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11.
Yuuki ISAJI Koki YOSHIDA Hiroshi IMAI Masayasu YAMADA 《The Journal of reproduction and development》2015,61(6):503-510
In mouse somatic cell nuclear transfer (SCNT), polyvinylpyrrolidone (PVP) is typically included in the
nuclear donor injection medium. However, the cytotoxicity of PVP, which is injected into the cytoplasm of
oocytes, has recently become a cause of concern. In the present study, we determined whether bovine serum
albumin deionized with an ion-exchange resin treatment (d-BSA) was applicable to the nuclear donor injection
medium in SCNT as an alternative to PVP. The results obtained showed that d-BSA introduced into the cytoplasm
of an enucleated oocyte together with a donor nucleus significantly enhanced the rate of in
vitro development of cloned embryos to the blastocyst stage compared with that of a conventional
nuclear injection with PVP in SCNT. We also defined the enhancing effects of d-BSA on the blastocyst formation
rate when d-BSA was injected into the cytoplasm of oocytes reconstructed using the fusion method with a
hemagglutinating virus of Japan envelope before oocyte activation. Furthermore, immunofluorescence experiments
revealed that the injected d-BSA increased the acetylation levels of histone H3 lysine 9 and histone H4 lysine
12 in cloned pronuclear (PN) and 2-cell embryos. The injection of d-BSA before oocyte activation also
increased the production of cloned mouse offspring. These results suggested that intracytoplasmic injection of
d-BSA into SCNT oocytes before oocyte activation was beneficial for enhancing the in vitro
and in vivo development of mouse cloned embryos through epigenetic modifications to nuclear
reprogramming. 相似文献
12.
Uemura A Watarai S Kushi Y Kasama T Ohnishi Y Kodama H 《Veterinary parasitology》2006,140(3-4):264-272
Neutral glycosphingolipids (GSLs) were isolated from Trypanosoma brucei and analyzed by thin-layer chromatography (TLC), TLC/secondary ion mass spectrometry (TLC/SIMS), and liposome immune lysis assay (LILA). Three species of neutral GSLs, designated as N-1, -2, and -3 were separated on TLC. N-1 GSL migrated very close to glucosylceramide (GlcCer) and N-2 GSL showed the same mobility as lactosylceramide (LacCer). On the other hand, the mobility of N-3 GSL on the TLC plate was slower than globotetraosylceramide (Gb4). In order to characterize the molecular species of neutral GSLs from T. brucei, N-1, -2 and -3 GSLs were analyzed by TLC/SIMS. The TLC/SIMS analysis of N-1 of the parasites revealed a series of (M–H)− ions from m/z 698 to 825 representing the molecular mass range of ceramide monohexoside (CMH) (GlcCer or galactosylceramide). On the other hand, the TLC/SIMS spectra of N-2 GSL revealed a series of (M–H)− ions from m/z 944–987 indicating the molecular mass range of LacCer. In the TLC/SIMS analysis of N-3 GSL, however, the characteristic molecular ions that can elucidate the structure of N-3 GSL were not obtained. In order to confirm the results obtained from TLC/SIMS, N-1, -2, and -3, GSLs were tested by LILA with specific antibodies against GlcCer, LacCer, and Gb4, respectively. N-1 GSL had reactivity to anti-GlcCer antibody and N-2 GSL reacted with the antibody against LacCer. However, N-3 GSL was not recognized by anti-Gb4 antibody. Using anti-GlcCer and anti-LacCer antibodies, furthermore, we studied the expression of GlcCer and LacCer in T. brucei parasites. Both GlcCer and LacCer were detected on the cell surface of T. brucei. 相似文献
13.
Acute hypotonic stress becomes a threat to the survival of bacteria in the environment. Mechanosensitive channels play an essential role in the maintenance of bacterial cell integrity during hypoosmotic shock. A database search suggested that Campylobacter jejuni, a major worldwide cause of bacterial gastroenteritis in humans, possesses two putative mechanosensitive channels, designated Cjj0263 and Cjj1025, in C. jejuni strain 81-176. Osmotic downshock experiments demonstrated that a mutant lacking Cjj0263 showed a severe defect in survival of hypoosmotic shock, while a mutant lacking Cjj1025 exhibited the same survival capacity as the wild type. We further examined the colonization ability of each mutant using the one-day old chick model. Cjj0263 or Cjj1025 mutants were able to colonize chick ceca at the same level as the wild type, but a Cjj0263 Cjj1025 double mutant revealed significantly reduced ability to colonize chick ceca. To examine whether C. jejuni that have grown in the digestive tract of chicks are protected against acute hypotonic stress, bacteria in ceca were directly exposed to water. The wild type was able to survive acute osmotic downshift, but the Cjj0263 mutant suffered a substantial loss of viability when subjected to a rapid osmotic downshock. Immunoblot analysis suggested that both Cjj0263 and Cjj1025 were glycosylated via the N-linked protein glycosylation pathway, but glycan modification of these proteins was unlikely to have a major effect on their function and stability. Our data suggest that Cjj0263, a mechanosensitive channel, has a pivotal role in protection against hypoosmotic stress experienced during environmental transmission. 相似文献
14.
The influence of proanthocyanidin (PA) structures contained in bark on color development in the vanillin-hydrochloric acid (V-HCl) method used widely as a quantitative method for measuring PA were examined. The maximal absorption wavelength was different in terms of the bark from which the PA was obtained. Phenyl nucleus (resorcinol, phloroglucinol) constituting the A-ring of PA reacts with vanillin to produce the color. The maximal absorption wavelengths of the solutions from synthesized procyanidin and profisetinidin were 500 and 540 nm, respectively, indicating that the color tone differs in the V-HCl method based on the hydroxylation patterns of the A-ring. The colored solution of (+)-catechin with vanillin was dialyzed, and the resulting product (C-VC) was analyzed by gel permeation chromatography and 1H nuclear magnetic resonance. It was found that C-VC was a polymer complex consisting of 9mol (+)-catechin moieties and 10mol vanillin moieties. It was presumed that the cationized vanillin molecules that do not combine with (+)-catechin play an important role on color development in the presence of C-VC.This study was presented at the 44th and 45th Annual Meetings of The Japan Wood Research Society, Nara and Tokyo, April 1994 and April 1995 相似文献
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17.
Hiromichi Mitamura Nobuaki Arai Yukiko Yamagishi Yuuki Kawabata Yasushi Mitsunaga Metha Khachaphichat Thavee Viputhanumas 《Fisheries Science》2009,75(1):175-182
The horizontal and vertical movements of eight immature hatchery-reared (F2) Mekong giant catfish Pangasianodon gigas were monitored using acoustic telemetry in the Mae Peum reservoir, Thailand, from September to December 2005. All tagged
fish were successfully monitored throughout the study period. All fish moved throughout the reservoir for approximately 1 month
after release. Subsequently, their utilized areas became small, and the fish utilized deep areas of the reservoir. The fish
displayed diel spatial movement patterns between deep areas in the day and shallow areas in the night. The vertical movements
of the fish were related to the environment declination such as existence of hypoxic water and thermocline. Our results suggest
that the establishment of a protected area in addition to conventional fisheries regulations may sustain the fish population
in this reservoir. 相似文献
18.
Shoji K Mizuno T Shiiba D Kawagoe T Mitsui Y 《Journal of agricultural and food chemistry》2012,60(10):2490-2496
It was previously reported that compared to triacylglycerol (TAG) oil, diacylglycerol (DAG) oil improves postprandial lipid response. However, the effects of DAG oil on postprandial hyperglycemia and incretin response have not yet been determined. In this study, the effects of DAG oil on both postprandial hyperlipidemia and hyperglycemia and the response to the glucose-dependent insulinotropic polypeptide (GIP) were studied. This randomized, double-blind, crossover study analyzed data for 41 individuals with high fasting triacylglycerol concentrations. The subjects ingested test meals (30.3 g of protein, 18.6 g of fat, and 50.1 g of carbohydrate) containing 10 g of DAG oil (DAG meal) or TAG oil (TAG meal) after fasting for at least 12 h. Blood samples were collected prior to and 0.5, 2, 3, 4, and 6 h after ingestion of the test meal. Postprandial TAG concentrations were significantly lower after the DAG meal compared with the TAG meal. Postprandial TAG, insulin, and GIP concentrations were significantly lower after the DAG meal compared with the TAG meal in 26 subjects with fasting serum TAG levels between 1.36 and 2.83 mmol/L. DAG-oil-based meals, as a replacement for TAG oil, may provide cardiovascular benefits in high-risk individuals by limiting lipid and insulin excursions. 相似文献
19.
Rin YANAI Yudai YAMASHITA Kohei UMEZU Yuuki HIRADATE Kenshiro HARA Kentaro TANEMURA 《The Journal of reproduction and development》2021,67(1):59
The structure of microtubules is essential for the fertilizing ability of spermatozoa. Acetylation of α-tubulin plays an important role in flagellar elongation and spermatozoa motility. Previous reports have suggested that alpha-tubulin N-acetyltransferase 1 (ATAT1) is the main acetyltransferase involved in the acetylation of α-tubulin. Although ATAT1 is reported to express in the testis, no information is available regarding its expression in elongated spermatids, epididymis, and mature spermatozoa. Hence, it remains unclear whether ATAT1 is involved in spermatozoa maturation and capacitation. Therefore, we evaluated the expression of ATAT1 in the mouse male reproductive system using immunostaining and western blotting. Our results showed that ATAT1 was expressed in spermatids during spermiogenesis in mouse testes, but its expression varied according to the seminiferous tubule stage. We observed ATAT1 in the cytoplasm of round spermatids, the flagella of elongated spermatids, and in the cytoplasm of step 16 spermatids, just before its release into the lumen. In addition, ATAT1 was expressed in epithelial cells of the epididymis. In spermatozoa of the cauda epididymis, ATAT1 expression was primarily observed in the midpiece of the spermatozoa. The localization of ATAT1 protein in the male germline was observed during spermiogenesis as well as during spermatozoa maturation. Our results suggest that ATAT1 may be involved in the formation of flagella and in the acetylation process, which has attracted attention in recent years regarding male infertility. 相似文献
20.
Takeharu KANEDA Yuuki KIDO Tsuyoshi TAJIMA Norimoto URAKAWA Kazumasa SHIMIZU 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(1):15-19
The effects of various
selective phosphodiesterase (PDE) inhibitors on carbachol (CCh)-induced contraction in the
bovine abomasum were investigated. Various selective PDE inhibitors, vinpocetine (type 1),
erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA, type 2), milrinone (type 3), Ro20-1724 (type
4), vardenafil (type 5), BRL-50481 (type 7) and BAY73-6691 (type 9), inhibited CCh-induced
contractions in a concentration-dependent manner. Among the PDE inhibitors, Ro20-1724 and
vardenafil induced more relaxation than the other inhibitors based on the data for the
IC50 or maximum relaxation. In smooth muscle of the bovine abomasum, we
showed the expression of PDE4B, 4C, 4D and 5 by RT-PCR analysis. In the presence of CCh,
Ro20-1724 increased the cAMP content, but not the cGMP content. By contrast, vardenafil
increased the cGMP content, but not the cAMP content. These results suggest that
Ro20-1724-induced relaxation was correlated with cAMP and that vardenafil-induced
relaxation was correlated with cGMP in the bovine abomasum. In conclusion, PDE4 and PDE5
are the enzymes involved in regulation of the relaxation associated with cAMP and cGMP,
respectively, in the bovine abomasum. 相似文献