Differentiation of canine bone marrow stromal cells into voltage- and glutamate-responsive neuron-like cells by basic fibroblast growth factor     

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   

Detecting endotoxin activity in bovine serum using an automated testing system     

Kazuyuki SUZUKI  Toshio SHIMAMORI  Ayano SATO  Kenji TSUKANO  Masakazu TSUCHIYA  Jeffrey LAKRITZ 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(8):977-979

The aim of the present study was to compare the ability of the commercially available portable test system (PTS^TM) to detect endotoxin activity in bovine serum, with that of the traditional LAL-kinetic turbidimetric (KT) and chromogenic (KC) assays. Prior to testing, serum samples, which were obtained from endotoxin-challenged cattle, were diluted 1:20 in endotoxin-free water and heated to 80°C for 10 min. The performance of the PTS^TM was not significantly different from that of the traditional LAL-based assays. The results using PTS^TM correlated with those using KT (r²=0.963, P<0.001) or KC assays (r²=0.982, P<0.001). Based on these findings, the PTS^TM could be applied as a simplified system to assess endotoxin activity in bovine serum.  相似文献   

Detection of the staphylococcal enterotoxin D-like gene from staphylococcal food poisoning isolates over the last two decades in Tokyo     

Yasunori SUZUKI  Makiko KOBAYASHI  Shigeru MATSUSHITA  Satomi UEHARA  Rei KATO  Yusuke SATO’O  Hisaya K. ONO  Kenji SADAMASU  Akemi KAI  Yoichi KAMATA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(8):905-911

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Pathogenicity of emerging Japanese type 1 porcine reproductive and respiratory syndrome virus in experimentally infected pigs     

Hiroshi ISEKI  Michihiro TAKAGI  Kenji KAWASHIMA  Tomoyuki SHIBAHARA  Yoshiko KURODA  Hiroshi TSUNEMITSU  Makoto YAMAKAWA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(12):1663-1666

To clarify the pathogenicity of Japanese type 1 porcine reproductive and respiratory syndrome virus (PRRSV) isolate in experimentally infected pigs, we evaluated clinical signs and monitored viremia for 21 days post-inoculation (dpi). Lungs were mottled, tanned and reddish in appearance; had lesions predominantly in the cranial, middle and accessory lobes; and failed to collapse at 10 dpi. Although microscopic lesions of lungs were reproduced using the Japanese emerging type 1 PRRSV isolate under experimental conditions, no significant differences were noted between the challenge and control groups regarding mean rectal temperature and daily weight gain. These results provide useful insights into the limited pathogenicity of single infection with the Japanese type 1 PRRSV isolate in piglets, which differ from findings in reported field cases.  相似文献   

Hypoxia inducible factor 1α expression and effects of its inhibitors in canine lymphoma     

Satoshi KAMBAYASHI  Masaya IGASE  Kosuke KOBAYASHI  Ayana KIMURA  Takako SHIMOKAWA MIYAMA  Kenji BABA  Shunsuke NOGUCHI  Takuya MIZUNO  Masaru OKUDA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(11):1405-1412

Hypoxic conditions in various cancers are believed to relate with their malignancy, and hypoxia inducible factor-1α (HIF-1α) has been shown to be a major regulator of the response to low oxygen. In this study, we examined HIF-1α expression in canine lymphoma using cell lines and clinical samples and found that these cells expressed HIF-1α. Moreover, the HIF-1α inhibitors, echinomycin, YC-1 and 2-methoxyestradiol, suppressed the proliferation of canine lymphoma cell lines. In a xenograft model using NOD/scid mice, echinomycin treatment resulted in a dose-dependent regression of the tumor. Our results suggest that HIF-1α contributes to the proliferation and/or survival of canine lymphoma cells. Therefore, HIF-1α inhibitors may be potential agents to treat canine lymphoma.  相似文献   

Different effects of an extended photoperiod treatment on growth,gonadal function,and condition of hair coats in Thoroughbred yearlings reared under different climate conditions     

Tsuyoshi SUZUKI  Hirotoshi MIZUKAMI  Yasuo NAMBO  Mutsuki ISHIMARU  Kenji MIYATA  Kentaro AKIYAMA  Kenji KOROSUE  Hiroshi NAITO  Kentaro NAGAOKA  Gen WATANABE  Kazuyoshi TAYA 《Journal of Equine Science》2015,26(4):113-124

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Investigation of a new natural adhesive composed of citric acid and sucrose for particleboard II: effects of board density and pressing temperature     

Kenji Umemura  Osamu Sugihara  Shuichi Kawai 《Journal of Wood Science》2015,61(1):40-44

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Papaya seed represents a rich source of biologically active isothiocyanate   总被引：1，自引：0，他引：1  

Nakamura Y  Yoshimoto M  Murata Y  Shimoishi Y  Asai Y  Park EY  Sato K  Nakamura Y 《Journal of agricultural and food chemistry》2007,55(11):4407-4413

In the present study, papaya (Carica papaya) seed and edible pulp were carefully separated and then the contents of benzyl isothiocyanate and the corresponding glucosinolate (benzyl glucosinolate, glucotropaeolin) quantified in each part. The papaya seed with myrosinase inactivation contained >1 mmol of benzyl glucosinolate in 100 g of fresh seed. This content is equivalent to that of Karami daikon (the hottest Japanese white radish) or that of cress. The papaya seed extract also showed a very high activity of myrosinase and, without myrosinase inactivation, produced 460 micromol of benzyl isothiocyanate in 100 g of seed. In contrast, papaya pulp contained an undetectable amount of benzyl glucosinolate and showed no significant myrosinase activity. The n-hexane extract of the papaya seed homogenate was highly effective in inhibiting superoxide generation and apoptosis induction in HL-60 cells, the activities of which are comparable to those of authentic benzyl isothiocyanate.  相似文献   

Identification of an allele attributable to formation of cucumber-like flavor in wild tomato species (Solanum pennellii) that was inactivated during domestication     

Matsui K  Ishii M  Sasaki M  Rabinowitch HD  Ben-Oliel G 《Journal of agricultural and food chemistry》2007,55(10):4080-4086

Carbon 6 (C6)-aldehydes formed by fatty acid 13-hydroperoxide lyase (13HPL) specific to fatty acid 13-hydroperoxides (13-HPO) are important flavor constituents in fresh tomato fruits. C9-aldehydes are usually formed by 9/13HPL showing dual specificity to 9- and 13-HPOs and are scarcely found in tomato fruits. Mature red fruits of one of the introgression lines, IL1-4, generated by hybridization of a cultivated tomato (Solanum lycopersicon) to its wild relative Solanum pennellii, form high amounts of C9-aldehydes upon homogenization. The IL1-4 fruits showed high 9/13HPL activity. One of the genes isolated from IL1-4 showed a high similarity to plant 9/13HPLs. Recombinant proteins expressed in Escherichia coli showed 9/13HPL activity. Cleaved amplified polymorphic sequence analyses indicated that the gene was specific to IL1-4 and S. pennellii. S. lycopersicon had a gene having high similarity to the S. pennellii gene. It was absent in IL1-4. Among the differences of amino acid residues found between the two genes, a Cys to Ser exchange may be responsible for the inactivation of resultant protein product of S. lycopersicon gene because the Cys is an essential amino acid residue for HPL activity. From these observations, it could be assumed that a tomato gene corresponding to S. pennellii 9/13HPL gene had been inactivated through domestication of tomatoes.  相似文献   

Pathogenicity of Pepper mild mottle virus Is Controlled by the RNA Silencing Suppression Activity of Its Replication Protein but Not the Viral Accumulation     

Tsuda S  Kubota K  Kanda A  Ohki T  Meshi T 《Phytopathology》2007,97(4):412-420

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