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1.
Osamu Yamato Hiroyuki Satoh Naoaki Matsuki Kenichiro Ono Masahiro Yamasaki Yoshimitsu Maede 《Journal of veterinary diagnostic investigation》2004,16(1):39-44
In the present study, laboratory techniques were used to diagnose canine GM2-gangliosidosis using blood and cerebrospinal fluid (CSF) that can be collected noninvasively from living individuals. Lysosomal acid beta-hexosaminidase (Hex) was measured spectrofluorometrically using 4-methylumbelliferyl N-acetyl-beta-D-glucosaminide and 4-methylumbelliferyl 7-(6-sulfo-2-acetamido-2-deoxy-beta-D-glucopyranoside) as substrates. Main isoenzymes A and B of Hex in leukocytes were also analyzed using cellulose acetate membrane electrophoresis. GM2-ganglioside in CSF was detected and determined quantitatively by using thin-layer chromatography/enzyme-immunostaining method with anti-GM2-ganglioside antibody. In normal dogs, Hex activities could be determined in leukocytes, serum, and CSF and the total activities were markedly reduced in all the enzyme sources in a dog with Sandhoff disease. Electrophoresis of a leukocyte lysate from a normal dog showed that the Hex A and Hex B were not separated distinctively with formation of a broad band, whereas there were no bands in electrophoresis of a lysate from a dog with Sandhoff disease, showing a deficiency in the total enzyme activity. GM2-ganglioside could be detected and determined quantitatively in as little as 100 microl of canine CSE GM2-ganglioside in CSF in a dog with Sandhoff disease increased to 46 times the normal level. In conclusion, the methods in the present study are useful for diagnosis of canine GM2-gangliosidosis. These techniques enable definitive and early diagnosis of canine GM2-gangliosidosis even if tissues and organs cannot be obtained. 相似文献
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Chie SUZUKI Yosuke SAKAGUCHI Hiroyoshi HOSHI Koji YOSHIOKA 《The Journal of reproduction and development》2016,62(1):79-86
The effects of lipid-rich bovine serum albumin (LR-BSA) on the development of porcine blastocysts produced
in vitro were examined. Addition of 0.5 to 5 mg/ml LR-BSA to porcine blastocyst medium
(PBM) from Day 5 (Day 0 = in vitro fertilization) significantly increased the hatching rates
of blastocysts on Day 7 and the total cell numbers in Day-7 blastocysts. When Day-5 blastocysts were cultured
with PBM alone, PBM containing LR-BSA, recombinant human serum albumin or fatty acid-free BSA, addition of
LR-BSA significantly enhanced hatching rates and the cell number in blastocysts that survived compared with
other treatments. The diameter, ATP content and numbers of both inner cell mass and total cells in Day-6 and
Day-7 blastocysts cultured with PBM containing LR-BSA were significantly higher than in blastocysts cultured
with PBM alone, whereas LR-BSA had no effect on mitochondrial membrane potential. The mRNA levels of enzymes
involved in fatty acid metabolism and β-oxidation (ACSL1, ACSL3,
CPT1, CPT2 and KAT) in Day-7 blastocysts were
significantly upregulated by the addition of LR-BSA. The results indicated that LR-BSA enhanced hatching
ability and quality of porcine blastocysts produced in vitro, as determined by ATP content,
blastocyst diameter and expression levels of the specific genes, suggesting that the stimulatory effects of
LR-BSA arise from lipids bound to albumin. 相似文献
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Tamura K Arai H Ueno E Saito C Yagihara H Isotani M Ono K Washizu T Bonkobara M 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2007,69(9):925-930
Dendritic cell (DC) vaccination is one of the most attractive immunotherapies for malignancies in dogs. To examine the differences in DC-mediated immune responses from different types of malignancies in dogs, we vaccinated dogs using autologous DCs pulsed with keyhole limpet hemocyanin (KLH) and cell lysate prepared from squamous cell carcinoma SCC2/88 (SCC-KLH-DC), histiocytic sarcoma CHS-5 (CHS-KLH-DC), or B cell leukemia GL-1 (GL-KLH-DC) in vitro. In vivo inductions of immune responses against these tumor cells were compared by the delayed-type hypersensitivity (DTH) skin test. The DTH response against SCC2/88 cells were observed in dogs vaccinated with autologous SCC-KLH-DC, while the response was undetectable against CHS-5 and GL-1 cells in dogs vaccinated with autologous CHS-KLH-DC and GL-KLH-DC. Skin biopsies taken from DTH challenge sites were then examined for immunohistochemistry, and recruitment of CD8 and CD4 T cells was detected at the site where SCC2/88 cells were inoculated in dogs vaccinated with SCC-KLH-DC. By contrast, neither CD8 nor CD4 T cell infiltration was found at the DTH challenge site in the dogs vaccinated with CHS-KLH-DC or GL-KLH-DC. These findings may reflect that the efficacy of immune induction by DC vaccination varies among tumor types and that immune responses could be inducible in squamous cell carcinoma. Our results encouraged further investigation of therapeutic vaccination for dogs with advanced squamous cell carcinoma in clinical trials. 相似文献
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Atsushi IDETA Yoshito AOYAGI Kanami TSUCHIYA Yuuki NAKAMURA Kou HAYAMA Atsushi SHIRASAWA Kenichiro SAKAGUCHI Naomi TOMINAGA Yoshiyuki NISHIMIYA Sakae TSUDA 《The Journal of reproduction and development》2015,61(1):1-6
Embryos obtained via superovulation are necessary for mammalian artificial reproduction, and viability is a key determinant of success. Nonfreezing storage at 4 C is possible, but currently used storage solutions can maintain embryo viability for only 24–48 h. Here we found that 10 mg/ml antifreeze protein (AFP) dissolved in culture medium 199 with 20% (v/v) fetal bovine serum and 25 mM HEPES could keep bovine embryos alive for 10 days at 4 C. We used a recombinant AFP isolated from the notched-fin eelpout (Zoarces elongatus Kner). Photomicroscopy indicated that the AFP–embryo interaction was enhanced at 37 C. Embryos pre-warmed with the AFP solution at 37 C for 60 min maintained high viability, whereas those that were not pre-warmed could live no longer than 7 days. Thus, short-term storage of bovine embryos was achieved by a combination of AFP-containing medium and controlled pre-warming. 相似文献
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Several lines of evidence implicate small spirochetes, presumably treponemata, as etiologic agents in the production of the maternally transmitted "sex ratio" condition (SR) in Drosophila nebulosa, in D. willistoni, and in strains of D. melanogaster into which the SR condition has been artificially transferred. The presence of treponemata in the hemolymph of adult females of these species is completely correlated with the production of unisexual progenies and like this condition is dependent on the genotype of the host and of the infectious agent. 相似文献
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Glucose uptake activity in murine red blood cells infected with Babesia microti and Babesia rodhaini
Ohmori T Adachi K Fukuda Y Tamahara S Matsuki N Ono K 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2004,66(8):945-949
The glucose uptake activity in Babesia rodhaini and B. microti - infected red blood cell (IRBC) was investigated in mice using 2-deoxy-D-glucose (2DOG) and L-glucose (L-Glc), a non-metabolizable analogue of D-glucose and non-incorporative glucose to non-infected RBC (NRBC), respectively. The uptake activities of both DOG and L-Glc were higher in IRBCs than those in NRBC. The concentration dependent uptake of 2DOG and L-Glc in both IRBC revealed a linear curve, indicating non-transporter mediated uptake. In addition, B. microti IRBC showed higher 2DOG uptake than B. rodhaini IRBC, whereas no difference was observed in L-Glc uptake. These results indicated that some new glucose uptake system, at least two systems, developed in both IRBC. The new systems were sodium independent, non-competitive to L-Glc, and sensitive to temperature. One of two systems had no kinetical difference between B. rodhaini and B. microti IRBC, however another one might have higher uptake activity in B. microti IRBC compared to that in B. rodhaini IRBC. 相似文献