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.

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.

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.

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.

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.

This study was undertaken to detemine the normal appearance of the postpartum uterine involution. Postpartum changes in uterine shape, architecture, echogenicity and diameter were monitored with ultrasonography in 11 Korea Jin-do dogs. Serial ultrasonographic examination was done daily during the first week, 3 days interval from 8 to 30 days, and weekly from 31 to 100 days postpartum, respectively. Of 11 postpartum bitches, 10 bitches(90.9%) had normal involution and 1 bitch (9.1%) had subinvolution of the placental sites (SIPS) by gross findings, vaginal discharges, and by ultrasographic findings, uterine shape and echogenicity. The excretory period of vaginal discharges in 10 normal bitches of uterine involution was finished completely at 20.2+_4.6 days(Mean+_SD) postpartum, but in 1 SIPS at 50 days postpartum. The short axis shape of the uterus was initially often flaccid-appearing. It varied from circular to crescent shaped to polygonal.This lasted until 16.5+_3.7 days postpartum, during which time the short axis uterine shape gradually changed to circular. Also, the long axis shape of the uterus was created a beaded appearance of the horns until 29.9+_3.2 days postpartum. After 30 days, it was appeared as tubular shape without distinguished between placental and interplacental sites. The ultrasonographic image of the postpartum uterus consisted of four echogenicity distinct layers. Ulterine wall was represented the very hyperechoic serosa, hypoechoic mycomtrium, hyperechoic endometrium and anechoic structures of fluid in the uterine cavity until 7 days postpartum. The individual uterine layers were most prominent during the first weeek postpartum,and they became progressively less distinct throughout the course of uterine involution. Anechoic structrues of fluid inthe uterinecavity was detected until 25.0+_6.4 days prostpartum, after which time it was not reliably detected. The uterine diameter was decreased not only in the placental sites from 24.1+_2.5mm at 1 day to 15.4+_1.4mm at 7 days postpartum, but also in the interplacental sites 14.9+_1.5mm at 1 day, 10.6+_0.8mm at 7 days postpartum. There was a general trend of decresing uterine diameter, whichoccurred more rapidly at the placental sites. At 31 days postpartum, these diameter reached almost same size, after that time, they could be not distinguished between placental and interplacental sites. At 87 days postpartum, the uterine diameter was 5.6+_0.6mm both placental and interplacental sites, and the uterine horns were uniform hypoechoic, tubular structures without enlargement. Therefore, complete involution of the uterus occurred at that time. It was concluded that normal postpartum uterine involution in Korea Jin-do bitches appeared to be completed around 87 days postpartum by gross findings such as vaginal discharges, and by ultrasonographic findings, uterine shape and echogenicity. Also ultrasonographic characteristics of the postpartum uterine involution were described. Therefore, these result suggest that ultrasonographic assessment is a reliable method for diagnosing the SIPS and uterine dysfunction, such as pyometra in bitch.

Magnesium (Mg2+) plays an important role in the regulation of a range of intracellular processes. Regulation of extracellular Mg2+ contents was studied in the anesthetized Sprague-Dwley (SD) rats. Animals were injected intraperitoneally with sodium nitrite (NaNO2), and circulating Mg2+([Mg2+]c) was observed in animals injected with NaNO2 at a dose of 10mg/kg or higher. Pretreatment with methylene blue preventd the NaNO2-induced increse in [Mg2+]c.[Mg2+]c displayed an inverse linear correlation with hemoglobin and exponential correlation during NaNO2 injection. Injection of KCN or rotenone also induced an increase in [Mg2+]c. An increase in [Mg2+]c was observed when respiration rate was reduced from 100/min (140ml/min) to 10/min (14ml/min) during 30 min. These results indicate that changes in [Mg2_]c inversely reflect alteration of ATP in a model of metabolic inhibition

Previous studies suggested that the inhibition of thyroxine (T4) release by alpha1-adrenoceptor and muscarinic receptor stimulation results in activated protein kinase C (PKC) from mouse and guinea pig thyroids. In the present study, the effect of carbachol, methoxamine, phorbol myristate acetate (PMA), and R59022 on the release of T4 from the mouse, rat, and guinea pig thyroids was compared to clarify the role of PKC in the regulation of the release of T4. The thyroids were incubated in the medium containing the test agents, samples of the medium were assayed for T4 by EIA kits. Forskolin, an adenylate cyclase activator, chlorophenylthio-cAMP sodium, a membrane permeable analoge of cAMP, and isobutyl-methylxanthine, a phospho-diesterase inhibitor, like TSH (thyroid stimulating hormone), enhaced the release of T4 from the mouse, rat, and guinea pig thyroids. Methoxamine, an alpha1-adrenoceptor agonist, inhibited the TSH-stimulated release of T4 in mouse, but not rat and guinea pig thyroids. In contrast, carbachol, a muscarinic receptor agonist, inhibited the release of T4 in guinea pig, but not mouse and rat thyroids. These inhibition were reversed by prazosin, an alpha1-adrenoceptor antagonist or atropine, a muscarinic antagonist or M1- and M3- muscarinic antagonists, in mouse or guinea pig thyroids. In addition, staurosporine, a PKC inhibitor, reversed methoxamine or carbachol inhibition of TSH stimulation. Furthermore, PMA, a PKC activator, and R59022, a diacylglycerol (DAG) kinase inhibitor, inhibited the TSH-stimulated release of T4 in mouse, rat, and guinea pig thyroids. These inhibition were blocked by staurosporine. these findings suggest that the activation of receptor or DAG inhibits TSH-stimulated T4 release through a PKC-dependent mechanism in thyroid gland.

Atrial natriuretic peptide(ANP) is a hormone with potent natriuretic, diuretic and relaxing properties on vascular smooth muscle. Specific chemical modulator in response for the ANP secretion has not been found yet. Therefore, we have investigated the role of Ca2+ responsible for the regulation of ANP induced by protein kinase C(PKC) on mechanically stretch-induced ANP secretion in the rat atria. The results obtained were as follows; 1. ANP secretion and ANP concentration were increased to more in Ca2+-free buffer than in the Kreb-Henseleit buffer on mechanically stretch-induced ANP secretion(p0.05), but extracellular fluid translocation(ECF) was not significant. Phorbol 12-myristate 13-acetate(PMA, 10-7M) induced ANP secretion and ANP concentration in Ca2+-free buffer shown to more accentuate on mechanically stretch-induced ANP secretion than in the Ca2+-free buffer(p0.05), but ECF translocation was not significant. 2. In the presence of ryanodine(3*10-6M), PMA(10-7M) induced ANP secretion and ANP concentration in the Kreb-Henseleit buffer were shown to more increase on mechanically stretch-induced ANP secretion than in the ryanodine(3*10-6M) with the Kreb-Henseleit buffer(p0.05), but ECF translocation was not significant. 3. In the presence of ryanodine(3*10-6M), PMA(10-7M) induced ANP secretion and ANP concentration in the Ca2+-free buffer was shown to more increase on mechanically stretch-induced ANP secretion than in the ryanodine(3*10-6M) with the Ca2+-free buffer on mechanically induced ANP secretion(p0.05), but ECF translocation was not significant. The results suggest that PKC-induced ANP secretion may not be related to the change of Ca2+ on mechanically induced ANP secretion in the rat atria.

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.