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941.
Shuji Udagawa Satoshi Sakami Takahiro Takemura Mikiya Sato Takahiro Arai Aiko Nitta Takumi Aoki Koji Kawai Tomokatsu Iwamura Seiji Okazaki Takehiro Takahashi Mie Kaino 《Bioorganic & medicinal chemistry letters》2013,23(6):1617-1621
A series of novel 5-trans-hydroxyadamantan-2-yl-5,6,7,8-tetrahydropyrazolo[4,3-c]azepin-4(1H)-ones that inhibit 11beta-hydroxysteroid dehydrogenase type 1 are described. We discovered these 7-membered cyclic amide derivatives by introducing a distinctive linker through pharmacophore analysis of known ligands included in X-ray co-crystal structures. Further optimization using docking studies led to highly potent inhibitors 15b and 27, which furthermore showed the potent efficacy in in vivo studies. 相似文献
942.
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944.
Shuhei Takizawa Ryoki Asano Yasuhiro Fukuda Yasunori Baba Chika Tada Yutaka Nakai 《Microbial biotechnology》2022,15(6):1729-1743
Treatment with rumen fluid improves methane production from non-degradable lignocellulosic biomass during subsequent methane fermentation; however, the kinetics of xylanases during treatment with rumen fluid remain unclear. This study aimed to identify key xylanases contributing to xylan degradation and their individual activities during xylan treatment with bovine rumen microorganisms. Xylan was treated with bovine rumen fluid at 37°C for 48 h under anaerobic conditions. Total solids were degraded into volatile fatty acids and gases during the first 24 h. Zymography showed that xylanases of 24, 34, 85, 180, and 200 kDa were highly active during the first 24 h. Therefore, these xylanases are considered to be crucial for xylan degradation during treatment with rumen fluid. Metagenomic analysis revealed that the rumen microbial community’s structure and metabolic function temporally shifted during xylan biodegradation. Although statistical analyses did not reveal significantly positive correlations between xylanase activities and known xylanolytic bacterial genera, they positively correlated with protozoal (e.g., Entodinium, Diploplastron, and Eudiplodinium) and fungal (e.g., Neocallimastix, Orpinomyces, and Olpidium) genera and unclassified bacteria. Our findings suggest that rumen protozoa, fungi, and unclassified bacteria are associated with key xylanase activities, accelerating xylan biodegradation into volatile fatty acids and gases, during treatment of lignocellulosic biomass with rumen fluid. 相似文献
945.
Paecilomyces variotii IRI017 was isolated as a formaldehyde-resistant fungus from wastewater containing formaldehyde. The fungus grew in a medium
containing 0.5% formaldehyde and had consumed formaldehyde completely after 5 days. Alcohol oxidase was purified from the
fungus grown on methanol. A 20-fold purification was achieved with a yield of 44%. The molecular mass of the purified enzyme
was estimated to be 73 and 450 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration chromatography,
respectively, suggesting that the enzyme consists of six identical subunits. The N-terminal amino acid sequence of the subunit
was TIPDEVDIII. The enzyme showed an absorption spectrum typical of a flavoprotein and had a noncovalently bound flavin different
from FAD, FMN, and riboflavin. The pH optimum of the enzyme activity was pH 6–10. The enzyme was stable in the pH range of
pH 5–10. The enzyme retained full activity after incubation at 50°C for 30 min. The enzyme oxidized not only methanol but
also lower primary alcohols and formaldehyde. The K
m values for methanol, ethanol, and formaldehyde were 1.9, 3.8, and 4.9 mmol l−1, respectively. 相似文献
946.
Kazuaki Nakano Masahito Watanabe Hitomi Matsunari Taisuke Matsuda Kasumi Honda Miki Maehara Takahiro Kanai Gota Hayashida Mirina Kobayashi Momoko Kuramoto Yoshikazu Arai Kazuhiro Umeyama Shuh-hei Fujishiro Yoshihisa Mizukami Masaki Nagaya Yutaka Hanazono Hiroshi Nagashima 《PloS one》2013,8(4)
Background
The development and validation of stem cell therapies using induced pluripotent stem (iPS) cells can be optimized through translational research using pigs as large animal models, because pigs have the closest characteristics to humans among non-primate animals. As the recent investigations have been heading for establishment of the human iPS cells with naïve type characteristics, it is an indispensable challenge to develop naïve type porcine iPS cells. The pluripotency of the porcine iPS cells can be evaluated using their abilities to form chimeras. Here, we describe a simple aggregation method using parthenogenetic host embryos that offers a reliable and effective means of determining the chimera formation ability of pluripotent porcine cells.Methodology/Significant Principal Findings
In this study, we show that a high yield of chimeric blastocysts can be achieved by aggregating the inner cell mass (ICM) from porcine blastocysts with parthenogenetic porcine embryos. ICMs cultured with morulae or 4–8 cell-stage parthenogenetic embryos derived from in vitro-matured (IVM) oocytes can aggregate to form chimeric blastocysts that can develop into chimeric fetuses after transfer. The rate of production of chimeric blastocysts after aggregation with host morulae (20/24, 83.3%) was similar to that after the injection of ICMs into morulae (24/29, 82.8%). We also found that 4–8 cell-stage embryos could be used; chimeric blastocysts were produced with a similar efficiency (17/26, 65.4%). After transfer into recipients, these blastocysts yielded chimeric fetuses at frequencies of 36.0% and 13.6%, respectively.Conclusion/Significance
Our findings indicate that the aggregation method using parthenogenetic morulae or 4–8 cell-stage embryos offers a highly reproducible approach for producing chimeric fetuses from porcine pluripotent cells. This method provides a practical and highly accurate system for evaluating pluripotency of undifferentiated cells, such as iPS cells, based on their ability to form chimeras. 相似文献947.
Feather corticosterone reveals stress associated with dietary changes in a breeding seabird 下载免费PDF全文
Alexis Will Yutaka Watanuki Dale M. Kikuchi Nobuhiko Sato Motohiro Ito Matt Callahan Katherine Wynne‐Edwards Scott Hatch Kyle Elliott Leslie Slater Akinori Takahashi Alexander Kitaysky 《Ecology and evolution》2015,5(19):4221-4232
Changes in climate and anthropogenic pressures might affect the composition and abundance of forage fish in the world's oceans. The junk‐food hypothesis posits that dietary shifts that affect the quality (e.g., energy content) of food available to marine predators may impact their physiological state and consequently affect their fitness. Previously, we experimentally validated that deposition of the adrenocortical hormone, corticosterone, in feathers is a sensitive measure of nutritional stress in seabirds. Here, we use this method to examine how changes in diet composition and prey quality affect the nutritional status of free‐living rhinoceros auklets (Cerorhinca monocerata). Our study sites included the following: Teuri Is. Japan, Middleton Is. central Gulf of Alaska, and St. Lazaria Is. Southeast Alaska. In 2012 and 2013, we collected “bill loads” delivered by parents to feed their chicks (n = 758) to document dietary changes. We deployed time–depth–temperature recorders on breeding adults (n = 47) to evaluate whether changes in prey coincided with changes in foraging behavior. We measured concentrations of corticosterone in fledgling (n = 71) and adult breeders' (n = 82) feathers to determine how birds were affected by foraging conditions. We found that seasonal changes in diet composition occurred on each colony, adults dove deeper and engaged in longer foraging bouts when capturing larger prey and that chicks had higher concentrations of corticosterone in their feathers when adults brought back smaller and/or lower energy prey. Corticosterone levels in feathers of fledglings (grown during the breeding season) and those in feathers of adult breeders (grown during the postbreeding season) were positively correlated, indicating possible carryover effects. These results suggest that seabirds might experience increased levels of nutritional stress associated with moderate dietary changes and that physiological responses to changes in prey composition should be considered when evaluating the effect of prey quality on marine predators. 相似文献
948.
Yasuka Toda Tomoya Nakagita Takashi Hayakawa Shinji Okada Masataka Narukawa Hiroo Imai Yoshiro Ishimaru Takumi Misaka 《The Journal of biological chemistry》2013,288(52):36863-36877
Umami taste perception in mammals is mediated by a heteromeric complex of two G-protein-coupled receptors, T1R1 and T1R3. T1R1/T1R3 exhibits species-dependent differences in ligand specificity; human T1R1/T1R3 specifically responds to l-Glu, whereas mouse T1R1/T1R3 responds more strongly to other l-amino acids than to l-Glu. The mechanism underlying this species difference remains unknown. In this study we analyzed chimeric human-mouse receptors and point mutants of T1R1/T1R3 and identified 12 key residues that modulate amino acid recognition in the human- and mouse-type responses in the extracellular Venus flytrap domain of T1R1. Molecular modeling revealed that the residues critical for human-type acidic amino acid recognition were located at the orthosteric ligand binding site. In contrast, all of the key residues for the mouse-type broad response were located at regions outside of both the orthosteric ligand binding site and the allosteric binding site for inosine-5′-monophosphate (IMP), a known natural umami taste enhancer. Site-directed mutagenesis demonstrated that the newly identified key residues for the mouse-type responses modulated receptor activity in a manner distinct from that of the allosteric modulation via IMP. Analyses of multiple point mutants suggested that the combination of two distinct determinants, amino acid selectivity at the orthosteric site and receptor activity modulation at the non-orthosteric sites, may mediate the ligand specificity of T1R1/T1R3. This hypothesis was supported by the results of studies using nonhuman primate T1R1 receptors. A complex molecular mechanism involving changes in the properties of both the orthosteric and non-orthosteric sites of T1R1 underlies the determination of ligand specificity in mammalian T1R1/T1R3. 相似文献
949.
Mari Nekohashi Mana Ogawa Takuo Ogihara Kyoko Nakazawa Hisanori Kato Takumi Misaka Keiko Abe Shoko Kobayashi 《PloS one》2014,9(5)
Niemann–Pick C1-Like 1 (NPC1L1) mediates cholesterol absorption, and ezetimibe is a potent NPC1L1 inhibitor applicable for medication of hypercholesterolemia. Epidemiological studies demonstrated that consumption of polyphenols correlates with a decreased risk for atherosclerosis due to their antioxidant effect. This activity can hardly be attributable to the antioxidant activity only, and we hypothesized that polyphenols inhibit intestinal transport of cholesterol. We elucidated the kinetic parameters of intestinal cholesterol absorption, screened several polyphenols for their ability to specifically inhibit intestinal cholesterol absorption, and determined the inhibitory effects of selected flavonoids in vitro and in vivo. The concentration-dependent uptake of cholesterol by Caco-2 cells obeyed a monophasic saturation process. This indicates the involvement of an active-passive transport, i.e., NPC1L1. Parameters of cholesterol uptake by Caco-2 cells were as follows: J
max, K
t, and K
d were 6.89±2.96 19.03±11.58 µM, and 0.11±0.02 pmol/min/mg protein, respectively. Luteolin and quercetin inhibited cholesterol absorption by Caco-2 cells and human embryonic kidney 293T cells expressing NPC1L1. When preincubated Caco-2 cells with luteolin and quercetin before the assay, cholesterol uptake significantly decreased. The inhibitory effects of these flavonoids were maintained for up to 120 min. The level of inhibition and irreversible effects were similar to that of ezetimibe. Serum cholesterol levels significantly decreased more in rats fed both cholesterol and luteolin (or quercetin), than in those observed in the cholesterol feeding group. As quercetin induced a significant decrease in the levels of NPC1L1 mRNA in Caco-2 cells, the in vivo inhibitory effect may be due to the expression of NPC1L1. These results suggest that luteolin and quercetin reduce high blood cholesterol levels by specifically inhibiting intestinal cholesterol absorption mediated by NPC1L1. 相似文献
950.