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51.
Toshihiro Miyazaki Masako Mori Carolina A. Yoshida Chizuru Ito Kenji Yamatoya Takeshi Moriishi Yosuke Kawai Hisato Komori Tetsuya Kawane Shin-ichi Izumi Kiyotaka Toshimori Toshihisa Komori 《Histochemistry and cell biology》2013,139(2):339-354
Galnt3 belongs to the GalNAc transferase gene family involved in the initiation of mucin-type O-glycosylation. Male Galnt3-deficient (Galnt3 ?/?) mice were infertile, as previously reported by Ichikawa et al. (2009). To investigate the involvement of Galnt3 in spermatogenesis, we examined the differentiation of germ cells in Galnt3 ?/? mice. Galnt3 mRNA was most highly expressed in testis, and Galnt3 protein was localized in the cis-medial parts of the Golgi stacks of spermatocytes and spermatids in the seminiferous tubules. Spermatozoa in Galnt3 ?/? mice were rare and immotile, and most of them had deformed round heads. They exhibited abnormal acrosome and disturbed mitochondria arrangement in the flagella. At the cap phase, proacrosomal vesicles of various sizes, which had not coalesced to form a single acrosomal vesicle, were attached to the nucleus in Galnt3 ?/? mice. TUNEL-positive cells were increased in the seminiferous tubules. The binding of VVA lectin, which recognizes the Tn antigen (GalNAc-O-Ser/Thr), in the acrosomal regions of spermatids and spermatozoa in Galnt3 ?/? mice was drastically reduced. Equatorin is a N, O-sialoglycoprotein localized in the acrosomal membrane and is suggested to be involved in sperm–egg interaction. Immunohistochemical and Western blot analyses showed a drastic reduction in the reactivity with MN9 antibody, which recognizes the O-glycosylated moiety of equatorin and inhibits sperm–egg interaction. These findings indicate that deficiency of Galnt3 results in a severe reduction of mucin-type O-glycans in spermatids and causes impaired acrosome formation, leading to oligoasthenoteratozoospermia, and suggest that Galnt3 may also be involved in the process of fertilization through the O-glycosylation of equatorin. 相似文献
52.
Ryushi Kawakami Haruhiko Sakuraba Toshihisa Ohshima 《Bioscience, biotechnology, and biochemistry》2013,77(12):2045-2050
We previously found a very large NAD-dependent glutamate dehydrogenase with approximately 170?kDa subunit from Janthinobacterium lividum (Jl-GDH) and predicted that GDH reaction occurred in the central domain of the subunit. To gain further insights into the role of the central domain, several single point mutations were introduced. The enzyme activity was completely lost in all single mutants of R784A, K810A, K820A, D885A, and S1142A. Because, in sequence alignment analysis, these residues corresponded to the residues responsible for glutamate binding in well-known small GDH with approximately 50?kDa subunit, very large GDH and well-known small GDH may share the same catalytic mechanism. In addition, we demonstrated that C1141, one of the three cysteine residues in the central domain, was responsible for the inhibition of enzyme activity by HgCl2, and HgCl2 functioned as an activating compound for a C1141T mutant. At low concentrations, moreover, HgCl2 was found to function as an activating compound for a wild-type Jl-GDH. This suggests that the mechanism for the activation is entirely different from that for the inhibition. 相似文献
53.
Yoshihisa Tanaka Masanobu Kirita Yuko Abe Satoshi Miyata Motoyuki Tagashira Tomomasa Kanda 《Bioscience, biotechnology, and biochemistry》2013,77(7):1140-1146
Seven new O-methylated theaflavins (TFs) were synthesized by using O-methyltransferase from an edible mushroom. Using TFs and O-methylated TFs, metabolic stability in pooled human liver S9 fractions and inhibitory effect on H2O2-induced oxidative damage in human HepG2 cells were investigated. In O-methylation of theaflavin 3′-O-gallate (TF3′G), metabolic stability was potentiated by an increase in the number of introduced methyl groups. O-methylation of TF3,3′G did not affect metabolic stability, which was likely because of a remaining 3-O-galloyl group. The inhibitory effect on oxidative damage was assessed by measuring the viability of H2O2-damaged HepG2 cells treated with TFs and O-methylated TFs. TF3,3′G and O-methylated TFs increased cell viabilities significantly compared with DMSO, which was the compound vehicle (p?<?0.05), and improved to approximately 100%. Only TF3′G did not significantly increase cell viability. It was suggested that the inhibitory effect on H2O2-induced oxidative damage was potentiated by O-methylation or O-galloylation of TFs. 相似文献
54.
Marcelo Magioli Katia Maria Paschoaletto Micchi de Barros Ferraz Eleonore Zulnara Freire Setz Alexandre Reis Percequillo Michelle Viviane de Sá Santos Rondon Vanessa Villanova Kuhnen Mariana Cristina da Silva Canhoto Karen Evelyn Almeida dos Santos Claudia Zukeran Kanda Gabriela de Lima Fregonezi Helena Alves do Prado Mitra Katherina Ferreira Milton Cezar Ribeiro Priscilla Marqui Schmidt Villela Luiz Lehmann Coutinho Márcia Gonçalves Rodrigues 《European Journal of Wildlife Research》2016,62(4):431-446
55.
Naohisa Kanda Mutsuo Goto Hidehiro Kato Megan V. McPhee Luis A. Pastene 《Conservation Genetics》2007,8(4):853-864
Bryde’s whales (Balaenoptera brydei) differ from other typical baleen whale species because they are restricted to tropical and warm temperate waters in major
oceans, and frequent trans-equatorial movement has been suggested for the species. We tested this hypothesis by analyzing
genetic variation at 17 microsatellite loci (N = 508) and 299 bp of mitochondrial DNA (mtDNA) control region sequences (N = 472) in individuals obtained from the western North Pacific, South Pacific, and eastern Indian Ocean. Combined use of microsatellite
and mtDNA markers allowed us to distinguish between contemporary gene flow and ancestral polymorphism and to describe sex-specific
philopatry. A high level of genetic diversity was found within the samples. Both nuclear and mtDNA markers displayed similar
population structure, indicating a lack of sex-specific philopatry. Spatial structuring was detected using both frequency-based
population parameters and individual-based Bayesian approaches. Whales in the samples from different oceanic regions came
from genetically distinct populations with evidence of limited gene flow. We observed low mtDNA sequence divergence among
populations and a lack of concordance between geographic and phylogenetic position of mtDNA haplotypes, suggesting recent
separation of populations rather than frequent trans-equatorial and inter-oceanic movement. We conclude that current gene
flow between Bryde’s whale populations is low and that effective management actions should treat them as separate entities
to ensure continued existence of the species. 相似文献
56.
Numerous delphacid planthopper species are major pests of economically important and widely cultivated crops (i.e. rice, corn, and sugarcane). These insects have the potential to become serious crop pests in areas where they have either naturally migrated or been newly introduced. The white-bellied planthopper, Stenocranus pacificus Kirkaldy, 1907, originally known from tropical South Pacific islands, appeared in tropical and subtropical Asia in the early years of the 21st century. Since then, S. pacificus has become a serious pest of corn in some Southeast Asian countries, although it also feeds on rice, sugarcane, sorghum, and other grasses. Here, we report the presence of S. pacificus in mainland Japan, representing the first record of this species in temperate Asia. Seven male and 17 female adult individuals collected in Kumamoto Prefecture in 2019 and 2020 were identified as S. pacificus based on their morphological characteristics and mitochondrial COI sequences. In addition, molecular phylogenetic analysis showed that S. pacificus formed a distinct clade from other Stenocranus species, indicating uncertainty in its generic assignment. Although crop damage by S. pacificus has not yet been reported from temperate regions, given its wide range of plant hosts and the potential for future range expansions, damaged crops in Asia, including in temperate regions, should be monitored for the presence of this species. 相似文献
57.
Localization of pp60c-src in growth cone of PC12 cell 总被引:2,自引:0,他引:2
By immunocytochemical and biochemical techniques, we observed the localization and expression of pp60c-src in nerve growth factor (NGF)-treated PC12 cells. Immunostaining of pp60c-src is detected in the neuronal soma and the tips of neurites (growth cones). Immunofluorescence in the neurites is less significant. High-resolution microscopy reveals that the location of pp60c-src in growth cone is in good agreement with the adhesive site of growth cone to the substratum. The pp60c-src kinase activity and the pp60c-src protein level increase 3.1- to 3.5-fold and 2.0-fold during differentiation of PC12 cells, respectively. The pp60c-src levels in the neurite fraction are also higher than those in the neuronal soma fraction. These results support the immunocytochemical finding that pp60c-src is localized in growth cones of differentiated PC12 cells. Furthermore, we discuss the possible role of pp60c-src in growth cone. 相似文献
58.
Thrombin is a potent mitogen for vascular smooth muscle cells. However, the signaling pathways by which thrombin mediates its mitogenic response are not fully understood. The ERK (extracellular signal-regulated protein kinase) and JNK (c-Jun N-terminal kinase) members of the mitogen-activated protein kinase (MAPK) family are reported to be activated by thrombin. We have investigated the response to thrombin of another member of the MAPK family, p38 MAPK, which has been suggested to be activated by both stress and inflammatory stimuli in vascular smooth muscle cells. We found that thrombin induced time- and dose-dependent activation of p38 MAPK. Maximal stimulation of p38 MAPK was observed after a 10-min incubation with 1 unit ml(-1) thrombin. GF109203X, a protein kinase C inhibitor, and prolonged treatment with phorbol 12-myristate 13-acetate partially inhibited p38 MAPK activation. A tyrosine kinase inhibitor, genistein, also inhibited p38 MAPK activation in a dose-dependent manner. p38 MAPK activation was inhibited by overexpression of betaARK1ct (beta-adrenergic receptor kinase I C-terminal peptide). p38 MAPK activation was also inhibited by expression of dominant-negative Ras, not by dominant-negative Rac. We next examined the effect of a p38 MAPK inhibitor, SB203580, on thrombin-induced proliferation. SB203580 inhibited thrombin-induced DNA synthesis in a dose-dependent manner. These results suggest that thrombin activates p38 MAPK in a manner dependent on Gbetagamma, protein kinase C, a tyrosine kinase, and Ras, that p38 MAPK has a role in thrombin-induced mitogenic response in the cells. 相似文献
59.
Leucine dehydrogenase (l-leucine: NAD+ oxidoreductase, deaminating, EC 1.4.1.9) has been purified to homogeneity from a moderate thermophilic bacterium, Bacillus stearothermophilus. Am improved method of preparative slab gel electrophoresis was used effectively to purify it. The enzyme has a molecular mass of about 300,000 and consists of six subunits with identical molecular mass (Mr, 49,000). The enzyme does not lose its activity by heat treatment at 70° C for 20 min, and incubation in the pH range of 5.5–10.0 at 55° C for 5 min. It is stable in 10 mM phosphate buffer (pH 7.2) containing 0.01% 2-mercaptoethanol at over 1 month, and is resistant to detergent and ethanol treatment. The enzyme catalyzes the oxidative deamination of branched-chain l-amino acids and the reductive amination of their keto analogs in the presence of NAD+ and NADH, respectively, as the coenzymes. The pH optima are 11 for the deamination of l-leucine, and 9.7 and 8.8 for the amination of -ketoisocaproate and -ketoisovalerate, respectively. The Michaelis constants were determined: 4.4 mM for l-leucine, 3.3 mM for l-valine, 1.4 mM for l-isoleucine and 0.49 mM for NAD+ in the oxidative deamination. The B. stearothermophilus enzyme shows similar catalytic properties, but higher activities than that from Bacillus sphaericus.Dedicated to Prof. Dr. G. Drews on the occasion of his 60th birthday 相似文献
60.
Satoshi Makishima Masahiro Mizuno Nobuaki Sato Kazunori Shinji Masayuki Suzuki Kouichi Nozaki Fumihiro Takahashi Takahisa Kanda Yoshihiko Amano 《Bioresource technology》2009,100(11):2842-2848
The semi-pilot scale of continuous flow type hydrothermal reactor has been investigated to separate hemicellulose fraction from corncob. We obtained the effective recovery of hemicellulose using tubular type reactor at 200 °C for 10 min. From constituent sugar analysis of corncob, 82.2% of xylan fraction was recovered as mixture of xylose, xylooligosaccharides and higher-xylooligosaccharide which has more than DP 10. During purification of solubilized fraction by hydrothermal reaction such as ultrafiltration and ion exchange resin, higher-xylooligosaccharide was recovered as the precipitate. This precipitate was identified as non-blanched xylan fraction which has from DP 11 to DP 21 mainly. In this system, only a small amount of furfural has been generated. This tubular reactor has a characteristic controllability of thermal history, and seems to be effective for sugar recovery from soft biomass like corncob. 相似文献