The association between Campylobacter jejuni infection and Guillain-Barre Syndrome (GBS) was investigated serologically in 52 GBS patients. The specific antibody titers of immunoglobulin G class to C. jejuni were measured in the patient' sera using an enzyme-linked immunosorbent assay with acid-extracted antigen. The analysis showed that 31 (59.6%) of the 52 patients had evidences of recent C. jejuni infections in contrast to 16 (8.9%) of 180 healthy persons. The seropositive patients were analyzed with respect to the nature of their antecendent events. C. jejuni antibodies were found in 22 (75.9%) of the 29 patients with diarrhea, one patient with ensophagus cancer, 4 (30.8%) of the 13 patients with upper respiratry infections, and 4 (50.0%) of the 8 patients without any symptoms, respectively. These findings support that there may be a close relationship between C. jejuni infection and the GBS with preceding diarrhea.
Heterosexual transmission of the human immunodefficiency virus (HIV) appears to occur readily all over the world. Recent reports have suggested an association between HIV and sexually transmitted disease (STD). We conducted a case study among foreign female prostitutes who were seropositive to HIV to determine the prevalence of STD. In this study, we checked the prevalence of STD infection in 19 HIV seropositive female prostitutes. Overall, 84 percent were seropositive for Chlamydia tracomatis, 37% for HBs, 32% for Treponema pallidum (Tp) and 6% for hepatitis C virus (HCV). The high frequency of a history of STD may be associated with an increased risk of HIV infection acquired through heterosexual contact. Prevention of heterosexually transmitted HIV infection will require an extensive educational program aimed specifically at the risk associated with the number and selection of sexual partners and at promoting safer sexual practices.
We performed a clinical study of 16 cases (18 episodes) of Pseudomonas pneumonia by transtracheal aspiration (TTA) from April 1983 to March 1993. The isolation rate of Pseudomonas aeruginosa (P. aeruginosa) among 235 episodes of pneumonias with positive microorganism by TTA was 7.7%. All patients had one or more underlying disease. The most frequent underlying disease was chronic lower respiratory tract infection, followed by lung epidermoid cell carcinoma. More than half of the cases had been given antibiotics prior to the occurrence of pneumonia, and administration of adrenal corticosteroid, heavy smoking habit, and aspiration were seen as other predisposing factors concerning the onset of pneumonias. Monomicrobial infection of P. aeruginosa was 61.1%, and polymicrobial infection containing P. aeruginosa was 38.9%. Community-acquired pneumonia was 61.1% and hospital-acquired pneumonia was 38.9%; the rates of polymicrobial infections and prior administration of antibiotics were higher in the latter group. Mortality due to Pseudomonas pneumonia was 27.8% and the levels of serum albumin and total protein at the onset of pneumonia were significantly lower in the fatal cases than in the recovered cases. It was considered that not only general underlying disease which weaken the immunological resistance of the host, but local bronchial lesion was also important to the onset of the Pseudomonas pneumonia. Also, nutrition is an important prognostic factor on the host's side.
Surveillance cultures of the stool were obtained from 55 patients with hematological malignancies, who have been receiving norfloxacin or tosufloxacin during their neutropenic periods, from May, 1991 to September, 1992. Cultures were performed using 5% blood agar with piperacilin 300μg/ml and amikacin 20μg/ml and bacteriae resistant to these antibiotics were analyzed further. Thirty-four over 55 patients were positive for enterococci and 22 strains of 23 examined were Enterococcus faecium. They are resistant to quinolones, aminoglycosides, cephalosporines and penicillines but not to vancomycin. Eighteen of them were also resistant to high-dose gentamicin. Appropriate measures should be used to prevent intrahospital spread of these resistant isolates.
Porins from wild type strains of Salmonella typhimurium are known to consist of three species of proteins named 34, 35 and 36K. It is understood that these three species of proteins construct porin trimers and the porin trimers from the wild type strain of S. typhimurium are constructed from homologous species of porin subunits. However, we have demonstrated previously that only porin trimers from wild type strains but not mixtures of homologous porin trimers from mutant strains of S. typhimurium could induce antigen-specific cell-mediated immunity (CMI). In this study, therefore, a mechanism of antigen-presentation of porin from a wild type strain of S. typhimurium to T cells was studied. Our data showed that porin proteins from the wild type strain received antigen-processing as well as other protein antigens in murine macrophages and were presented with Ia antigen to T cells. These results suggested that porin trimers from wild type strains of S. typhimurium were constructed from heterologous species of porin subunits and only heterologous porin trimers held epitopes for a CMI-eliciting antigen.
We studied the method for detection of bacterial contamination. Polymerase chain reaction (PCR) was used to amplifity the 888 base pair of 16S ribosomal RNA (rRNA) gene fragment of various strains of bacterial species. The supernatant of bacterial suspension after treatment for 10 min at 100°C was used for a template DNA. A total of 151 strains of 16 genus of Gram-negative rod and of one genus of Gram-positive cocci were confirmed and were divided into five categories by comparing digestion patterns resulting from restriction endonuclease (Mlu I, Eco RI and Hind III) cleavage of target rDNA fragment. These five types, such as Shigella spp. and E. coli group (Group I), other nine genera of Enterobacteria excluding Group I and Aeromonas spp. (Group II), Vibrio spp. (Group III), Campylobacter spp. and P. aeruginosa (Group IV), and S. aureus (Group V), were recognized. The group I was digested by three enzymes used, group II was by Mlu I and Eco RI but not by Hind III, group III was only by Mul I, and group IV was not digested with all enzymes. The group V was sensitive to Eco RI and Hind III but was resistant to Mlu I. This method is a widely applicable technique for detection of bacterial contamination.
In the presence of 40% human serum plus polymorphoneuclear leukocytes (PMNs, 107 cells/ml), changes in the surface properties of clinical isolates of P. aeruginosa were investigated by determining their serotypes and pyocin types as the markers. Furthermore, three isolates were tested for their susceptibility to anti-pseudomonal drugs and profiles of outer membrane proteins by the SDS-PAGE analysis. P. aeruginosa No.21 which did not change in serotype and pyocin type after exposure to serum plus PMNs did not alter their susceptibility to all the drugs tested or their profiles of OMPs. In the case of P. aeruginosa No.1-S, the variants with different serotypes were formed after the exposure, and increased their susceptibilities to some β-lactams and norfloxacin which could penetrate into the bacterial cells through the porin channels of the outer membrane. Furthermore, two of the three type variants formed decreased their susceptibilities to gentamicin and polymyxin B which penetrated into the cells by the self-promoted uptake pathway. P. aeruginosa No.1-R formed the serotype A and G variants with different pyocin types, only when exposed to serum plus PMNs for 24 hours, and the results were accompanied by the applarance of the porin D2 which was not detected in the parent cells. A small number of P. aeruginosa formed the variants with different serotypes and pyocin types owing to the alteration of their surface structures, when the cells were exposed to serum plus PMNs. The alterations were accompanied by the changes in some outer membrane proteins and the drug susceptibility to anti-pseudomonal drugs. In these cases, there were differences between β-lactams (through the porin channels) and poly-cationic drugs (through the self-promoted uptake pathway) in patterns of changes in drug susceptibilities of variants.
As part of the investigation on the source and route of infection with Verocytotoxin producing Echerichia coli (VTEC) in human beings, isolation of VTEC was attempted using fresh feces collected from healthy livestock (cattle, swine and goat) raised in Sagamihra, Yokohama and Hiratsuka cities from October, 1991 to March, 1992. VTEC could be isolated from 1 (1.0%) of 105 swine, 2 (3.6%) of 55 cattle and 2 (15.4%) of 13 goats. VTEC was isolated for the first time from goats in Japan. The combinations of the serotype and toxin type of the isolated strains were 0116: H21 (VT2) and O163: H19 (VT2) for those isolated from the cattle, OUT: H19 (VT2vp) for that from the swine and OUT: H21 (VT1) for those from the goats. Since VTEC isolated from the cattle and goats were found to produce VT of the same serotypes as human VTEC, domestic animals were considered to be involved as a source of VTEC infection in human beings.
We investigated the prophylactic and therapeutic effects of biological response modifiers (rHGCSF, M-CSF, rhIL-2) on pulmonary candidiasis in neutropenic mice. Cyclophosphamide treated mice were injected by the intratracheal route with 5×106Candida yeast cells. Prophylactic treatment with rhG-CSF afforded significant protection against pulmonary candidiasis in neutropenic mice. Treatment with rhG-CSF also increased the number of peripheral blood neutrophils. The histopathological investigations in our experiments showed that the assembly of PMNs to the infected lung at 24 hrs after bacterial challenge was more remarkable in the rhG-CSF treated mice than that in the vehicle alone. Number of viable candida cells in the infected lung in the rhG-CSF treated mice were significantly decreased. The combination of rhG-CSF and fluconazole was more effective than those of each monotherapy. Prophylactic treatment with M-CSF or rhIL-2 had no influence on pulmonary candidiasis. These results show the possibility that rhG-CSF could be of help for treating human deep candidiasis not successfully treated with antimicrobial agents alone.
Based on our previous study using monoclonal antibodies against three Japanese encephalitis (JE) virus strains, Nakayama-RFVL, Beijing 1 and Kamiyama, twenty-five JE virus strains isolated between 1935 and 1979 were classified into four or five serotypes. In the present study, monoclonal antibodies against Muar and 691004 strains which showed different reactivities from the three above strains were produced to analyze immunological characteristics of JE virus in detail and identify isolated viruses accurately. The ten anti-Muar (MUAMA 1-10) and the fourteen anti-691004 (69-MA 1-14) monoclonal antibodies which reacted specifically with the JE virus by the hemagglutination inhibition (HI) test were obtained. In order to clarify the immunological characteristics of MUAMAs and 69-MAs, the HI reactivity of each was tested against the twenty-five JE virus strains. Of the ten MUAMAs, one strain-specific (MUAMA 1), two intermediately reactive and seven JE species-specific antibodies were recognized. Of the fourteen 69-MAs, five intermediately reactive and nine JE speciesspecific antibodies were recognized, but 691004 strain-specific antibody was not obtained. The serological classification of the 25 JE virus strains using MUAMAs and 69-MAs basically corresponded with our previous results. However, 691004 strain would be classified into a subtype of Nakayama serotype according to the reactive pattern. Consequently, the 25 JE virus strains all fell into four serotypes: Nakayama, Beijing 1, Kamiyama and Muar. All twenty-four monoclonal antibodies produced in this study showed neutralization activities and belonged to the IgM class, kappa type.
Antigenic comparison of the twenty-two Japanese encephalitis (JE) virus strains, which were isolated from Taiwan, Singapore, Thailand and India between 1963 and 1984, was carried out by the hemagglutination inhibition (HI) test using the 15 monoclonal antibodies characterized by the different reactivities against Nakayama-RFVL, Beijing 1, Kamiyama, Muar or 691004 strain. Of these twenty-two strains, the seventeen strains reacted with the Kamiyama type-specific monoclonal antibody (KAMIMA 6), but anti-Nakayama, anti-Beijing 1 and anti-Muar type-specific antibodies showed no reactivities with any of the strains. This suggested that the currently prevalent JE virus strains in these areas belonged to the Kamiyama serotype. The other five strains (ThCMP 1982, KE083, KE093, 733913 and Ling) did not react with the above four type-specific monoclonal antibodies. Of these five strains, however, all except the KE093 strain showed a similar pattern to the Kamiyama strain on the basis of the HI reactivities against the other eleven antibodies. The KE093 isolated from Thailand in 1983 showed immunologically outstanding different from the other strains. This result showed the immunological diversity of the JE virus.
A 76-year-old male with multiple myeloma, who was born and had lived in Ehime Prefecture was admitted to our hospital because of high fever. The chest X-p film showed right middle lobal pneumonia. Eosinophilia was detected and a stool smear examination revealed rhabditis form larvae of the nematode. The filaria form larvae of Strongyloides stercoralis were detected by stool culture. Antibotics and pyruvinium pamoate were administered to bacterial pneumonia and hyperinfection of the strongyloides, respectively. Consequently, pneumonia and strongyloidiasis were promptly improved. It was considered that he was infected with strongyloides in south-east Asia during World-War II and a little autoinfection of this nematode had continued about 50 years. In addition, it was suggested that hyperinfection of strongyloides resulted from the immunosuppressive state induced by the chemotherapy for multiple myeloma.
A 42-year-old male was admitted to our hospital because of high grade fever on October 6, 1992. He had no history of cardiac and underlying disease. For the past 10 days, he had complained of high grade fever and noticed arthralgia on his left shoulder. Physical examination on admission revealed that there was a body temperature of 39.0°C and tenderness in the left shoulder. There were no abnormal findings for the chest or abdomen. On the second hospital day, he developed a diastolic murmur which had not been present on admission. And blood culture was positive for Streptococcus agalactiae. Ultrasonic-cardiogram indicated the presence of vegetation. He was diagnosed as infective endocarditis and treated with PCG 20 million units/day, IPM/CS 2 g/day and ISP 400 mg/day. But he was not responding to the chemotherapy. Aortic valve replacement was done on 22nd, October. Valve surgery succeeded and he became well after that time. Endocarditis caused by S. agalactiae is extremely rare, and is an important condition which carries a high mortality. Only seven cases of S. agalactiae endocarditis have been reported in Japan. It is difficult to treat these cases with antibiotic therapy alone. Therefore, we suggest that early surgery should be considered in infective endocardiits caused by S. agalactiae.