Development and characterization of monoclonal antibodies against canine trypsin     

Takaki Waritani  Yoko Okuno  Yoshinori Ashida  Ryo Tsuchiya  Kosaku Kobayashi  Takatsugu Yamada   《Veterinary immunology and immunopathology》2001,80(3-4):333-338

Canine cationic trypsin was purified by salting-out, gel filtration and affinity chromatography. Purity was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight was ca. 28 kDa by SDS-PAGE.

Thirty hybridomas were obtained which produced mAb to canine cationic trypsin by the cell fusion technique. Twenty-two of these recognized cationic trypsin only, while eight hybridomas recognized both cationic and anionic trypsin. Several of the anti-canine cationic trypsin mAb were purified by salting-out and DEAE ion-change chromatography using ascites fluid of immunized BALB/c mice. The mAb proved to have very high specificity to canine cationic trypsin as shown by immunoblotting and it may be possible to use them to develop clinical assays.  相似文献   

Localization of extracellular matrix receptors in ICGN mice, a strain of mice with hereditary nephrotic syndrome.     

K Uchio-Yamada  N Manabe  M Yamaguchi  N Akashi  Y Goto  Y Yamamoto  A Ogura  H Miyamoto 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2001,63(11):1171-1178

Fibrotic degeneration was examined in the kidneys of ICR-derived glomerulonephritis (ICGN) mice, a novel inbred mouse line with a hereditary nephrotic syndrome of unknown etiology considered to be a good model of human idiopathic nephrotic syndrome. In the present study, we histochemically revealed changes in accumulation of extracellular matrix (ECM) components and in localization of integrins, cellular receptors for ECM, in the kidneys of ICGN mice with the progression of renal failure. Excessive accumulation of basement membrane (laminin and collagen IV) and interstitial (type III collagen) ECM components were demonstrated in the glomeruli and tubulointerstitum of ICGN mice. Marked deposition of type I collagen and tenascin was seen only in the glomeruli of ICGN mice but not in those of ICR mice as normal controls. Increased expression of integrin alpha1-, alpha2-, alpha5- and beta1-subunits in glomeruli with fibrotic degeneration and abnormal distribution of alpha6-subunit were noted in the kidneys of ICGN mice. Excessive laminin, a ligand of alpha6beta1-integrin, was demonstrated on the tubular basement membrane, but alpha6-subunit diffusely disappeared on the basal side of the tubular epithelial cells. We presumed that abnormal integrin expression in renal tubules causes epithelial cell detachment, and consequently tubular nephropathy, and results in disorder of ECM metabolism causing excessive accumulation of ECM components in the kidneys of ICGN mice.  相似文献   

Participation of H1-receptors in histamine-induced contraction and relaxation of horse coronary artery in vitro.     

T Obi  A Miyamoto  M Matumoto  S Ishiguro  A Nishio 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》1991,53(5):789-795

The mechanisms of histamine-induced contraction and relaxation were investigated in rings isolated from a middle part of the left descending coronary arteries of horses. Intact and endothelium-denuded preparations were compared. Rings of horse coronary arteries contracted in response to histamine in a concentration dependent manner, but some of them relaxed with lower concentrations and contracted with higher concentrations. Removal of the endothelium abolished the relaxation and potentiated the contraction. The pD2 values were 4.70 +/- 0.08 in the rings with intact endothelium and 4.95 +/- 0.08 in endothelium-denuded rings. Histamine-induced contractions in intact and denuded preparations were not affected by an H2-antagonist, cimetidine, but were inhibited by an H1-antagonist, diphenhydramine in non-competitive manner in the rings with endothelium and in competitive manner in denuded rings. After precontraction with PGF2 alpha or norepinephrine, histamine relaxed preparations with intact endothelium (pD2 value, 7.80 +/- 0.11), although histamine-induced relaxations were not observed in denuded preparations. The relaxation was competitively inhibited by diphenhydramine. Relaxing response was significantly attenuated by methylene blue, quinacrine, L-nitro-arginine, gossypol and AA861 but not by indomethacin. These results suggest that the histamine-induced contraction and relaxation in horse coronary arteries are mediated mainly by H1-receptors in the smooth muscle and endothelium, respectively, and H1-receptor activation of endothelial cells may liberate vasodilator substances.  相似文献   

Purification and properties of pyrethroid carboxyesterase in rat liver microsome     

Takashi Suzuki  Junshi Miyamoto 《Pesticide biochemistry and physiology》1978,8(2):186-198

Pyrethroid carboxyesterase which hydrolyzes the esters of chrysanthemumic acid was purified from rat liver microsome by cholic acid solubilization, ammonium sulfate fractionation, heat treatment, and DEAE-Sephadex A-50 column chromatography. The 45-fold purified enzyme (38% yield) is likely to consist of single protein, as evidenced by polyacrylamide gel disc electrophoresis and Sephadex G-100 column chromatography, and had a molecular weight of approximately 74,000 and a K_m of 0.21 mM. It is susceptible to inhibition by organophosphates and carbamate insecticides and insensitive to pCMB, mercuric ion, and cupric ion. It is capable of hydrolyzing trans isomers of synthetic pyrethroids much more rapidly (five to ten times) than the cis counterparts. The purified pyrethroid carboxyesterase is apparently identical in nature with malathion carboxyesterase and with p-nitrophenyl acetate carboxyesterase.  相似文献   

Whole-Cell Fatty Acid Composition to Characterize and Differentiate Isolates of Rhizoctonia Species Associated with Turfgrass Diseases in Japan     

Achmadi PRIYATMOJO  Ryo YAMAUCHI  Koji KAGEYAMA  Mitsuro HYAKUMACHI 《Journal of General Plant Pathology》2002,68(1):1-7

Cellular fatty acids were analyzed to characterize and differentiate 34 isolates of Rhizoctonia species representing binucleate Rhizoctonia AG-D (I), AG-D (II), R. solani AG 2-2 IIIB, AG 2-2 LP, R. circinata var. circinata and var. oryzae associated with turfgrass diseases in Japan. Myristic, pentadecanoic, palmitic, palmitoleic, stearic, oleic, linoleic and linolenic acids were consistently present in varying quantities in all isolates. Heptadecanoic and 9-heptadecenoic acids were present in isolates of Rhizoctonia AG-D (I), AG-D (II), R. solani AG 2-2 IIIB and AG 2-2 LP but not in isolates of R. circinata var. circinata and var. oryzae. Palmitic, oleic and linoleic acids were the major fatty acids found, constituting 88.30-98.37% of the whole-cell fatty acid content. The remaining fatty acids were present in smaller amounts. Isolates within a single group were closely clustered, whereas isolates from different groups were clearly distinguishable based on average linkage cluster analysis of cellular fatty acids. Principal component analysis, based on all fatty acids detected, confirmed the distinct separation of isolates representing the six groups of Rhizoctonia species obtained from turfgrasses. These results suggested that fatty acid analysis is useful for the characterization and differentiation of isolates of Rhizoctonia species associated with turfgrass diseases. Received 21 May 2001/ Accepted in revised form 28 September 2001  相似文献   

Effect of dasatinib in a xenograft mouse model of canine histiocytic sarcoma and in vitro expression status of its potential target EPHA2          下载免费PDF全文

K. Ito  R. Miyamoto  H. Tani  S. Kurita  M. Kobayashi  K. Tamura  M. Bonkobara 《Journal of veterinary pharmacology and therapeutics》2018,41(1):e45-e48

Canine histiocytic sarcoma (HS) is an aggressive and highly metastatic tumor. Previously, the kinase inhibitor dasatinib was shown to have potent growth inhibitory activity against HS cells in vitro, possibly via targeting the EPHA2 receptor. Here, the in vivo effect of dasatinib in HS cells was investigated using a xenograft mouse model. Moreover, the expression status of EPHA2 was examined in six HS cell lines, ranging from insensitive to highly sensitive to dasatinib. In the HS xenograft mouse model, dasatinib significantly suppressed tumor growth, as illustrated by a decrease in mitotic and Ki67 indices and an increase in apoptotic index in tumor tissues. On Western blot analysis, EPHA2 was only weakly detected in all HS cell lines, regardless of sensitivity to dasatinib. Dasatinib likely results in the inhibition of xenograft tumor growth via a mechanism other than targeting EPHA2. The findings of this study suggest that dasatinib is a targeted therapy drug worthy of further exploration for the treatment of canine HS.  相似文献   

Identification of the core motif of the BRCA2 C-terminal RAD51-binding domain by comparing canine and human BRCA2     

Yasunaga YOSHIKAWA  Masami MORIMATSU  Kazuhiko OCHIAI  Toshina ISHIGURO-OONUMA  Ryo MORIOKA  Kento OKUDA  Koichi ORINO 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2021,83(5):759

Mammary tumors are the most common tumors in women and non-spayed female dogs. One of the reasons for mammary tumors is mutations of the tumor suppressor gene, BRCA2. BRCA2 participates in homologous recombination repair by interacting with the RAD51 recombinase. BRCA2 has two RAD51-binding domains, consisting of BRC repeats and the C-terminal RAD51-binding domain, respectively. Although several studies have addressed the function of the C-terminal RAD51-binding domain of human BRCA2, the amino acid sequences required for the RAD51-interaction activity remain unclear. In this study, the C-terminal RAD51-binding domains of canine and human BRCA2 were compared; the canine domain displayed a weaker interaction with RAD51. This difference was attributed to the C-terminal portion of the domain via a comparison between canine and human domains. Furthermore, peptides shorter than those previously reported displayed RAD51-interacting activity, and a core motif of this domain consisting of 25 amino acids was identified. Since a mutation (S3323N) was reported in the core motif of this domain, the effect of this mutation was evaluated. The mutant exhibited similar RAD51-binding activity as that of the wild-type protein, suggesting that the mutation was functionally neutral. These data suggested that the C-terminal portion of the BRCA2 C-terminal RAD51-binding domain influenced its RAD51-interaction activity, and a minimum core motif of 25 amino acids was identified in this domain. These data may help clarify BRCA2 function, as well as the tumorigenic effects of BRCA2 mutation.  相似文献   

Chronic low-dose exposure to imidacloprid potentiates high fat diet-mediated liver steatosis in C57BL/6J male mice     

Collins NIMAKO  Yoshinori IKENAKA  Yuko OKAMATSU-OGURA  Jussiaea V. BARIUAN  Atsushi KOBAYASHI  Ryo YAMAZAKI  Kumiko TAIRA  Nobuhiko HOSHI  Tetsushi HIRANO  Shouta M. M. NAKAYAMA  Mayumi ISHIZUKA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2021,83(3):487

  相似文献   

Identification of the core rumen bacterial taxa and their population dynamics during the fattening period in Japanese Black cattle     

Hiroto Miura  Takuya Hashimoto  Yukiko Kawanishi  Hiroki Kawauchi  Ryo Inoue  Noriaki Shoji  Kunihiko Saito  Mario Sekiya  Yosuke Saito  Jumpei Yasuda  Chiemi Yonezawa  Tetsushiro Endo  Hirotaka Kasuya  Yutaka Suzuki  Yasuo Kobayashi  Satoshi Koike 《Animal Science Journal》2021,92(1):e13601

The rumen microbiota comprises a vast range of bacterial taxa, which may affect the production of high-quality meat in Japanese Black cattle. The aim of this study was to identify core rumen microbiota in rumen fluid samples collected from 74 Japanese Black cattle raised under different dietary conditions using 16S rRNA gene amplicon sequencing. In the rumen of fattening Japanese Black cattle, 10 bacterial taxa, showing >1% average relative abundance and >95% prevalence, irrespective of the dietary conditions and the fattening periods, were identified as the core rumen bacterial taxa, which accounted for approximately 80% of the rumen microbiota in Japanese Black cattle. Additionally, population dynamics of the core rumen bacterial taxa revealed two distinct patterns: Prevotella spp. and unclassified Bacteroidales decreased in the mid-fattening period, whereas unclassified Clostridiales, unclassified Ruminococcaceae, Ruminococcus spp., and unclassified Christensenellaceae increased during the same period. Therefore, the present study reports the wide distribution of the core rumen bacterial taxa in Japanese Black cattle, and the complementary nature of the population dynamics of these core taxa, which may ensure stable rumen fermentation during the fattening period.  相似文献   

A review of okara (soybean curd residue) utilization as animal feed: Nutritive value and animal performance aspects     

Mohammad Mijanur Rahman  Khairiyah Mat  Genki Ishigaki  Ryo Akashi 《Animal Science Journal》2021,92(1):e13594

Year by year, huge quantities of by-products are generated during the manufacturing process of soybean-based products. Okara is one of the by-products, and it is an insoluble portion of the soybean. It consists of high moisture (8.4–22.9%); on dry matter basis, it contains high metabolizable energy (9.0–14.2 MJ/kg) and other components that include crude protein (20.9–39.1%), crude fiber (12.2–61.3%), crude fat (4.9–21.5%), and ash (3.4–5.3%). Fermentation of okara improves its nutritional quality and reduces its anti-nutrient contents. Due to animals' palatability, okara can be used to replace the soybean meal/concentrate feed partially or completely in ruminant's diet and partially in nonruminant's diet. Okara feeding does not depress the intake, digestibility, growth, milk production, blood metabolic profiles, and meat quality of animals. However, this by-product decays quickly due to its high moisture content, and its heavy weight and sticky nature make it difficult to process and expensive to dry using conventional methods. This paper thoroughly summarizes the utilization of okara as animal feed in the cause of developing a general guideline with favorable levels of inclusion in the diets of animals for its exploitation and valorization. This review will encourage further research to develop eco-friendly and value added feed for animals.  相似文献