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   

Bovine Posterior Pituitary Extract Stimulates Prolactin Release from the Anterior Pituitary Gland In Vitro and In Vivo in Cattle     

T Hashizume  T Sasaki  S Nonaka  T Hayashi  M Takisawa  M Horiuchi  T Hirata  E Kasuya 《Reproduction in domestic animals》2005,40(2):184-189

It has been reported that the posterior pituitary (PP) gland contains a potent, unknown prolactin (PRL)-releasing factor (PRF) in rats. PRFs are assumed to be produced in neurones located within the hypothalamus, and to be peptidergic in nature. However, little is known about PRFs in domestic animals. To characterize the PRF in the PP of domestic animals, the present study examined the PRL-releasing activity of an acidic extract from bovine PP (bPP) in vitro and in vivo in cattle. First, the PRL-releasing effect of bPP extract was compared with that of PRL-releasing peptide (PrRP), and thyrotropin-releasing hormone (TRH) from cultured bovine anterior pituitary cells. The extract significantly increased PRL concentrations in the culture medium, at doses of 0.002 and 0.02 eq./ml (one eq. is the PP extract from one animal), compared with the control (p < 0.05). PrRP failed to stimulate the release of PRL. TRH significantly increased PRL concentrations in the culture medium, at doses from 10(-9) to 10(-7) M, compared with the control (p < 0.05). The rate of increase in the PRL concentration, by 0.02 eq./ml bPP extract, was significantly greater than that in TRH (p < 0.05). Secondly, plasma PRL responses to the intravenous (i.v.) injection of bPP extract (0.5 eq./head), PrRP [3.59 mug/kg body weight (BW)], TRH (1 mug/kg BW), and a dopamine receptor antagonist (sulpiride, 0.1 mg/kg BW), were examined in calves. PrRP failed to stimulate PRL release; however, plasma PRL increased immediately following the injection of bPP extract, TRH and sulpiride. The PRL-releasing effect of i.v. injections of TRH and sulpiride was more potent than that of bPP extract. Finally, plasma PRL responses to the intra-hypothalamic injection of bPP extract were examined in calves. The intra-hypothalamic infusion (arcuate nucleus) of 0.0625 eq./head of bPP extract strongly stimulated PRL release in calves (p < 0.05). The present results show that PP contains a physiologically potent PRF in cattle.  相似文献   

Pathological analysis of batch safety testing of veterinary vaccines using small laboratory animals     

Kasumi Sudo  Mariko Ochiai  Naoyuki Aihara  Noriyuki Horiuchi  Atsushi Yamamoto  Sachiko Matsumoto  Koji Oishi 《Journal of toxicologic pathology》2021,34(2):137

Batch safety tests (BSTs) of veterinary vaccines are conducted using small laboratory animals to assure the safety of vaccines according to several criteria, including clinical signs and change in body weight. Although the latter is used as an evaluation index in BSTs, there have been no reports on the internal changes that affect body weight during the test period. Therefore, we analyzed BST via pathological examination of the tested animals. Here, BSTs were performed for 176 batches using mice and 126 batches using of guinea pigs. Most of the gross findings could be classified into four lesion types (nodules, adhesions, ascites, no apparent signs), with only one vaccine inducing lesions that could not be classified into any of these four types. Histopathological examination revealed that the reactions caused by BST were pyogenic and/or granulomatous inflammation. Nodular or adhesive lesions comprised more severe pyogenic granulomatous inflammation than ascites or cases with no apparent gross lesions. These nodular or adhesive lesions were more frequently induced by vaccines that contained an adjuvant than by vaccines that did not contain an adjuvant. The cases with “exceptional” gross findings histologically presented severe necrosis of the hematopoietic system. Additional testing showed that these “exceptional” lesions were induced when a specific type of light liquid paraffin was injected along with other vaccine additives. Our results show that body weight loss and/or lesions during BST were induced by proinflammatory properties of the tested vaccines and that BST is a sensitive method for detecting unexpected effects of vaccine components.  相似文献   

Flow cytometric analysis of chicken NK activity and its use on the effect of restraint stress     

Kushima K  Fujita M  Shigeta A  Horiuchi H  Matsuda H  Furusawa S 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(9):995-1000

Immune system is organized by the influence of both neural and endocrine systems. NK activity plays an important role in the innate immunity. In this study, we observed the effects of restraint stress on chicken peripheral blood NK activity. Viability of FITC-labeled RP9 was measured with PI after treatment with the effector cells. Chicken peripheral blood CD8alpha+ cells expressed strong cytotoxic activity, in contrast to thrombocytes, while peripheral blood CD3+ CD8alpha+ cells and CD4+ cells had little cytotoxic activity. Con A supernatant enhanced the cytotoxic activity of CD8alpha+ cells. Therefore, it is considered that these cytotoxic activities measured by flow cytometry (FCM) analysis are NK activity. When chickens were exposed to restraint stress, the levels of serum corticosterone increased transiently over a short period of time while the NK activity decreased. The decreased NK activity, however, did not recover to the intact levels for a long time, even once the serum corticosterone levels had recovered. These data indicate that chicken NK activity is able to be measured by flow cytometric analysis and that restraint stress causes severe damage to the chicken NK activity.  相似文献   

Effect of ethephon (2-chloroethylphosphonic acid) on the fruit ripening characters of rabbiteye blueberry     

Takuya Ban  Mihoko Kugishima  Tsuneo Ogata  Shuji Shiozaki  Shosaku Horiuchi  Hisafumi Ueda 《Scientia Horticulturae》2007

Effect of ethephon (2-chloroethylphosphonic acid) application on rabbiteye blueberry fruit quality during the growth period was investigated. Ethephon treatment stimulated the decrement of titratable acidity, anthocyanin accumulation and fruit softening 4 days after treatment and the promoting effects continued through the investigation period. The ripening promotion effect of ethephon on total soluble solids content was observed only 8 days after treatment. Ethephon treatment did not affect the fruit enlargement during the investigation period. From these results, it is concluded that ethephon application for rabbiteye blueberry promote the fruit ripening, but the stimulatory effects of ethephon on fruit ripening were different in degree on each ripening characters.  相似文献   

Growth,Reproductive Performance,Carcass Characteristics and Meat Quality in F1 and F2 Progenies of Somatic Cell-Cloned Pigs     

Noritaka ADACHI  Daisuke YAMAGUCHI  Akiyuki WATANABE  Narumi MIURA  Seiji SUNAGA  Hitoshi OISHI  Michiko HASHIMOTO  Takatsugu OISHI  Masaki IWAMOTO  Hirofumi HANADA  Masanori KUBO  Akira ONISHI 《The Journal of reproduction and development》2014,60(2):100-105

The objective of this study was to examine the health and meat production of cloned sows and their progenies in order to demonstrate the application of somatic cell cloning to the pig industry. This study compared the growth, reproductive performance, carcass characteristics and meat quality of Landrace cloned sows, F1 progenies and F2 progenies. We measured their body weight, growth rate and feed conversion and performed a pathological analysis of their anatomy to detect abnormalities. Three of the five cloned pigs were used for a growth test. Cloned pigs grew normally and had characteristics similar to those of the control purebred Landrace pigs. Two cloned gilts were bred with a Landrace boar and used for a progeny test. F1 progenies had characteristics similar to those of the controls. Two of the F1 progeny gilts were bred with a Duroc or Large White boar and used for the progeny test. F2 progenies grew normally. There were no biological differences in growth, carcass characteristics and amino acid composition among cloned sows, F1 progenies, F2 progenies and conventional pigs. The cloned sows and F1 progenies showed normal reproductive performance. No specific abnormalities were observed by pathological analysis, with the exception of periarteritis in the F1 progenies. All pigs had a normal karyotype. These results demonstrate that cloned female pigs and their progenies have similar growth, reproductive performance and carcass quality characteristics and that somatic cell cloning could be a useful technique for conserving superior pig breeds in conventional meat production.  相似文献   

Pathological changes of tracheal mucosa in chickens infected with infectious laryngotracheitis virus   总被引：1，自引：0，他引：1  

S Hayashi  Y Odagiri  T Kotani  T Horiuchi 《Avian diseases》1985,29(4):943-950

Six-week-old chickens were inoculated via the posterior thoracic air sac with infectious laryngotracheitis virus. Chickens were sacrificed on various days through day 16 postinoculation (PI), and the trachea was examined by scanning electron microscopy (SEM) and light microscopy (LM). The pathological changes observed on day 1 PI were hypertrophy and hyperplasia of goblet cells. From day 3 PI, the epithelial cells protruded collectively and fused to form syncytia, which contained many intranuclear inclusion bodies. Subsequently, epithelial syncytia desquamated, one after another, and connective tissues were exposed in places. Serofibrinous exudate and detritus were abundant on the surface of the exposed connective tissues and seemed to form a pseudomembrane. On day 5 PI, the remaining epithelial cells began to repair the devastated mucosa just under the pseudomembrane. On day 6 PI, microvillus-rich regenerating epithelial cells were arranged like paving stones. On day 8 PI, the epithelial cells proliferated extensively and formed folds with cyst-like structures. By day 16 PI, the tracheal epithelium was covered with cilia and regained its normal histologic appearance.  相似文献   

Preliminary evidence of growth factor(s) from chicken thrombocytes--growth effects on chicken embryo fibroblasts culture     

H Horiuchi  H Matsuda  M Murata 《Nippon juigaku zasshi. The Japanese journal of veterinary science》1990,52(3):559-565

The thrombocyte extracts (TE) and thrombocyte-secretion products (TSP) prepared from chicken peripheral blood were examined for the growth activity of chicken embryo fibroblasts (CEF). When the cells were incubated with TE and TSP, the enhancement of cell spreading was observed within one hour after seeding, but the control culture showed little spreading. Three to six days after cell seeding, the increase of the cell densities in the culture supplemented with TE and TSP was noticed in comparison with the culture without thrombocyte materials. The results strongly indicate the possibility for the presence of thrombocyte-derived growth factor(s) to CEF.  相似文献   

Similarity of synthetic peptide from human tumor to parathyroid hormone in vivo and in vitro   总被引：18，自引：0，他引：18  

N Horiuchi  M P Caulfield  J E Fisher  M E Goldman  R L McKee  J E Reagan  J J Levy  R F Nutt  S B Rodan  T L Schofield 《Science (New York, N.Y.)》1987,238(4833):1566-1568

One mechanism considered responsible for the hypercalcemia that frequently accompanies malignancy is secretion by the tumor of a circulating factor that alters calcium metabolism. The structure of a tumor-secreted peptide was recently determined and found to be partially homologous to parathyroid hormone (PTH). The amino-terminal 1-34 region of the factor was synthesized and evaluated biologically. In vivo it produced hypercalcemia, acted on bone and kidney, and stimulated 1,25-dihydroxy-vitamin D3 formation. In vitro it interacted with PTH receptors and, in some systems, was more potent than PTH. These studies support a long-standing hypothesis regarding pathogenesis of malignancy-associated hypercalcemia.  相似文献   

Bacteriological and genetic assessment of game meat from Japanese wild boars     

Naya Y  Horiuchi M  Ishiguro N  Shinagawa M 《Journal of agricultural and food chemistry》2003,51(2):345-349

Bacterial tests were used to assess bacterial contamination of game meat from Japanese wild boars. The bacterial contamination of wild boar meat was less than that of domestic pork, as determined by aerobic plate counts (APC) and coliform counts. None of the meat examined in this study was contaminated by Salmonella or E. coli O-157. To detect adulteration by domestic pig meat or European wild boar meat, 46 samples of game meat sold as Japanese wild boar were examined genetically. A total of 17 samples showed genetic haplotypes of European and Asian domestic pigs in the D-loop of mitochondrial DNA (mtDNA), and 16 samples showed nuclear glucosephosphate isomerase-processed pseudogene (GPIP) genotypes of European domestic pigs. The European GPIP genotypes of these samples were confirmed by PCR-RFLP analysis. These results indicate that some game meat sold as Japanese wild boar is adulterated by cross-breeding between pigs and wild boars or by contamination with meat from domestic pigs or European wild boars.  相似文献