Although there are some studies concerning tuberculosis among the homeless persons in Japan, almost no detailed analysis is available about non-tuberculous lung infections in this population .
We retrospectively studied the clinical features and microbial etiology of pneumonia among the homeless. This series included twenty so-called homeless persons, 19 male and one female, who were admitted to our hospital with a clinical diagnosis of pneumonia from April 1993 to July 1996. The average age was 56 years-old. All patients were transferred by ambulance. Most of them were admitted night in the winters, and had respiratory failure together with othoer serious complications such as gastrointestinal disease, liver failure and cerebrovascular disease. Chest X ray revealed so-called lobar pneumonia in 8 of 20 cases (40%). Microbial etiologic agents were found in 14 patients (70%). Pneumococcus amounted to 80% of all the speculated pathogens. Despite intensive care after admission, 6 of 20 patients (30%) died subsequently. Although fatal pneumococcal pneumonia were reported to have decreased in the general population in Japan, morbidity and mortality of pneumococcal pneumonia was very high in this series.

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Growth of 10 strains of VTEC O157, which were isolated from faecal specimens, was evaluated in conventionally recommended enrichment broth media Trypticase soy broth (TSB) and Davis's Minimal Medium (DMM) at different temperatures.
Five strains of VTEC O157 used in the test proliferated well in TSB and reached to 10⁹ CFU/ml at 36°C in 24 hours incubation and to the same population densities at 25°C in 48 hours. Whereas in DMM at 36°C there was no prolifiration in 6 hours, but grew to 10⁶ CFU/ml in 24 hours and 108 CFU/ ml in 48 hours. Three strains of VTEC O157 tested at 25°C grew to 1.52×10⁴ CFU/ml in 24 hours, 1.67×10⁸ CFU/ml in 48 hours and 6.80×10⁸ CFU/ml in 72 hours. No proliferation was observed at 4°C in 72 hours in groth TSB and DMM.
The growth in glucose free DMM was found in 7 out of 10 strains, and 4 out of these 7 strains grew profusely to 10⁵ CFU/ml in 48 hours. One out of 3 strains showed no prolifiration but survived 72 hours postinoculation, decreasing from 179 CFU to 29 CFU in 100μl of medium. The other 2 strains became completely extinct in 48 hours after inoculation with 263 CFU and 2, 420 CFU in 100g1 of medium respectively. However, these 3 strains which showed no growth in glucose free DMM could proliferate in DMM containing 1/10 of the usual glucose concentration, at a rate depending on the concentration.
In conclusion, it is suggested that the finding of increase of VTEC O157 in glucose free DMM would be a useful solution for the food poisoning problem caused by VTEC O157.

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In 14 subjects whose chest radiographs showed abnormal shadows during the two ears from January 1995 until December 1996, no definite diagnosis could be obtained because sputum, smears and cultures all gave negative results for mycobacteria. Bronchoscopy was therefore performed, revealing atypical mycobacteria in cultures of the bronchial washing fluid for mycobacteria, and the significance of bronchoscopic examinations in cases diagnosed an atypical pulmonary mycobacteriosis was investigated. Most of the subjects (9) were women. Nine subjects had been informed that they had abnormal chest shadows; five had subjective symptoms; bloody sputum, 3 and cough, 2. The characteristics of the shadows were as follows: in the plain radiographs, the main shadows had a mottled or granular appearance in the majority of the patients (9) and there were infiltrative shadows in 3 patients and nodular shadows in another 3. In computed tomograms, the shadows in the vicinity of the pleura appeared as micronodular conglomerates in 12 patients, in 11 of whom bronchiectasis was also present near the shadows. Alveolar infiltrative shadows were present in four cases, and a cavity was seen in only one. Smears of the bronchial washing fluid for mycobacteria were positive in 7 patients, and cultures of this fluid yielded at least 100 colonies in 8 of the 14 subjects for whom the results were positive. By culture, Mycobacterium avium complex (MAC) was identified in 13 patients, but eleven of these in whom the bronchial washing fluid was concurrently tested for MAC by the polymerase chain reaction, only four were MAC-positive. Transbronchial lung biopsies were performed in 11 cases, in which the histological findings of mycobacterial infections showed granuloma in four, and caseation in three.
Bronchoscopy is making possible initial-stage diagnosis, which are normally difficult, among the recently growing number of cases of the bronchial form of atypical pulmonary mycobacteriosis and is also useful for reaching definite diagnosis in the early stage.

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To investigate sensitivity to rubella virus (RV) in healthy individuals, we examined levels of antibodies to RV in sera by an indirect immunofluorescence assay (IFA) and compared levels of antibodies by IFA with those by a hemagglutination inhibition (HI) assay. Of 114 healthy individuals, we detected antibodies to RV in serum specimens from 103 (90.3%) by IFA and in those from 109 (95.6%) by HI assay. The peak value of levels of antibodies by HI assay was 4 fold higher than that by IFA. When levels of antibodies by IFA were less than 32, levels of antibodies by HI assay ranged from<8 to 1024. We did not detect anti-rubella antibodies of IgM class in all serum specimens and detected anti-rubella antibodies of IgA class in serum from only 1 individual by IFA.
We detected antibodies to rubella in sera from 51 (94.4%) by IFA and in sera from 52 (96.3%) by HI assay of 54 individuals who reported having had rubella, and in sera from 23 (88.5%), by IFA and in sera from 26 (100%) by HI assay of 26 individuals reported having been vaccinated. Also, we detected anti-rubella antibodies in sera from 13 (76.5%) by IFA and in sera from 15 (88.2%) by HI of 17 individuals who reported having had neither rubella nor vaccination.
In serum from 1 individual who reported having had rubella, we detected antibodies to rubella by IFA but not by HI assay. In serum specimens from 2 individuals who reported having had rubella vaccination, from 3 having had vaccination, from 2 having had neither rubella nor vaccination, we detected anti-rubella antibodies by HI assay but not by IFA. On the other hand, by both assays, we detected antibodies to RV in all sera of individuals who reported having had rubella and been vaccinated.
The serodiagnosis, at least, by two methods is necessary to prevent individuals from rubella virus infection, because of following results: 1) influence of an inhibitor in serum specimens was thought to be variable. 2) The results measured by IFA were differed from those by HI assay in some individuals. 3) It is difficult in diagnosis of rubella from clinical symptoms alone.
Also, it might be required to use vaccine to the individual who lacks detectable antibodies to rubella in serum by any method to prevent rubella infection.

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The clincal features of invasive deep mycosis in the critical care center was studied and the usefulness for determinations of plasma (1-3)-β-D-glucan, one of the major structural components of fungi, in making the diagnosis of deep mycosis was evaluated in comparison with that of blood culture and candida antigen titer using CAND-tec kit. A total of 92 febrile patients (mean age=54.5yr., M/F=70/22) in our critical care center were enrolled in this study.
Seventeen out of the 92 febrile patients (18.5%) were those with deep mycosis. In the deep mycosis group, there were 10 patients with fungal panperitonitis, 5 with fungaemia, one with candidal pneumonia and one with candidal empyema.
A total of 52 blood samples were obtained from 17 patients with deep mycosis. Forty five out of the 52 blood samples (86.5%) were positive for serum (1-3)-β-D-glucan while only 10 were culturepositive. In contrast, six (15.0%) out of the 40 blood samples were obtaind from 17 patients with deep mycosis were positive for candida antigen by CAND-tec kit.
In the critical care center, deep mycosis is a common infection and determination of serum concentration of (1-3)-β-D-glucan is found to be a very useful examination in screening of deep mycosis with high sensitivity and specificity.

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The simple and rapid identification method of coxsackie virus A16 (CA16) and enterovirus 71 (Ev71), the main cause of hand foot and mouth disease, was described in this report. This method was consists of three steps, those were virus isolation, amplification by RT-PCR, and digestion by restriction enzyme Taq I and Eco T22 I. In 1990, many virus strains were isolated in Vero cell line. But after 1994 the number of isolated viruses in Caco-2 cell line increased instead of isolated in Vero cell line. Concerning to isolation of CA16, in 1998, MRC-5 cell line was also used and it's sensitivity was same as Caco-2 cell line. Cytopathic effects were first observed in MRC-5 cell line among these three lines.
RNAs of CA1-10, poliovirus 1-3, echovirus 1-7, 9, 11, 14, 16, 17, 18, 24, 25, 27, 30, Ev71, and isolated viruses were extracted by using QIAamp viral RNA kit^® (QIAGEN). Then two series of reverse transcription using two down stream primers (E31 and E33) were performed. In PCR, the same upper stream primer (primer 2) was used. CA6, CA16 and Ev71 were the only viruses those were not amplified by RT-PCR using primer 2/E31 but amplified by RT-PCR using primer 2/E33. After PCR, PCR products of isolated viruses using primer 2/E33 were digested by Taq I and Eco T22 I. All of Ev71 products were not digested but all of CA16 products were digested. The band pattern of PCR products (CA16) digested by Taq I were divided into three groups.
And Eco T22 I digestion pattern is only one. These results were in accord with Taq I and Eco T22 I digestion sites on sequences of CA16 and Ev71.
This method should be useful for the rapid identification of CA16 and Ev71.

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Since 1961, recovery of Shigella bachilli from healthy food handlers in Tokyo has been carrying out, and detection of Salmonella carriers has also been adding from 1980. Recovery rate of Shigella has decreased from 0.28% (589 cases) in 1961 to 0.01% (9 cases) in 1969, and 7 cases between 1971 and 1975 and only 3 carriers since 1976 have been detected. On the other hand, Salmonella has been detected from about 9, 000 cases (0.07%) during 18 years. The isolates were typed into 150 serovars, in which the most frequent one was S. Enteritidis, following S. Litchfield, S. Thompson, S. Hadar, S. Typhimurium, S. Infantis, S. Tennessee, S. Montevideo, S. Agona and S. Braenderup. These serovars except S. Agona caused in 90% of 1, 650 Salmonella food poisoning outbreaks which had occurred between 1980 and 1996. Recovery of S. Enteritidis from healthy subjects increased year by year since 1989, and this tendency was well consistent with the increase of food poisoning outbreak caused by this serovar. These results indicate that the recovery of carrier with enteropathogen from food handlars is significant as preventive measures or food hygiene.

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A study was made to clarify to what degree the booster phenomenon was present when the employees in a community hospital in Japan received two-step tuberculin testing(PPD). Of the seventyfive employees, most of all BCG-vaccinated subjects, twenty-three showed strongly positve of more than 30 mm of induration at the first test (PPD/T1), and the remaining fifty-two subjects received a second(PPD/T2)after 2 weeks. The second PPD showed a marked increase in reacitivity, sixteen subjects newly became strongly positive and six of eight who were initially PPD negative(<10 mm)were converted positive, and the mean reaction size changed from 14.7±5.6 mm(PPD/T1)to 31.5 ±15.5 mm(PPD/T2). The degree of boosting, measured by the change in millimeters induration size between PPD/T1 and PPD/T2(PPD/T2-PPD/T1), was correlated with neither the size of PPD/T1 nor the age of the study participants, and was not associated with the difference of the position in the hospital. Thus, we could not explain the factor why such a marked boosting was observed in this study. However, these results indicated that the two-step tuberculin testing is an essential means of distinguishing new tuberculous infection from booster phenomenon. We agree with the current recommendation for routine two-step testing of new emplyees in the hospital.

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The antibodies against LPS of enterohemorrhagic Escherichia coli (EHEC) O157 in sera from patients infected with this organism and sera from healthy control subjects were examined by ELISA to investigate their immune response. Analysing these data, we found some heterogeneity of antibody response between adults and infants.
In healthy adults and children, the cutoff values for IgM antibodies to O157 LPS were calculated as 0.85, 0.40 and for IgG were as 0.57, 0.39 respectively. In patients, the ELISA values of adult were lower and decreased earlier than those of children. Therefore sero diagnosis for EHEC O157 infection for adult patients the timing of blood collection is very important. On the other hand, in some patients the sera of children showed a high level of IgM only 3 days from the onset. This tendency seemed to be related to severity of the disease. This immunological test was proved to be useless for healthy carriers because the antibodies against O157 LPS were not observed in their sera.

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A three-year old girl was hospitalized in a semi-conscious state following a febrile convulsion. She did not recover despite treatment and died 16 days after admission. Influenza A virus (H3N2) was detected from a throat swab from the patient, and serum hemagglutinin-inhibiting antibodies to the virus elevated from less than 8 to 256. Brain CT revealed bilateral thalamic hemorrhage and peripheral low density. Subarachnoid hemorrhage was also observed thereafter. Based on clinical manifestations and neuroimaging, this patient was diagnosed as an atypical case of acute necrotizing encephalopathy associated with influenza A virus infection. Such rapid progressive encephalopathies may occur due to intracranial vascular injury including vasculitis or spasms. Although it is clear that influenza A virus triggered this case, we cannot confirm that it was a pathogen. Also, it might be advisable to consider other possible contributing factors such as drugs administered before hospitalization.

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[in Japanese]

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A previously healthy 15-year-old female was admitted to our hospital complaining of nausea and vomiting. She did not complain of diarrhea. A physical examination revealed a lower right quadrant abdominal tenderness without rebound or spontaneous pain and a knocking pain of the costovertebral angle. A high fever, knocking pain of costovertebral angle, and urinary findings including Gram's stain, lead us to suspect a urinary tract infection, cefotiam was administered intravenously. Spiking fever with shaking chills continued for three days, and three sets of blood cultures were positive for Salmonella Oranienburg, but her urine culture was negative. Her history was taken again, revealing an intake of a processed squid product. The product was confirmed by the local public health center to be Salmonella Oranienburg. Finally food poisoning by Salmonella Oranienburg with sepsis was diagnosed. With cefotiam she became better and was discharged from the hospital on the 10th hospital day. During admission to the hospital she did not experience any diarrhea, and her stool culture was negative.
Epidemics of Salmonella Oranienburg food poisoning are relatively rare in the literature. In Japan, one has arisen as a result of contamination of a processed squid product in March 1999. However, there have been no cases without so-called gastroenteritic symptoms (abdominal pain and diarrhea) who were previously healthy and developed sepsis caused by Salmonella Oranienburg, reported in Japan. Even in previously healthy patients, with an epidemic situation of non-typhoidal salmonellosis, salmonella sepsis must be ruled out. Among such cases, those who present with spiking fever and shaking chills should be given antibiotic therapy after taking appropriate cultures.

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