We performed the in situ hybridization (ISH) study of the smear cells taken from the patients of herpes zoster. We used the biotinylated EcoRI-B, H fragment of varicella-zoster-virus (VZV) DNA as the probe. In the control study of ISH VZV-infected cells showed strong staining, but no staining was observed in the infected cells by other herpes virus. We examined the smear cells of 19 patients of herpes zoster and 2 patients of herpes simplex by ISH. 30 of the 35 specimens of herpes zoster was positive by ISH. We detected VZV DNA in the smear cells of all cases of the bullos stage, but the sensitivity was about 75% in the laterstage. Smear cells of herpes simplex was not stained by ISH. We also performed virus isolation of 13 cases of herpes zoster. But the sensitivity of ISH was higher than that of virus isolation in the other stages of herpes zoster. This method is very simple and can be completed in about 1 hour. It also has high specificity and sensitivity. So this method is very useful for the rapid diagnosis of VZV infection.

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The nationwide DPT vaccine program was started in 1969 for infants under 12 months of age. In order to estimate the tetanus immune status among the general population of various age groups, we measured the serum tetanus antibody level in randomly selected outpatients from Metropolitan Tokyo (n=211, 6 months-60 years) and Hamamatsu, a city of 510 thousand population (n=128, 3 year-80 years) between January 1987 and June 1989.
Among the 211 subjects from Tokyo, the antibody value exceeding effective level of 0.01 HAU/ml was observed in 102 subjects (48.3%). The antibody positive rate was 90.8% in subjects of 3 to 21 years and was 27.7% in subjects of 22 years or older. The positive rate was significantly higher in subjects of 21 years or younger (p<0.005). Among 128 subjects from Hamamatsu, 60 (46.1%) had a positive antibody and the antibody positive rare was 96.9% in subjects of 3 to 21 years and was 29.5% in subjects of 21 years or younger (p<0.005).
The above findings indicate that our DPT vaccination program has been functioning well for the last 20 years and that the immunized population is adequately protected against tetanus.

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Antigen of Entamoeba histolytica for complement fixation (CF) test was applied to counterimmunoelectrophoresis (CIE), and the result was compared with that of CF test and enzyme-linked immunosorbent assay (ELISA) in regard to sensitivity and specificity. Besides, we studied antibody titers to E. histolytica in sera from acute and convalescent patients, and from patients with amoebic colitis and amoebic liver abscess.
Sera used were from patients with amoebic colitis, amoebic liver abscess and uninfected control subjects. The CIE was less sensitive than the CF test and ELISA. Most CIE-negative samples had low CF and ELISA titers, but a few samples had higher CF and ELISA titers. On the other hand, all uninfected controls were negative by CIE and ELISA, and 98% were negative by the CF test.
The level of antibodies tended to decrease with time after clinical recovery, but CF test and ELISA results remained positive in all patients 6 months after clinical cure. CIE converted negative in only one patient after 6 months.
The level of antibodies of CF test and ELISA was significantly higher in amoebic liver abscess than amoebic colitis.
These results suggested that all these three methods were so specific, and antibody titers by the CF test and ELISA were well correlated to clinical manifestations. The CIE with CF-antigen was less sensitive as compared with CF test or ELISA.

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The auxotypes of 592 strains of Neisseria gonorrhoeae isolated in Sapporo between 1980 and 1989 were investigated. The serotypes of some of these strains were also studied, and the relation between the auxotype and the sensitivity to Amoxicillin (AMPC) was analyzed. The results were as follows.
1. The auxotypes of Neiseria gonorrhoeae isolated in Sapporo were similar to those distributed in Southeast Asia.
2. In a study of the auxotypes by year, the frequency of proto and Pro decreased, while the frequency of AHU-increased.
3. The serotypes of all strains of Neisseria gonorrhpeae investigated were WII/WIII, there was no WI serotype.
4. With respect to the relation between auxotypes and sensitivity to AMPC, the AHU-strains were more sensitive than proto or Pro^-.

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Twenty-two patients with polymyositis and dermatomyositis (PM-DM) were retrospectively studied with regard to development of herpes zoster.
Herpes zoster occurred with high frequency in patients with PM-DM. The clinical courses of zoster infections were uneventful; no severe complications nor deaths occurred, and only one patient had postherapeutic neuralgia. No specific therapy for this infection was necessary. Zoster tended to occur in the inactive stage of PM-DM. PM-DM patients with herpes zoster had a significantly higher incidence of antinuclear antibody. There seemed to be no relationship between steroid therapy and herpes zoster infection.

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For the past two years, five strains of new quinolone resistant Enterobacter cloacae have been isolated from three patients in our hospital; strain A was isolated from patient A, strain B from patient B, and strains C₁, C₂, C₃ from patient C. These five strains were resistant to new quinolones and other antimicrobial agents including ampicillin, piperacillin, methicillin, cefotaxime, cefoperazone, kanamycin, gentamicin, tetracycline, minocycline, and chloramphenicol. Plasmid DNA profiles on agarose gel indicated that strain B and C₃ carried completely the same plasmid pattern, but strain A gave a different plasmid pattern. Furthermore chromosomal DNAs extracted from strains A, B and C₃ were digested with restriction endonucleases EcoR I, BamH I and Sma I. They were separated by pulsefield gel electrophoresis. The banding profiles of strains B and C₃ showed the same pattern.
It is, therefore, highly suggested that strains B and C₃ are the same E. cloacae strain, although isolated from different patients at different time. The possible route of nosocomial infection between these two patients who were hospitalized in the same room after an interval of 4 months was discussed.

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Although Strongyloides stercoralis (S. stercoralis) infection rate in Okinawa Prefecture was less than 2% by the traditional method, it has been proven to be 6.2% by the new technique-agar plate method-Thiabendazole has strong activity to eradicate the organism, but it is well known that the rate of severe side effects is extremely high. Therefore, we attempted to evaluate the new treatment for the infection by mebendazole and its combination with thiabendazole.
The reason for use of the drug is based on the reports of successful treatment of S. stercoralis infection in humans with the mild and infrequent side effects produced by the drug.
Thirty three patients were orally given mebendazole 100 mg twice daily for 28 days. Twenty six patients were given thiabendazole 500 mg thrice daily for 5 days and after that, mebendazole 100 mg twice daily for 9 days. This combination therapy was repeated twice.
The following results were obtained:
1) Out of a total of 59 patients, the cure rate was 83.3%(20/24) in single use of mebendazole and 100.0%(22/22) in the combination therapy.
2) Constipation (9.1%) and headache (9.1%) were of relatively high incidence in the mebendazole group, but they were mild. Nausea (19.2%) and headache (15.4%) were observed in the combination therapy group and the drug was discontinued in 2 patients.
3) The incidence of the elevation of S-GOT, S-GPT was noted in 71.4%(20/28) for the mebendazole group and 52.2%(12/23) for combination therapy group. All 13 patients of the mebendazole group were negative in lymphocyte stimulation test for mebendazole.4) Positive rate of HTLV-1 antiboty was 40.0% in the patients.
As described above, although combination therapy was effective for the eradication of the parasite, its side effects were too high and severe to continue the treatment. Although liver injury developed much more in single use of mebendazole, it seemed to dose-dependent.
From these results, it will be concluded that mebendazole can be used for treatment of strongyloidiasis because of lack of severe side effects and a changing of the treatment period would prevent liver injury.

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There is concerned that the new quinolone-resistant strains have increased along with its widespread usage.
We analysed the annual changes in frequency of ofloxacin-resistant strains isolated from urine in the past four years at two different types of hospitals, department of urology in Sapporo Medical College and Muroran City Hospital, since the usage of these agents seem to be related to the annual changes of the frequencies.
The results were summarised as follows:
1) In the two hospitals, drug volume of the new quinolones had been increased, in particular, the past six years from 1984.
2) The annual changes in frequency of ofloxacin-resistant Staphylococcus aureus have been increasing from 0 to 41.2 percent in Sapporo Medical College and 16.7 to 96.7 percent in Muroran City Hospital. The frequency of ofloxacin-resistant Pseudomonas aeruginosa also have been increasing from 24 to 66.7 percent in Sapporo Medical College and 37.5 to 81.8 percent in Muroran City Hospital.
3) The frequency of ofloxacin-resistant indole positive Proteus spp. and Serratia marcescens for four years at Sapporo Medical College (indole positive Proteus spp.: 0-1.8 percent, Serratia macrcescens: 10-43 percent) was very different from that at Muroran City Hospital (indole positive Proteus spp.: 65-82 percent, Serratia marcescens: 71-100 percent). The difference seems to be caused by the hospital acquired infection.

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Fecal flora of the patients without remarkable gastrointestinal diseases were studied. All patients were treated with antimicrobial drugs with or without Miya-BM (the preparation of Clostridium butyricum M588). The stools were examined before and after (during) antimicrobial treatment.
Fecal flora of 69 patients before medication was almost the same with that of healthy adults as has been reported by Mitsuoka et al. After giving antimicrobials, most bacterial genus except Enterococcus and Yeasts in the stool decreased their detection rate and their population. This floral change was not much influenced by co-administration of Miya-BM.
The detection rate of Clostridium difficile and/or the toxin A from the stool were markedly increased after giving antimicrobials. When Miya-BM was co-administered with antimicrobials, however, the detection of C. difficile and/or toxin A was very rare.
C. butyricum M588 was recovered from 7 cases out of 10 patients treated with antimicrobials and Miya-BM. Non-spore form of C. butyricum was dominant in the feces of 3 cases, and spore form was dominant in the other 4. This result showed that administrated C. butyricum M588 germinated in intestinal tract.

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We studied the polymicrobial infection by transtracheal aspiration (TTA) on pneumonia in the elderly.
Incidence of mixed isolation of organisms from TTA was 35% or 39 out of 111 episodes.
The principal organisms in polymicrobial infection detected from TTA were S. aureus (N.23), P. aeruginosa (N.15), K.pneumoniae (N.12) and S. pnemoniae (N.11).
The principal combination of mixed infection were S. aureus+P. aeruginosa (N.7), S. aureus+K. pneumoniae (N.5), S. aureus+S. marcescens (N. 5), S. aureus+S. pneumoniae (N.5), S. aureus+Anaerobes (N.4) and K.pneumoniae+P. aeruginosa (N.4).
Hospital acquired pneumonia was of a higher rate of mixed infection than the nursing home acquired pneumonia.
In the mixed infection on pneumonia in the elderly, normal upper airway flora was highly detected with pathogens from transtrcheal aspirates.
From these results, we realize that aspiration and superinfection were important factors in mixed infection in the elderly.

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Macrolide antibiotics at concentrations by far lower than their MICs proved to inhibit the production of alginate, elastase and protease by mucoid Pseudomonas aeruginosa. The morphological study of mucoid P. aeruginosa under the electron microscope revealed the slime-like structures common to the cell morphology of the organism in cultured colonies and foci in a model for respiratory tract infection in mice, and the strains of mucoid P. aeruginosa which had been allowed to get in contact with the drugs proved to produced obviously fewer slime-like structures than control strains. The effect of a 14-membered macrolide, erythromycin, against mucoid P. aeruginosa was also observed with a 16-membered macrolide, rokitamycin this effect appeared to be common to the macrolide.

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The detection of C. trachomatis antigen in first-voided urine sediments has recently been achieved by means of IDEIA CHLAMYDIA^® (IDEIA, Novo Nordisk), an EIA kit using monoclonal antibodies. Therefore, this kit was used as a screening test to examine the infection rate of C. trachomatis in young adult men without symptoms. The titers of serum IgA and IgG antibodies to C. trachomatis were also determined.
1. Antigen detection from first-voided urine sediments of young adult men without urogenital symptoms (141 cases): The detection rate by IDEIA was 5.0%(7/141). Three of the 7 cases which were positive for antigen in first-voided urine sediments were reaffirmed as having asymptomatic C. trachomatis urethritis, since they were also revealed to have C. trachomatis in the urethra.
2. The positive rates of serum antibodies: The titers of serum antibody were determined in 128 cases out of the 141 cases. The positive rates of IgA and IgG were 6.3%(8/128) and 35.9%(46/128) respectively. The positive rates of IgA and IgG antibodies were significantly higher in cases with positive antigen in first-voided urine sediments than in those which were negative. These results indicate that this kit is useful for antigen detection.
3. The screening test revealed asymptomatic C. trachomatis infections in 5% of young adult men, suggesting the extensive spread of the infection.
The screening test using first-voided urine sediments will be useful in public health.

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The enzyme-linked immunosorbent assay (ELISA) has been evaluated for the detection of antibodies against Legionella pneumophila.
Three-grade antigens were prepared from Legionella pneumophila serogroup I. Crude antigen was made by enzyme digestion, sonication and centrifugation and then became half pure by ammonium sulfate precipitation. It was purified to form 60-kDa protein antigen by size-exclusion chromatography on a Sephacryl S400 column and ion-exchanged chromatography on a DEAE-5PW column.
60-kDa protein antigen was the most sensitive of the three antigens, but more cross reactive to K pneumoniae type II than the other two antigens.
It is suggested that crossreaction occurs on the grounds whether 60-kDa protein is antigen common to L. pneumophila serogroup I and K pneumoniae type II or antigens of such two bacteria co-exist on 60-kDa protein.

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Yersinia pseudotuberculosis (Y. pseudotuberclosis) infection is an intestinal infectious disease comarable in importance as those with Campylobacter or Salmonella. Clinical symptoms of Y. pseudotuberclosis infection vary. In this report, we will describe the clinical symptoms and immunological conditions of the patients with Y. pseudotuberclosis infection, including 2 or our own cases.
Case 1 was a 4 years old male infant admitted to the hospital with major complaints of fever, diarrhea, and vomiting. Kawasaki disease was the most suspected diagnosis from the clinical viewpoint. These symptoms improved by symptomatic treatments. Serum examination during hospitalisation revealed the infection of Y. pseudotuberclosis 4a. Case 2 was a 7 months old male baby with psychomotor developmental delay. The patient was admitted to hospital with major complaints of fever and eruptions. The patient was diagnosed to have a severe infectious disorder based on the clinical symptoms and findings of laboratory tests. Treatments with antibiotics improved the conditions. Serum examination during hospitalisation also revealed the Y. pseudotuberclosis 5a infection. Both of these cases showed decreased cellular immunity during the acute phase of the infection which was normalized with the improvement in clinical conditions.
It was thus suggested that Y. pseudotuberclosis had a possibility to influence the cellular immunity of hosts transiently but significantly.

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We reported five cases of listeriosis (sepsis and meningitis) in the elderly in our hospital during the last 4 years, where no cases of listeriosis had been found. These 5 cases had diabetes mellitus, lung cancer, chronic respiratory failure, gastric ulcer and aplastic anemia respectively as their underlying diseases. At the onset of listeriosis, 3 cases received corticosteroid and 3 cases received H2-blocker. 2 patients were cured and 3 patients died. Three autopsy cases had meningitis or meningoencephalitis and 2 cases of these autopsy cases had granulomatous changes in these spleens. In serotypes of Listeria monocytogenes (L. monocytogenes), 4 cases were 4b and 1 cases was 1b. All 5 strains were resistant to 3rd generation cephems. Wide uses of 3rd generation cephems and H2-blocker may be one of the reasons for the recent increase of listeriosis. Ingestion of contaminated food is the pathogenetic mechanism for initiating L. monocytogenes infections. And following the change of eating habits and the increase of imported foods, food-born listeriosis may increase.
We suppose the increase of L. monocytogenes infections and must give attention to L. monocytogenes infections.

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