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Biomedical subjects

H Hara

Publications and source records attributed to H Hara.

At least 307 records · Page 17Linked to original sources
PubMed1997

Molecular basis of the altered gag p19 protein (MA) of the transformation-defective mutant of Rous sarcoma virus, tdPH2010.

The transformation-defective mutant of Rous sarcoma virus (RSV), tdPH2010, has a gag p19 (matrix; MA) protein which migrates on SDS-polyacrylamide gels faster than that of the parental (tsNY68) as well as other wild-type strains of RSV. To study the molecular basis of this altered migration pattern and its biological significance, the nucleotide sequence of the p19 region of tdPH2010 was determined. Comparison of the nucleotide sequence of tdPH2010 with that of the Schmidt-Ruppin A strain of RSV revealed a point mutation at nucleotide 755 (G to A), resulting in an amino-acid substitution at residue 126 of p19 (Glu to Lys). Acidic-methanol esterification of free carboxyl groups suppressed the difference in electrophoretic mobility of p19 between tdPH2010 and the wild-type virus. Recombinant virus constructs having the mutated gag region of tdPH2010 produced a p19 with the same electrophoretic mobility as the p19 of tdPH2010. We concluded that the point mutation caused the altered electrophoretic behavior of p19 of tdPH2010. The mutation had no effect on the growth of infected cells.

Amino Acid Sequence↗
PubMed1996

Cloning and expression of a rat brain basic helix-loop-helix factor.

We cloned two rat cDNAs of brain basic helix-loop-helix factor 1 (BHF1). These have an identical coding region, contain 357 amino acids and exhibit 94.6% identity to MATH-2/NEX1 in the basic helix-loop-helix region. BHF1mRNAs are dominantly expressed in the brain particularly in the cerebellum, in the adult bovine, rat and mouse. Two shorter BHF1mRNAs (1.6 kb and 1.8 kb) were also detected in the mouse embryo, and these decreased in the developmental process. These results suggest that BHF1 may play important roles in cerebellum-specific functions and development of neurons.

Amino Acid Sequence↗
PubMed1996

Characterization of single chain urokinase-type plasminogen activator with a novel amino-acid substitution in the kringle structure.

ECV304 is a cell line established by a spontaneous transformation of endothelial cells of a human umbilical vein. It was shown that ECV304 secretes single chain urokinase-type plasminogen activator (scu-PA). A subclone, ECV304 clone 15, was obtained by acclimatization of parental clone to serum-free medium followed by limiting dilution. The clone was found to produce approximately five times as much scu-PA (approximately 20 IU/10(6) cells per day) as the parental clone after a 40 days' culture. Though the biochemical characteristics of the purified scu-PA were indistinguishable from those of the native scu-PA, it had a lower affinity for fibrin clots under the employed conditions. Molecular cloning of a cDNA encoding the scu-PA has identified a novel substitution from C to T in the nucleotide sequence encoding the kringle structure. The substitution resulted in an alteration from Pro (CCG) to Leu (CTG) at amino-acid position 121, which may be directly or indirectly involved in the decrease in the apparent affinity.

Amino Acid Sequence↗
PubMed1996

Case of myoepithelioma originating from the palate: collagenogenesis in myoepithelial tumor cells.

We report a case of myoepithelioma of the palate in a 53-yr-old man. Myoepithelioma cells consisting of both plasmacytoid type and spindle type were obtained. A reticular pattern composed of a mosaic arrangement of hyaline-like abundant cytoplasm, and scant matrix and considerable fiber formation were characteristic features of the plasmacytoid type; granular cytoplasm and a snake-like shape were characteristic features of the spindle type. These are important points in differentiating this tumor from cellular type pleomorphic adenoma. The present study shows that tumorous myoepithelial cells can synthesize and secrete extracellular components such as proteoglycans, basal lamina, collagen fibers, and elastic fibers.

Adenoma, Pleomorphic↗
PubMed1996

In vitro differentiation of murine Sca-1+Lin- cells into myeloid, B cell and T cell lineages.

Hematopoietic progenitor cells were shown to be capable of differentiating into myeloid, B cell and T cell lineages. We used a two-step culture system in which enriched murine hematopoietic progenitors in bone marrow were first plated in viscid culture medium containing methylcellulose, erythropoietin (EPO), stem cell factor (SCF) and interleukin (IL)-7. One thousand enriched murine marrow cells formed 53.5 +/- 12.1 (mean +/- SD) primary colonies. Cells from a single blast colony were separated into two aliquots and replated in secondary methylcellulose cultures containing SCF and IL-7 for B cell lineage and SCF, IL-3, G-CSF, GM-CSF and EPO for myeloid lineage. Next, cells from five to ten primary blast colonies were cultured again in embryonal thymus (25 Gy irradiated). One aliquot of blast colonies in a primary culture contained four colony forming units (CFU) of granulocytes, erythroblasts, macrophages and megakaryocytes, eight CFU-granulocytes and macrophages, and 28 BFU-E in a representative secondary myeloid culture. Another aliquot formed a few B cell colonies (2-10) in a secondary B cell culture. B lymphoid colonies were composed of blast-like cells with B-220 antigen. T cells in a secondary T cell culture consisted of 16% L3T4+, 16% CD8+ and 11% CD3+ of bone marrow origin in the thymus. From these results, we concluded that cells in the primary colonies from Sca-1+Lin- hematopoietic stem cells could differentiate into B cell, T cell and myeloid lineages.

Animals↗
PubMed1996

Presence of a hypervariable region within the hr2 domain of the host range determining sequences of the envelope protein gp85 (SU) of subgroup-A avian sarcoma-leukosis viruses.

The nucleotide sequence of the gene for the envelope protein gp85 (SU) of the Schmidt-Ruppin subgroup A (NY) strain of Rous sarcoma virus [SRA(NY)] was determined, and the deduced amino acid sequence was compared with those of other avian sarcoma-leukosis viruses. Among the five host-range determinant sequences (vr1, vr2, hr1, hr2, and vr3), the host-range determinant sequence hr2 of SRA(NY) showed a significant deviation from the hr2 sequences of other subgroup A viruses namely, RAV-1 and SRA(SF). A phylogenetic analysis of the amino acid sequence of this region indicated that this intra-subgroup diversity was as great as or even greater than the inter-subgroup diversity found among other subgroups of ASLV. Within the hr2 region, we found a short hypervariable segment that differs in length and sequence from hr2 of other subgroup-A viruses. The difference in the hr2 amino acid sequence between SRA(NY) and SRA(SF) is reflected in the predicted protein secondary structure of this region.

Amino Acid Sequence↗
PubMed1996

Brain distribution of nitric oxide synthase in neuronal or endothelial nitric oxide synthase mutant mice using [3H]L-NG-nitro-arginine autoradiography.

The regional distribution of nitric oxide synthase in the central nervous system was assessed by quantitative autoradiography using [3H]L-NG-nitro-arginine binding in wild-type mice (SV-129 and C57black/6) and in mice lacking expression of the neuronal (type 1) and endothelial (type 3) nitric oxide synthase gene. The distribution of nitric oxide synthase binding sites in wild-type mice was similar to that described for rat brain by nicotinamide adenine dinucleotide phosphate-diaphorase staining and immunohistochemistry, and as determined by quantitative autoradiography. In the wild-type mice, the densest labelling was observed in the granular layer of the olfactory bulb, tenia tecta, rhinal fissure, amygdaloid complex and molecular layer of cerebellum. The islands of Calleja, the hippocampal CA1 and CA3 subfields, dentate gyrus, cortical layers I-II, the superficial gray layer of superior colliculus and the granule layer of cerebellum displayed intermediate binding. Cortical layers III-VI, the striatum and the thalamus were only weakly labelled. Binding was saturable and of high affinity, and was displaced by 7-nitroindazole (100 microM), a potent and selective inhibitor of type 1 nitric oxide synthase, and by unlabelled L-NG-nitro-arginine (10 microM). The density of [3H]L-NG-nitro-arginine binding was dramatically reduced in all brain regions in type 1 mutant mice, whereas there were no detectable binding differences between wild-type and type 3 nitric oxide synthase mutant mice. Hence, type 1 nitric oxide synthase is the major source of [3H]L-NG-nitro-arginine binding in the mouse brain. [3H]L-NG-Nitro-arginine autoradiography may be a useful tool to quantify nitric oxide synthase in different brain areas after pharmacological or physiological manipulations.

Animals↗
PubMed1996

Delayed remission after thymectomy for myasthenia gravis of the purely ocular type.

Twenty-two cases of purely ocular myasthenia gravis were reviewed to evaluate the long-term effects of thymectomy. Remission rate increased gradually with time (11.8% at 3 years, 23.1% at 5 years, and 33.3% at 10 years). Analysis of factors influencing remission with time showed that patients with short duration of illness attained remission significantly earlier (p = 0.035 at 5-year follow-up). One of 22 patients with purely ocular myasthenia gravis (4.5%) had disease progression. Because ocular myasthenia gravis often progresses to the generalized type and because duration of illness before operation is one of important factors influencing remission, we conclude that thymectomy in the earlier stages of the disease is the preferred treatment for ocular myasthenia gravis, just as for generalized myasthenia gravis.

Adolescent↗
PubMed1996

Spatial and temporal expression of P-glycoprotein in the congenitally hydrocephalic HTX rat brain.

The purpose of the present study is to examine spatial and temporal expression of P-glycoprotein in the brain of congenitally hydrocephalic HTX rats. P-glycoprotein has been reported not only as a drug efflux pump but also one of the factors that restricts brain edema. We examined the rat brain from postnatal day 1 to 60 using light and electron microscopy, immunohistochemistry, Western immunoblot and reverse transcriptase-polymerase chain reaction (RT-PCR) methods with monoclonal antibody specific for P-glycoprotein. Immunohistochemically, the positive anti-P-glycoprotein reactivity was found in capillaries of the normal control rat cerebrum. In the hydrocephalic HTX rat brains, it was also found in the capillaries, but only very weak to no reactivity was found in the capillaries of the spongy changes and cystic wall in the subcortical and lateral periventricular white matter. Immunoelectron microscopically, the reaction product was found exclusively on the luminal surface of the capillary endothelium in control rats. A tracer study with intracardiac perfusion of lanthanum chloride showed that lanthanum penetrated the tight junctions and passed through the intercellular space. In the Western immunoblot analysis, P-glycoprotein of 170 kDa was detected clearly in most normal control rat brains but it was not found in the hydrocephalic HTX rat brains. Moreover, mdr1 P-glycoprotein gene expression in the subcortical white matter was examined by RT-PCR. It was detected in all normal control rat brains, but not found in the hydrocephalic HTX rat brains. The results suggested that the absence of P-glycoprotein expression in the capillaries of deep subcortical and lateral periventricular white matter of hydrocephalic HTX rats led to a deficiency of the blood-brain barrier and might be related to vasogenic edema and to the formation of the spongy changes and cystic cavities.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1996

mdm2 gene mediates the expression of mdr1 gene and P-glycoprotein in a human glioblastoma cell line.

The overexpression of the multidrug resistance (mdr1) gene and its product, P-glycoprotein (P-gp), is thought to limit the successful chemotherapy of human tumours. The mechanism by which mdr1 gene and P-gp are overexpressed in human tumours, however, is not yet clear. In this report, we show that the mdm2 (murine double minute 2) gene induced the expression of the mdr1 gene and P-gp in human glioblastoma U87-MG cells, which did not express the MDM2 protein or P-gp. The mdm2 gene, in addition, conferred the resistance of U87-MG cells to the apoptotic cell death induced by etoposide (VP-16) or doxorubicin. Furthermore, treatment with mdm2 antisense oligonucleotides inhibited the expression of P-gp in MDM2-expressing U87-MG cells. These findings suggest that the mdm2 gene may play an important role in the development of MDR phenotype in human tumours.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1996

Increases in calcium absorption with ingestion of soluble dietary fibre, guar-gum hydrolysate, depend on the caecum in partially nephrectomized and normal rats.

Effects of feeding soluble dietary fibre on apparent Ca absorption and the contribution of the caecum to Ca absorption were examined in five-sixths nephrectomized (NPX) and normal rats with or without caecectomy in four experiments. It is known that Ca absorption is lowered by renal failure. In the first experiment the amounts of femur Ca increased linearly with increasing dietary Ca up to 3.0 g Ca/kg diet in intact rats. Partial nephrectomy decreased apparent Ca absorption in rats fed on diets containing 3-0 and 4.5 g Ca/kg diet. In the NPX groups, Ca absorption in rats fed on the diet containing guar-gum hydrolysate (GGH; 50 g/kg diet; 3.0 g Ca/kg diet) was significantly higher than that in rats fed on a fibre-free diet, and the increase in Ca absorption with GGH feeding was completely abolished by caecectomy. Also, ingestion of GGH increased Ca absorption in normal rats, but not in normal, caecectomized rats. Mg absorption was also increased with GGH feeding and was decreased with caecectomy in NPX and normal rats. In experiments which used caecectomized rats, coprophagy was prevented with an anal cup to avoid re-ingestion of faecal Ca. We conclude that ingestion of the soluble dietary fibre, GGH, increased apparent Ca absorption in NPX and non-NPX rats, and the caecum was responsible for these increases in Ca absorption.

Animals↗
PubMed1996

Overproduction of penicillin-binding protein 7 suppresses thermosensitive growth defect at low osmolarity due to an spr mutation of Escherichia coli.

Escherichia coli delta prc mutants lacking periplasmic protease Prc, which was originally found involved in the C-terminal processing of penicillin-binding protein (PBP) 3, show thermosensitive growth at low osmolarity. We isolated thermoresistant revertants containing extragenic suppressor (spr) mutations. In the prc+ background the mutations also caused thermosensitivity at low osmolarity. They were all mapped at about 48 min on the chromosome and most probably allelic to one another. From this chromosomal region we cloned a gene that could correct the thermosensitive defect of an spr mutant, which turned out to be a multicopy suppressor of spr. Analysis of the nucleotide sequence predicted that the gene would code for a low-molecular-weight PBP, and penicillin-binding experiments revealed the product to be PBP 7. Disruption of the gene on the chromosome caused no apparent growth defect. PBP 7 seemed to be degraded by protease Prc. Overproduction of mutant PBP 7 that had the active site serine residue replaced with alanine did not correct the spr thermosensitivity, suggesting importance of the DD-endopeptidase activity in the multicopy suppression.

Alleles↗
PubMed1996

Oxidized ethyl linoleate induces mucosal hypertrophy of the large intestine and affects cecal fermentation of dietary fiber in rats.

Oxidized ethyl linoleate (OEL) was prepared by aeration at low temperature. Peroxide value (POV, mEq/kg lipid) of OEL was 1400; the major oxidized compounds were 9-hydroperoxy-cis, trans- and 13-hydroperoxy-trans, cis-octadecadienoate. Rats fed fiber-free or sugar-beet fiber (SBF, 100g/kg diet) diets were divided into three groups for each diet, and administered OEL (high OEL group), OEL diluted with ethyl linoleate (low OEL group, POV 700) and nonoxidized ethyl linoleate (EL group) through gastric tubes each day at 1400-1600 h (2.5 g/kg body wt) for 16 d. The relative wet weight, and DNA and protein contents of the cecal mucosa were higher in the high OEL groups than in the low OEL and EL groups in rats fed the fiber-free diet and in rats fed the SBF diet except for mucosal protein content. Spermidine concentration in cecal mucosa of rats fed the fiber-free diet was greater in the high OEL group than in the EL group. These results suggest that metabolism related to mucosal proliferation of the cecum was affected by the high dose of OEL. The total short-chain fatty acid (SCFA) concentration in the cecal contents of SBF-fed rats was 100% higher than the concentration in rats fed the fiber-free diet in the EL group, but the administration of low dose and high dose OEL lowered the SCFA concentration in fiber-fed rats to that of rats fed the fiber-free diet. Butyric acid concentration was markedly lowered by ingestion of OEL in a dose-dependent manner in rats fed the SBF diet. In contrast, the isobutyric acid concentration was higher in the OEL-treated groups than in the EL groups. We conclude that a low dose of OEL depresses cecal fermentation of dietary fiber with changes in SCFA composition, and that a high dose of OEL induces mucosal hypertrophy in the cecum. These data show that dietary oxidized lipids affect cecal metabolism and may be associated with colon cancer.

Animals↗
PubMed1996

Reduced brain edema and infarction volume in mice lacking the neuronal isoform of nitric oxide synthase after transient MCA occlusion.

Infarct volume and edema were assessed after transient focal ischemia in mice lacking neuronal nitric oxide synthase (NOS) gene expression. With use of an 8-0 coated monofilament, the middle cerebral artery (MCA) of mutant (n = 32) and wild-type mice [SV-129 (n = 31), C57Black/6 (n = 18)] were occluded for 3 h and reperfused for up to 24 h. Regional CBF (rCBF), neurological deficits, water content, and infarct volume were examined in all three strains. rCBF, blood pressure, and heart rate did not differ between groups when measured for 1 h after reperfusion. Neurological deficits were less severe in mutant mice after MCA occlusion. Brain water content at 3 h after reperfusion and infarct volume at 24 h after reperfusion were greater in wild-type mice. These data indicate that genetic deletion of neuronal NOS confers resistance to focal ischemic injury in a reperfusion model. The findings agree with previous studies showing that tissue injury is less extensive after both permanent MCA occlusion and global ischemia in mice lacking neuronal NOS gene expression. Hence, NO may play a pivotal role in the pathogenesis of ischemic brain damage.

Animals↗
PubMed1996

Establishment and characterization of choroid plexus carcinoma cell lines: connection between choroid plexus and immune systems.

Murine choroid plexus cell lines were produced from choroid plexus carcinoma generated in transgenic mice harboring the viral oncogene simian virus 40 large tumor antigen under transcriptional control of an intronic enhancer region from the human immunoglobulin heavy chain (IgH) gene. Two morphologically distinct cell lines have been cloned. These established cell lines retained the characteristics of choroid plexus cells in that they expressed such choroid plexus cell marker or related proteins as transthyretin and alpha2-macroglobulin. They were tumorigenic in nude mice. In the cell lines, the muA and muB (HE2) motifs within the IgH intronic enhancer were active and we also demonstrated the existence of the proteins binding to these motifs, suggesting a potential link between the choroid plexus and immune systems. It is considered that these binding proteins act as trans-activators for the enhancer and may belong to the class of ETS-related proteins. These cell lines and xenografts should be useful materials for analyses of choroid plexus functions.

Animals↗
PubMed1996

Proliferation marker MIB-1 correlates well with proliferative activity evaluated by BrdU in breast cancer: an immunohistochemical study including correlation with PCNA, p53, c-erbB-2 and estrogen receptor status.

The proliferative activity of 30 cases of non-treated invasive ductal breast carcinoma was evaluated by bromodeoxyuridine (BrdU), proliferation marker (MIB-1) and proliferating cell nuclear antigen (PCNA), and the relation between these proliferation markers and histological subtype and histological grade were investigated. In addition, the association of these proliferation markers with overexpression of p53 protein, c-erbB-2 oncoprotein, estrogen receptor (ER) status and clinicopathologic findings were also examined. The BrdU labeling index (LI), MIB-1 score and PCNA labeling rate (LR) correlated with the histological grade. However, there was no statistical difference in proliferative activity among the histological subtypes. A linear strong correlation was demonstrated between BrdU LI and MIB-1 score (r = 0.732). Significant correlation was also found between BrdU LI and PCNA LR (r = 0.446); however, the relation between MIB-1 score and PCNA LR was weak. BrdU LI and MIB-1 score correlated positively with tumor size, TNM stage and overexpression of p53, and negatively with the presence of ER. PCNA LR correlated only with p53. These results indicate that MIB-1 is closely associated with BrdU in clinicopathologic findings and is a more useful tool for evaluating cell proliferation than PCNA. However, it will be necessary to consider the clinical significance of MIB-1 immunohistochemistry cautiously until further widespread clinical and pathological studies are performed.

Adult↗
PubMed1996

Histological grade in invasive ductal carcinoma of breast correlates with the proliferative activity evaluated by BrdU: an immunohistochemical study including correlations with p53, c-erbB-2 and estrogen receptor status.

Thirty cases of invasive ductal carcinoma of the breast were classified to histological subtype according to the General Rules for Clinical and Pathological Recording of Breast Cancer of the Japanese Breast Cancer Society and histologically graded using the Nottingham method and the correlation of histology with proliferative activity was investigated using bromodeoxyuridine (BrdU). In addition, the overexpression of p53 protein, c-erbB-2 oncoprotein and estrogen receptor (ER) were immunohistochemically examined in order to discuss the relationship with histological subtype and histological grade. Histological grade correlated positively to the BrdU labeling index (LI) and overexpression of p53. High grade carcinoma demonstrated c-erbB-2 more frequently and exhibited a low incidence of ER. However, no significant relationship was found between BrdU LI, overexpression of p53 and c-erbB-2 and histological subtype. These results suggest that the histological grade does represent the proliferative activity of tumor cells and that adding the histological grade to the pathological diagnosis in invasive ductal breast carcinoma may be useful from the clinicopathological aspect concerning tumor behavior.

Adult↗