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51.
52.
Azuma A Udo Y Sato A Mitani N Kono A Ban Y Yakushiji H Koshita Y Kobayashi S 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2011,122(7):1427-1438
Skin color is one of the most important fruit traits in grape, and has become greatly diversified due to hybridization and human selection. Many studies concerning the genetic control of grape color in European species (Vitis vinifera L.), especially the role of MYB-related genes, have been reported. On the other hand, there have been few studies of the MYB-related genes in grapes belonging to V.?×labruscana L.H. Bailey, a subgroup of grapes that originated from the hybridization of V. labrusca with V. vinifera. In the present study, we found a novel functional haplotype, HapE2 (consisting of the genes VlMYBA2 and VlMYBA1?C3), in diploid V.?×labruscana. Moreover, we developed a method to determine the haplotype compositions of tetraploid grapes by means of quantitative real-time PCR, and investigated the relationship between haplotype composition and skin color. The color locus in V.?×labruscana grapes usually consists of functional haplotypes (HapE1 and/or HapE2), and non-functional haplotype HapA. The number of functional haplotypes in the genome was found to be correlated with the level of anthocyanin in the skin. Anthocyanin contents of grapes that contained HapE2 were significantly higher than those containing HapE1. These results suggest that the number and kind of functional haplotypes at the color locus are the major genetic factors that determine skin color variation. These findings provide new knowledge about the unique genetic control of color in V.?×labruscana grapes, and should contribute to development of new cultivars that have the desired color and anthocyanin content. 相似文献
53.
Kazuo Sugiyama Hirotoshi Ebinuma Nobuhiro Nakamoto Noriko Sakasegawa Yuko Murakami Po-sung Chu Shingo Usui Yuka Ishibashi Yuko Wakayama Nobuhito Taniki Hiroko Murata Yoshimasa Saito Masayoshi Fukasawa Kyoko Saito Yoshiyuki Yamagishi Takaji Wakita Hiroshi Takaku Toshifumi Hibi Hidetsugu Saito Takanori Kanai 《PloS one》2014,9(4)
Most of experiments for HCV infection have been done using lytic infection systems, in which HCV-infected cells inevitably die. Here, to elucidate metabolic alteration in HCV-infected cells in a more stable condition, we established an HCV-persistently-infected cell line, designated as HPI cells. This cell line has displayed prominent steatosis and supported HCV infection for more than 2 years, which is the longest ever reported. It enabled us to analyze metabolism in the HCV-infected cells integrally combining metabolomics and expression arrays. It revealed that rate-limiting enzymes for biosynthesis of cholesterol and fatty acids were up-regulated with actual increase in cholesterol, desmosterol (cholesterol precursor) and pool of fatty acids. Notably, the pentose phosphate pathway was facilitated with marked up-regulation of glucose-6-phosphate dehydrogenase, a rete-limiting enzyme, with actual increase in NADPH. In its downstream, enzymes for purine synthesis were also up-regulated resulting in increase of purine. Contrary to common cancers, the TCA cycle was preferentially facilitated comparing to glycolysis pathway with a marked increase of most of amino acids. Interestingly, some genes controlled by nuclear factor (erythroid-derived 2)-like 2 (Nrf2), a master regulator of antioxidation and metabolism, were constitutively up-regulated in HPI cells. Knockdown of Nrf2 markedly reduced steatosis and HCV infection, indicating that Nrf2 and its target genes play important roles in metabolic alteration and HCV infection. In conclusion, HPI cell is a bona fide HCV-persistently-infected cell line supporting HCV infection for years. This cell line sustained prominent steatosis in a hypermetabolic status producing various metabolites. Therefore, HPI cell is a potent research tool not only for persistent HCV infection but also for liver metabolism, overcoming drawbacks of the lytic infection systems. 相似文献
54.
Akt protein kinase inhibits non-apoptotic programmed cell death induced by ceramide. 总被引:4,自引:0,他引:4
Toshihiro Mochizuki Akio Asai Nobuhito Saito Sakae Tanaka Hideki Katagiri Tomoichiro Asano Makoto Nakane Akira Tamura Yoshiyuki Kuchino Chifumi Kitanaka Takaaki Kirino 《The Journal of biological chemistry》2002,277(4):2790-2797
A growing body of evidence now suggests that programmed cell death (PCD) occurs via non-apoptotic mechanisms as well as by apoptosis. In contrast to apoptosis, however, the molecular mechanisms involved in the regulation of non-apoptotic PCD remain only poorly understood. Here we show that ceramide induces a non-apoptotic PCD with a necrotic-like morphology in human glioma cells. Characteristically, the cell death was not accompanied by loss of the mitochondrial transmembrane potential, cytosolic release of cytochrome c from mitochondria, or the activation of the caspase cascade. Consistent with these characteristics, this ceramide-induced cell death was inhibited neither by the overexpression of Bcl-xL nor by the pan-caspase inhibitor zVAD-fmk. However, strikingly, the ceramide-induced non-apoptotic cell death was inhibited by the activation of the Akt/protein kinase B pathway through the expression of a constitutively active version of Akt. The results for the first time indicate that the Akt kinase, known to play an essential role in survival factor-mediated inhibition of apoptotic cell death, is also involved in the regulation of non-apoptotic PCD. 相似文献
55.
Gene transfection of H25A mutant heme oxygenase-1 protects cells against hydroperoxide-induced cytotoxicity. 总被引:4,自引:0,他引:4
Rio Hori Misato Kashiba Tomoko Toma Akihiro Yachie Nobuhito Goda Nobuya Makino Akinori Soejima Toshihiko Nagasawa Kimimasa Nakabayashi Makoto Suematsu 《The Journal of biological chemistry》2002,277(12):10712-10718
Heme oxygenase (HO)-1 is a stress-inducible enzyme protecting cells against oxidative stress, and mechanisms have been considered to depend exclusively on its enzyme activity. This study aimed to examine if the protein lacking catalytic activities could also display such resistance against oxidative stress. Stable transfectants of rat wild type HO-1 cDNA (HO-1-U937) and those of its H25A mutant gene (mHO-1-U937) were established using human monoblastic lymphoma cell U937. HO-1-U937 and mHO-1-U937 used in the study exhibited similar levels of the protein expression, while only the former increased enzyme activities. HO-1- and mHO-1 U937 cells became more and less sensitive to H(2)O(2) than mock transfectants, respectively; such distinct susceptibility between the cells was ascribable to differences in the capacity to scavenge H(2)O(2) through catalase and to execute iron-mediated oxidant propagation. On the other hand, both cell lines exhibited greater resistance to tert-butyl hydroperoxide than mock transfectants. The resistance of HO-1-U937 to hydroperoxides appeared to result from antioxidant properties of bilirubin, an HO-derived product, while that of mHO-1-U937 was ascribable to increased contents of catalase and glutathione. These results provided evidence that gene transfection of the activity-lacking mutant HO-1 protects cells against oxidative stress through multiple mechanisms involving up-regulation of catalase and glutathione contents. 相似文献
56.
Hanako Morita Masahiko Yasuda Masafumi Yamamoto Yurina Tomiyama Ritsuki Uchida Yuyo Ka Tomoyuki Ogura Kenji Kawai Hiroshi Suemizu Nobuhito Hayashimoto 《Experimental Animals》2021,70(3):355
Astroviruses are often associated with gastrointestinal diseases in mammals and birds. Murine astrovirus (MuAstV) is frequently detected in laboratory mice. Previous studies on MuAstV in mice did not report any symptoms or lesions. However, little information is available regarding its pathogenicity in immunodeficient mice. Therefore, in this study, we experimentally infected germ-free NOD.Cg-PrkdcscidIl2rgtm1Sug/ShiJic (NOG) mice, which are severely immunodeficient, with MuAstV. Germ-free mice were used for experimental infection to eliminate the effects of intestinal bacteria. Mice in each group were then necropsied and subjected to PCR for MuAstV detection, MuAstV RNA quantification in each organ, and histopathological examination at 4 and 28 days post inoculation (DPI). Tissue samples from the small intestine were examined by transmission electron microscopy. No symptoms or abnormalities were detected in any mice during necropsy. The MuAstV concentration was highest in the lower small intestine, where it increased approximately 8-fold from 4 to 28 DPI. Transmission electron microscopy revealed circular virus particles of approximately 25 nm in diameter in the cytoplasm of the villous epithelial cells of the lower small intestine. Histopathological examination did not reveal any abnormalities, such as atrophy, in the intestinal villi. Our results suggest that MuAstV proliferates in the villous epithelial cells of the lower small intestine and has weak pathogenicity. 相似文献
57.
Sone N Fukuda M Katayama S Jyoudai A Syugyou M Noguchi S Sakamoto J 《Biochimica et biophysica acta》2003,1557(1-3):125-131
Structural genes encoding quinol-cytochrome c reductase (QcR) were cloned and sequenced from nocardia-form actinomycete Rhodococcus rhodochrous. QcrC and qcrA encode diheme cytochrome cc and the Rieske Fe-S protein, respectively, while the qcrB product is a diheme cytochrome b. These amino acid sequences are similar to those of Corynebacterium and Mycobacterium, the members of high G+C content firmicutes. The presence of diheme cytochrome cc subunit as a sole c-type cytochrome in these organisms suggests the direct elecron transfer to cytochrome c oxidase. The N-terminal half of the Rieske Fe-S proteins of these bacteria has a unique structure with three transmembrane helices, while the C-terminal half sequence is conserved. A phylogenetic tree using the latter region showed that high G+C firmicutes form a clear clade with Thermus, but not with low G+C firmicutes. 相似文献
58.
Goto K Horiuchi H Shinohara H Motegi K Hashimoto K Hongo S Gemma N Hayashimoto N Itoh T Takakura A 《Journal of microbiological methods》2007,69(1):93-99
We developed a microfabricated electrochemical DNA chip for detection of polymerase chain reaction (PCR) products from 16S rRNA sequences of Clostridium piliforme (Cp), Helicobacter bilis (Hb) and Helicobacter hepaticus (Hh), and the nucleocapsid protein gene of mouse hepatitis virus (MHV). This chip does not require DNA labeling, and the hybridization signal can be detected as an anodic current. The average anodic currents of 9 (Cp), 5 (Hb), 8 (Hh) and 7 (MHV) PCR positive samples derived from feces of spontaneously infected mice (Cp, Hb and Hh) and MHV-contaminated tumor cells were 27.9+/-7.2, 31.9+/-8.1, 29.3+/-10.1, and 27.6+/-3.0 nA, respectively. On the other hand, the average anodic currents of 19 (Cp), 27 (Hb), 18 (Hh), and 13 (MHV) PCR negative samples were 0.3+/-2.9, 3.7+/-2.4, -1.0+/-1.7, and -2.3+/-2.7 nA, respectively. The anodic current increased with increasing concentrations of pathogens. For experimentally infected samples, the results of PCR/electrophoresis were in complete accord with those of this system when anodic currents of 6.1 (Cp), 8.5 (Hb), 2.4 (Hh), and 3.1 nA (MHV) were taken as the cut-off value. The results suggested that the electrochemical DNA chip system is useful for specific and quantitative detection of PCR products. 相似文献
59.
Rieko Urakawa Nobuhito Ohte Hideaki Shibata Ryunosuke Tateno Yoshiyuki Inagaki Tomoki Oda Hiroto Toda Karibu Fukuzawa Tsunehiro Watanabe Takuo Hishi Nobuhiro Oyanagi Makoto Nakata Keitaro Fukushima Asami Nakanishi 《Ecological Research》2017,32(2):279-285
We tested the potential of estimating in-field (in situ) nitrogen (N) transformation rates based on soil temperature data and N transformation parameters (Q10 and N transformation rates at standard temperature) obtained through laboratory incubations at three constant temperatures for 4 weeks. This test was conducted based on a comparison between in situ measurements and estimates using soils from 16 sites across 9 regions within the Japanese archipelago. The actual in situ N mineralization and nitrification rates measured using the buried-bag method at 0–50-cm-soil depth were 111 ± 34 and 106 ± 45 kg N ha?1 year?1, respectively, and estimates of both the rate and the amount were largely accurate. For rate alone, estimates were accurate in the 0–10-cm soil layer for annual and seasonal averages (except for spring–summer) whereas for amount alone, estimates were accurate to depths of 50 and 30 cm for N mineralization and nitrification, respectively. Thus, estimates of the rates and amounts were approximately equal to the actual in situ rate/amount, given the wide range of prediction intervals of the field measurement data. The differences between the estimates of N transformation rates derived from hourly measured and monthly average soil temperatures were negligible. Therefore, in situ soil N transformations, which are laborious to measure in the field, have the potential to be estimated from a combination of monthly average soil temperatures and N transformation parameters, which are relatively straightforward to obtain through laboratory incubation. 相似文献
60.
Chisato Kubota Seiji Torii Ni Hou Nobuhito Saito Yuhei Yoshimoto Hideaki Imai Toshiyuki Takeuchi 《The Journal of biological chemistry》2010,285(1):667-674
Reactive oxygen species (ROS) are involved in several cell death processes, including cerebral ischemic injury. We found that glutamate-induced ROS accumulation and the associated cell death in mouse hippocampal cell lines were delayed by pharmacological inhibition of autophagy or lysosomal activity. Glutamate, however, did not stimulate autophagy, which was assessed by a protein marker, LC3, and neither changes in organization of mitochondria nor lysosomal membrane permeabilization were observed. Fluorescent analyses by a redox probe PF-H2TMRos revealed that autophagosomes and/or lysosomes are the major sites for basal ROS generation in addition to mitochondria. Treatments with inhibitors for autophagy and lysosomes decreased their basal ROS production and caused a burst of mitochondrial ROS to be delayed. On the other hand, attenuation of mitochondrial activity by serum depletion or by high cell density culture resulted in the loss of both constitutive ROS production and an ROS burst in mitochondria. Thus, constitutive ROS production within mitochondria and lysosomes enables cells to be susceptible to glutamate-induced oxidative cytotoxicity. Likewise, inhibitors for autophagy and lysosomes reduced neural cell death in an ischemia model in rats. We suggest that cell injury during periods of ischemia is regulated by ROS-generating activity in autophagosomes and/or lysosomes as well as in mitochondria. 相似文献