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Production and identification of antisera against mu-opioid receptor usign synthetic peptide epitope
1999
Lee, J.H. | Kwon, Y.B. (Seoul National University, Suwon (Korea Republic). Department of Veterinary Physiology, College of Veterinary Medicine) | Han, H.J. (Chonnam National University, Kwangju (Korea Republic). Department of Veterinary Physiology, College of Veterinary Medicine)
In the present study we have analyzed the characteristics and distribution of the mu-opioid receptor(MOR) by raising anti-peptide antisera to the C-terminal peptide of MOR. The antisera against MOR was produced in New Zealand White rabbit against 15 residue corresponding to amino acids, 384-398of the cloned rat MOR. The antigenic peptide was synthesized using an Applied Biosystems 432 solid-phase peptide aynthesizer. The specificity and identification fo the antisera were tested by analysisi fo transfected cells, epitope mapping and immunohistochemical method. COS-7 cells electroporated with MOR cDNA were used to evaluate the characteristics and subcellular distribution of MOR.MOR immunoreactivity was prodominent in the plasmalemma and subcellular compartments such as encoplasmicreticulum, Golgi apparatus and vesicle like structure. Furthermore, both tissue sections and transfected cell lines could be immunostained with these antisera and the immunoreactivity ws abolished when anti-MOR sera were preincubated with the peptide against which they wer raised. Based on epitope mapping analysis, all antisera appeared to have a similar epitope, which ws detemined to be within the last amino acid,391-398. Moreover, immunohistochemistry showed that MOR immunoreactivity was ovserved in many brain areas including cerebral cortex, striatum, hippocampus, locus coeruleus and the superficial laminae of the dorsal horn. These stained spinal cord and brain aras showed themirrored pattern observed in autoradiographic studies of mu-opioid binding as well as a pattern similar to that seen by in situ hybridization for MOR. Thus, several lines of evidence support the conclusion that the antisera produced in the present study most likely recognize mu-opioid receptor. These results suggest that MOR antisera may be utilized as useful tool to analyze the physiological and pharmacological studies for mu-opioid receptor in the future
Show more [+] Less [-]Clinical investigations of canine superficial pyoderma
1999
Oh, T.H. (Seoul National Universtiy, Suwon (Korea Republic). College of Veterinary Medicine)
Fifteen dogs with superficial pyoderma were investigated clinically. Dermatological signs were mainly consisted of papule (66.6%), pustule (86.6%), epidermal collrarette and patchy (40%), and hyperpigmentation (53.3%). Distribution of skin lesion were consisted of back (35.5%), abdomen (29.0%), axillary (6.4%), leg (3.2%), neck (3.6%) and foot (16.1%), respectively. In pustular cytology PMN cells and cocci were examined. Cephalexin was very effective antibiotics on superficial pyoderma at administration of 30mg/kg bid P.O.for 3 weeks. Hyperadrenocorticism and atopy were diagnosed as a primary cause on pyoderma in 2 dogs.
Show more [+] Less [-]Diagnosis of canine distemper by in situ hybridization
1999
Cho, H. | Park, N.Y. | Kim, Y.H. | Cho, K.O. | Park, H.S. | Park, Y.S. | Lee, B.J. | Chung, C.Y. | Im, H.H. (Chonnam National University, Kwangju (Korea Republic). College of Veterinary Medicine)
We have developed in situ hybridization(ISH) technique for rapid diagnosis of canine distemper(CD) which is the major infectious disease in dogs. In our experiment, we rapidly detected distribution of the specific canine distemper viral genome without disrupting morphology of tissues of cells. Two oligonucleotide probes for ISH were synthesized chemically and labelled 5' end with nonisotopic biotin by DNA synthesizer. The whole procedures of ISH was completed within 1~2 hours using the Microcapillary action system. On histological study, typical cytoplasmic or intranuclear inclusion bodies wer observed in the trachea, bronchiole, brain, and urinary bladder with the presence of prominent red positive signals on ISH, indicating specific CDV genome from the paraffin-embedded tissues of infected 13 cases. The results showed ISH can be used as a rapid and effective diagnostic method for diagnosis of CD.
Show more [+] Less [-]The reasons of drug residues in bulk milk
1999
Kang, J.H. | Kim, J.S. | Lee, W.C. (Konkuk University, Seoul (Korea Republic). School of Veterinary Medicine) | Choi, P.S. (Konkuk Dairy, Seoul (Korea Republic). Department of Dairy Control)
The 102 farms received a positive result of the milk drug residue test were selected to investigate the reasons of drug residues in bulk milk. The most frequent causes of drug residues were milker or producer mistakes (28.4%), failure to observe withdrawal time (21.5%), and withholding milk from treated quarters only (19.6%). Milker or producer mistakes occurred high at the farms having a parlor system (4 cases out of 11 farms), and related to the inadequate records and marking of treated cows. The lack of knowledge on the absorption of antibiotic from treated quarters and its excretion from untreated quarters caused mainly withholding milk from treated quarters only. Among the 91 farms identified the cause of drug residues, most of the route of drug administration was intramammary infusion (81.3%), and mostly drug used for the treatment of cows was beta-lactam antibiotics (57.1%).
Show more [+] Less [-]Dissemination of Borrelia burgdorferi and immunological responses after experimental infection in rabbits
1999
Kim, J.B. | Park, S.U. | Song, H.W. | Park, S.W. | Kim, Y.M. (Yonsei University, Wonju (Korea Republic). Department of Medical Technology, College of Health Science)
The visceral dissemination of Borrelia burgdorferi in New Zealand White rabbits was evaluated following intradermal inoculation of 1*10 8 spirochetes. We inoculated Borrelia burgdorferi B31, B garinii KW1 and B afzelii S13, respectively, and monitored the dissemination in the experimentally infected rabbits for 28 days. In the B burgdorferi B31-challenged group, the spirochetes were com;oetely cleared in rabbits at day 1 and visceral dissemination was not demonstrated. However, B garinii KW1 and B afzelii S13 were found to successfully disseminate in visceral organs of rabbits during the experiment period of 28 days. And experimentally infection-derived immunological responses in rabbits were identified with enzyme-linked immunosorbent assay and immunoblot analysis. Based on these results, the differences in the virulence of Lymeborrelial strains were proved in rabbit model.
Show more [+] Less [-]An immunohistochemical study on the gastrointestinal endocrine cells in the bean goose, Anser fabalis Latham
1999
Park, K.D. | Lee, J.H. | Ku, S.K. (Kyungpook National University, Taegu (Korea Republic). Department of Histology, College of Veterinary Medicine) | Lee, H.S. (Kyungsan University, Kyungsan (Korea Republic). Department of Biology, Faculty of Basic Science)
The regional distributions and relative frequencies of the gastrointestinal endocrine cells inthe gean goose (Anser fabalisLatham) were investigated by immunohistochemical methods using bovineSp-1/chromogranin (CG), serotonin, gastrin, cholecystokinin (CCK)-8, somatostatin and glucagon antisera. BCG-immunoreactive cells were widespread throughout the gastrointestinal tract (GIT) with moderated frequencies except for the gizzard and proventriculus which were a few frequencies. Serotonin-immunoreactive cells were detected throughout the GIT except for the proventriculus and gizzard. These cells wer observed inthe pylorus with rare frequencies but numerous cells wee cetected in the intestinal tract. Gastrin-immunoreactive cells were restricted to the gizzard, pylorus and duodenum. These cells were most predominant in the pylorus and a few or rare in the gizzard and duodenum, respectively. CCK-8-immunoreactive cells were observed from the gizzard to ileum. the highest frequencies of endocrine cells were observed in the duodenum. These cells were increased from the gizzard to duodenum but thereafter decreased. Somatostatin-immunoreactive cells were detected in the GIT except for the large intestine. In the proventriculus and pylorus, numerous immunoreactive cells were demonstrated but a few cells were present in the other regions. Glucagon cells were observed in the gizzard, pylorus, ileum, colon and rectum with a few or moderated numbers.
Show more [+] Less [-]An immunohistochemical study on the endocrine pancreas of the bean goose, Anser fabalis, Latham
1999
Lee, J.H. | Ku, S.K. (Kyungpook National University, Taegu (Korea Republic). Department of Histology, College of Veterinary Medicine) | Lee, H.S. (Kyungsan University, Kyungsan (Korea Republic). Faculty of Basic Science, College of Natural Science)
The regional distribution and relative frequency of the endocrine cells in the pancreas of the bean goose were investigated by immunohistochemical methods using 6 types of the specific antisera. Spindle shpaed serotonin-immunoreactive cells were detected in the exocrine portions. Spherical or spindle shpaed glucagon-immunoreactive cells were observed in the exocrine and dark and mammalian type islets. In the dark type islets, numerous cells were dispersed throughout whole islets but they were located in the peripheral regions of the mammalian type islets. No glucagon-immunoreactive cells were detected in light type islets. Round or spherical shpaed insulin-immunoreactive cells were observed in the exocrine and dark, light and mammalian type islets. They were observed in the exocrine regions with a few numbers. Extremely rare cells were detected in central portion of the dark type islets but moderate to numerous cells were found in the central regions of the mammalian and light type islets, respectively. Spherical or spindle shaped somatostatin-immunoreactive cells were observed in the exocrine and dark, light and mammalian type islets. A few single cells were detected in the exocrine portions. In the dark type islets, numerous cells were dispersed throughout whole islets but a few to moderate numbers of cells were located in the peripheral regions of the light and mammalian type islet. Moderate numbers of the bovine pancreatic polypeptide-immunoreactiv ecells were found in the exocrime portions with round, spherical or spindle shape. But no bovine Sp-1/chromogranin-immunoreactive cells were observed in this study.
Show more [+] Less [-]Pathological evaluation of renal changes induced by myltiple nephropathogenic factors in SPF chickens
1999
Kang, K.I. | Hahn, T.W. | Han, J.H. (Kangwon National University, Chuncheon (Korea Republic). Department of Veterinary Medicine) | Mo, I.P. | Kwon, Y.K. | Kang, M.S. (National Veterinary Research and Quarantine, Anyang (Korea Republic).)
Renal failure is one of the main causes of economic impacts in the poultry industry and complex syndrome with different severity of clinical signs caused by multiple nephropathogenic factors such as infectious bronchitis viral infection and excess salt and calcium in diet. To evaluate the correlation between severity of renal failure and the causative nephropathogenic factors, one-day-old specific pathogen free chickens were treated with either single causative factor or multiple causative factors described as above. Each group was designed as control for non-treated control, IB for infectious bronchitis virus (IB virus) infection, IBHNa for IB virus infection with high diet salt, IBHCa for IB virus infection with high diet calcium, IBHNC for IB virus infection with high diet salt and calcium, HNa for high diet salt, HCa for high diet calcium and HNC for high diet salt and calcium. Chickens were inoculated with IB virus at 1-day-old and remained on their respective diets until 21 day of age. Plasma Na+, Cl-, BUN, creatinine, calcium and uric acid values were examined. The results obtained were as follows; IB virus and high dietary calcium combined treatment showed elevated plasma uric acid. BUN and creatinine values were not characteristic on chicken renal failure. But plasma uric acid values were increased according to renal lesion. Hypercalcemia and hyperuicemia did not induce urate deposition and mineralization in the kidney.
Show more [+] Less [-]Effects of xylazine-ketamine combination on serum ACTH, corticosterone and glucose concentrations in rabbits
1999
Park, K.M. | Li, L.H. | Han, S.K. | Ryu, P.D. (Seoul National University, Suwon (Korea Republic). College of Veterinary Medicine)
Anesthetic agents are useful in inducing the anesthesia for surgical operations and various biological experiments, but they can disturb the body homeostasis and cause the stress in animal. Much efforts have been directed on reducing such side effects of anesthesia. In this work, we measured the serum ACTH, corticosterone and glucose concentration in rabbits to compare the degree of stress induced by two commonly-used anesthetics, ketamine, xylazine, and the combination of xylazine and ketamine. 1. The anesthesia was induced in about 10 min in the rabbits treated with xyalzine, ketamine and xylazine-ketamine. The duraion of complete loss of righting reflex were 12, 13 and 115 min in the groups treated with xylazine, ketamine and xylazine-ketamine, respectively. 2. Serum ACTH concentrations in all treatment groups were gihger than those in control group. At 30 min after the administration of the drugs, serum ACTH levels in ketamine-treated group were significantly higher than those in control, xylazine- and xylazine-ketamine-treated groups. However, at 1, 2, 5 and 9 hours after the drug administration, serum ACTH levels in xylazine-treated-group were gihger than those in control. 3. Serum corticosterone levels in xylazine- and xylazine-ketamine-treated groups were lower than those in control or ketamine=treated groups at 0.5 and 1 hour after the administration. However, at 5 and 9 hours after the administration, serum corticosterone levels inxylazine- and xylazine-ketamine-treated groups were significantly higher than those in ketamine-treated group or control. 4. Serum glucose levels transiently increased to 3 tives of the pre-injection levels at 0.5 and 1 hours after the administration in xylazine or xylazine-ketamine-treated groups, but were not changed in control and ketamine-treated group. These results indicate that xylazine-induced stress lasts longer than ketamine-induced, suggesting that the difference in stress-related hormone levels during anethesia could be due to the differences in modes of actions of individual drugs used and the depth of anesthesia.
Show more [+] Less [-]Anesthetic and cardiovascular effects of xylazine/fentanyl/azaperone and medetomidine/midazolam as preanesthetics and combinations with their antagonists in halothane-anesthetized dogs
1999
Yang, H.S. | Kweon, O.K. | Woo, H.M. | Nam, T.C. (Seoul National University, Suwon (Korea Republic). College of Veterinary Medicine)
This study was performed to evaluate anesthetic and cardiovascular effects of xylazine/fentanyl/azaperone and medetomidine/midazolam as preanesthetics and their combinations with antagonists in halothane-anesthetized dogs. Eight clinically healthy dogs(4.54+_2.16kg) were used at the interval of more than 14 days between experiments in turn for propionyl promazine(PP 0.3mg/kg, IM), xylazine/fentanyl/azaperone(XFA 2mg/kg, 0.0137mg/kg, 0.11mg/kg, IM), medetomidine/midazolam(MM 0.02mg/kg, 0.3mg/kg, IM), combination of XFA and their antagonists (yohimbine 0.05mg/kg, naloxon 0.0005 mg/kg, IV) and combination of MM and their antagonist(atipamezole 0.08mg/kg IM). The sedation induction times in XFA(2.56+_1.01 min) and MM(5.44+_2.07 min) groups were sighificantly better than that of PP group(10.75+_2.38 min)(p0.05). The thiopental sodium dose required for tracheal intubation in XFA(2.38+_3.38mg/kg) and MM(3.91+_3.47mg/kg)groups were significantly less than that of PP group(12.57+_2.13mg/kg)(p0.05). All time indices expressing the recovery(pedal refles recurrence time, extubation time, arousal time, standing time and walking time) were significantly shorter in the combination groups of XFA or MM with their antagonistis than in PP, XFA and MM groups(p0.05). The suppressions of cardiovascular function of XFA and MM were more than that of PP. Heart rate and cardiac output were recovered by the antagonists of XFA and MM, but mean arterial pressure were not recovered by the antagonists.
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