Using a new endotoxin-specific chromogneic limulus assay (Endospecy test), endotoxin concentrations were measured in 93 specimens of cerebrospinal fluid (CSF) from 66 pediatric patients. Eighteen patients were diagnosed as having menigitios. Of these, 6 cases (group A) with gram-negative meningitis proven by culture had high CSF endotoxin concentrations of 115.3, (82-133) (median, range) pg/ml. Ten cases (group B) with gram-positive or aseptic meningitis had endotoxin concentrations of 2.15 (0.1-3.6) pg/ml. Other 2 cases with bacterial meningitis (group C), in whom no pathogen was detected, had CSF endotoxin concentrations of more than 100pg/ml. Four cases with encephalitis (group D) and 45 cases with non-meningitis or non-encephalitis (group E), had CSF endotoxin concentrations of less than 5 pg/ml. Despite a negative culture after antibiotic treatment in group A patients, endotoxin was cleared slowly from the CSF. A clearing of endotoxin from CSF was followed by alleviation of fever with a more gradual decline in CRP values. In 2 cases of group C, the negative bacterial culture appeared to be attributable to the previous treatment with antibiotics. However, these patients had high CSF endotoxin levels, indicating gram negative baceterial meningitis. In 17 CSF specimens from 5 patients of group A, in whom Haemophilus influenzae was detected on admission, an additional a latex agglutination test for the detection of H. influenzae polysaccharide antigen was performed. Only 3 specimens from 3 patients with CSF endotoxin concentrations of more than 80 pg/ml had a positive aggultination test.
These results suggest that quantitation of endotoxin in CSF is useful for the diagnosis of gram-negative meningitis. And also, the clearance of endotoxin from CSF during treatment appears to be useful in determining the timing of when antibiotic should be stopped.

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Various antiseptics, commonly used in hospital, were tested against methicillin-resistant Staphylococcus aureus (MRSA) at high concentrations (∼10⁹CFU/ml) and in short time exposures.
The antiseptic solutions of 0.05% alkyldiaminoethylglycine hydrochloride, 0.05% benzalkonium chloride, 0.2% povidone-iodine gargle and 0.03% dominophen bromide produced 1/ 1, 000 to 1/100, 000 reduction after a 30 sec exposure. 7.5% povidone-iodine scrub and 0.2% benzalkonium chloride in 83% ethanol were most effective, reducing MRSA under detection limits. (less than 10 CFU/ml) 0.05% chlorhexidine gluconate was less effective, producing no evident reduction even after 1 min of exposure.
Though 0.2% povidone-iodine solution reduced MRSA under detection limits at 37, 24 hrs incubation, colonies of several strains were detected after 48 hrs extended incubation. This suggests the existence of strains resistant to the antiseptic solution of povidone-iodine.
We recommed that the disinfection for these strains should be repeated short time disinfection, because repeated short time exposure (15 sec 2 times) of antiseptics was more effective than one long time exposure (30 sec once).

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The prevalence of influenza in Kyushu-Okinawa District in April 1994-March 1995 was studied as the prevalence of influenza virus, to determine the sero-type of influenza viruses isolated in Kyushu-Okinawa District prefectures and cities. As a result, three sero-types of infuluenza viruses, i.e., type A/H1N1, type A/H3N2 and type B, were isolated in Kyushu Okinawa District in this season, but most of the isolates were type A/H3N2 and type B. Weekly changes of reported influenza patients and period of virus isolation at local public health institutes revealed that influenza epidemics of the earlier part in this season was caused by type A/H3N2 and the latter part due to type B. Type A/H3N2 spread all over Kyushu-Okinawa District in a shorter period (about 2 weeks) through the westside of Kyushu and down south, and type B stayed about one month in northern Kyushu and took about 7 weeks to spread all over Kyushu-Okinawa District. Based on these results, the spread of influenza viruses in Kyushu Okinawa District was visualized on the isopleth maps.

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Isolated pathogenic bacteria from sputum of the patients with pulmonary emphysema whowere admitted in our hospital from 1984 to 1994 were examined to elucidate the relationship between isolated bacteria from sputum and pulmonary functions including vital capacity (VC), forced expiratory volume (FEV1.0), PaO₂ and PaCO₂. VC of the patients from whom MSSA (methicillin-sensitive Staphylococcus aureus) or Enterococcus faecalis (E. faecalis) were isolated was signfificantly lower than that of the patients from whom Streptococcus pneumoniae (S. pneumoniae), Branhamella catarrhalis (B. catarrhalis) or Haemophilus influenzae (H. influenzae) were isolated. FEV_1.0 had a similar tendency as VC in terms of isolated organisms from the patients with emphysema. Similarly, PO₂ of the patients from whom MSSA or E. cloacae were isolated was significantly lower than that of the patients from whom S. pneumoniae, B. catarrhalis or H. influenzae were isolated, and PCO₂ of the patients from whom E. faecalis were isolated was significantly higher than that of the patients from whom S. pneumoniae, B. catarrhalis or H. influenzae were isolated. There was also impaired respiratory function in the patients from whom MSSA, Escherichia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa), Xanthomonas maltophilia (X. maltophilia) or Enterobacter cloacae (E. cloacae) were isolated, compared with those in the patients from whom S. pneumoniae, B. catarrhalis or H. influenzae were isolated. These results suggest that isolated pathogenic bacteria are shifted from S. pneumoniae, B. catarrhalis or H. influenzae to MSSA, E. coli, P. aeruginosa, X. maltophilia or E. cloacae in the course of impairment of respiratory function in pulmonary emphysema. Thetreatment and prophylaxis for acute exacerbation in pulmonary emphysema should be based on these results.

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In early 1980's methicillin-resistant Staphylococus aureus (MRSA) was reported as amajor pathogenic organism of geriatric hospital infection in Japan. At the same time in the A geriatric hospital MRSA infection was prevalent. To decrease nosocomial infections some active preventive measures against hospital infection were taken since Oct. 1991.
After introduction of preventive measures of hospital infection in the geriatric ward (190 beds) nosocomial bacteremia and pneumonia were markedly decreased in comparison to the episode number before introduction of prevention.
However several patients with MRSA colonization were observed every month. The aim of this clinical study was to clear how frequent MRSA was isolated from the gastric juice and stool.
Any MRSA was not observed in 63 cultured stool, but just one MRSA was isolated in patients with MRSA colonization. On the other hand gram-negative organisms, which were E. coli, P. aeruginosa, P. rnirabilis etc., were frequently observed in cultured stool.
In conclusion we considered frequency of MRSA colonization in gastrointestinal space was not so high but rather very low.

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We reported five patients with purple urine bag syndrome (PUBS). Four patients had indicanuria, and, in three of them, purple pigmentation was reproduced by inoculating urinary isolates in the broth with indoxyl sulfate. Klebsiella pneumoniae, Pseudomonas aeruginosa and Enterococus avium were considered to produce the purple pigment in three patients. However, attempts to reproduce the purple pigment failed in two patients, and one of them did not have indicanuria. These results suggest that indicanuria is not necessarily required for the development of PUBS.

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Recently, nosocomial outbreaks of MRSA have become an serious social problem in Japan. To examine the routes of transmission of MRSA, the establishment of an accurate MRSA typing system is essential. Previously, we reported that the DNA fingerprinting by AP-PCR might be a useful method to differentiate MRSA strains. In this study, we tried to investigate the clinical usefulness of DNA fingerprinting by AP-PCR using clinically-isolated MRSA strains. Twentyfour MRSA strains (12 with coagulase type IV, and 12 with coagulase type II) isolated from patients in our department were used. Other typing methods (the sensitivity of antibiotics, pulsed-field gel electrophoresis, and plasmid analysis) were also performed. As a result, the typing pattern by AP-PCR correlated well with other typing methods. MRSA strains with coagulase type IV showed almost the same pattern, suggesting that these strains were nosocomially transmitted. On the other hand, MRSA strains with coagulase type II showed various patterns, suggesting these strains were not nosocomially trasmitted. In conclusion, the typing by AP-PCR seemed to be a useful tool to evaluate a nosocomial MRSA transmission.

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Among human immunodeficiency type 1 viruses (HIV-1) isolated during long-term 3'-azido-3'-deoxythymidine (AZT) therapy, 4 condons (Asp67, Lys70, Thr215, and Lys219) in the HIV-1 reverse transcriptase (RT)-coding region have been considered to be related to the AZT resistance of HIV-1. Therefore we determined these mutation patterns in HIV-1 isolates from patients undergoing long-term AZT treatment. In 41 clones of HIV-1 from 7 patients, the Thr215 mutation was the most predominant (97.6%), and more frequent than Asp67 (48.8%), Lys70 (31.7%) and Lys219 (9.8%) mutations.
All 22 clones from isolates cultured in the presence of 1, u M AZT showed Thr215 mutation; such a high frequency was not found for the other 3 codon mutations.
In a clinical follow-up study, Thr215 mutation appeared in the late stage of AZT treatment in parallel with the emergence of AZT insusceptibility. It is worthnoting that this Thr215 mutation to Tyr or Phe is a 2-nucleotide mutation in contrast to the 1-nucleotide mutations seen in the other 3 codons. Isolates with the single amino acid change of Thr215 of the RT-coding region obtained after long-term AZT treatment grew in the presence of 1 & mu;M AZT. This single amino acid mutation in the Thr215 codon is the most important factor in AZT resistance.

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Six sporadic cases of VTEC infection have been confirmed in Akita Prefecture from Jun. 1991 to Nov. 1994. Six VTEC strains isolated in these cases were examined for their serotype, Vero toxin type, existance of eae gene and 60 MDa plasmid, and CVD 419 probe reactivity.
Of the 6 siolates, 5 were O157: H7. Two isolates of which possessed TV-1 and VT-2 genes and the rest of 3 possessed VT-1 and VT-2vh genes, VT-2 gene and VT-2vh gene, respectively. One isolate was O26: NM possessing VT-1 gene.
A primer pair, designated as EA-1 and EA-2, was designed for detecting eae gene in VTEC. Examination of the 6 VTEC isolates by PCR using EA-1 and EA-2 primers indicated that all of the isolates were eae positive. This was further confirmed by dot blot hybridization using a ^eae probe. All of the 6 isolates also harbored approximately 60 MDa plasmid, which hybridized with the CVD419 probe in southern blot analysis.
These results supported the hypothesis that ^eae gene and the 60 MDa plasmid might play a role in the mechanism for VTEC to cause diseases including diarrhea, hemorrhagic colitis and hemolytic uremic syndrome in humans.
Diversity observed in plasmid profile and VT type of the 6 isolates suggested that they were originated from independent sources, which could not be identified.

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A total of 1242 strains of Salmonella isolated from patients with sporadic diarrhea during the period from April 1985 to December 1994 in Yamanashi Prefecture, were tested for their serovars and drug susceptibility.
The results can be summerized as follows:
1) The isolates were serologically classified into 54 different serovars. The predominant serovars were S. Enteritidis (39.6%), S. Typhimurium (16.6%), S. Oranienburg (4.8%), S. Hadar (4.6%), S. Litchfield (4.4%).
2) Serovars of S. Stanleyville, S. Hartford, S. Rissen, S. Chincol, S. Gaminara, S. Poona and S. Hvittingfoss were isolated from human sources for the first time in Yamanashi Prefecture.
3) Yearly frequency of isolates was 19.2% in 1989, 15.5% in 1994, 11.8% in 1993.
4) Monthly frequency of isolates was 21.2% in August, 17.1% in September, 12.0% in October, 11.2% in July.
5) The predominant ages of isolates were 2 years of age (99 strains: 8.0%), 1 year of age (92 strains: 7.5%), 4 years of age (68 strains: 5.5%), 3 years of age (64 strains: 5.2%) and under 1 year of age (62 strains: 5.0%).
6) The rate of isolates from males was higher than females (Male: 58.0%, Female: 42.0%).
7) The frequency of resistant isolates was 62.5% in the period. The most predominant resistance pattern was SM single resistance because of the increase of S. Enteritidis.
8) The number of resistant strains of S. Enteritidis was 424 out of 492 strains (86.2%).

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A 60-year-old male who had been suffering from liver cirrhosis was admitted to our hospital with high grade fever accompanied by right chest pain. Chest X-rays revealed a moderate amount of pleural fluid suggesting pleuritis. P. multocida was isolated from the blood culture as well as the pleural fluid. Antibiotic therapy was initiated according to the drug susceptibility of the isolates. Ten days treatment was effective on the cessation of both septicemia and the clinical symptoms.
Since the patient had been bitten several times by his own pet cats, their mouth swabs were taken for pathogenic investigations. Serotypes of the cats' isolates coincided with that of the patient's which consequently indicated the route of infection.
P. multocida is a Gram negative coccobacillary organism that resides as normal flora in the oral cavity of animals, including dogs and cats. It has been originally known to be a causative agent for hemorrhagic septicemia in domestic animals. However, recently, reports of P. multocida infections in man has been increasing due to the enlargement of pet populations. Although outbreaks of septicemia is rare, it occurs most ofetn in immunologically compromised hosts, including patients with liver cirrhosis as in this case.
Therefore, it is important to initiate an urgent antibiotic therapy in such cases. Overall, it is of utmost importance to instruct immunosuppressed patients to avoid excessive exposure to animals including pets.

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A seventy-one year old male with pemphigus vulgaris and treated with steroid therapy for long periods of 7 months was suffered from invasive pulmonary aspergillosis.
He complained of a productive cough and the chest X-p indicated a mild infiltration shadow in his both lungfields. He was treated by intravenous antibiotics and no clinical improvement was observed.
Two days after the onset of the clinical respiratory symptoms, he was transferred to the division of respiratory diseases and the diagnosis of aspergillosis was confirmed by serological and histopathological studies on the same day. Intravenous amphotericin B and oral itraconazole administrations were started immediately after the diagnosis was made. However, the progression of the disease was so rapid and his immune condition was too weak to respond to the treatment. The overall clinical course of the case was extremely short only 5 days.

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We report a case of dermatomyositis (DM) in a 15-year-old female with toxoplasmosis after ingestion of raw bovine liver. Facial erythema and cervical lymphadenopathy preceded myalgia and muscle weakness of the extremities. The diagnostic criteria of DM was fulfilled because of symmetrical and proximal dominant muscle weakness, elevation of myogenic enzyme (CPK, GOT, LDH, myoglobin, aldorase), myogenic pattern of electromyogram, skeletal muscle biopsy showing interstitial myositis with mild destruction of muscle fiber, and facial erythema. Immunological findings showed IgG anti-toxoplasma antibody to be 1340 IU/ml and IgM to be 7.0 (Cut off index 0.7), suggesting acute toxoplasmosis. Treatment with prednisolone for DM and acetylspiramycin for toxoplasmosis was successful. Toxoplasmosis should be considered as a possibility in patients with myositis.

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