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Biomedical subjects

M Kohara

Publications and source records attributed to M Kohara.

At least 55 records · Page 3Linked to original sources
PubMed1997

Effects of cyclosporin A on hepatitis C virus infection in bone marrow transplant patients. Bone Marrow Transplantation Team.

The hepatitis C virus (HCV) infection has, in general, been considered not to affect liver function severely during the course of bone marrow transplantation (BMT) except for late hepatitis which coincided with a decrease in immunosuppressive therapy. We examined serial sera of two patients with positive HCV antibody who underwent allogeneic BMT and found that while the dose of cyclosporin A tapered off, the serum concentration of HCV core protein increased before the occurrence of hepatitis. This suggests that viral reactivation and growth might be one of the important mechanisms of hepatitis after BMT in patients with positive HCV antibody.

Adult↗
PubMed1996

Detection of hepatitis C virus specific core protein in serum of patients by a sensitive fluorescence enzyme immunoassay (FEIA).

A protein-capture fluorescence enzyme immunoassay (FEIA) was developed using monoclonal antibodies (mAbs) against recombinant hepatitis C virus (HCV) core protein. Four hybridoma cell lines (5E3, 5F11, 515S, 1080S) were established and characterized. These monoclonal antibodies (mAbs) each had IgG1 and OgG2 isotypes, and recognized major B cell epitopes within the immunodominant nucleoprotein amino terminal subregion. Using mAb 5F11 as the first antibody to the solid phase and beta-D-galactosidase-conjugated mAb 5E3 as the second antibody to the protein, we established a specific HCV core protein capturing FEIA capable of detecting as little as 20 pg/ml of recombinant HCV core protein. HCV core protein in serum was detectable after treatment with 4.0% polyethyleneglycol, 0.5 NaOH, and 5% Triton X-100. The results of a peptide inhibition assay indicated that this FEIA is specific for HCV RNA positive sera. The quantity of HCV core protein detected in serum was significantly correlated to the level of HCV RNA. The detection limit for HCV core proteins was an HCV RNA per titer of approximately 10(4)/ml. Using this FEIA system, the detection ratio of HCV core protein in patients with chronic HCV infection was 92.3% (70/76).

Animals↗
PubMed1996

Diurnal variations in resting-active cycles in full-term fetal heart rate changes.

To elucidate the mechanism of the resting-active cycles (RAC) of fetal heart rates (FHR), in the resting and non-resting phases (RP and NRP), 24-h FHR recordings were made on 16 normal full-term pregnant women. RP, NRP, RAC-1 (NRP-NRP cycle), and RAC-2 (RP-RP cycle) were defined based on the criteria of Nijhuis et al. Frequency distributions were plotted separately for the entire 24-h period as well as for the day-time (07:00-21:00 h) and night-time (21:00-07:00 h), and were compared using Kolmogorov-Smirnov two-sample tests. The mean durations (+/- S.D.) (min) of RP, NRP, RAC-1, and RAC-2 were 22.7 +/- 11.2, 67.3 +/- 47.2, 90.0 +/- 47.6, and 89.9 +/- 48.6 during 24-h periods, 20.1 +/- 7.7, 68.3 +/- 52.3, 88.6 +/- 53.1, and 88.4 +/- 53.0 during the day-time, and 25.4 +/- 13.2, 66.2 +/- 41.2, 91.4 +/- 41.0, and 91.5 +/- 43.4 during the night-time. Length of RP was the only factor significantly different during the day and night (P < 0.05). We propose that there are different mechanisms controlling RP and NRP.

Activity Cycles↗
PubMed1996

Hepatitis C virus: detection of intracellular virus particles by electron microscopy.

We previously demonstrated that a human T-cell line, HPBMa10-2 derived from HPBALL, was capable of supporting a productive infection of hepatitis C virus (HCV). We subsequently found Daudi cells, a human B-cell line, to be susceptible to HCV infection. Employing these cell lines infected with HCV as well as liver obtained during the acute phase of hepatitis C from a chimpanzee, we observed intracellular HCV particles by electron microscopy (EM). We detected viruslike particles with a diameter of approximately 50 nm in the cytoplasmic vesicles. In Daudi cells harvested 15 days after virus inoculation, the appearance of cytoplasmic tubular structures which we originally described for chimpanzee hepatocytes in association with HCV infection was noted. Immunoperoxidase EM using antibodies against HCV core and envelope demonstrated that the particles contained the viral antigens.

Animals↗
PubMed1996

Quantification of serum hepatitis C virus core protein level in patients chronically infected with different hepatitis C virus genotypes.

BACKGROUND/AIM: A novel fluorescent enzyme immunoassay (FEIA) for the detection and quantification of serum hepatitis C virus (HCV) core protein was developed. The aim of this study was to evaluate the relation among serum HCV core protein level, HCV RNA level, and HCV genotype in patients with chronic HCV infection. PATIENTS AND METHODS: Serum HCV core protein, HCV RNA, HCV genotype were determined in 175 patients using the FEIA, branched DNA assay (Quantiplex HCV RNA ver 1.0), and serologically defined genotyping assay, respectively. For the specificity, all 13 patients seronegative for anti-HCV were negative for serum core antigen and HCV RNA by FEIA and bDNA, respectively. RESULTS: FEIA assay seems to be more sensitive than bDNA for patients with HCV type 2 infection (detection: 83.4% v 63.4%, p < 0.01). There was a good overall correlation between the FEIA and bDNA results. However, when the patients were stratified into their HCV types, a correlation was observed in HCV type 1 but not in type 2 infection. Patients with HCV type 2 infection had a lower serum HCV core protein level (median 56 RFI) compared with type 1 infection (median 149 RFI, p < 0.01). Thirty seven patients subsequently received interferon alpha therapy, patients who showed a complete and sustained response had a lower pretreatment serum HCV core protein level compared with patients who had a relapse and nonresponders (36 v 338 RFI, p < 0.01). CONCLUSIONS: This study showed that FEIA (1) is a good assay for the detection and quantification of serum HCV core protein level, (2) is also very sensitive in detecting HCV core protein in patients with HCV type 2 infection, and (3) may have a role as a predictor of subsequent response to interferon therapy.

Female↗
PubMed1995

Poly(U) binding activity of hepatitis C virus NS3 protein, a putative RNA helicase.

A non-structural protein of the hepatitis C virus (HCV), NS3, contains amino acid sequence motifs characteristic of serine-proteinases and RNA helicases. RNA binding activity of the NS3 protein with an apparent dissociation constant of 2 x 10(-7) M was detected using a poly(U)-Sepharose resin. Competitive RNA binding analysis suggested that the NS3 protein binds preferentially to the poly(U) sequence, which is located at the 3' end of HCV RNA. Mutational analysis of NS3 protein revealed the possibility that both the RNA helicase region and the serine-proteinase region were necessary for full RNA binding activity.

Amino Acid Sequence↗
PubMed1995

Significance of hepatic expression of hepatitis C viral antigens in chronic hepatitis C.

To determine the significance of hepatic expression of hepatitis C viral (HCV) antigens, HCV core and NS4 antigens were detected by immunohistochemistry in 46 patients with chronic HCV infection. Serum HCV RNA was quantitated by branched DNA assay in 41 and HCV genotype determined in 30 patients. HCV core and NS4 antigens were detected exclusively in the cytoplasm of hepatocytes in 83% and 61% of patients, respectively. There was no correlation between the expression of HCV antigens and clinical, biochemical, histological parameters and HCV genotype. Hepatic expression of HCV antigens was positively associated with serum HCV-RNA levels (P < 0.02). At the end of interferon-alpha (IFN) therapy, expression of HCV antigens remained either unchanged or decreased in 11/12 patients studied (undetectable in all four patients who had complete and sustained response). We conclude that hepatic expression of HCV core and NS4 antigens parallels serum HCV-RNA levels and IFN therapy reduces hepatic expression of these viral antigens.

Female↗
PubMed1995

Hepatitis C virus genotypes 1 and 2 respond to interferon-alpha with different virologic kinetics.

Responses of patients infected with chronic hepatitis C virus (HCV) to interferon-alpha (IFN-alpha) treatment were studied. HCV genotypes were determined by molecular and serologic techniques. Levels of HCV viremia were determined by quantitative reverse transcription-polymerase chain reaction. Infection with HCV genotype 2 or low pretreatment HCV viremia levels in subjects infected with genotype 1 were associated with favorable (complete and sustained) responses (CR-SR; P < .001) to IFN-alpha treatment. HCV viremia levels in genotype 2 infection were significantly lower (P < .05) than in genotype 1 infection. The reduction rates of serum HCV RNA levels were four times higher in patients with genotype 2 infection than in those infected with genotype 1. The proportion of patients with CR-SR who experienced virologic relapse after completion of IFN-alpha treatment was higher for those infected with genotype 1. The proportion of patients with CR-SR and genotype 1 infection increased linearly in accordance with increases in single or total IFN-alpha dose.

Adult↗
PubMed1995

Chronological changes in subjective symptoms during pregnancy in nulliparous and multiparous women.

BACKGROUND: To investigate chronological changes of subjective symptoms during a normal pregnancy in nulliparous and multiparous women. METHODS: Prospective data were collected using a 20-item questionnaire (general fatigue, headache, palpitation, nausea/vomiting, fever, insomnia, edema, abnormal abdominal size, urinary frequency, lumbago, fetal descent, genital bleeding, watery discharge, high frequency of uterine contraction, increase in frequency of uterine contraction, strong intensity of uterine contraction, increase in intensity of uterine contraction, no change of uterine contraction at rest, high frequency of fetal movements, strong intensity of fetal movements) at Tsushima Izuhara Hospital in Nagasaki Prefecture, Japan. Seven hundred and twenty-nine nulliparous and 588 multiparous normal pregnant women were questioned from 1988 to 1992. A simple (chi-square test) analysis of appearance percentages in each item for every term of pregnancy was made. RESULTS: Each symptom showed different chronological patterns. In the simple analysis, there were significant differences (p < 0.05) in 10 symptoms (headache, palpitation, fever, insomnia, lumbago, fetal descent, watery discharge, increase in frequency of uterine contraction, strong intensity of uterine contraction, strong intensity of fetal movements) between nulliparous and multiparous subjects. In the multiparous group, there was a higher severity in eight of the ten symptoms, except for fever and watery discharge. CONCLUSIONS: The multiparous group had more complaints than did the nulliparous subjects. These normal patterns are of practical clinical use concerning subjective symptoms of pregnancy.

Chronology as Topic↗
PubMed1995

[Searching for hepatitis C virus by immunoelectron microscopy and its morphology].

Many investigators have searched for the causative agent(s) of blood-borne non-A, non-B hepatitis in sera and liver from patients and experimental animals, and have reported various virus-like particles, morphologically resembling parvovirus, togavirus, picornavirus, hepadnavirus, paramyxovirus, papovavirus, retrovirus or bunyavirus. Despite extensive effort, none of these virus-like particles has been confirmed universally as an etiologic virus because of the absence of immunological identification. The frustration was solved by the success in cloning cDNA of the genome of hepatitis C virus (HCV), a major etiologic agent of human non-A, non-B hepatitis, in 1989, but the morphology of HCV remained in riddles. We attempted to visualize HCV by indirect immunogold electron microscopy, using specific antibodies to the putative HCV envelope 1 protein and succeeded in identifying HCV particle in HCV-RNA-rich plasma. Our study showed that HCV particles are 55-65 nm spherical particles with 6 nm fine surface spike-like projections, and have a 30-35 nm inner core.

Hepacivirus↗
PubMed1995

Mechanism of translation initiation on hepatitis C virus RNA.

In vitro translation experiments involving reticulocyte and HeLa cell lysates have demonstrated that translation initiation on hepatitis C virus (HCV) RNA occurs by entry of ribosomes to the internal sequence (internal ribosomal entry site [IRES]) within the 5' noncoding region (5'NCR). Specific binding factors that bind to the HCV IRES were examined by UV cross-linking tests. A protein p57 (polypyrimidine tract binding protein [PTB]), known to be a binding factor that binds to picornavirus IRES, was found to bind to the HCV IRES. To investigate functions of this PTB in IRES-dependent translation initiation, hyperimmune antibodies to PTB (anti-PTB) were prepared by immunizing rabbits with purified recombinant PTB produced in E. coli. Anti-PTB IgGs were purified and used to deplete PTB in a cell-free protein synthesis system prepared from HeLa cells. In vitro translation experiments using PTB-depleted lysates suggested that host factor dependency of IRES is different in individual IRESs. As we showed previously, function of the IRES typical of group II HCV is more efficient than the IRES typical of group I HCV in a cell-free protein synthesis system from HeLa cells. Recombinant HCV IRESs were constructed and tested for their function in the in vitro system. Most recombinant IRESs showed an initiation activity of the more efficient typical group II IRES level or higher. This observation, together with the fact that group I HCV is the most common isolate of HCV worldwide, suggests that the efficiency of the IRES function of HCV may be generally suppressed in nature, and that the replication ability of HCV that permits its persistent infection in humans may require a delicate balance at a molecular level.

Animals↗
PubMed1994

Neonatal assessment using the Apgar fuzzy expert system.

We applied fuzzy theory to the Apgar scoring system (APG) to devise an Apgar fuzzy expert system (AFES). Three AFESs were determined separately by each one of three doctor groups (four inexperienced obstetricians, four experienced obstetricians and four expert neonatologists) in order to demonstrate that the AFES reflected the examiner's expertise and situation. Two-hundred and sixty-seven neonates were assessed 1 min after birth by an experienced obstetrician using the APG and the three AFESs. Statistical analysis of the relationship between the APG and the three AFESs in the acidosis group showed that the AFES determined by four expert neonatologists had the highest sensitivity and were significantly different (p < 0.05) from the APG. The AFES determined by the four expert neonatologists was revealed to be a valuable tool.

Acidosis↗
PubMed1994

Adaptation of hepatitis C virus for persistent infection in patients with acute hepatitis.

Nucleotide sequencing was used to analyze amino acid substitutions in the putative envelope 1 (E1) and envelope 2/nonstructural 1 (E2/NS1) regions of hepatitis C virus (HCV) to clarify a viral mechanism of persistent infection in three patients with acute hepatitis C who developed chronic hepatitis and two patients with chronic hepatitis. The HCV RNA titer in serum decreased markedly after the onset of acute hepatitis and then re-elevated. During this period, the substitution rate in the E2/NS1 region (especially in the hypervariable region located in the N-terminus) was significantly higher in patients with acute hepatitis than in patients with chronic hepatitis (P < 0.05). When patients with acute hepatitis C became persistent HCV carriers, the substitution rate decreased to the level seen in patients with chronic hepatitis. The amino acid substitution rate in the E1 region in the acute phase was similar to that found in the chronic HCV carrier state. These observations suggest that rapid substitution of the amino acid sequence in the hypervariable region of the E2/NS1 region may be one of the mechanisms of persistent HCV infection.

Acute Disease↗