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291.
The Native Form and Maturation Process of Hepatitis C Virus Core Protein 总被引:23,自引:5,他引:23
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Kohichiroh Yasui Takaji Wakita Kyoko Tsukiyama-Kohara Shin-Ichi Funahashi Masumi Ichikawa Tadahiro Kajita Darius Moradpour Jack R. Wands Michinori Kohara 《Journal of virology》1998,72(7):6048-6055
The maturation and subcellular localization of hepatitis C virus (HCV) core protein were investigated with both a vaccinia virus expression system and CHO cell lines stably transformed with HCV cDNA. Two HCV core proteins, with molecular sizes of 21 kDa (p21) and 23 kDa (p23), were identified. The C-terminal end of p23 is amino acid 191 of the HCV polyprotein, and p21 is produced as a result of processing between amino acids 174 and 191. The subcellular localization of the HCV core protein was examined by confocal laser scanning microscopy. Although HCV core protein resided predominantly in the cytoplasm, it was also found in the nucleus and had the same molecular size as p21 in both locations, as determined by subcellular fractionation. The HCV core proteins had different immunoreactivities to a panel of monoclonal antibodies. Antibody 5E3 stained core protein in both the cytoplasm and the nucleus, C7-50 stained core protein only in the cytoplasm, and 499S stained core protein only in the nucleus. These results clearly indicate that the p23 form of HCV core protein is processed to p21 in the cytoplasm and that the core protein in the nucleus has a higher-order structure different from that of p21 in the cytoplasm. HCV core protein in sera of patients with HCV infection was analyzed in order to determine the molecular size of genuinely processed HCV core protein. HCV core protein in sera was found to have exactly the same molecular weight as the p21 protein. These results suggest that p21 core protein is a component of native viral particles. 相似文献
292.
Shigeki Yamaguchi Alexander Balbir Brian Schofield Judith Coram Clarke G Tankersley Robert S Fitzgerald Christopher P O'Donnell Machiko Shirahata 《Journal of applied physiology》2003,94(4):1536-1542
In a previous study, DBA/2J and A/J inbred mice showed extremely different hypoxic ventilatory responses, suggesting variations in their carotid bodies. We have assessed the morphological and functional differences of the carotid bodies in these mice. Histological examination revealed a clearly delineated carotid body only in the DBA/2J mice. Many typical glomus cells and glomeruli appeared in the DBA/2J but not in the A/J mice. The size of the carotid body in the DBA/2J and A/J mice was 6.3 +/- 0.5 x 10(6) and 1.5 +/- 0.3 x 10(6) micro m(3), respectively. The area immunostained for tyrosine hydroxylase, an estimation of the glomus cell quantity, was four times larger in the DBA/2J mice than in the A/J mice. The individual data points in the DBA/2J mice segregated from those in the A/J mice. ACh increased intracellular Ca(2+) in most clusters (81%) of cultured carotid body cells from the DBA/2J mice, but only in 18% of clusters in the A/J mice. These data suggest that genetic determinants account for the strain differences in the structure and function of the carotid body. 相似文献
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294.
Toshiyuki Takeuchi Machiko Ogawa Chie Furihata Takashi Kawachi Takashi Sugimura 《Biochimica et Biophysica Acta (BBA)/General Subjects》1977,497(3):657-662
In rats amylase activity in the pancreas increased greatly from day 15 of gestation to a maximum on day 21. Then it decreased to less than one-tenth of this maximum value on about day 5 after birth. It increased again about 15 days after birth and reached the adult level about 30 days after birth.No amylase activity was in the parotid gland before birth: it appeared about 12 days after birth and reached the adult level, which was higher than that in the pancreas, about 30 days after birth.The serum corticosterone level was as high as the adult level before birth. Then it decreased to less than one-tenth of the adult level 5 days after birth and increased again from 15 to 25 days after birth to the adult level. The developmental change in the serum corticosterone level seemed to influence amylase activity in the pancreas both before and after birth, and that in the parotid gland only after birth.The serum contained both pancreatic and paratoid type isozymes of amylase until 1 day after birth but only the parotid type from 3 days after birth. 相似文献
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296.
Nakajima M Yamashita T Takahashi M Nakano Y Takeda T 《Applied and environmental microbiology》2012,78(16):5682-5689
A glycoside hydrolase responsible for laminarin degradation was partially purified to homogeneity from a Ustilago esculenta culture filtrate by weak-cation-exchange, strong-cation-exchange, and size-exclusion chromatography. Three proteins in enzymatically active fractions were digested with chymotrypsin followed by liquid chromatography-tandem mass spectrometry (LC/MS/MS) analysis, resulting in the identification of three peptide sequences that shared significant similarity to a putative β-1,3-glucanase, a member of glucoside hydrolase family 16 (GH16) from Sporisorium reilianum SRZ2. A gene encoding a laminarin-degrading enzyme from U. esculenta, lam16A, was isolated by PCR using degenerate primers designed based on the S. reilianum SRZ2 β-1,3-glucanase gene. Lam16A possesses a GH16 catalytic domain with an N-terminal signal peptide and a C-terminal glycosylphosphatidylinositol (GPI) anchor peptide. Recombinant Lam16A fused to an N-terminal FLAG peptide (Lam16A-FLAG) overexpressed in Aspergillus oryzae exhibited hydrolytic activity toward β-1,3-glucan specifically and was localized both in the extracellular and in the membrane fractions but not in the cell wall fraction. Lam16A without a GPI anchor signal peptide was secreted extracellularly and was not detected in the membrane fraction. Membrane-anchored Lam16A-FLAG was released completely by treatment with phosphatidylinositol-specific phospholipase C. These results suggest that Lam16A is anchored in the plasma membrane in order to modify β-1,3-glucan associated with the inner cell wall and that Lam16A is also used for the catabolism of β-1,3-glucan after its release in the extracellular medium. 相似文献
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298.
Background
We examined how the prevalence of individuals diagnosed with diabetes differs by age and sex using the diagnostic criteria of fasting plasma glucose (FPG) and/or glycated haemoglobin (HbA1c) in a large Japanese population.Methods
We conducted a cross-sectional study using a dataset of 33,959 people (16,869 men and 17,090 women) without known diabetes who underwent health checkups from 1998 to 2006. We divided the age range of the participants into six groups of similar numbers. We compared the prevalence of diabetes using the criteria of FPG ≥7.0 mmol/l (126 mg/dl), HbA1c ≥48 mmol/mol (6.5%), or both, in men and women in each age group.Results
Men had higher prevalence of diabetes than women using the criterion of either FPG or HbA1c (7.5% men vs. 3.4% women, P<0.001), or both (4.3% men vs. 1.8% women, P<0.001). HbA1c increased steadily in women through the six age groups. In the oldest group (≥66 years), the proportion of women among those diagnosed with diabetes was as high as 42.3% (215/508) using the criterion of either FPG or HbA1c, and 41.6% (116/279) using both criteria.Conclusions
Using either FPG or HbA1c, the prevalence of people diagnosed with diabetes would almost double compared to using the criterion of both scores, and this would include more elderly women than men. The impact of introducing HbA1c for diabetes diagnosis should be considered in terms of age and sex. 相似文献299.
300.
Mimura S Kimura N Hirata M Tateyama D Hayashida M Umezawa A Kohara A Nikawa H Okamoto T Furue MK 《The International journal of developmental biology》2011,55(2):181-187
Human bone marrow-derived mesenchymal stem cells (hMSCs) are potential cellular sources of therapeutic stem cells as they have the ability to proliferate and differentiate into a wide array of mesenchymal cell types such as osteoblasts, chondroblasts and adipocytes. hMSCs have been used clinically to treat patients with graft vs. host disease, osteogenesis imperfect, or alveolar cleft, suggesting that transplantation of hMSCs is comparatively safe as a stem cell-based therapy. However, conventional culture medium for hMSCs contains fetal bovine serum (FBS). In the present study, we developed a growth factor-defined, serum-free medium for culturing hMSCs. Under these conditions, TGF-beta1 promoted proliferation of hMSCs. The expanded hMSC population expressed the human pluripotency markers SSEA-3, -4, NANOG, OCT3/4 and SOX2. Furthermore, double positive cells for SSEA-3 and a mesenchymal cell marker, CD105, were detected in the population. The potential to differentiate into osteoblasts and adipocytes was confirmed. This work provides a useful tool to understand the basic biological properties of hMSCs in culture. 相似文献