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The ability of in vitro cultured bovine oviduct epityelial cells in binding and maintaining motility of bull sperm
1999
Roh, S.H. (Hankyong National University, Ansung (Korea Republic). Department of Animal Life Resources) | Lee, B.C. | Hwang, W.S. (Seoul National University, Suwon (Korea Republic). College of Veterinary Medicine)
The aim of these experiments was to investigate the effects of bovine oviduct epithelial cells(OEC) derived from different segments to bind sperm binding and maintain their motility in vitro. In experiment 1, the number of sperm attached to OEC derived from isthmus or ampulla, the motility of unattached sperm during co-culture and fertilizing ability were assessed. In experiment 2, heparin treated sperm 9hsp) or no treated sperm (nsp) were used to evaluate OEC binding ability ofcapacitated sperm. In experiment 1, regardless ofthier origin, approximately 65% of the sperm were attached to OEC within 2h. From 6h of co-culture, the numbers of unattached sperm on ampullary OEC were significantly higher than those on isthmic OEC (p0.005). From 12h of co-culture, the motility of unattached sperm on isthmic OEC were significantly higher than those on ampullary OEC(p0.05). The cleavage rate of oocytes inseminated on OEC derived from isthmic segment was also significantly higher than those from ampullary segment (p0.01). In experiment 2, the numbers of unattached hsp on OEC were significantly higher than those of controls(p0.01), between 2~24h examination. From 12h of co-culture, the motility of unattached nsp were sighificantly greater than those of hsp(p0.01). These results show that bovine OEC derived from the isthmus play more important role(s) for sperm binding, maintaining motility and fertilization in vitro than those from the ampulla, and heparin induced capacitation may change sperm binding ability on OEC in vitro.
Show more [+] Less [-]Histological and immunohistochemical effects of Jengjengamiyjintang on the duodenal ulcer induced by HCI-aspirin
1999
Ku, S.K. (Dong-Wha Pharmaceutical Industry Company, Anyang (Korea Republic). Parmacol & Toxicol Laboratory, Central Research Laboratories) | Lee, H.S. (Kyungsan University, Kyungsan (Korea Republic). Department of Biology, Faculty of Basic Science) | lee, J.H. (Kyungpook National University, Taegu (Korea Republic). Laboratory of Histology, College of Veterinary Medicine)
In order to study the effects of Jengjengamiyjintang on the duodenal ulcer induced by HCI-aspirin in rats, the changes of histological profiles, goblet cells(PAS-positive cells), and the distribution and frequency of cholecystokinin(CCK)-8 and serotonin-producing gastro-entero-endocrine cells were observed after oral administration of Jengjengamiyjintang. Histologically, very severe injury to duodenal epithelium were observed in control groups and theses injuries were increased with time intervals. But in the Jengjengamiyjintang administrated groups, no gross lesion of ulcer were demonstrated and histologically minor injury to the mucosal epithelium were observed. PAS-positive cells were increased in the Jengjengamiyjintang administrated groups compared to that of control groups. Severe degranulation of CCK-8- and serotonin-immunoreactive cells were observed in control groups but these phenomenon was seldom in the Jengjengamiyjintang administrated groups. Serotonin-immunoreactive cells were significantly decreased in control groups but increased in Jengjengamiyjintang administrated groups compared with control groups. According to these result, it is suggested that Jengjengamiyjintang would accelarat the healing of the duodenal ulcer but the functional mechanisms were unknown.
Show more [+] Less [-]Isolation and comparison of somatic and excretory-secretory antigens for serological diagnosis in Trichuris suis
1999
Jee, C.H. | Lee, C.S. | Park, S.J. (Chungbuk National University, Cheongju (Korea Republic). College of Veterinary Medicine)
Swine shipworm(Trichuris suis) is cosmopolitan nematode which can cause serious pathology in immature stage(larva2~larva5) of infected pigs, such as anorexia, diarrhea, anemia, and death in heavy infections. In this larval stages, it is very difficult to diagnose the infection of whipworm and to differentiate from other common swinegstrointestinal disorders such as 21 day scours which are associated with TGE virus, rota virus, coccidium, and the stress of weaning. In this experiment, the isolated antigens of Trichuris spp. were carried out to examine the structure and specificity of antigens and to select the resonable antigens which would be used in serological diagnosis by electrophoresis, Western blotting, ELISA. The results of this experiment were as follews: 1. The common fractions of each Trichuris suis antigen were identified 28,32,45, 80kDa by SDS-PAGE with silver stain and four major fractions could be detected in positive swine sera by Western blot analysis. 2. The OD(optical density) values of somatic and excretory-secretory antigens which were reacted against positive(negative) sera from pigs infected with Trichuris suis by ELISA reader were; 1) OD values(mean+_SD) of adult somatic antigen against positive(negative) sera were O.30+_0.12(0.09+_0.006) and third-stage larva of somatic antigen were 0.28+_0.038(0.10+_0.005). And OD values of excretory-secretory antigens of adult and third-stage larva were 0.24+_0.031(0.11+_0.005) and 0.08+_0.013(0.10+_0.003), respectively. 2) OD values of adult somatic, larval somatic antigen and adult excretory-secretory antigen response to positive sera were significantly (p0.01) associated with negative swine sera. And the Cut-off OD values(minimum positive value) were determined to be mean negative value plus 3 SD that would minimized the risk of false positives. 3. The OD values of somatic antigens of T suis and T vulpis against swine positive(negative) sera were 0.30+_0.120(0.09+_0.006) and 0.25+_0.141(0.09+_0.003). These data mean that the somatic antigens of T suis and T vulpis were able to diagnose T vulpis infection in dogs as well as T suis infection in pigs. These results suggest that somatic antigen of third-stage larva and excretory-secretory antigen of adult T suis could be used the diagnostic antigen by serological test(ELISA) in immature Trichuris spp. infection.
Show more [+] Less [-]Specific detection of Salmonella serogroup D1 by polymerase chain reaction(PCR) for sefA gene
1999
Jun, M.H. | Kim, T.J. | Chang, K.S. | Kang, K.I. | Kim, K.H. | Kim, H.S. | Shin, K.S. | Kim, C.J. (Chungnam National University, Taejon (Korea Republic). College of Veterinary Medicine) | Kim, K.S. (National Veterinary Research and Quarantine Service, Anyang (Korea Republic).) | Yoo, S.S. (Taejon City Institute of Health and Environment, Taejon (Korea Republic).)
Sal enteritidis thin fimbriae, SEF14, were found to be restricted to the predominantly poultry-associated members of the Salmonella serogroup D1 that are considered as the important pathogens in poultry industry. SefA together with sefB and sdfC encode the proteins involved in SEF14 biosynthesis. In order to develop the rapid and specific detection methods for Salmonella serogroup D1, a PCR technique for the am;lification of sefA gene was established, and its specificity and sensitivity were investigated with various microorganisms. The bacterial genomic DNA was extracted by colony-picking and rapid boiled-lysate technique. In comparison of SefI and SefII primers used in the PCR. SefI primer for sefA gene of 513bp showed higher specificity than that of SefII. The established PCR was s sensitive as to detect 1pg of Sal enteritidis DNA. When 73 strains in 28 genera including the reference strains and the field isolates of various Salmonella serotypes, Bacillus subtilis, Bordetella bronchiseptica, E coli, Listeria spp., Micrococcus luteus, Rhodococcus equi, Staphylococcus spp., Streptococcus spp., Vibrio parahemolyticus, Yersinia spp. were studied, the established PCR yielded specifically positive results with only Salmonella serogroup D1. The results suggested that the PCR for sefA gene could be a potential candidate among the specific detection methods for Salmonella serogroup D1.
Show more [+] Less [-]Immunogold studies on the gonadotropes in adenohypophysis of the Korean native goat
1999
Lee, I.S. | Lee, H.S. | Song, S.H. | Yoon, S.T. | Hwang, I.K. (Seoul National University, Suwon (Korea Republic). College of Veterinary Medicine) | Seo, J.H. (Seoul National University, Seoul (Korea Republic). College of Medicine | Kang, T.C. | Won, M.H. (Hallym University, Chuncheon (Korea Republic). College of Medicine)
There have been a number of studies of gonadotropes secretin LH and FSH in the adenohypophysis, but the pattern of hormone storage and secretion of these cells still remains a controversial matter. In this study, we examined whether gonadotropes contained both of LH and FSH, and if so, how these hormones were distributed within the secretory granules. Hypophyseal sections of Korean native goat were simultaneously immunostained for LH and FSH antisera by protein A-gold technique. It was found that most gonadotropes contained both FSH and LH, but hormone storages in the secretory granules were some different among cells. Three types of gonadotropes were identified by the shape and size of the secretory granules and their hormone storage patterns. One type(I) of gonadotropes contained oval secretory granules, which immunoreactivity for FSH and LH were very weak. The size of secretory granules ranged from 160 to 310nm in diameter. Most granules contained both FSH and LH, but some contained only one of them. In another type(II)of gonadotropes, the immunreactivity and hormone storage patterns of the secretory granules were similar to those of type I cells. However, the secretory granules were round in shape and larger in size than those of type I. The other gonadotropes(type III) were distinctly distinguished by plenty of hormones in their secretory granules which were densely packed with numerous immunolabelled gold particles. These data are some inconsistent with other results that have been obtained in other ruminants like as cattle and sheep.
Show more [+] Less [-]Morphogenetical characteristics of Korean wild rat(Rattus norvegicus)
1999
Seong, J.K. | Yun, Y.M. | Park, J.Y. | Oh, S.H. (Yonsei University, Seoul (Korea Republic). Department of Laboratory Animal Medicine, Medical Research Center, College of Medicine) | Do, S.G. | Jin, H.K. | Suh, J.G. | Oh, Y.S. (Hallym University, Chuncheon (Korea Republic). Experimental Animal Center, College of Medicine) | Hyun, B.H. (Korea Institute of Science and Technology, Taejon (Korea Republic). Bioresource Program, Korea Research Institute of Bioscience and Biotechnology)
The morphometrical characteristics such as external measurements and mandible size assessment in mice and rats have to be highly heritable and sufficiently variable between strains in order to calculate a strain specific profiles. The coat color of Korean wild rats were observed and morphometric analysis of external measurements were carried out on Korean wild rats compared to laboratory strains in order to clarify the genetic characteristics of Korean wild rats and to establish background data as a domestication of Korean wild rats for new laboratory strain. Korean wild rats were captured from Chunchon and Hoengsong. 4 inbred and 1 outbred strains of rats were used in this study for the comparison of genetic characteristics of Korean wild rats. Total body length, head length, tail length, hind foot length and ear length were measured and then statistical analysis were carried out by discrimiant analysis. The coat color of Korean wild rat showed golden white in ventral portion and dark agouti in dorsal portion. Korean wild rats could be distinguished from the other laboratory strains distinctly by morphogenetical analysis. There was sighificant variations among Korean wild rat compared to those of the other laboratory strains of rat. This study may provide that Korean wild rats have a unique genetic characterization compared to those other inbred strains of rats based on morphogenetical characteristics by external measurements.
Show more [+] Less [-]Therapeutic effects of 1alpha,25 dihydroxycholecalciferol on osteoporotic fracture in a rat model
1999
Bae, C.S. (Konkuk University, Seoul (Korea republic). Department of Surgery, Faculty of Veterinary Medicine)
Osteoporosis is defined as a decrease in bone mass that leads to an increased risk of fracture. The therapeutic effect of 1alpha,25 dihydroxycholecalciferol, the hormonal form of vitamin D3 that mediates calcium translation in intestine and bone, on the healing process of fracture has still been controversial. These studies were designed to understand the healing process of normal fibular fracture, the osteoporotic changes after ovariectomy, and the theraqeutic effects of 1alpha,25 dihydroxycholecalciferol on the osteoporotic fracture in rats. The simple transverse fractures of rat fibulae were produced with a rotating diamond saw. The changes of the biochemical and mechanical indices of rats were investigated. The mechanical study based on bending test revealed the healing of the fibular fracture in the 5th week after simple transverse fracture. The osteoporosis impaired more the healing of osteoporotic fibular fracture than normal non-osteoporotic fibular fracture. The healing process of osteoporotic fracture was facilitated by the treatment with 1alpha,25 dihydroxycholecalciferol, however was delayed more than the healing process of normal fracture. The bone strength based on the bending test also confirmed this tendency. The bone strengths in the 5th week after fracture of normal bone, osteoporotic bone, and 1alpha,25 dihydroxycholecalciferol-treated osteoporotic bone were 75%, 41% and 67%, respectively, in comparison with those of intact bone. In conclusion, 1alpha,25 dihydroxycholecalciferol was effective in promoting the osteroporotic fracture healing.
Show more [+] Less [-]Immunohistochemical study onthe expression of bcl-2 protein during follicular development and atresia in the rat ovary
1999
Koh, P.O. | Kwak, S.D. (Gyeongsang National University, Chinju (Korea Public). Institute of Animal Medicine, College of Veterinary Medicine) | Jeong, S.Y. | Cho, G.J. | Choi, W.S. (Gyeongsang National University, Chinju (Korea Public). Department of Anatomy, College of Medicine)
In the mannalian ovary, follicular development and stresia continuously occur during the reproductive cycles. Follicular atresia occurs through granulosa cell apoptosis. Apoptosis is known as the physiological cell death, which is regulated by bcl-2 gene family. In the vcl-2 gene family, bcl-2 and bcl-xLong are known as inhibitors of apoptosis, whereas bax and bcl-xShort are known as inducer of apoptosis. We thought that bcl-2 protein is associated with follicular development and atresia. But it is not known that the distribution of cells containing bcl-2 protein during follicular development and atresia. Therefore, to examine the distribution of cells with bcl-2 protein during ovarian follicular development and atresia. Therefore, to examine the distribution of cells with bcl-2 protein during ovarian follicular development and atresia, the immunohistochemistry was used ih the rat ovary. Bcl-2 immunorectivity was localized in the intestitial cells, theca externa cells and granulosa cells around of antrum. All positive sighals were observed in the cytoplasm of these cells. Positive sighals were strongly observed in the interstitial and theca externa cells of growing antral follicles. While, positive signals were weakly observed in these cells from atretic antral follicles. Positive sighals were very weakly observed in the granulosa cells of growing and atreticantral follicles. According to these data, we suggested that bcl-2 proteins which were wtrongly expressed in the interstitial cells and theca externa cells of growing antral folicles inhibit follicular atresia. And wer purposed that bcl-2 proteins regulated follicular development and atresia through the action of acl-2 gene family.
Show more [+] Less [-]The expression of interleukin-1beta converting enzyme in experimental autoimmune encephalomyelitis
1999
Moon, C.J. | Kim, S.J. | Lee, Y.D. | Shin, T.K. (Cheju National University, Cheju (Korea Republic). Department of Veterinary Medicine)
To elucidate the involvement of interleukin-1beta converting enzyme (ICE) in the courseof experimental autoimmune encephalomyelitis (EAE), we induced EAE by immunizing rats with an emulsion of rat spinal cord homogenate with complete Freund's adjuvant supplemented with Mycobacterium tuberculosis(H37Ra, 5mg/ml) and then examined the expression of ICE in the spinal cord of rats with EAE. In normal rate spinal cords, ICE is constitutively, but weakly, expressed in ependmal cells, neurons, and some neuroglial cells. In EAE, many inflammatory cells are positive for ICE, and the majority of ICE+ cells were identified as ED1+ macrophages. During this stage of EAE, the number of ICE+ cells in brain cells, including neurons and astrocytes, increased and these cells also had incresed ICE immunoreactivity. These findings suggest that the upregulation of ICE in both brain cells and invading hematogenous cells is stimulated by a secretory product from inflammatory cells, and that this enzyme is involved in the pathogenesis of EAE via the production of IL-1 beta.
Show more [+] Less [-]A study on recovery of deep hypothermia in rabbits
1999
Lee, B.H. | Chun, W.C. | Kim, J.Y. | Chung, B.H. (Kon-Kuk University, Seoul (Korea Republic). School of Veterinary Medicine) | Kim, J.S. | Park, J.H. | Park, J.S. | Han, J.S. (Samsung Biomedical Research Institute, Seoul (Korea Republic). Department of Laboratory Animal Research)
The studies were carried out to investigate the effect of recovery in rewarming using the esophageal thermal tube in the deep hypothermia(25+_1 degrees centigrade ; rectal temp) in rabbits. Fifteen rabbits were divided into control group(n=6), peritoneal dialysis group(n=5) irrigated with dialysate at 42+_1 degrees centigrade, and esophageal rewarming group(n=6), peritioneal dialysis group(n=5) irrigated with dialysate at 42+_1 degrees centigrade, and esophageal rewarming group(n=4) perfused with circulating water at 38+_1 degrees centigrade. Rewarming of the rabbits was performed for 5 hours. MAP, HR, RR, esophageal temp, rectal temp, pH, pCO2, pO2, Na+, and K+ were observed. The results obtained in these experiments were summarized as follows:Esophageal rewarming group(38+_1 degrees centigrade) had more effect on esophageal temperature than other groups. Peritoneal dialysis group(42+_1 degrees centigrade) had more effect on rectal temperature and pO2 than other groups. The both groups also had more effects on MAP, HR, RR, and pCO2 than control group. Three groups had no significant effect on pH, Na+, and K+. In conclusion, we found that the simple, safe, and non-invasive esophageal rewarming method had an effect on the treatment of profound hypothermia as well as the peritoneal dialysis method in spite of the temperature difference between the dialysate and the circulating water, and the circulating water at 38+_1 degrees centigrade for esophageal rewarming also had an effect on the recovery of deep hypothermia.
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