Agar dilution and semi-solid agar dilution were used to determine the MIC of faropenem (FRPM) against 24 H. pylori isolates. FRPM was active against clarithromycin resistance H. pylori isolates. And, the MICs obtained by both methods were in agreement. These result suggest that FRPM was not affected by pH and is a clinically useful oral antibiotic for the eradication therapy of H. pylori infections.

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We examined the bactericidal activity of catechin, an astringent ingredient of tea, on enterohemorrhagic Escherichia coli(EHEC) O157: H7 and the anti-toxin activity of catechin on vero toxin (VT), the main pathogenic factor of EHEC O157: H7. To examine bactericidal activity, we added 1×104 CFU/ml bacteria to 1.25 to 20 W/V% of green tea extract or the PBS solution containing 25 to 400μg/ml of (-) epigallocatechin gallate (EGCg), which is the main catechin ingredient of gree tea leaf, and counted the number of live bacteria at various intervals. After 3 to 5 hours, no live bacteria were seen in 1.25 to 2.5 (regular drinking concentration)% green tea extract. In the high concentrations of 100 to 400μg/ml EGCg the number of live bacteria decreased with time and after 24 hours no survivors were seen. In the low concentrations of 25 to 50μg/ml EGCg, however, no change was observed in the number of live bacteria during 5 hours. After 24 hours the bacteria in 50μg/ml were killed and the number of bacteria in 25 μg/ ml decreased to one tenth of that at the start. To examine the anti-toxin activity, we mixed equal volumes of 2 ng/0.1 ml VT₂ and 0.5 to 2 mg/0.1 ml catechin in vitro and incubated them at 37°C for various times. Then we inoculated 0.2 ml of the mixture intraperitonealy to BALB/c mice. One mg of catechin inhibited by 100% the lethal toxicity of 2 ng of VT2 (LD 100) to mice. The inhibition of lethal toxicity of VT2 by catechin depended on the incubation time. The rate of inhibition was 0, 40 and 100% for 9, 12 and 18-24 hours incubation, respectively. These results suggest that catechin has not only bactericidal activity on EHEC O157: H7 but also anti-toxin activity on vero toxin.

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To assess the existence of Vibrio cholerae non-O1 in the environmental water system during the cold season, the organism was incubated in both river sediment or terrestrial soil as particle matter-water (1: 1) suspension. The low temperature condition was set to 5° and 10°C At 5°C, V. cholerae non-O1 did not grow in any medium, sediment and soil. At 10°C, the decrease of the number of organisms was delayed in river sediment as compared with peptone medium. At 30°C, the organism grew in river sediment and its extract solution. In soil, the organism was decreased immediately under low temperature. Though the number of organisms was decreased temporarily in soil, its number was maintained within 14 days at 30°C. But, in the extract solution from soil, the organism was decreased immediately under 5°, 10°, 20°, 30°C. It showed that survival of V. cholerae non-01 was maintained in particle substances contained water. These results suggested that V. cholerae non-O1 would be able to exist in river sediment during the cold season.

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Recently, Beutin et al have studied hemolysin production in a large number of serologically diverse verotoxin (VT) producing Escherichia coli (VTEC) strains and found a close association between hemolysin production and VT production. This study was to examine whether hemolysin production on sheep blood agar can be used as an epidemiological marker for microbiological screening of enterohemorrhagic Escherichia coli (EHEC). In comparison of hemolysis on washed blood agar plates prepared in our laboratory and commercially available blood agar, all VT producing strains exhibited a typical hemolytic phenotype on the former, on the other hand, VT non-producers were less hemolytic on the former than the latter.
On the washed blood agar, 11 VT1 producing strains belonging to 4 different serotypes, 1 strain of VT2 producer, 5 VT1 & 2 producers belonging to 2 different serotypes, all the 17 strains of VTEC were positive for hemolysis. The 85 VT non-producing strains belonging to 18 different serotypes isolated from faecal specimens from patients with diarrhea were tested, 10 strains (12%) belonging to 4 different serotypes (06, 18, 26, UT: untypable) produced hemolysin. Twohundred four (10%) of the 2, 036 strains isolated from faecal specimens from 507 healthy persons exhibited typical hemolysis.
As a result of identification of these 204 hemolytic strains, 148 straisn of E. coli, 13 strains of Citrobacter and 10 strains of other enterobacteria, and 13 strains of Pseudomonas, 12 strains of Aeromonas, 5 strains of Proteus and 3 strains of Vibrio were detected. 148 hemolytic strains of E. coli were carried out O-serotyping with commercially available pathogenic E. coli antisera, 57 strains (39%) were classified into 11 different serotypes, but 91 strains were not typable. Most frequent serotype was 018 (24 strains), next 06 (18 strains), 01 (3 strains), and other 8 different serotypes (025, 26, 28ac, 29, 55, 146, 152, 159) were less than 2 strains respectively.

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When murine resident peritoneal macrophages were treated in vitro with overnight culture supernatants of clinical isolates of S. aureus (21 strains) (S. aureus-CS) for 4 hrs at 37°C in a 5% CO₂-air humidified incubator, TNFα induction from the treated cells was observed in 20/21 strains. The amount of TNFα dispersed from 88.4 to 1726.5 pg/ml but more or less amount of TNFα did not relate with the presence of TSST-1 and enterotoxin production of S. aureus. Recombinant protein A induced TNFα production by macrophages dose-dependently, but the relationship between the amount of protein A in each S. aureus CS and that of TNFα induced from macrophages treated with them were not satisfactory (r =0.69). When each strain was injected (×10⁹ bacterial cells) interperitonealy to mice, 13/21 strains indicated lethal activity. Relation between TNFα inducibility in vitro and lethal activity, however, was not found (r= 0.4). These results suggest that TNFα inducibility is the basic biological activity of S. aureus, although there is a difference in the induction amount and in component (s) to induce TNFα on each strain, and then because the strain is strong in TNFα indicibility, it does not necessarily follow that pathogenicity is strong.

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The prevalence of mycobacterial infection in AIDS patients has increased in Japan. This report describes details of the clinical and radiological features of eight AIDS patients with mycobacterial disease (6 with M. tuberculosis infection and 2 with M. kansasii infection) i our hospital during the period from October 1995 through February 1997. Six of the 8 were men, and two were women. The mean age was 36.5 years. Six were Japanese, one was from Myanmar, and one was Malaysian. The median CD4 positive T lymphocyte counts (CD4 count) at the time of diagnosis of the M. tuberculosis was 75.5 (range 14-569)/microliter, and of the M. kansasii was 21.5 (range 19-24)/microliter. Clinical findings and symptoms of all patients were non-specific, but almost all patients had a cough and fever. In the radiographic findings, the patients of the M. tuberculosis group presented multiple hilar and mediastinal lymphadenopathy, miliary shadow, and obstructive pneumonia. Both M. kansasii patients showed a multiple infiltration shadow. There were no drug resistant strains in M. tubeculosis except one isolate with moderate resistant to Streptomycin.
These observations suggest that AIDS-associated mycobacterial disease shows atypical clinical and radiological features in some cases, especially in advanced stages of AIDS. Therefore, we need to recognize the characteristics of the clinical and radiological features of the patients with mycobacterial diseases and AIDS.

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The measurement of serum (β3)-β-D-glucan (β-glucan) is commonly used in the clinical stage. Although it has been considered very useful in the carly diagnosis of deep mycosis, it is influenced by several factors, one of which is hemodialysis. Therefore, we decided to investigate clinically the influence of hemodialysis membranes on serum β-glucan. Plasma samples were collected from 26 patients receiving hemodialysis in Kawasaki Medical School Hospital. The kinds of hemodialysis membranes examined were four synthetic polymers and three natural polymers. We also measured β-glucan in five washing liquids used in the hemodialysis circuit, five dialyzates, two sodium heparin and two filling liquids for wet type dialyzers to investigate the influence of other factors associated with hemodialysis on serum β-glucan. Elevations of β-glucan were observed in some cases treated by hemodialysis apparatus using a saponified cellulose acetate membrane. But, in all cases treated by hemodialysis using a cellulose triacetate membrane, slight elevations of β-glucan were observed. No significant increases in β-glucan after hemodialysis were observed in the cases treated by hemodialysis using a synthetic polymer membrane. Elevations of β-glucan in sodium heparin and in one of the filling liquids for the wet type dializer were observed. We, therefore, concluded that there are some other factors having an effect on serum β-glucan besides the hemodialysis membrane in the patients treated by hemodialysis.

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We have evaluated a new kit, PEPTIDE Chlamydia (Meiji Milk Products Co., Ltd, Tokyo), for detecting species specific antibodies to Chlamydia trachomatis with synthetic peptides as an antigen. Serum samples from women with C. trachomatis cervicitis (n=45), healthy pregnant women (n=100), and children suffering from C. pneum oniae lower respiratory tract infection (n=9) were used. We have measured the serum Ig or Ig antibodies to C. trachomatis of those sera with PEPTIDE, Sero IPALIZA Chlamydia (Savyon Diagnostics, Israel), and HITAZYME Chlamydia (Hitachi Chemical Co., Ltd, Hitachi).
Serum samples with discrepant results were further analyzed by a microimmunofluorescence test and immunoblotting (western blotting). Ig and Ig serum positive rate to C. trachomatis of sera from women with C. trachomatis cervicitis were similar in three kits, that is, 91% and 80% in PEPTIDE, 89% and 82% in Sero IPALIZA, and 84% and 76% in HITAZYME, respectively. Ig and Ig serum positive rate to C. trachomatis of sera from healthy pregnant women were 18% and 9% in PEPTIDE, 12% and 15% in Sero IPALIZA, and 15% and 13% in HITAZYME, respectively. Serum antibodies to C. trachomatis in serum samples from children with C. pneumoniae infection were all negative in PEPTIDE, but falsely positive in several cases in Sero IPALIZA or HITAZYME. In sera with discrepant results, PEPTIDE gave more identical results with a micro-IF test and immunoblotting analysis than Sero IPALIZA or HITAZYME.
These results indicate that PEPTIDE is an useful kit to detect more species specific antibodies to Chlamydia trachomatis than former kits.

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A survey was made on the situation of Group A Streptococcal toxic shock syndrome (STSS) based on questionnaires. The survey was divided into two parts. The first survey was done by sending out an outline of the STSS inquiring if any STSS cases were observed by mail to university hospitals, residence training hospitals and other major hospitals totaling 2512 institutes. The second survey was subsequently done to the institutes that had STSS cases asking for the clinical course, data and sampling of the bacteria.
The diagnosis of STSS was confirmed based on the diagnostic criteria induced by the working group of the United State. We have found 97 cases of STSS which 48.5% had fatal outcomes. There was no significant sex difference in the onset or the mortality rate. It occurred more in the older population, and occurred throught out Japan but was not found to be epidemic. The first case was backed in 1978 and it began to increase since 1993, reaching its peak in 1994 and now decreasing in number. Most of the isolated Group A streptococcus were of type Ml and M3. We have modified the United States diagnostic criteria creating a new Japanese criteria, which includes the symptoms of the central nervous system in the term MOF. The aim for the Japanese criteria is to search for the etiology of the disease. The Japanese criteria requires that the disease progresses rapidly and that the patient be free from any conditions that might suppress the immunal system.

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An outbreak of Escherichia coli O157: H7 infection occurred in July 1996 in Sakai City. About 5000 children were infected, 122 of whom developed hemolytic uremic syndrome (HUS). In this outbreak, almost all patients were administrated some type of antibiotics. The effects of antibiotics on E. coli O157 associated hemorrhagic colitis (HC) have been controversial. In this study, we focused on the effects of antibiotics on development of HUS in the Sakai outbreak.
We retrospectively determined the antibiotics administrated within three days after the onset of HC, clinical courses, and laboratory data of 301 patients who were hospitalized and identified as Escherichia coli O157 infection by stool culture, from results of questionnaires sent by the Osaka Prefecture Medical Association to hospitals in Osaka Prefecture. The antibiotics used could be identified for 216 patients. The incidence of HUS among these patients was 11.6%. They were divided into 19 groups based on the type of antibiotics administrated. The incidence of HUS in the new quinolone (3.7%) group was low, but was high in the intravenous cephalosporin (18.2%) group. The differences in the incidence of HUS among the 19 antibiotic groups was significant (p<0.05) on analysis of covariance which eliminated the contributions of variables including age, sex and laboratory data.
These findings indicate that the suitable antibiotics can prevent the development of E. coli O157-associated HUS.

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An optical biosensor to monitor molecular intractions was developed. This system made possible to observe reactions within a few minutes. We tried to apply this system for rapid diagnosis of viral infections. Measles virus propagated in our laboratory and crude rabbit anti-measles antiserum were used. A commercially available human immunoglobulin was used as a representative of patient sera.
A crude rabbit anti-measles antiserum was fixed on the reaction surface of aminosilane coated cuvette, and specific binding of measles virus to this antibody was monitored. Specific purifed IgG antibody was known to give as high sensitivity as the conventional culture method. But the sensitivity by crude antibody was found to be 1/10 in the sensitivity. This might be caused by the masked effects on specific antibody to the contaminated high amounts of non-specific proteins. None purifed polyclonal antisera for most of the viruses are commercially available, and 10-20 times highly concentrated antibody solutions could be used for this titrations. Estimated anti-measles antibody titer of the commercial available human immunoglobulin by biosensor system was found to be the same as that by the conventional culture method. It was suggested that 1×10³ virus particles/ml in the test solutions could be detectable and titerable by using comercially available specific anti-viral antibody in real time, and that viral infections could be diagnosed within an hour.

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A clinical analysis on 127 cases out of 140 cases presenting with fungemia at Fukuoka University Hospital between 1984 and 1994 was done. The number of the positive blood cultures during the same period was 1188 and the rate of fungemia was 11.8%. The rates generally increased in recent years, 9.0%(1984-1986), 10.2%(1987-1989) and 13.6%(1990-1994), but decreased after 1992. C. albicans has shown a tendency to decrease while C. parapsilosis and C.glabrata have increased in recent years thus suggesting the effect of the prevalence of intravenous hyperalimentation (IVH) and azole antifungal agents. Only 3.9% of the fungemia were preceded by bacteremia. In addition to the known risk factors for fungemia such as IVH (89.0%) and the antibacterial agents, H2-receptor antagonists were used in 58.3% of the cases. 14 cases of the fungemia were observed during the prophylactic use of antifungal drugs. Therefore, fungemia should always be kept in mind regarding the differential diagnosis for bacteremia when an indwelling venous catheter is used. The prompt extubation of the catheter in addition to the adjustment of the dosage of antifungal drugs in response to the status of the host defense system are thus considered to be important in the treatment of fungemia.

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Legionellosis is an important cause of severe pneumonia in the community. Inadequate therapy will lead to respiratory distress syndrome, disseminated intravascular coagulation (DIC) and finally fatal multiple organ failure. We encountered a rare case in which early manifestation included septic shock and DIC complicated by acute myocardial infarction (AMI) suspected to be derived from Legionnaires' disease. A 54-year-old healthy female complained of lumbago, high fever and dry cough 10 days after visiting a hot spring spa. She was emmergently admitted due to shock. Physical examination demonstrated hypotension, high fever, course creakle in the right lower lung. Hepatosplenomegaly, lymphadenopathy and eruption were not found. WBC count was 34600/, u1 with nuclear shift. CRP elevated. FDP, D dimer and TAT also elevated. CPK elevated with dominance of the MB isozyme. Chest roentogenography revealed congestive heart failure, pleural effusion and obscure pneumonic shadow and EKG showed ST segment elevation in leads I, II, III, aVF, V4, V5 and V6. The patient was diagnosed as having septic shock, DIC and AMI. She was treated with gabexate mesilate, high dose methyl prednisolone and dopamine hydrochloride as well as piperacillin, meropenem, isepamycin and fluconzaole. Despite intensive care, the blood pressure fell again and pneumonia had progressed on the 8th hospital day. These antibotics appeared to be ineffective. Erythromycin was then administered and a dramatic effect was obtained as the patient recovered. Serum titer of Legionella pneumophila (serogroup 1) rose to 128-fold 2 weeks after the onset. Other serum titers such as Chlamydia psittaci, Rickettsia, Mycoplasma were all negative. Cultures obtained from the sputum, thorat swab, urine and blood did not yield any microorganisms. Although the diagnosis could not be confirmed because the titer did not elevate over 256-fold or 4-fold within 2 weeks after the onset, Legionella infection was hightly suspected from the clinical features. This is a rare case in which septic shock and DIC with AMI preceded pulmonary symptoms in a non-immunocompromised patient.

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