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Biomedical subjects

J Osuga

Publications and source records attributed to J Osuga.

31 records · Page 2Linked to original sources
PubMed1998

Clinical features associated with the homozygous Trp64Arg mutation of the beta3-adrenergic receptor: no evidence for its association with obesity in Japanese.

To characterize the clinical features associated with the Trp64Arg mutation of the beta3-adrenergic receptor (beta3-AR), the effects of this mutation, in particular the homozygous state (Arg/Arg), on obesity, blood pressure, and plasma lipoproteins were investigated in 2 populations: subjects residing on a small isolated island (group 1; n=746) and patients residing in Tokyo who attend a clinic for metabolic diseases (group 2; n=371). The allelic frequency of the Trp64Arg mutation was 23.4% in group 1 and 18.3% in group 2. No significant difference in the body mass index was observed between subjects with 3 different genotypes in each group. There was a trend that the Arg/Arg had higher systolic blood pressure than the Trp/Trp in both groups, but the differences were not statistically significant. The plasma LDL cholesterol levels were significantly lower in Arg/Arg than in Trp/Trp in men from the group 1 cohort (2.82+/-0.84 versus 3.19+/-0.7 mmol/L, P<0.05). These results suggest that the homozygous Trp64Arg mutation is not a major contributing factor for obesity, but potentially contributed to higher systolic blood pressure and low plasma levels of LDL cholesterol in Japanese men.

Arginine↗
PubMed1998

A truncated species of apolipoprotein B (B-38.7) in a patient with homozygous hypobetalipoproteinemia associated with diabetes mellitus.

Familial hypobetalipoproteinemia is caused by mutations in the apolipoprotein (apo) B gene. We identified a 57-year-old woman whose plasma total cholesterol and apoB levels were 2.17 mmol/L and 0.03 g/L, respectively. Separation of plasma lipoproteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the absence of apoB-100 and the presence of a faster-migrating form of apoB with an apparent Mr of 195 kDa. Direct sequencing of a polymerase chain reaction-amplified fragment of the patient's apoB gene DNA revealed a single C-->T transition at nucleotide 5472 that converts glutamine 1755 (CAA) to a stop codon (TAA). We predict this novel nonsense mutation of the apoB gene to produce a truncated protein that contains 1754 amino-terminal amino acid residues of apoB-100. We designated this mutant form of apoB apoB-38.7 by following the centile nomenclature of the apoB species. The same mutation was found in both of her children. The proband revealed clinical findings of retinitis pigmentosa, acanthocytosis, and loss of deep tendon reflexes that are characteristic of severe hypobetalipoproteinemia. In addition, the proband had type II diabetes mellitus with nephropathy, anemia, cholelithiasis, hepatic hemangioma, bronchiectasis, and extensive calcification of major arteries including, the celiac, splenic, and renal. In summary, we have found a novel truncated apoB, apoB-38.7, in a patient with an unusual presentation of hypobetalipoproteinemia that includes diabetes mellitus and extensive arterial calcification.

Apolipoproteins B↗
PubMed1997

Effects of apo E deficiency on plasma lipid levels in mice lacking APOBEC-1.

Apolipoprotein (apo) B100 mRNA undergoes site specific C to U editing, generating a stop-translation codon of apo B48 in the small intestine. This reaction is catalyzed in an editosome which contains APOBEC-1, a catalytic subunit. To clarify the functional significance of the apo B mRNA editing in lipoprotein metabolism, we have generated APOBEC-1 knockout mice and double knockout mice which are deficient in both APOBEC-1 and apo E. The apo B mRNA editing activity was markedly reduced and complete elimination of apo B48 from the plasma was observed in APOBEC-1(-/-) mice. Plasma triglyceride levels significantly increased in the double knockout mice (APOBEC-1(-/-);apo E(-/-)) as compared to apo E(-/-) mice. These results suggest that APOBEC-1(-/-) mice are a valuable model for experiments designed to understand a role of apo B mRNA editing.

APOBEC-1 Deaminase↗
PubMed1997

Bcl-2 protein inhibits oxysterol-induced apoptosis through suppressing CPP32-mediated pathway.

Oxysterols are presumed to mediate cytotoxicity of oxidized LDL in atherosclerotic lesions. To elucidate its molecular mechanism, we established murine macrophage-like P388-D1 cells which over-express Bcl-2 protein by retrovirus-mediated gene transfer. Oxysterols (7-ketocholesterol, 25-hydroxycholesterol) induced nuclear condensation and oligonucleosomal DNA fragmentation, which were partially inhibited by Bcl-2 over-expression. Though CPP32 inhibitor suppressed the cell death in control cells, it showed no additive protection in the cells over-expressing Bcl-2. These findings indicate that oxysterols induce apoptosis via Bcl-2-inhibitable and -uninhibitable pathways, and the former depends on CPP32 activation.

Animals↗
PubMed1997

Suppression of neutral cholesterol ester hydrolase activity by antisense DNA of hormone-sensitive lipase.

In order to investigate the role of the hormone-sensitive lipase (HSL) gene in the activity of neutral cholesterol ester hydrolase (NCEH) from a molecular perspective, Chinese hamster ovary (CHO) cells were transfected with rat antisense hormone-sensitive lipase cDNA, and three different cell lines, designated as anti-HSL, were established. NCEH activity in anti-HSL cells was reduced to approximately 50% of that in control CHO cells. The concentration of cellular esterified cholesterol increased and the concentration of free cholesterol decreased in the anti-HSL cell lines. These results suggest that the HSL gene has a function of NCEH as well.

Animals↗
PubMed1997

Apoptotic cell death in atherosclerotic plaques of hyperlipidemic knockout mice.

In an attempt to understand the roles of apoptosis in the development of atherosclerosis, we classified lesions developed in the aortas of apo E- and LDL receptor-deficient mice, murine models of atherosclerosis, and determined frequency, spatial distribution and cell types of apoptotic cells in each lesion. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and nuclear staining with propidium iodide were used to demonstrate apoptotic cells. Mean frequencies of TUNEL-positive cells were as follows: 0%, in Type I, 0.3% in Type II, 0.05% in Type III, 0.06% in Type IV and 0.06% in Type V lesions. Most of the TUNEL-positive cells were filled with fat and distributed in close proximity to lipid pools. The TUNEL-positive cells in the intimal side of the lipid cores were macrophages, while some of those in the adventitial side were smooth muscle cells. In conclusion, apoptosis is involved in the active turn-over of foam cells of both macrophage- and smooth muscle cell-lineage especially in the early atherosclerotic lesions of the hyperlipidemic mice.

Age Factors↗
PubMed1997

Apolipoprotein E accumulates with the progression of A beta deposition in transgenic mice.

To study the role of apolipoprotein E (apoE) in vivo in deposits of amyloid beta protein (A beta), a major component of senile plaque amyloid in the brain of patients with Alzheimer disease, the transgenic mice were examined by apoE immunostaining. The mice were systemically overexpressing signal peptide and 99 amino acid residues of the carboxy-terminal fragment of human amyloid beta protein precursor (betaAPP) under control of the powerful cytomegalovirus enhancer/chicken beta-actin promotor. A beta deposits appeared at 4 months and increased with aging in the acinar cells of the transgenic pancreas. Similarly, apoE deposits appeared in the pancreatic acinar cells at 4 months old. The number and size of apoE deposits increased with aging and correlated with the progression of A beta deposits. Interstitial macrophages labeled by apoE immunostaining appeared at 8 months after birth and their number increased with aging. On serial section of the pancreata of 24-month-old mice, approximately 70% of A beta deposits were labeled with the apoE antiserum. ApoE was detected in the highly insoluble formic acid fraction of the transgenic pancreas by an immunoblot study. The Northern blot study revealed no increase in synthesis of endogenous apoE mRNA. These findings indicate that apoE is closely related to progression of A beta deposits with aging and suggest that A beta deposition in the transgenic pancreas is similar to that in the senile plaque of Alzheimer brains. Therefore, our experimental system using transgenic mice will provide a useful tool to analyze the molecular mechanism of A beta deposition in association with apoE in vivo.

Aging↗
PubMed1997

Skipping of exon 14 and possible instability of both the mRNA and the resultant truncated protein underlie a common cholesteryl ester transfer protein deficiency in Japan.

Among the Japanese population, a G-to-A mutation at the beginning of intron 14 of the human cholesteryl ester transfer protein (CETP) gene is a frequent cause of CETP deficiency characterized by markedly increased HDL cholesterol. The resulting abnormalities responsible for null CETP deficiency were studied in detail. The CETP mRNA transcripts amplified by polymerase chain reaction from the monocyte-derived macrophages of two homozygous patients were both found to be normal except for the whole deletion of exon 14. The deletion causes a shift of reading frame and introduces a premature termination codon downstream. Examination of the macrophage RNA from heterozygotes suggested the increased instability of the abnormal mRNA in the cytoplasm, because the amount of the aberrant transcript was nearly one third that of a normal transcript in the cytoplasm, while they were equal in the nucleus. Although this indicated the synthesis of a mutant CETP that lacks about 15% at its carboxy terminus, immunoblot analysis demonstrated that the abnormal CETP was virtually absent in both the media and cell lysates of transfected COS-1 cells, which massively expressed the mutant CETP mRNA. These results elucidate the primary abnormality due to the common CETP splicing mutation to be the exon skipping of mRNA, which decreases the level of mRNA and produces a truncated protein that should be rapidly degraded intracellularly.

Alternative Splicing↗
PubMed1996

Synergistic effects of transforming growth factor-beta on the expression of c-fms, macrophage colony-stimulating factor receptor gene, in vascular smooth muscle cells.

Vascular smooth muscle cells (SMC) transform to foam cells in the process of atherosclerosis. We have reported that SMC derived from the intima of atherosclerotic lesions express c-fms, macrophage colony-stimulating factor receptor gene, which is not normally expressed in medial SMC. In the present study, we demonstrated that transforming growth factor-beta (TGF-beta) synergistically induced expression of c-fms in the presence of platelet-derived growth factor-BB in human medial SMC, a level comparable to that observed in the intima. The induction of c-fms was not inhibited by protein kinase C (PKC) inhibitor, suggesting that TGF-beta induces c-fms via a PKC-independent pathway. These results suggest that TGF-beta plays an important role in the phenotypic change of smooth muscle cells to macrophage-like cells in the process of atherosclerosis.

Becaplermin↗
PubMed1996

Hyperinsulinaemia accelerates accumulation of cholesterol ester in aorta of rats with transplanted pancreas.

Hyperinsulinaemia may play a role in the development of atherosclerosis; however, the direct effect of endogenous insulin on the atherosclerotic process is not well understood. To clarify this situation we performed pancreas transplantation with systemic venous drainage in Wistar Shionogi (WS) and Spontaneous Hypertensive (SHR) rats. Both rats received syngeneic pancreaticoduodenal transplants from donor rats. SHR rats were used to observe the additive effects of both hypertension and hyperinsulinaemia on the atherosclerotic process. Peak blood insulin levels after a glucose load were approximately two times higher in transplanted rats than in non-transplanted WS and SHR rats. By contrast, there was no difference in plasma glucose responses between transplanted and non-transplanted rats. Hyperinsulinaemia was not related to dyslipidaemia and hypertension in transplanted rats. Nine months after transplantation, the cholesterol ester contents of the aortas of both WS and SHR transplanted rats were significantly higher than in the control rats (WS: 1.9 +/- 1.0 vs 3.8 +/- 2.1 mg/g dry tissue, p < 0.01; SHR: 1.7 +/- 1.3 vs 3.7 +/- 1.4 mg/g dry tissue, p < 0.05). No differences were demonstrated in the thickness of the intima or in the histology of the aortas of transplanted and control rats. To study the mechanism for cholesterol ester accumulation in the arterial wall, we measured neutral cholesterol ester hydrolase activities in vascular medial smooth muscle cells. Insulin significantly suppressed neutral cholesterol ester hydrolase activities in medial smooth muscle cells. Our results indicate that endogenous hyperinsulinaemia contributes to the development of atherosclerosis by accelerating cholesterol ester accumulation in the arterial wall.

Animals↗
PubMed1995

Cloning and structural analysis of the mouse apolipoprotein B mRNA editing protein gene.

Mouse apolipoprotein B mRNA editing protein gene has been isolated and characterized. The amino acid sequences deduced from the exonic sequences of the gene showed 90, 70 and 68% identity to those of rat, human and rabbit, respectively. RNA blotting experiments demonstrated high expression in small intestine especially in the jejunum as a major 2.4 kb mRNA species. mRNA in other tissues including the liver is smaller in amounts and larger in size (3.1 kb) than the intestinal mRNA. The gene spans 12 kb and contains 5 exons and 4 introns. Sequencing of the 5'-flanking region revealed that a canonical TATA box is absent and the consensus recognition sequences for several transcription factors have been identified.

APOBEC-1 Deaminase↗
PubMed1994

Regulation of low density lipoprotein receptor activity in Chinese hamster ovary cells transfected with the c-fms gene.

Chinese hamster ovary (CHO) cells were transfected with the human c-fms gene, which encodes the receptor for macrophage colony-stimulating factor, to examine the effects on the low density lipoprotein (LDL) receptor activity. Degradation of [125I]LDL was significantly reduced in the transfected CHO cells as compared to non-transfected cells when incubated in lipoprotein-deficient serum. Quantitative analysis of LDL receptor mRNA using a competitive PCR method demonstrated that LDL receptor suppression occurred at transcription. These findings suggest that expression of the c-fms gene is involved in the regulation of LDL receptors on macrophages.

Animals↗