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1.
Rei NAKANO Kazuya EDAMURA Tomohiro NAKAYAMA Kenji TESHIMA Kazushi ASANO Takanori NARITA Ken OKABAYASHI Hiroshi SUGIYA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(1):27-35
We investigated the in
vitro differentiation of canine bone marrow stromal cells (BMSCs) into voltage-
and glutamate-responsive neuron-like cells. BMSCs were obtained from the bone marrow of
healthy beagle dogs. Canine BMSCs were incubated with the basal medium for neurons
containing recombinant human basic fibroblast growth factor (bFGF; 100
ng/ml). The viability of the bFGF-treated cells was
assessed by a trypan blue exclusion assay, and the morphology was monitored. Real-time
RT-PCR was performed to evaluate mRNA expression of neuronal, neural stem cell and glial
markers. Western blotting and immunocytochemical analysis for the neuronal markers were
performed to evaluate the protein expression and localization. The Ca2+
mobilization of the cells was evaluated using the Ca2+ indicator Fluo3 to
monitor Ca2+ influx. To investigate the mechanism of bFGF-induced neuronal
differentiation, the fibroblast growth factor receptor inhibitor, the phosphoinositide
3-kinase inhibitor or the Akt inhibitor was tested. The bFGF treatment resulted in the
maintenance of the viability of canine BMSCs for 10 days, in the expression of neuronal
marker mRNAs and proteins and in the manifestation of neuron-like morphology. Furthermore,
in the bFGF-treated BMSCs, a high concentration of KCl and L-glutamate induced an increase
in intracellular Ca2+ levels. Each inhibitor significantly attenuated the
bFGF-induced increase in neuronal marker mRNA expression. These results suggest that bFGF
contributes to the differentiation of canine BMSCs into voltage- and glutamate-responsive
neuron-like cells and may lead to the development of new cell-based treatments for
neuronal diseases. 相似文献
2.
HEISUKE NAKAGAWA TETSUYA UMINO TAKASHI SEKIMOTO IRFAN AMBAS W LINN MONTGOMERY TAKETO NAKANO 《Fisheries Science》2002,68(2):341-346
ABSTRACT: The gut and gut contents of ayu Plecoglossus altivelis caught in the Ohta River in Hiroshima prefecture, Japan were examined. Relative intestine length (intestine length/body length) was almost constant. Except for relative intestine length, characteristics of the gut of wild ayu were not significantly different from those of cultured ayu. The height of intestinal folds increased and their width declined from stomach to anus. Cyanobacteria ( Homoeathrix sp. and Calothrix sp.) and diatoms ( Cymbella sp., Gomphonema sp., Melosira sp., Navicula sp., and Synedra sp.) were well digested during passage through the gut despite a lack of apparent destruction. In contrast, green algae ( Dictyosphearium sp., Coelastrum sp., and Pediastrum sp.) exhibited little degradation. The pH of stomach contents ranged from 2.8 to 7.4 (mean ± SD = 4.1 ± 1.0), whereas that of the posterior intestine was 7.0–8.5 (7.9 ± 0.4). Gut contents of wild ayu increased with bodyweight, and were more than threefold greater than those of cultured ayu, suggesting that wild ayu compensate for low nutrient content of algae through the continuous ingestion of large quantities of feed organisms. 相似文献
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4.
WULIJIDELIGEN Takayuki ASAHINA Kazushi HARA Kensuke ARAKAWA Hiroyuki NAKANO Taku MIYAMOTO 《Animal Science Journal》2012,83(10):704-711
The purification and characterization of a bacteriocin produced by Leuconostoc mesenteroides strain 406 that was isolated from traditional Mongolian fermented mare's milk, airag, were carried out. Leuconostoc mesenteroides strain 406 was identified on the basis of its morphological and biochemical characteristics and carbohydrate fermentation profile and by API 50 CH kit and 16S ribosomal DNA analyses. The neutral‐pH cell‐free supernatant of this bacterium inhibited the growth of several lactic acid bacteria and food spoilage and pathogenic organisms, including Listeria monocytogenes and Clostridium botulinum. The bacteriocin was heat‐stable and not sensitive to acid and alkaline conditions, but was sensitive to several proteolytic enzymes such as pepsin, pronase E, proteinase K, trypsin, and α‐chymotrypsin, but not catalase. Optimum bacteriocin production (4000 activity units/mL) was achieved when the strain was cultured at 25°C for 24–36 h in Man Rogosa Sharpe medium. The bacteriocin was partially purified by ammonium sulfate precipitation (80% saturation), dialysis (cut‐off MW: 1000), and gel filtration chromatography. Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis revealed that the bacteriocin had a molecular weight of approximately 3.3 kDa. To our knowledge, this is the first report of the isolation of a bacteriocin‐producing Leuconostoc strain from airag. An application to fermented milks would be desired. 相似文献
5.
MICHIO AOYAMA HIROHITO GOTO HITOMI KAMIYA IKUO KANEKO SATOSHI KAWAE HIROKI KODAMA YASUSHI KONISHI KENN-ICHI KUSUMOTO HISATO MIURA EIICHI MORIYAMA KIYOSHI MURAKAMI TOSHIYA NAKANO FUTOSHI NOZAKI DAISUKE SASANO TAKAO SHIMIZU HIROKI SUZUKI YASUSHI TAKATSUKI AKITO TORIYAMA 《Fisheries Oceanography》2008,17(3):206-218
Impact of climate change on marine biogeochemical parameters and ecosystem is one of the important issues of our environment. Direct evidence of marine pelagic ecosystem changes is found with warming of sea water and sea‐level rise in the main stream of the Kuroshio in the East China Sea and the western North Pacific during these three decades based on the analysis of long‐term comprehensive hydrographic observations. In terms of annual mean, the warming rate of surface air temperature and sea surface temperature ranged from 0.15 to 0.21°C per decade in and around the main stream of the Kuroshio in the East China Sea, which exceed the global mean warming rate of 0.128 ± 0.026°C per decade during the period from 1956 to 2005 reported in IPCC 2007. One of the features in this rapid warming region is an increase of number of Pterosagitta draco, a cosmopolitan warm‐water zooplankton. Biogeochemical parameters, such as wet weight of zooplankton, plant pigment and nutrients concentration in the upper 200 m have been decreasing while dissolved oxygen content and seawater temperature have been increasing in the upper 200 m in the main stream of the Kuroshio in the East China Sea. These observed linear trends of the biogeochemical parameters would be foresights for temperate oceans in the future. 相似文献
6.
Hitomi MATSUNARI Toshihiro KOBAYASHI Masahito WATANABE Kazuhiro UMEYAMA Kazuaki NAKANO Takahiro KANAI Taisuke MATSUDA Masaki NAGAYA Manami HARA Hiromitsu NAKAUCHI Hiroshi NAGASHIMA 《The Journal of reproduction and development》2014,60(3):230-237
The development and regeneration of the pancreas is of considerable interest because of the role of these processes in
pancreatic diseases, such as diabetes. Here, we sought to develop a large animal model in which the pancreatic cell lineage
could be tracked. The pancreatic and duodenal homeobox-1 (Pdx1) gene promoter was conjugated to Venus, a
green fluorescent protein, and introduced into 370 in vitro-matured porcine oocytes by intracytoplasmic
sperm injection-mediated gene transfer. These oocytes were transferred into four recipient gilts, all of which became
pregnant. Three gilts were sacrificed at 47–65 days of gestation, and the fourth was allowed to farrow. Seven of 16 fetuses
obtained were transgenic (Tg) and exhibited pancreas-specific green fluorescence. The fourth recipient gilt produced a litter
of six piglets, two of which were Tg. The founder Tg offspring matured normally and produced healthy first-generation (G1)
progeny. A postweaning autopsy of four 27-day-old G1 Tg piglets confirmed the pancreas-specific Venus expression.
Immunostaining of the pancreatic tissue indicated the transgene was expressed in β-cells. Pancreatic islets from Tg pigs were
transplanted under the renal capsules of NOD/SCID mice and expressed fluorescence up to one month after transplantation. Tg
G1 pigs developed normally and had blood glucose levels within the normal range. Insulin levels before and after sexual
maturity were within normal ranges, as were other blood biochemistry parameters, indicating that pancreatic function was
normal. We conclude that Pdx1-Venus Tg pigs represent a large animal model suitable for research on
pancreatic development/regeneration and diabetes. 相似文献
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Kazuhiro UMEYAMA Kasumi HONDA Hitomi MATSUNARI Kazuaki NAKANO Tatsuro HIDAKA Keito SEKIGUCHI Hironori MOCHIZUKI Yasuhiro TAKEUCHI Tsukasa FUJIWARA Masahito WATANABE Masaki NAGAYA Hiroshi NAGASHIMA 《The Journal of reproduction and development》2013,59(6):599-603
Somatic cell nuclear transfer (SCNT) is a useful technique for creating pig strains
that model human diseases. However, production of numerous cloned disease model pigs
by SCNT for large-scale experiments is impractical due to its complexity and
inefficiency. In the present study, we aimed to establish an efficient procedure for
proliferating the diabetes model pig carrying the mutant human hepatocyte nuclear
factor-1α gene. A founder diabetes transgenic cloned pig was generated by SCNT and
treated with insulin to allow for normal growth to maturity, at which point
epididymal sperm could be collected for cryopreservation. In vitro
fertilization and intrafallopian insemination using the cryopreserved epididymal
sperm resulted in diabetes model transgenic offspring. These results suggest that
artificial reproductive technology using cryopreserved epididymal sperm could be a
practical option for proliferation of genetically modified disease model pigs. 相似文献
9.
ABSTRACT: We examined the stomach contents of 26 Baird's beaked whales caught off the coast of Japan by small-type coastal whalers. The main prey for these whales was rat-tails and hakes in the western North Pacific. Pollock and squids were also important food in the whales collected from the southern Sea of Okhotsk. The prey species found in the stomachs of the whales were almost identical to those caught in bottom-trawl nets at depths greater than about 1000 m in the western North Pacific, which suggests that the Baird's beaked whale forages for prey at depths of about 1000 m or more. Baird's beaked whales in the western North Pacific migrate to waters of 1000–3000 m in depth, where demersal fish are abundant. This implies that Baird's beaked whales migrate to waters where demersal fish, especially rat-tails and hakes, are abundant. Although there is limited information on the feeding habits of ziphiid whales, they are generally thought to prefer squid. The present data suggest that demersal fish are also important prey for ziphiid whales. 相似文献
10.
Masahito WATANABE Mirina KOBAYASHI Masaki NAGAYA Hitomi MATSUNARI Kazuaki NAKANO Miki MAEHARA Gota HAYASHIDA Shuko TAKAYANAGI Rieko SAKAI Kazuhiro UMEYAMA Nobuyuki WATANABE Masafumi ONODERA Hiroshi NAGASHIMA 《The Journal of reproduction and development》2015,61(3):169-177
Monomeric Plum (Plum), a far-red fluorescent protein with photostability and photopermeability, is potentially suitable for in vivo imaging and detection of fluorescence in body tissues. The aim of this study was to generate transgenic cloned pigs exhibiting systemic expression of Plum using somatic cell nuclear transfer (SCNT) technology. Nuclear donor cells for SCNT were obtained by introducing a Plum-expression vector driven by a combination of the cytomegalovirus early enhancer and chicken beta-actin promoter into porcine fetal fibroblasts (PFFs). The cleavage and blastocyst formation rates of reconstructed SCNT embryos were 81.0% (34/42) and 78.6% (33/42), respectively. At 36–37 days of gestation, three fetuses systemically expressing Plum were obtained from one recipient to which 103 SCNT embryos were transferred (3/103, 2.9%). For generation of offspring expressing Plum, rejuvenated PFFs were established from one cloned fetus and used as nuclear
donor cells. Four cloned offspring and one stillborn cloned offspring were produced from one recipient to which 117 SCNT embryos were transferred (5/117, 4.3%). All offspring exhibited high levels of Plum fluorescence in blood cells, such as lymphocytes, monocytes and granulocytes. In addition, the skin, heart, kidney, pancreas, liver and spleen also exhibited Plum expression. These observations demonstrated that transfer of the Plum gene did not interfere with the development of porcine SCNT embryos and resulted in the successful generation of transgenic cloned pigs that systemically expressed Plum. This is the first report of the generation and characterization of transgenic cloned pigs expressing the far-red fluorescent protein Plum. 相似文献