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Biomedical subjects

T Miyamura

Publications and source records attributed to T Miyamura.

At least 145 records · Page 8Linked to original sources
PubMed1994

[Bone marrow transplantation for a patient with ALL from her 75-year-old mother using cryopreserved bone marrow cells].

We report a 42-year-old woman with acute lymphoblastic leukemia who received allogeneic bone marrow transplantation (BMT) in the first remission from her 75-year-old, HLA-identical, MLC-non-reactive mother. Considering the difficulty to obtain a sufficient number of bone marrow cells from such an old donor, we harvested the cells (2.31 x 10(8)/kg) on day -37 and cryopreserved them until use. BMT was performed on June 3rd, 1993 after conditioning regimen with total body irradiation, high-dose AraC and cyclophosphamide. Graft-versus-host disease (GVHD) prophylaxisis was attempted by cyclosporin A and short-term methtrexate. Her hematopoietic recovery was favorable with no signs and symptoms of GVHD as far as day 218.

Adult↗
PubMed1994

[The relationship of amniotic fluid volume and fetal renal function to fetal distress in intrauterine growth retardation].

The Hourly fetal urine production rate (HFUPR) was measured by ultrasound to evaluate the relationship between qualitative amniotic fluid volume (vertical depth of amniotic cavity) and fetal renal function. A positive correlation was found between HFUPR and neonatal birth weight (r = 0.625, p < 0.01), and between HFUPR and the vertical depth of the amniotic cavity (r = 0.615, p < 0.01). In three cases of IUGR complicated by fetal distress, the cyclicity of micturition disappeared and the vertical depth of the amniotic cavity was less than 30mm. The values for urinary beta 2-microglobulin were significantly higher in three neonates with fetal distress (15.00 +/- 1.00mg/L) than in healthy neonates (1.51 +/- 1.48mg/L) (p < 0.01). The mean value for vertical depth of the amniotic cavity in normal pregnancies was stable during pregnancy (mean +/- 2SD; 30-80mm). Thus patients with an amniotic cavity vertical depth less than 30mm were classified as decreased. In the IUGR group, the vertical depth of the amniotic cavity was normal in 81 patients, 14 of whom gave birth to a fetus with fetal distress. In contrast, the vertical depth of the amniotic cavity was decreased in 34 patients, 26 of whom gave birth to a fetus with fetal distress. The difference was significant (76.5% vs 17.3%, p < 0.01). The incidence of fetal distress was significantly related to the vertical depth of the amniotic cavity in the IUGR group.

Amniotic Fluid↗
PubMed1994

Establishment of a human myeloma cell line with growth-promoting activity for bone marrow-derived fibroblastoid colony-forming cells.

A human myeloma cell line, PCM6, was newly established from peripheral blood of a patient with advanced IgG myeloma by addition of recombinant interleukin-6 (IL-6) in culture. PCM6 cells had a morphology typical of mature plasma cells. Cytogenetic and surface marker studies confirmed that PCM6 cells were identical to fresh myeloma cells. Coculture of PCM6 cells with normal bone marrow mononuclear cells resulted in increased colony size of bone marrow-derived fibroblastoid colony-forming cells (CFU-F). Conditioned medium of PCM6 (PCM6-CM) cells increased the CFU-F colony size in a dose-dependent manner. The activity was labile to trypsin treatment but was heat stable (60 degrees C, 30 minutes). Molecular weight of the activity was approximately 165 kd by Sephacryl S-300 gel filtration. Platelet-derived growth factor (PDGF), epidermal growth factor (EGF), transforming growth factor-beta (TGF-beta), and IL-1 beta were not detectable in the conditioned medium. These findings suggest that in some myeloma cases, bone marrow stroma may be affected by CFU-F growth-promoting activity.

Antigens, Surface↗
PubMed1994

High-level expression of hepatitis B virus HBx gene and hepatocarcinogenesis in transgenic mice.

We studied the development of liver tumors in male HBx gene transgenic mice. Of two lineages studied, in the lineage with the lowest HBx gene expression liver tumors developed only in an incidence comparable with that in normal CD-1 strain, whereas 84% of male mice with a high level of the HBx gene product succumbed to liver neoplasia, indicating that continued HBx gene expression higher than a certain threshold level may be necessary for the development of hepatic neoplasia. Sixty-five mice from a lineage with a high level of HBx expression were then followed throughout their 24-mo lifespan. The livers of transgenic mice showed foci of cellular alteration with cytoplasmic vacuolations around the central veins from the age of 2 mo, but these foci did not expand progressively by the age of 12 mo. Immunostaining demonstrated such hepatocytes had higher expression of HBx protein than surrounding cells. Neoplastic lesions including liver cell adenomas and hepatocellular carcinomas developed from the age of 13 mo. By bromodeoxyuridine labeling analysis, hepatocytes in altered foci were found to have increased DNA synthesis, whereas no labeling was observed in age- and sex-matched nontransgenic littermate controls. Furthermore, DNA content analysis revealed the existence of several small aneuploid peaks in the transgenic liver before the age of tumor development. These results suggest that the continued expression of HBx gene may initiate a complex process to hepatocellular carcinoma by inducing DNA synthesis and placing large numbers of hepatocytes subjective to secondary events for transformation.

Adenoma, Liver Cell↗
PubMed1993

An efficient expression vector for stable expression in human liver cells.

The expression of the bacterial neomycin resistance (NmR)-encoding gene was examined under control of the promoter region of the gene encoding the human polypeptide chain elongation factor 1 alpha (EF-1 alpha). In a human liver cell line, HepG2, the plasmid pEF321-neo directed higher expression of the bacterial neo gene than the NmR gene expression vectors using other promoters, like the SV40 early region or thymidine kinase of Herpes simplex virus and SV40 early region promoter linked to human T-cell leukemia virus-1 enhancer sequences.

Cell Line↗
PubMed1993

Clinical significance of antibodies to nonstructural and core proteins of hepatitis C virus in posttransfusion hepatitis patients during long-term follow-up.

To clarify the long-term clinical significance, antibody to hepatitis C virus (HCV) was examined using core (p22) and nonstructural (C100-3) protein assays in sera of 18 patients with non-A,non-B posttransfusion hepatitis (PTH-NANB) who were selected retrospectively. Each patient had been followed for more than 5 years after the development of the disease. They were divided into three groups according to clinical outcome: acute hepatitis that resolved within 1 year, group 1 (n = 3); chronic hepatitis that resolved within 1-4 years, group 2 (n = 4); and chronic hepatitis that persisted for 5 years or longer, group 3 (n = 11). Sixteen of the 18 were positive for anti-C100-3 and anti-p22, one was positive for anti-p22 alone, and one was negative for both. In ten of the 16 (62.5%), anti-p22 appeared before anti-C100-3. The anti-C100-3 titer peaked about 12 months after disease onset in all cases and thereafter declined gradually, finally becoming negative in groups 1 and 2, while the titer fluctuated in group 3. The mean titer in group 3 at 12 months (69.2 units) significantly exceeded that of groups 1 (4 units) and 2 (8.2 units). Group 1 was seronegative for HCV antibodies and HCV RNA at the last examination, suggesting the cessation of HCV replication. Group 3 remained positive for those markers, indicating the continued replication of HCV.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1993

Structural proteins of hepatitis C virus.

Despite the lack of an in vitro replication system, genes coding for viral proteins of hepatitis C virus have been identified. The putative nucleocapsid (p22) and envelope (gp35 and gp58) proteins have been expressed in cells by different vectors under various foreign promoters. Detection of antibodies to these proteins in patient sera suggests that they are indeed synthesized during the course of viral replication.

Hepacivirus↗
PubMed1993

Parvovirus B19-associated haemophagocytosis in Evans syndrome: aplastic crisis accompanied by severe thrombocytopenia.

A 36-year-old man who had been treated for Evans syndrome (ES) developed an aplastic crisis caused by acute human parvovirus B19 (HPV) infection. Profound thrombocytopenia (8.0 x 10(9)/l) followed with a sudden increase in platelet-associated IgG (PAIgG) (1376.9 ng/10(7) plts). Bone marrow examination revealed a considerable number of haemophagocytic histiocytes without any disturbance of megakaryopoiesis. To our knowledge this is the first case of aplastic crisis with virus-associated haemophagocytosis in a patient with ES, which provides an interesting insight into the mechanisms for thrombocytopenia in HPV infection.

Adult↗
PubMed1993

Follow-up study of anti-hepatitis C virus antibodies in blood donors implicated in post-transfusion non-A, non-B hepatitis.

We retrospectively examined the antibodies to p22, a hepatitis C virus (HCV) nucleocapsid protein, and to c100-3, a HCV nonstructural protein, in donors whose blood was transfused to patients who later developed post-transfusion non-A, non-B hepatitis. Of 13 such blood donors, three seroconverted and three seroreverted with the anti-c100-3 test. In contrast, 12 of the 13 blood donors showed the same results at transfusion and follow-up, and one donor showed seroconversion with the anti-p22 assay. The follow-up study shows that the anti-p22 antibody test provides consistent results and is far more suitable for screening blood than the anti-c100-3 test.

Antigens, Viral↗
PubMed1993

Suppression of hepatitis B virus expression and replication by hepatitis C virus core protein in HuH-7 cells.

Hepatitis B and C viruses (HBV and HCV, respectively) are associated with acute and chronic liver diseases and hepatocellular carcinoma. To elucidate the molecular status of superinfection with these two hepatitis viruses, we cotransfected the full-length or truncated version of HCV structural genes (core and envelope 1) together with the cloned HBV DNA into a human hepatoma cell line (HuH-7). Expression of HBV-specific major transcripts (3.5 and 2.1 kb), as well as HBV antigens (hepatitis B surface antigen and hepatitis B e and core antigens), was reduced about two- to fourfold by the presence of the HCV structural genes. In addition, the secretion of HBV viral particles, including the viral nucleocapsid and mature virion, was drastically suppressed about 20-fold. Analysis of the intracellular HBV core protein-associated nucleic acid indicated that the encapsidated HBV pregenomic RNA was similarly reduced about 14-fold. Deletion analysis of the HCV structural genes demonstrated that the core gene alone or the fragment containing the core protein's N-terminal 122 amino acid residues conferred the same level of suppressive activity as the full-length structural genes. By indirect immunofluorescence, we found that the core protein of HCV was located in the cytoplasm of transfected HuH-7 cells at day 3 posttransfection and was targeted to the nucleus at day 6. Thus, the kinetics of the suppressive effect exerted by HCV constructs matched the timing of core protein entrance into the nucleus. Our results substantiate the clinical finding that HBV markers are suppressed by superinfection with HCV and further imply that this inhibitory effect may occur in the processes of transcription and encapsidation of HBV pregenomic RNA and may be mediated by the core protein of HCV. The deduced amino acid sequence of the HCV core protein has revealed that it is a basic protein which contains a putative DNA-binding motif (SPRG), as well as triplicate nuclear localization signals and several putative protein kinase A and C recognition sites. These characteristics imply that the HCV core protein can also function as a gene-regulatory protein.

Amino Acid Sequence↗
PubMed1993

Immortalization of differentiated human hepatocytes by a combination of a viral vector and collagen gel culture.

Adult human and chimpanzee hepatocytes, which have no proliferative potential in vitro, were immortalized by introducing an oncogene encoding simian virus 40 large tumor(T) antigen by means of infection with a recombinant adenovirus vector. The frequency of immortalization was enhanced by a suspension method for virus adsorption and especially by using a collagen gel culture. The replicating hepatocyte colonies were isolated and selected by albumin secretion which is specific for parenchymal hepatocytes, then continuously subcultured. The immortalized hepatocyte lines also secreted other hepatocyte-specific proteins (complement C3, transferrin, fibrinogen). However, the amount of protein secretion declined with subculture. The subline RY5, cloned from a human immortalized line, showed relatively stable albumin secretion. Therefore, chromosomal studies and anchorage-independent growth properties were discussed and compared with its original line.

Adenoviruses, Human↗
PubMed1993

[A case of primary non-Hodgkin lymphoma of the adrenal gland presenting with Addison's disease].

A 69-year-old male was admitted to our hospital complaining of skin pigmentation. Masses in bilateral adrenal glands were noted on computed tomography. An exploratory laparotomy revealed malignant lymphoma confined to the bilateral adrenal glands. Histological diagnosis was diffuse mixed-cell type, B-cell lymphoma. Laboratory tests disclosed an elevated ACTH level and decreased urinary 17KS and 170HCS secretion. Based on hypofunction of adrenal glands and skin pigmentation, the patient was diagnosed as having Addison's disease. After complete remission with combination chemotherapy, adrenal function recovered normal and pigmentation disappeared, Addison's disease is an extremely rare complication of primary malignant lymphoma of the adrenal gland.

Addison Disease↗
PubMed1992

Seroprevalence of hepatitis C virus nucleocapsid antibodies in patients with cryptogenic chronic liver disease.

The serological responses to two different hepatitis C virus antigens were studied by enzyme-linked immunosorbent assay in a variety of chronic liver diseases and in healthy blood donors. The study population comprised 97 cases of cryptogenic chronic liver disease (40% with a history suggestive of parenterally transmitted non-A, non-B hepatitis and 60% without such a history), 87 cases of other well-characterized chronic liver diseases and 96 voluntary blood donors. The commercially available C100-3 assay and a new assay utilizing a 22 kD recombinant protein (c22) from the nucleocapsid region of the virus were used for antibody detection. Overall in the non-A, non-B hepatitis group, 77% were positive for anti-c22, 55% were positive for anti-C100-3 and 24% were negative by both tests. In the parenterally transmitted chronic liver disease group, 82% were positive for anti-C100-3 and 90% were positive for anti-c22 (not significant). In the cryptogenic chronic liver disease cases 36% were positive for anti-C100-3 and 67% were positive for anti-c22 (p less than 0.001). Only in one case (a patient with hepatitis B virus infection) was anti-C100-3 detected without concomitant anti-c22. None of the voluntary blood donors had detectable hepatitis C virus antibodies. The new enzyme-linked immunosorbent assay test for anti-c22 would appear to be a more sensitive indicator of chronic hepatitis C virus infection than the existing commercial test, suggesting a useful diagnostic role in both cases of cryptogenic chronic non-A, non-B hepatitis liver disease and for the screening of blood products for prevention of hepatitis after transfusion.

Base Sequence↗
PubMed1992

Improved serodiagnosis of non-A, non-B hepatitis by an assay detecting antibody to hepatitis C virus core antigen.

We examined sequential serum samples from 12 patients with well-characterized posttransfusion non-A, non-B hepatitis who had an acute, resolving self-limited type of clinical course for the presence of antibody to the hepatitis C virus nucleocapsid (core) protein (p22) expressed by a recombinant baculovirus. These sera were simultaneously examined for antibody to the hepatitis C virus nonstructural protein (C100-3) that is presently used for blood screening worldwide. In three patients, both anti-p22 and anti-C100-3 antibodies were detected, but anti-p22 was detected much earlier. In four patients, only anti-p22 was detected. Two other patients were considered to be hepatitis C virus carriers who had been already infected with hepatitis C virus. In one patient, only anti-C100-3 was detected, and it was transient. In two patients, neither antibody was detected. Anti-p22 was detected in at least one of eight samples of transfused blood. Of the nine samples of donated blood that were positive for anti-p22, only four were positive for anti-C100-3. This new assay detecting the antibody to the p22 protein is thus useful for the serodiagnosis of non-A, non-B hepatitis in the acute phase and for blood screening.

Antigens, Viral↗
PubMed1992

Correlation between detection of anti-viral antibody and histopathological disease activity in an epidemic of hepatitis C.

There was an epidemic of non-A non-B hepatitis in a small area of a town in the central part of Japan, which began with an outbreak of several patients in 1981 and then spread extensively with the result that about one third of the inhabitants showed abnormality in serum liver function tests at the health check performed in 1985. We determined histological diagnoses on that occasion for 167 individuals of the abnormal population and recently assayed antibodies against hepatitis C virus (HCV) for most of their sera left available. Histologically, chronic active hepatitis (CAH) was the major pattern, accounting for 59.3% (99 cases) of the total. Others were chronic persistent hepatitis (CPH) (13.2%), chronic lobular hepatitis (CLH) (16.2%), liver cirrhosis (LC) (6.6%) and fatty liver (4.8%). In the serological studies, the newly developed system to detect antibodies against the viral core protein p 22 was found to be much more sensitive than the conventional system to detect anti C 100-3 antibodies. By using these two methods in combination, we found that 82% were antibody-positive, indicating strong implication of HCV in this epidemic. This was further supported by direct detection of the viral genome in patients' sera by polymerase chain reaction following reverse transcription. We further found a strong correlation between the histological inflammatory activity and the antibody prevalence, since nearly all (97.6%) of the CAH cases were antibody-positive by at least either of the antibody assays, while only about 50% were positive in the less active cases such as CPH and CLH.

Base Sequence↗