C. jejuni from being excreted in the stools for several weeks.

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Two bacterial strains (ME448 and ME474) isolated from stool cultures of imported cases in Tokyo in 1987 had typical biochemical characteristics of Shigella dysenteriae. The results of antigenic analyses showed that they were serologically identical to each other, but did not belong to any of the established Shigella serovars.
These strains were positive for Sereny test in guinea pig eye and cell-invasion test in HeLa cells. The strains also had virulence-plasmid encoding outer membrane proteins, indicating that they were pathogenic.
We then contacted the Centers for Disease Control in the United States and the Central Public Health Laboratory in the United Kingdom and arranged serological examinations of the strain ME448. From the results, the strain was confirmed to have provisional S. dysenteriae serovar E23507.
Although the serovar had been isolated from a Swedish patient who developed diarrhea while in India, this is thought to be the first report of its isolation in Japan.

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We treated 70 patients with strongyloidiasis (41 males and 29 females) with ivermectin (IVM), and obtained the following results:
1. The eradication rates at 1-2 months, 3-4 months and 5-6 months after treatment were 90.7%(49 of 54 patients), 100.0%(47 of 47 patients) and 95.7%(45 of 47 patients), responsively.
Twelve patients were resistant (non-responsive) to treatment.
2. When compared to patients whose parasites were completely eradicated, the resistant patients showed the following results:
1) Incidence of symptoms observed before treatment was significantly lower (50.0% vs. 84.5%).
2) Positive rate of anti-HTLV-I antibody was significantly higher (66.7% vs. 20.7%).
3) Blood eosinophil counts before treatment were significantly lower (266.6 ± 117.2/mm3 vs.533.2 ± 429.7/mm3).
4) Serum IgE levels before treatment were significantly lower (217.2 ± 442.9 IU/ml vs. 1, 076.8 ± 2, 108.0 IU/ml).
5) There were no significant differences in age, sex and dosage of ivermectin.
3. Comparing anti-HTLV-I antibody positive and negative patients, the following results were obtained:
1) Eradicated patients;
a) Eosinophils and IgE levels before and after the first administration of medicine in anti-HTLV-I antibody positive patients were significantly lower than those of negative patients.
b) Gammaglobulin levels before treatment and after both administrations of the drug, IgG before therapy and OKT4/OKT8 after therapy were significantly higher than in anti-HTLV-I antibody positive patients.
2) Resistant patients;
Eosinophils after treatment were significantly lower in anti-HTLV-I antibody negative patients than in positive patients.
Our results suggest that when the patients with strongyloidiasis have either positive anti-HTLV-I antibody, low blood eosinophils or low serum IgE levels, they have a tendency to resist treatment with ivermectin.

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We studied the detection of the HA gene of human influenza viruses in throat swabs obtained from the outbreakes of influenza in school children utilizing the polymerease chain reaction (PCR) method. Sensitivity and specificity of the PCR method was compared to conventional virus isolation using MDCK cells.
Three pairs of primers for PCR in detecting the HA genes of AH1, AH3, and B influenza viruses showed both subtype and type specificity. The dilution experiments showed that influenza viruses, as few as 1.1-3.5 plaque-forming units per 50 p1, were sufficient for the detection of HA genes by PCR method and the detection rate by PCR method was 2-3 fold higher than that by conventional method.
Our results showed that the PCR method was a fast, sensitive and reliable method for the diagnosis of influenza infections.

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Two hundred and sixty-six piglets with diarrhea (from 4 farms), 73 healthy pregnant pig (from 2 farms), 27 calves with diarrhea (from 9 farms) and 47 healthy milk cows (from 1 farm) were examined for Verocytotoxin-producing Escherichia coli (VTEC), and 52, 11, 15 and 67 strains of VTEC were isolated from 17 piglets, 11 pregnant pigs, 6 calves and 23 milk cows, respectively. All VTEC strains from the piglets produced only VT2vp, while the strains from the healthy pigs did not produce VT2vp, but did VT1 and/or VT2. Most VTEC strains from calves and cows produced VT2vhb and some produced VT2 and VT1. Serotyping of the isolated strains showed that many strains from the piglets belonged either O 139: H1, O 141: H4 or. O 141: HUT, but the strains from the pigs were either R-form or O-untypable. Many strains from the calves and cows were serotyped into O 116 or O 113, but there were several R-form and O-untypable. From these results, it is suggested that VTEC strains, especially from the pregnant pigs, calves with diarrhea and healthy milk cows, which produced the same type of Verotoxins to that produced by human isolates, may become sources of human infections.

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Eighty-seven oropharyngeal isolates of Haemophilus influenzae were obtained by two time cultures six months apart from a total of 288 children who attend a kindergarten. We analyzed the strains by comparing their serotypes, biotypes, β-lactamase activity and by performing electrophoresis of outer membrane proteins on polyacrylamide gels. Only nineteen strains were not identical, the rest of the 68 strains were classified into 23 types. During 6 months at least 15 types of strains lost from this group and 21 types of new strains were obtained. There were no children who had identical pairs of H. influenzae in their oropharynx during the 6 months.
The classification of strains in oropharynx suggested that person-to-person transmission of nontypable H. influenzae can occur. We concluded that oropharyngeal colonization by nontypable H. influenzae is not a normal flora in children.

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We studied the local immune response in a mouse experiment with acute ascending cystitis and pyelonephritis. The experimental infections were induced in BALB/c female mice by transurethral instillation of Escherichia coli O6. Immune response cells were stained, including Ia-positive cells, macrophages, neutrophils, T cells (CD4⁺ and CD8⁺) and B cells (IgA, IgM, IgG-positive B cell). They were stained by the immunohistochemical method (ABC method) using monoclonal antibodies against lineage specific antigens except for neutrophils that were readily identified by the standard hematoxylin-eosin.
Even in the control mice having no evidence of the infection, mucosa associated lymphoid tissue (MALT) consisted of macrophages, Ia-positive cells and T cells that were sparely found in the urinary tract tissue and renal parenchyma.
Ia-positive cells, macrophages, neutrophils, T cells (CD4⁺, CD8⁺) and IgA positive B cells were signigicantly infiltrated in the bladder submucosa from 6 hours after bacterial inoculation.
The infiltration of similar immune response cells was found in the submucosa of the renal pelvis, except for IgA positive B cells that appeared one day after the induction of the infection.
In renal parenchyma, Ia-positive cells appeared at 6 hours after introduction of the infection, followed by an infiltration of neutrophils, macrophages and T cells (CD4⁺, CD8⁺) at the first day, and IgA positive B cells at the third day.
These results are summarized as follows. When microbes invaded the urinary tract tissue, a significant number of Ia-positive cells infiltrated, which were initially present in normal urinary tract tissue.
Subsequently, neutrophils, macrophages and T cells (CD4⁺, CD8⁺) appeared in the lesion followed by a delayed occurrence of IgA positive B cells.

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Of 34 non-bacterial gastroenteritis outbreaks which occurred at day-care centers, kindergartens, elementary and secondary schools in Tokyo during the period from February, 1985 to June, 1991, 28 outbreaks from which small round structered viruses (SRSV) were detected in the patients' stool specimens by electron microscopy were subjected to an epidemiological investigation. The outbreaks tended to occur frequently in the cold season; twenty-two (79%) of these outbreaks from November through April. Though detailed epidemiological informations was not obtained from all outbreaks, the common source of infection were presumed to be present in many of the outbreaks, judged from the incidence as to time course of patients. Food doubted to be incriminated as transmission vehicles in these outbreaks was served at schools, kindergartens, and lodgings. In some outbreaks, SRSV was detected from stool specimens of food handlers, or they were seroconverted to SRSV, suggesting that food was incriminated as a transmission vehicle.
The symptoms of patients differ slightly from age to age: in the age range of 0 to 6 years, vomiting 90%, fever 41% and diarrhea 32%; in the 6 to 12 year-olds, nausea 61%, vomiting 48%, abdominal pain 65%, diarrhea 20% and fever 29%; and in the 12 to 15 year-olds, nausea 69%, vomiting 42%, abdominal pain 60%, diarrhea 30% and fever 34%. The lower the age of patient vomiting was more frequently observed. In these lower age groups, the frequency of nausea and vomiting tended to exceed that of diarrhea.

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A new micro particle enzyme immunoassay technique (MEIA, IMx HAVAB, Abbott) has been recently introduced for the detection of antibody to hepatitis A virus (anti-HA).
To evaluate the feasibility of the IMx HAVAB, we carried out comparison tests between MEIA, RIA and EIA. Furthermore, we investigated the prevalence of and-HA in Fukuoka City and Yonaguni Island, Okinawa, Japan using this method.
Results obtained were as follows:
In the test of 514 sera, 254 (49.0%) were positive by the three methods, and the remaining 260 (50.6%) were negative by the three methods.
Examination of diluted sera using IMx revealed that IMx can be used as efficiently as RIA and EIA.
Quantitative linear correlations were found between IMx and RIA (r=0.973), and IMx and EIA (r=0.969).
Anti-HA was assayed within 45 minutes by IMx, and the procedure was simple because of the auto analyser used in this method.
On Yonaguni Island, a significant decrease in the overall prevalence of and-HA from 75.2% in 1980 to 61.1%, in 1990 (p<0.05) was found, the prevalence of and-HA on Yonaguni Island in 1990 was significantly higher than in Fukuoka City (p<0.001).
These results indicate that the sensitivity of IMx is equivalent to those of RIA and EIA, that it is easier to use than either RIA or EIA and that hepatitis A virus infection in Okinawa, has significantly decreased during the past 10 years, but is still significantly more frequent than in Fukuoka City.

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[in Japanese]

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We experimenced a 5-year-old male case of Mycoplasma pneumoniae pneumonia accompanying Adenosin Deaminase (ADA) activity in pleural effusion. Chest roentgenograms revealed the infiltration in the left upper lung field and the left pleural effusion. In serum, the M. peumoniae CF titer increased to 1: 512. The pleural effusion was yellowish in color, with a specific gravity of 1.030, protein 3.7 g/dl, gulcose 101 g/dl, and ADA 50 IU/l.
Pleural effusion acconpanying M. pneumoniae pneumonia is rare, and the high ADA activity in this case has been reported only in one other case. This is a report of a high activity of ADA in the pleural fluid by M. pneumoniae pneunonia.

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We report a case of eosinophilic meningitis caused by Angiostrongylus cantonesis.
This patient, a 50-year-old male, had been eating uncooked slugs for 40 years. His chief complaints on admission were headache, fever and general fatigue. Neurological examination and CT findings were normal, but the CSF contained increased cells, most of which were eosinophilic cells. The presence of eosinophilic cells in the CSF is by itself abnormal. We therefore suspected eosinophilic meningitis and performed immunological tests. Since the geletin particle method and immunological antigen antibody reaction were positive. We diagnosed the patient as having eosinophilic meningitis caused by Angiostrongylus cantonesis.
There are only 27 reported cases of this disease in Japan, and most of them have been reported in Okinawa-Prefecture.

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