Primer pairs for PCR were designed from the gene encoding the 17, 000-molecular-weight genus-common antigen of Rickettsia japonica, Rickettsia rickettsii, Rickettsia conorii, Rickettsia typhi and Rickettsia prowazekii. Primers R1, R2 were designed for amplifying the genomic DNA from spotted fever group (SFG) rickettsiae and epidemic typhus rickettsiae. Primers Rj5, RjlO were designed for amplifying the genomic DNA from only R. japonica. Using the primers R1, R2, about a 540-bp fragment was observed by amplifying the genomic DNA from R. japonica, R.rickettsii, R. conorii, Thai tick typhus TT-118, Rickettsia sibirica, Rickettsia rnontana, Rickettsia askari, R. typhi, R. prowazekii and Katayama strain isolated from the patient infected with SFG rickettsiae. Using the primers Rj5, RjlO, the 357-bp fragment was observed by amplifying the genomic DNA from R. japonica and Katayama strain. Therefore, the Katayama strain was identified to belong to R. japonica. With primers R1, R2 and Rj5, Rj10, 537 bp and 357 bp bands were amplified from blood of the patients infected with SFG rickettsiae in Kanagawa prefecture.
These findings indicate that the causative agent of SFG rickettsiosis in these two patients was R. japonica. The ticks, Ixodes ovatus and Haernaphysalis flava, were collected by out field research in Kanagawa prefecture. With primers R1, R2 and Rj5, Rj 10, 537 bp and 357 bp were amplified from these ticks.
This indicates that I. ovatus and H. flava were the vector of R. japonica in Kanagawa prefecture. Also, with the primers R1, R2, about a 540 bp fragment was amplified but with primers Rj5, Rj10, no fragments were amplified from I. ovatus and H. flava. Therefore, these ticks may have SFG rickettsiae other than R. japonica and epidemic typhus rickettsiae.

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A PCR method was developed to detect enteropathogenic Escherichia coli (loccalized adherence) and enteroaggregative Escherichia coli. A portion of the structural gene (bfpA) encoding bundle-forming pilus (BFP), was amplified by the PCR for the detection of enteropathogenic Escherichia coli. It proved to be sensitive and specific in detecting enteropathogenic Escherichia coli.
For the detection of enteroaggregative Escherichia coli, a portion of aggR (a transcriptional activator of aggA coding a major fimbrial subunit) was amplified. Although few strains showing non-aggregative adherence gave positive result, the PCR method was shown to have greater sensitivity than the probe assay in detecting enteroaggregative Escherichia coli.

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A single intraperitoneal (ip) inoculation of heat-killed Actinobacillus suis ATCC 15557 (AS 15557) into normal and immunosuppressed (dexamethasone-treated) mice led to remarkable nonspecific resistance to ip challenge with lethal doses of opportunistic pathogens such as Pseudomonas aer-uginosa, methicillin-resistant Staphylococcus aureus and Candida albicans. The duration of this enhanced protective action and the minimal effective dose, in normal mice, induced by AS 15557 were superior to those induced by other bacterial immunostimulants such as heat-killed Lavtobacillus caseiYIT 9018 (LC 9018) and penicillin-treated Streptococcus pyogenes, Su (0K-432). In immunosuppressed mice, the reduced in vivo killing activity of peritoneal exudate cells (PECs) against P. aeruginosainfection was markedly augmented by ip injection of AS 15557. The degree of PEC augmentation induced by AS 15557 was higher than that induced by LC 9018 or by OK-432. The toxicity and histopathological changes associated with AS 15557 were very low, as compared with those by produced by LC 9018 and OK-432. The results suggest that AS 15557, which showed a strong resistance-enhancing capacity against opportunistic bacterial infections, may be a useful bacterial immunostimulant.

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The susceptibility of 10 antimicrobials against P. aeruginosa isolated from complicated UTI during 1992-1995 was determined, and the yearly trend was examinated. The drug tested included 3 cephems (ceftazidime [CAZ], cefpirome [CPR], cefclidin [CFCL]), 1 monobactam (aztreonam [AZT]), 2 carbapenem (imipenem/cilastatin [IPM/CS], biapenem [BIPM]), 2 aminoglycosides (netilmicin [NTL], gentamicin [GM]) and 2 new quinolones (ofloxacin [OFLX], ciprofloxacin [CPFX]). A total number of isolates of which MIC were determined was 77 in 1992/1993 and 70 in 1994/1995. MIC₅₀/MIC₉₀ (pμg/ml) on the isolates were as follows (1992/1993 1994/1995); 3.13/100 12.5/50 in CAZ, 12.5/100 12.5/100 in CPR, 3.13/25 1.56/25 in CFCL, 6.25/50 12.5/100 in AZT, 6.25/25 3.13/25 in IPM/CS, 1.56/6.25 0.78/50 in BIPM, 12.5/100 6.25/100 in NTL, 6.25/50 6.25/100< in GM, 25.100< 25/100< in OFLX, 6.25/100 6.25/100< in CPFX. When the susceptibility of the yearly trend is compared, no significant changes were detected among the drugs tested expect the decrease of susceptibility on CAZ, AZT and increase of it on IPM/CS. From the data obtained, against complicated UTI infected by P.aerugunosa, CFCL, IPM/CS and BIPM are considered to drugs of first choice in the treatment.

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Although macrolide antibiotics have now been widely used in the treatment of chronic airway infections including diffuse panbronchiolitis and chronic bronchitis, the mechanism of the efficacy remains uncertain.
Because the increased mucus glycoprotein secretion from airway goblet cells may play a significant role in the development of such diseases, to determine the effects of macrolides on airway goblet cell secretion, we studied guinea pig airways by a semiquantitative morphometric method. The goblet cell secretion was assessed in histological sections of the trachea and main bronchi stained with Alcian blue and PAS by determining mucus score, which is inversely related to the magnitude of mucus discharge.
Intravenous IL-8 decreased mucus score in a dose-dependent manner and increased the number of neutrophils present in bronchoalveolar lavage fluid. Oral administration of clarithromycin at a daily dose of 1-10 mg/day for 2 weeks dose-dependently inhibited IL-8 (5 mg/kg)-induced decrease in mucus score, with the maximal inhibition being 54±11%(p<0.001) in the trachea and 48±8%(p <0.01) in the main bronchi. This effect was accompanied by the inhibition of neutrophil accumulation into bronchoalveolar lavage fluid. Erythromycin produced similar inhibitory effects on IL-8-induced goblet cell secretion and neutrophil accumulation, whereas amoxicillin and cefaclor had no effect. These results suggest that macrolides protect against goblet cell hypersecretion probably through an inhibition of recruitment of neutrophils into the airway mucosa.

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The aim of this study was to examine the effects of interferon on cirrhotic patients with hepatitis C and the incidence of adverse reactions.
The subjects were 35 cirrhotic patients, and 29 chronic active hepatitis patients without cirrhosis (CAH) served as controls. The cirrhotic patients received 3 or 6 million units of human lymphoblastoid interferon daily for one or two weeks and then three times a week for 22 or 23 weeks, while the CAH patients received 6 million units daily for 2 weeks and then three times a week for 14 or 16 weeks. Discontinuation of interferon treatment or dose reduction was required in the 7 cirrhotic patients. The most frequent reason was thrombocytopenia. Dose reduction alone was necessary in two CAH patients. Five cirrhotic patients (14.3%) and nine CAH patients (31.0%) were classified as complete responders to interferon treatment. In all five complete responders with cirrhosis, the hepatitis C virus RNA level before treatment was less than 5 log copies/50μl. The results of this study confirm the beneficial effect of interferon in selected patients with cirrhosis on basis of pre-treatment virus levels and platelet count.

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Pseudomonas aeruginosa strains isolated from patients with different types of the infections each consisted of LPSs different in chain sizes. The drug-susceptibility patterns of these strains of P. aeruginosa were investigated to clarify the relationship between the LPS-compositions and susceptibility to some kinds of anti-pseudomonal drugs.
The susceptibilities of nineteen strains (seven long-LPS strains, four short-LPS strains and eight LPS-deficient strains) to piperacillin, ceftazidime, gentamicin, norfloxacin and polymyxin-B were determined and these strains were classified into six types (Types I-VI) according to their drug-resistance patterns. Six of the eight LPS-deficient strains were found to be highly resistant to gentamicin alone (Type IV). Four of the seven strains with the long-LPS and one strain with the short-LPS were resistant to three drugs such as piperacillin, ceftazidime and norfloxacin, and classified into Type I. These results indicated that the major part of the LPS-deficient strains and the considerable part of the long-chain LPS strains of P. aeruginosa tested had each characteristic profile in the drug resistance.
The outer membrane proteins of thirteen strains, consisting of different types of LPS compositions, were analyzed by SDS-PAGE. The strains belonging to the same types of the drug-resistance patterns were found to have similar OMP-profiles, although a few exceptions were found.
β-lactamase and gentamicin-inactivating activities were determined for piperacillin-resistant and gentamicin-resistant strains, respectively. Of the piperacillin-resistant strains tested, the activity of βg-lactamase was high in one (No.8) only, low in four and not found in four. The results showed that degrees of resistance of P. aeruginosa strains tested to piperacillin did not correlate to their producibility of β-lactamase except one strain.
Of the nine gentamicin-resistant strains tested, the gentamicin inactivating activity was high in one (No.30) only, moderate in six and low in two.
These results suggested that the significant levels of piperacillin-or gentamicin-resistance ofP. aeruginosa isolated tested might be expressed each due to their decreased abilities for drug-permeabilities in addition to drug-inactivating activities such as β-lactamase or gentamicin-modifying enzyme.
In the case of some resistant strains, the resistance to piperacillin or gentamicin was not explained by the results of the present study. Therefore, we must investigate the possibility that other mechanisms participate in the resistance of these strains.

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Second generation antibodies of HCV, HCV-RNA, genotype and data of liver function were investigated with the purpose of better understanding the condition of the 32 hemophiliac patients currently in our hospital.
The results being:
1) The positivity ratio of second generation antibodies was 96.9% and a negative example occurred in only one infant born in 1990.
2) In the positive example of second generation antibodies, HCV-RNA positivity was 77.4%, which is extremely high result.
3) There are several kinds of genotypes, such as type I, type II, type II + III and type III. Type I is most commonly found in the younger generation. The possibly for this presence was suggested to be from imported blood products.
4) There were few findings of liver disorder in cases of HCV-RNA negative patients. Meanwhrle, the presence of liver disorder was found in about 70% HCV-RNA patients.
There is evidence of correlation between the level disorder and age of the patient. In general, initial infection for hemophilac patients takes place early in life. Compared to non-hemophila hepatitis C patients, the liver disorder occurs at all ages but is becoming more prevalent younger patients. Although not common, there were occasionally a few cases of liver disorder turning into cirrhosis of the liver in patients in their 30's.

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We report two cases of sepsis and meningoencephalitis with listeriosis. They died in despite of administrating sensitive antibiotics.
A 2 day old girl was admitted to our hospital because of fever and cyanosis. Listeria monocytogenes type 4b was cultured from blood, CSF, throat, urine, ear. She was treated with twice exchange transfusion and sensitive antibiotics (ABPC, TOB), but died from DIC.
A 48 year old man suddenly experiented an unconscious condition. A CSF culture grew L.monocytogenes type l/2a. He was treated with sensitive antibiotics (ABPC, CEZ etc), but went bad conditions.
Listeria infection of this cases developed as unfortunate infection.

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A 40-year-old male was admitted to the hospital with acquired immunodeficiency syndrome (AIDS) and Pneumocystis carinii pneumonia (PCP). Two months before admission the patient had a high fever, general fatigue and mild epigastic tenderness. On admission, physical exsamination revealed numerous small tumors on the head, gingiva, neck, nsaal ala, anterior forehead, anterior thoracic, bilateral sole and bilateral lower limbs. At that time, the CD4 cell count was 130/μl. Upper GI endoscopy was performed because of sever epigastralgia and hematemesis. The gastric mucosa was diffusely nodular and erythematous with bleeding. This bippsy showed Kaposi's sarcoma, and the same findings were obtained from the duodenum, rectum and skin, AIDS with related cutaneous and gastrointenstinal KS and PCP was diagnosed. We performed a combination of chemotherapy and Interferon-α therapy, and the KS almost completely disappeared within 3 months.

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We encountered a case which proved to be a mixed infection of herpes simplex virus (HSV) type 1 and type 2 in retrospective terms by in situ hybridization (ISH) and polymerase chain reaction (PCR).
The case was a male. The gestational age was 39 weeks and 2 days. The birth body weight was 3024 g. A fever developed fromthe age of 6 days and he was admitted to the neonatal intensive care unit at the age of eight days. AST was 1042 IU/L, and ALT 206 IU/L.In spite of treatment, the patient died at the age of 12 days.
Using paraffin embedded tisues, we performed the ISH and PCR on the cerebrum, lungs, liver, spleen, bone marrow, adrenal gland, and kidneys.
With the ISH, the lungs, liver, spleen and adrenal gland were both HSV type 1 and type 2. With the PCR, only the liver was positive for type 1, and the lungs, liver, spleen, and adrenal gland were positive for type 2. In the ISH, a probe showing a cross reaction between type 1 and type 2 was used for type 1 probe this time. But a type 2 probe and PCR did not show a cross reaction.
We concluded that this case comfirmed the presence of mixed infection (HSV type 1 and type 2) in neonatal HSV infection.

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A 67-year-old Japanese male, suffering from liver cirrhosis with hepatoma, was admitted to the Yokohama National Hospital because of ascites retention. On physical examination, his abdomen was massively distended with ascites and his lower extremities were edematous. Laboratory findings on admission revealed hypoalbuminemia, moderate icterus, pancytopenia and hepatitis C virus antibody positivity. After admission, abdominal distention and edema were improved with the use of diuretics.
On the 15th day of hospitalization, the patient noted diarrhea and bowel movements that occurred 10 times a day. On the following day, his body temperature rose to over 39°C. On the morning of the 17th day, he complained of severe pain in the right lower extremity. Swelling and erythema over his right lower leg were evident. The skin lesion spread rapidly over the knee and became necrotic. His right leg became increasingly swollen with the development of edema and hemorrhagic bullae. About 4 hrs after the emergence of the skin lesion, his blood pressure fell to less than 60 mmHg. Laboratory findings suggested disseminated intravascular coagulation and multiple organ failure due to serious bacterial infection. In spite of vigorous treatment including administration of antibiotics, dopamine, gabexate mesilate and plasma, he did not recover from the state of shock and died about 14 hrs after the appearance of leg pain.
Bacterial culture of the blood and contents of the bullae grew a gram negative rod identified as Edwardsiella tarda (K tarda) . Histological findings showed necrotizing fasciitis. E. tarda has recently become recognized as a pathogenic bacteria, particularly in patients with an underlying illness. This is the first reported case of E. tarda septicemia with necrotizing fasciitis.

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A 53-year-old female was admitted to our hospital complaining of chest pain and gait disturbance. Examinations on admission showed that she was immunocompetent except the negative tuberculin test. The chest X-ray showed infiltrative shadows with old tuberculous lesions in the bilateral upper lung fields. In CT, a mass lesion was revealed in the lesion, which destructed the fifth thoracic vertebra and invaded into the epidural space. She died of meningitis on the 18th day after admission. On autopsy, it was made clear that the mass lesion was caused by Aspergillus furnigatus, and that the meningitis was the result of the invasion of the fungus into the epidural space.

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A 25-year-old male Japanese who had resided in Ethiopia, presented to our department with eosinophilia, which had been present for about 1 year. Stool examination revealed eggs of Schistosoma mansoni containing miracidia with flame cell activity, and he was diagnosed as having an infestation with this organism. He was treated with praziquantel, and a good parasitological therapeutic result was obtained. Although schistosomiasis mansoni is unfamiliar to Japanese doctors because the infecting organisms are not indigeneous to Japan, doctors should be aware of this disease when they encounter patients with eosinophilia who have visited or resided in tropical developing countries.

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