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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 [-]Observations of pneumonia inslaughtered pigs according to season
1999
Lee, S.K. | Han, J.H. (Kangwon National University, Chuncheon (Korea Republic). Department of Beterinary Medicine) | Jeong, H.K. (Dodram Pig Farmers Cooperation, ichon (Korea Republic).)
From 2,373 slaughtered pigs examined, 1,899 of them had visible evidence of pulmonary lesion. The lungs with pulmonary lesion were examined by grossly, histopathological technique to investigate the prevalence accordint to season, severity ofpulmonary lesion, types of the pneumonia and relationship between rearing management and prevalence of pulmonary lesion. The results were as follows: 1. Prevalence of pneumonia according to season was 72.3 % to 85.9% and the higher prevalence was in winter. 2. In the severity of pulmonary lesion, rates mild, moderate and severe lesions were 38.3%, 47.0% and 14.7%, respectively. Prevalence of mild lesion was the highest in autumn. Prevalence of moderate and severe lesions was the hihgest in winter. 3. In the type of pneumonia, rates of interstitial pneumonia and bronchopneumonia fibrinous pneumonia were 23.6%, 13.0%, and 3.4%, respectively. Prevalence of interstitial pneumonia was high in spring and autumn while that of broncho-pneumonia and fibrinous pneumonia was high in winter and summer, respectively. 4. In relationship of pulmonary lesions according to severityo fpulmonary lesion and types of pneumonia, similarity was observed in prevalence of mild lesion and bronchopneumonia.
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 [-]Production and partial purification of Staphylococcus aureus alpha toxin
1999
Park, H.M. | Oh, T.H. | Han, H.R. (Seoul National University, Suwon (Korea Republic). Department of Veterinary Internal Medicine, College of Veterinary Medicine)
Alpha toxin of S aureus has cytolytic activity respectively. This antigen has been received the most attention since it is a major virulence factor in pathogenesis of staphylococcal mastitis. Thus, alpha toxin has been focused as potential candidate ofvaccine tominimize mastitis in cows. the purpose of this study was to develop a simple, efficient production and purification methods of sufficient amount of alpha toxin antigen from S aureus. Alpha toxin production measured by hemolytic activity was the highest at 18 hrs postinoculation in yeast extract culture medium supplemented with thiamine, nicotinic acid and casamino acid. Alpha toxin was purifed by ammonium sulfate precipitation (65%) and ultrafiltration. Molecular weight of the toxin was 33 kDa in the analysis with SDS-PAGE. Conclusionally, when alpha toxin was included in the vaccine, the optimal harvest time of alpha toxin was at 18 hrs after inoculation in yeast extract medium supplemented with thiamine and nicotinic acid.
Show more [+] Less [-]Ultrastructure of hemal node and hemolymph node in Korea native goat
1999
Yoon, Y.S. | Lee, J.S. (Seoul National University, Suwon (Korea Republic). Department of Veterinary Histology, College of Veterinary Medicine) | Shin, J.W. (Kangwon National University, Chuncheon (Korea Republic). Department of Veterinary Medicine)
Hemal nodes and hemolymph nodes are lymphoid organs that share morphologic and functional characteristics of lymph nodes and spleens. The aim of the present study was to obtain new informations on the distinct morphological structures of hemal nodes and hemolymph nodes according to ages, andto get the basic data for their functions in Korean native goats. Goats were divided into 5 groups, consisting of 3 animals aged 1, 3, 6, 10 and 12 months, respectively. Ultrastructural features of the organs were observed by transmission and scanning electron microscopes. The sinuses of hemal nodes and hemolymph nodes were lined by enodthelial-like reticular cells which had euchromatin-rich nuclei and many cytoplasmic processes, surrounding collagen fibrils. Macrophages containing phagocytosed erythrocytes were often noted in the diffuse lymphatic tissues of hemal nodes and hemolymph nodes. Some mast cells were in contact with the plasma cells near the blood vessel. Hemal nodes and hemolymph nodes had venous sinusal-like vessels which were different from the deep sinus. The lymph vessels with valves were observed in the capsule of the hemolymph node. There were no ultrastructural differences of the organs in the age different groups of the animals. These results suggest that hemal nodes and hemolymph nodes may take part in hemopoiesis, blood filtration and immune reaction in Korean native goats.
Show more [+] Less [-]Effects of diazepam on fetal development in rats
1999
Kim, C.J. | Kim, Y.J. | Yu, I.J. (Chonbuk National University, Chonju (Korea Republic). Department of Obstetrics, College of Veterinary Medicine)
To investigate the effect of diazepam on fetal development in pregnant rats, this experiment was performed in eighty Sprague-Dawley female rats which were 8 weeks old and grouped into two according to different diazepam treatment period during 5-9 days of gestation and 10-14 days of gestation. Both experimental groups were included by saline treated groups (control) and diazepam-treated groups (6mg, 12mg and 24mg), respectively. Diazepam was injected to pregnant rats subcutaneously, which were wacrified on 20 days of gestation and mean litter size, fetal body weight, fetal crown-rump length (CRL) and pathological findings were examined. 1. Concerning mean litter size, diazepam-treated groups showed lower mean litter size than control in both 5-9 days and 10-14 days of gestation groups(p0.05) without difference according to dosage of diazepam and day of gestation. 2. Concerning fetal body weight, diazepam-treated groups during 5-9 days of gestation showed lower fetal body weight than control and the other treated group during 10-14 days(p0.01) without difference according to dosage of diazepam. Diazepam-treated group during 10-14 days of gestation showed no difference among experimented groups. 3. Concerning fetal crown-rump length (CRL), diazepam-treated groups during 5-9 days of gestation showed shorter CRL than control and the other treated group during 10-14 days of gestation(p0.01) without difference according to dosage of diazepam. 4. Reduction of mean litter size, fetal body weight and CRL was shown from when treated by the dosage of 6mg/kg of diazepam. 5. Maternal mortality according to dosage of the 20mg/kg of diazepam were 30% and 20% in the treated group during 5-9 days and 10-14 days of gestation, respectively. These results indicated that diazepam treatment in pregnant rats caused considerable reduction of mean litter size, fetal body weight and fetal crown-rump length when treated during 5-9 days of gestation.
Show more [+] Less [-]Localizations of substance P, CGRP and calcium binding proteins in Korean native goat duodenum
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) | Kang, T.C. | Won, M.H. (Hallym University, Chuncheon (Korea Republic). College of Medicine) | Seo, J.H. (Seoul National University, Seoul (Korea Republic). College of Medicine)
The localization of substance P(SP), calcitonin gene-related peptide(CGRP) and three calcium binding proteins, calbindin D-28k(CB), calretinin(CR) and parvalbumin(PA) was immunohistochemically examined in the myenteric and submucous plexuses of Korean native goat duodenum. In the neurons of myenteric and submucous plexuses of duodenum, immunoreactivities of SP, CGRP and CB were confirmed in both nerve cell bodies and fibers. In contrast, CR immunoreactivity was found only in nerve fibers of myenteric plexuses, while PA immunoreactivity was found only in nerve cell bodies of submucoun plexuses. In the inner circular muscle layer, dense SP-like immunoreactive fibers were prominent but only a few CGRP-like immunoreactivities were observed. Most of SP-and CGRP-like immunoreactive neurons of both plexuses colocalized with CB. This result showed that SP and CGRP may have a important role for the movement of small intestine. The colocalizations of CB with SP or CGRP in myenteric and submucous plexuses suggest that CB may serve neuromodulatory role for SP-and CGRP-immunoreactecd neurons on the movement of intestinal wall.
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 [-]Detection of bovine viral diarrhea virus by In situ hybridization
1999
Park, N.Y. | Hong, K.K. | Chung, C.Y. | Cho, K.O. | Lee, B.J. | Park, Y.S. | Park, H.S. (Chonnam National Univerisyt, Kwangju (Korea Republic). College of Veterinary Medicine) | Kweon, C.H. (National Veterinary Research and Quarantine Service, Anyang (Korea Republic).)
Detection and distribution of bovine biral diarrhea virus(BVDV) was studied in formalin-fixed, paraffin-embedded tissues from two naturally infected cattle by in situ hybridization with a non-radioactive biotinylated probe. A 600 base pair cDNA probe from BVDV B-25 strain ws used for probe. The whole procedure of ISH to diagnose was carried out within 1~2 hours in MicroprobeTM capillary action system. The biotin-labelled probe was demonstrated after hybridization under standard conditions by the application of streptoavidin and biotinylated alkaline phosphatase. Alkaline phosphatase was visualized using a fast red TR/naphthol phosphatase and the sections were counterstained with hematoxylin. We have obtained the result of positive reactions in digestive tract(sm1.all intestine and colon) and epidermis of tongue in the state of the intract tissues. The result suggested that in situ hybridization method can be considered as a useful diagnostic technique for detection of specific nucleic acid sequences of BVDV.
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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