Reverse transcriptase (RT) inhibiting antibody in a series of plasma of HIV-1-seropositive subjects was quantitatively measured by poly A-linked colorimetric microtiter plate assay. The plasma were obtained from 6 asymptomatic carrier (AC) s and from 3 patients who progressed to AIDS. They had been followed 29-51 months. RT inhibiting antibody levels in the plasma were measured by inhibition assay against HTLV-IIIB RT activity.
In five of the 6 AC cases, RT inhibiting antibodies in the serial plasma maintained high levels, and 50% inhibiting titers of the serial plasma did not decrease throughout the observation periods (45-51 months). HIV isolation from peripheral blood mononuclear cell (PBMC) of these 5 ACs did not succeed, and HIV p24 antigens were not detected in the plasma. In one AC case (046) RT inhibiting antibody levels gradually decreased after 48 months. In this case, HIV p24 antigen was not detected in the serial plasma throughout the observation period (48 months), but HIV was isolated from PBMC after 27 months.
On the other hand, RT inhibiting antibody levels in the serial plasma of all 3 patients who progressed to AIDS gradually decreased in observation periods (29-35 months). HIV strains were isolated from these 3 cases.
These results suggest that reduction of RT inhibiting antibody levels correlate well with the success of HIV isolation and with progression of clinical manifestation.

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To investigate HIV-1 transmission from mother to child, samples taken from 3 HIV-1 sero-positive pregnant mothers, 3 children born by caesareans from their mothers and therapeutic abortion from 2 HIV-1 sero-positive mothers were tested for HIV-1 antiboy by westernblot, p24 antigen by antigen caputured ELISA, proviral DNA by Nested-PCR and isolation of HIV-1 virus from peripheral blood, cord blood, amniotic fluid and placenta. HIV-1 proviral DNA was detected in the peripheral blood of all mothers, but p24 antigen and virus isolation were not detected. None of the HIV-1 markers, except for antibody, was detected in the samples from their children and placenta.
These facts strongly suggest that there was no HIV-1 transmission from motherto child.

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Recently, it has been suggested that macrolide antibiotics act as immunomodulators. In this study, we evaluated the effect of clarithromycin (CAM) on macrophage function. We used the mouse macrophage cell line, J774.1. The following direct effects of CAM on macrophage function were evaluated: chemotaxis to CAM, chemokinetic effect of CAM, and the effect of CAM on macrophage growth. In order to examine the indirect effects of CAM on macrophage functions, we preincubated macrophages with several concentrations of CAM and then removed the CAM. Thereafter, the phagocytosis of beads, cytocidal activity against Candida albicans, and chemotaxis to lipopolysaccharide were evaluated. In addition, the indirect effects of CAM on endoxan (4 mg/ml) treated macrophage phagocytosis, cytocidal activity, and chemotaxis were evaluated. CAM (at the concentration between 0.04 and 0.2 μg/ml) directly stimulated macrophage chemotaxis and chemokinesis. In addition, CAM dose-dependently stimulated the growth of macrophages. CAM pretreatment (for 4 hours at the concentrations between 0.04 and 0.2 μg/ml) stimulated macrophage phagocytosis, cytocidal activity against Candida albicans, and chemotaxis to lipopolysaccharide. In addition, CAM recovered macrophage phagocytosis, cytocidal activity, and chemotaxis which were decreased after endoxan exposure. These results suggest that CAM has direct and indirect effects on macrophage functions.

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We have encountered six cases with toxic shock-like syndrome (TSLS) by Group A Streptococcus since 1978. Every patient had from the onset flu-like symptoms or high fever and immediately developed some of the following clinical signs: hypotension, acute renal failure, disseminated intravascular coagulation (DIC), and adult respiratory distress syndrome (ARDS). Three of the six patients died with rapid clinical courses and three survived. As for the surviving patients, early administration of antibiotics and anti-DIC therapy were effective. Streptococcus pyogenes were isolated from the blood in five of the patients, ascites from another, and necrotizing tissue in one patient. The serovars of the isolated strains were typed as Tl/Ml in three, T8 in one, and T28 in another. The toxin-type of the strains were typed as B in four and B+C in one.
Though there have been few reports of TSLS before 1992 in Japan, it is supposed that many TSLS cases may have potentially occurred.

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For bacteriological examinations of whooping cough patients, nasopharyngeal specimens were directly inoculated on the cyclodextrin solid medium (CSM) supplemented with 5 μg of cephalexin (CEX) per ml. The inoculated plates were cultured in an incubator at 35°C for 3 to 7 days. During the period from 1990 to 1993, B. pertussis (43 strains) and B. parapertussis (1 strain) were isolated from 145 whooping cough patients and 34 relatives. B. pertussis were isolated sufficiently in June to December during the year. It was suggested that during this period epidemics of whooping cough occurred. All isolates of B. pertussis had K antigen consisting of 1, 3, and 6.
The determination of minimal inhibitory concentrations (MIC) of antibiotics to B. pertussis was carried out by the micro dilution technique modified from the standard methods. The Stainer-Scholte broth, supplemented with heptakis (2, 6-O-dimethyl)β-cyclodextrin, was used for dilution of the drug, and also for cultivation of bacteria. For determining susceptibility of bacteria to antibiotics, twenty seven isolates of B. pertussis and one of B. parapertussis were precultured on CSM at 35°C for 2 days. The turbidity of broth cultures was adjusted to that of McFarland no. 0.5, and diluted to the concentration of 10⁶ CFU/ml. One hundred, μl of these suspensions were added to 25, μl of each antibiotic solution. After incubation for 3 days at 35°C, the turbidity of the bacteria was read macroscopically for determining the MICs. On the other hand, we compared the agar dilution MICs of 23 antibiotics to the broth microdilution MICs for 27 B. pertussis isolates. MICs determined by the agar dilution method were approximately the same or 2 fold of MICs determined by the broth microdilution method. Therefore the in vitro data from the present studies confirm the excellent inhibitory activity of piperacillin, erythromycin and cefoperazone, MICs which ranged from 0.001-0.156, 0.01-0.078 and 0.002-0.078 μg/ml, respectively. Cephaloridine and cefaclor MICs for 90% of B. pertussis isolates were 10 and 80 μg/ml, respectively.

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Eight cases of infection caused by Streptococcus milleri were studied clinically, and a comparative study was made of the backgrounds and factors influencing the development of infectious diseases caused by Streptococcus milleri and Streptococcus pneumoniae.
The patients (six males and two females) with infectious diseases caused by Streptocossus milleri ranged in age from 21 to 81 years old with an average age of 62.0 years. The infectious diseases encountered among the patients were pyothorax in three patients, and in one patient each pyothorax and lung abscess, pleurisy, a secondary infection of pulmonary emphysema, skin infection related to a tracheostomy, a subdural abscess and a brain abscess, respectively. Six out of the eight cases had infections which formed pus.
An underlying disease existed in seven cases, and, in five of these seven cases, it influenced the general condition of these patients. Six serum antibody titers for the S. milleri groups were examined in the First Department of Internal Medicine, University of the Ryukyus. Three cases for S. anginosus, two cases for S. intermedius, and one case for S. constellatus showed high titers of × 1024 or × 2048. Therefore, the examination of the serum antibody titer seems useful for the diagnosis of infections caused by the S. milleri group. We also carried out a comparative investigation of the backgrounds and factors of infectious diseases caused by S. mitten and S. pneumoniae. There are more underlying diseases in infections caused by S. milleri than in those caused by S. pneumoniae. There were more smokers among the patients with infections caused by S. milleri than among those with S. pneumonia infections. In addition, the average Brinkmann Index value was higher in S. milleri than S. pneumoniae. Hemoglobin and albumin values were lower in S. milleri than in S. pneumoniae. From the above results, it was concluded that there may be a tendency for infections caused by S. milleri to appear more frequently than ones caused by S. pneumoniae in patients with an underlying disease, habit of smoking and/or low nutrition.

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Fungi were isolated from 113 (14.2%) of 789 patients with positive blood cultures at Oita Medical University Hospital between 1981 and 1992. The rates of fungemia increased in recent years; 13.9%(1981-1985), 12.1%(1986-1988) and 16.9%(1989-1992). The isolated fungi were Candida parapsilosis (25.7%), C. albicans (24.8%), C. tropicalis (14.2%), Trichosporon beigelii (10.6%), C. glabrata (8.0%) and so on. The major fungi were T. beigelii and C. glabrata in patients with hematologic malignancies, whereas they were C. albicans and C. parapsilosis in patients with non-hematologic diseases and C. glabrata increased in both groups. Prophylactic or emiric administration of antifungal agents probably influenced the difference of the causative organisms in the two groups.

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Acinetobacter spp. are non-fermented gram-negative rods that are widespread in the environment and colonize in the human skin. They are known to be a nosocomial pathogen causing, pneumonia, menigitis and bacteremia. Recently, they have been found increasingly in catheterrelated infections (CRI). Thirty-seven cases of bateremia were developed in our hospital during the past five years. Of these 27 cases were chosen out of the medical records for discussion in this paper.
Twenty-three cases are blood positive for Acinetobacter anitratus and 4 cases for A. lwoffii. Most cases have an underlying disease like hematological malignancy, solid tumor and infantile congenital abnormality. There were also some clinical signs ; high fever, hypotention, tachycardia, tachypnea, peripheral cyanosis. A central venus catheter was inserted in 22 cases, and in 13 of these, the catheter was removed after the bacteremic episode. Nine cases became afebrile after the removal of the catheter and A. anitratus was isolated from the catheter tip in four cases. Heparin was administered through the catheter in 7 cases.
Formerly Acinetobacter spp. were not recognized as a major pathogen, but recently found increasingly in CRI. We also found 9 cases which were definitely diagnosed or suspected as CRI, and were successfully treated by removal of the central venus catheter. Association between administration of heparin and bacteremia of Acinetobacter spp. was reported, we actually detected such association in 7 cases, but the potential role of heparin has not been clarified yet.
Compared with A. lwoffii, A. anitratus were resistant to many drugs, but had good susceptibility to imipenem, minocycline, aminoglycoside, and fluoroquinolone.
We have to pay attention to clinical signs of sepsis in patients with indwelling catheter, and appropriate treatment is required when CRI is suspected. Acinetobacter spp. have been identified as one of the important pathogens of CRI.

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A total of 300 extraintestinal clinical isolates of Bacteroides fragilis group organisms were assayed for enterotoxin using HT29/C₁ cells. Of the 179 strains of B. fragilis, 36 (20.0%) were positive for cell culture assay, a positivity which was neutralized by anti-B. fragilis enterotoxin serum; all 36 strains were confirmed to be enterotoxigenic B. fragilis (ETBF). Among B. fragilis isolated from blood, 18 (31%) of 59 strains were ETBF while 18 (15.0%) of 120 strains isolated from non-blood specimens were ETBF (p <0.05). None of the 121 strains of the B. fragilis group other than B. fragilis were positive for cell culture assay. These results demonstrated that isolation of ETBF from extraintestinal clinical specimens is not uncommon and that B. fragilis enterotoxin, a metalloprotease, may play a role as a pathogenic factor in blood stream infection and sepsis.

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We found that pediatricians have enhanced specific cellular immunity to varicella-zoster virus (VZV) compared with the general population, which may be due to reexposure to VZV from children with chickenpox. There have been some reported that the varicella vaccine enhance the specific cellular immunity.
To estimate the efficacy of varicella vaccine for protection against herpes zoster in the elderly, we investigated the incidence of herpes zoster in 500 pediatricians and family practitioners with their fifties and sixties, and history of reexposure to VZV in 61 patients with herpes zoster by questionnaires retrospectively.
Thirty-four of 352 pediatricians had a past history of herpes zoster. The incidence per 100, 000 person-years of herpes zoster was 65.2 in those in their fifties and 158.2 in those in their sixties, which are 1/2 to 1/8 of other reports regarding the general population. Among 61 immunocompetent patients with herpes zoster, only 4 patients (6.6%) had the chance for reexpose to VZV before their herpes zoster. Only 7 (17.5%) of the 40 patients older than 50 years of age lived with their children less than 14 years of age. Twenty-three (57.5%) of them lived without their children and grandchildren.
They are thought to be less chance to reexpose to VZV through children. We may think that the booster effect by reexposure to VZV plays an important role to prevent herpes zoster. Therefore, we can speculate that the varicella vaccine may protect aganist herpes zoster in the elderly by the enhanced specific celluar immunity due to the booster effect.

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The formation of experimental bladder calculus was studied. The calculus was formed by the uptake of ethylene glycoiwater (1%) and retaining the silk thread in rat bladder with high frequency. The components of the calculus were calcium oxalate and calcium phosphate from the results of the electron prove micro analysis (EPMA) and ion chromatography.
On the 7th day after the beginning of experiment, Pseudomonas aeruginosa was inoculated to the rat bladder via the urethra. Seven days after the infection, P. aeruginosa adhered to the surface of the calculus such as an aspect of a biofilm. It was considered that this experimental model was useful to study the adherence of bacteria, biofilm formation and its chemotherapy by antibacterial agents.

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The following results were obtained from a study of Enterococci isolated at the Okayama University Hospital from 1990 to 1994.
1) The isolation frequency of Enterococci was about 6% for each year.
2) The percentage of Enterococcus faecalis (E. faecalis) among Enterococci decreased each year, but the percentage of Enterococcus faecium (E. faecium) , and Enterococcus avium (E. avium) increased each year.
3) Results of drug sensitivity tests revealed that the incidence of ofloxacin (OFLX) resistant E. faecalis and E. avium increased each year. Also, the resistance of E. faecalis to ampicillin (ABPC), the primary medication choice for treating E. faecalis, increased each year.
4) A few vancomycin (VCM) non-sensitive Enterococci were isolated. These bacteria showed low-sensitivity toward ABPC, imipenem (IPM), gentamicin (GM), and OFLX, and the extent of multiple drug resistance increased each year.
5) One hundred strains of Enterococci were selected to examine the expression of β-lactamase using the broth method and the iodine-starch method. However, β-lactamase producing strains were not detected.

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Fifty clinical isolates of Pseudomonas aeruginosa were tested for inhibition of growth of clinical isolates of Escherichiacoli, Salmonella infantis, Klebsiella pneumoniae and other Gram-negative bacteria in the authors' laboratory. Pseudomonas aeruginosa was strongly active against both E. coli and Enterobacter cloacae, with 89.4% and 94.7% inhibition respectively, but weakly active against S. infantis, K. pneumoniae and Proteus mirabilis with 56.3%, 48.8% and 23.8% inhibition, respectively. The pigmented strains were found to have stronger antimicrobial activity than the unpigmented strains.
Pyocyanin, the major metabolite of Pseudomonas aeruginosa, has been shown to inhibit Escherichia coli, Proteus spp. and other Gram-negative bacteria, by research with a few strains of P. aeruginosa and a single inhibited strains. However, little attempt has been made to determine the inhibitory action of many strains of P. aeruginosa against a large number of clinical isolates such as Escherichia spp., Klebsiella spp., and Salmonella spp., up to now. For this reason, in this study we examined 50 randomly selected clinical isolates of P. aeruginosa for inhibition of growth of a wide range of Gram-negative bacteria, including 30 strains of E. coli, 30 of K. pneumoniae, 30 of S. infantis, 6 of Enterobacter cloacae and 9 of Proteus mirabilis.

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Human parvovirus B19 (B19) has been shown to be the cause of erthema infectiousum (EI). Recently the various clinical manifestations with B19 infection are coming to light. The relationship between B19 and anaphylactoid purpura (Scholein-Henoch purpura: SHP) was investigated in a retrospective study.
There were sixteen patients (male 9, female 7, average age 6.0 ± 1.7) who were diagnosed as SHP. Serum specimens were used in the present study. Specific IgM and IgG were assayed by enzyme immunoassay, B19 DNA was assayed by polymerase chain reaction (PCR).
Five of the sixteen patients were positive for B19 DNA assayed by PCR, three were positive for B19 IgM in the acute phase. Kidney involvement appeared in one case with B19 infection and in two cases without infection. Abdominal involvement appeared in one case with infection and in three cases without.<P></P>In this study, five of the sixteen patients had evidence of recent B19 infection by the existence of antibodies and PCR method. In conclusion, it is necessary to investigate the B19 infectin with or without EI in the cases of SHP.

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A 62-year-old male was admitted to our hospital after an abnormal shadow was pointed out by Chest X-ray in June, 1994, even though he was asymptomatic otherwise (first episode). The serum IgG antibody against Chlamydia pneumoniae was elevated up to 512-fold in the acute phase and decreased to 32-fold in the convalescent phase using the micro-immunofluorescence (MIF) test, indicating a C. pneumoniae acute infection. In fact, no other micro-organisms were detected. The patient recovered from the pneumonia without any treatment.
He was admitted to our hospital again after having right chest pain in December, 1994 (second epidose). An X-ray examination revealed a slight infiltration of the right lower lung field and pleural effusion. In this episode, he received therapy with carbapenem anti-bacterial agent and he recovered from the pneumonia 9 days after the administration of the antibiotic. The serum IgG titer against C. pneumoniae was elevated up to 1024-fold on admission, indicating mixed infection with bacteria and C. pneumoniae. It was concluded that both of these episodes indicated a spontaneous cure of the pneumonia which had developed from C. pneumoniae.

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[in Japanese]

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The Gen-Probe^® Amplified Mycobacterium Tuberculosis Direct Test (MTD) is a rapid direct specimen assay method for identification of M. tuberculosis by amplification of ribosomal RNA and detection of the product with a specific chemiluminescent DNA probe.
We report two cases of tuberculous meningitis which were positive in the MTD. Since the cerebrospinal fluid showed negative smears and positive cultures in both cases, we could diagnose tuberculous meningitis by MTD about 2 months earlier than by conventional smear and culture methods.
In one patient, the fever didn't subside immediately after staring chemotherapy. In the other patient, an intracerebral focus appeared after administration of antituberculosis drugs. However, we did not hesitate to continue the therapy because of the MTD positive findings.
MTD is considered to be a very useful test especially in diseases like tuberculous meningitis which need rapid diagnosis and rapid therapy, and is useful for improvement of convalescence and is economical.

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