We performed this study to evaluate the potential clinical marker of urinary trypsinogen-2 together with amylase, lipase and urinary amylase creatinine clearance ratio (ACCR) for the diagnosis of acute pancreatitis in dogs. In the experiment on daily changing pattern of amylase, lipase and ACCR measurements in experimentally induced pancreatitis dogs, compared to values measured in pre-induction state, significant difference was seen in amylase until 5th day of induction, and for lipase significant difference was found during the 7th day of observation period (p0.05). No significant difference was found in ACCR for the study period (p0.05). On SDS-PAGE analysis of urine from experimentally induced pancreatitis dog, The 26kd band was markedly increased compared with that of normal state and that band was confirmed trypsinogen-2 using substrate interaction and isoelectric focusing assay after being eluted. When assessing the appearance of 26kd band on urine SDS-PAGE 87.1% (range:50~100%) of experimentally induced pancreatitis dogs showned positive results, whereas no corresponding band was seen in dog without pancreatic disorders. With this result, determination of urinary trypsinogen-2 assay was found to have a high diagnostic value with a 70% of sensitivity and 100% of specificity as a routine test for pancreatitis, although the detection of trypsinogen-2 in urine can be varied on the progressionstage of pancreatitis at the initial visit to animal clinic. We therefore suggest that the promising results in this study be used for the development of dipstick test for detecting acute pancreatitis in the future research.

The regional distributions and relative frequencies of the gastrin, secretin and pancreatic polypeptide(PP)-immunoreactive cells in the gastrointestinal tract of the fetus(180 days of gestation) of Korean native goat were sutdied with immunohistochemical(ABC) methods. Gastrin-immunoreactive cells were detected in fundus, pylorus and duodenum and these cells were most predominant in pylorus. Secretin-immunoreactive cells were observed in pylorus, duodenum and ileum. PP-immunoreactive cells were restricted to fundus. These immunoreactive cells were situated in surfact epithelium and mucosal gland regions. The regional distribution and relative frequency of PP-immunoreactive cells was somewhat different to the adult Korean native goat. Immunoreactive cells in thesurface epithelial regions were open typed cells which were spindle shaped cells but closed typed cells which were round or/to spherical shaped cells were observed in the mucosal gland regions.

A species-specific 760 base pair(bp) BamHI to EcoRI DNA fragment(fMG-2) of lipoprotein gene was isolated from a Mycoplasma gallisepticum (M gallisepticum) genomic library. Based on the DNA sequence data of fMG-2, a pair of 25bp primers was synthesized. When used in the polymerase chain reaction(PCR), 732bp DNA products were amplified from 6 standard strains and 10 field isolates of M gallisepticum, but not from 2 Mycoplasma synoviae and 7 other Mycoplasma species. The lower detection limit was 100fg of the genomic DNA. Identity of the PCR products was confirmed by comparison of patterns of restriction endonuclease analysis with AseI, DraI, EcoRV and SspI.

This study was carried out (a) to investigate the variations of blood chemistry and (b) to examine the secretion trend of endocrine substances in a dog model after electroacupuncture and laser stimulation at different time period(9 to 11a.m. and 6 to 8p.m.). Two acupuncture points; Da Zhui(GV-14) and Shen Shu(BL-23) were electroacupunctured for 20 minutes with 2Vol, 20Hz and irradiated for 5 minutes with 8,000Hz. Before stimulation and after a lapse of time(10-minutes, 30-minutes and 60-minutes) all dogs were checked the following parameters; cortisol, ACTH, RBC, hemoglobin, hematocrit, WBC, Ca, P, SGPT, SGOT and creatinine. The results were as follow:The levels of cortisol and ACTH have been increased 10 minutes after the stimulation of the electroacupuncture and laser. The higher levels of cortisol and ACTH have been decreased to keep the normal levels from 30 minutes after the stimulation of the electroacupuncture and laser. The RBC, hemoglobin, hematocrit and WBC showed the physiological phenomena in the electroacupuncture and laser stimulation. In sero chemical analysis, calcium, phosphate, SGOT, SGPT and creatinine levels were within normal physiological ranges.

The present study was carried out to investigate the blood markers of Cheju horses. The red cell types (blood groups) were tested from 73 Cheju native horses and 118 Cheju racehorses by serological procedures with 23 reagents. The blood group phenotypes observed with high frequency were Pb(34.3%), Qc(56.2%), Qb(15.1%) and genotypes Dbcm/dghm(12.3%), Dde/dghm(9.6%), Dad/bcm(6.8%), Dcgm/de(6.8%) in Cheju native horses, while Aa(63.6%), Pa(44.9%), P-(28.8%), Qabc(36.4%), Dbcm/cgm(14.4%), Dbcm/bcm(10.2%), Dbcm/de(7.6%), Dbcm/dghm(5.1%), Dde/dk(5.1%) in Cheju racehorses. Alleles observed with high frequency were Ab(0.128), Ac(0.169), Dad(0.103), Dadn(0.075), Ddghm(0.226), Pb(0.316), Qc(0.494) in Cheju native horses and Aa(0.529), Dbcm(0.306), P-(0.531), Qabc(0.197), Q-(0.504) in Cheju racehorses. No specific variation of blood groups and allele frequencies of C, K, U system were observed in Cheju native horses and Cheju racehorses. The mean heterozygosity in Cheju native horses and Cheju racehorses was observed 0.5344 and 0.5102, respectively.

Ontogeny, distributions and relative frequencies of somatostatin-immunoreactive cells were investigated in the proventriculus of the chicken embryos with incubation periods. Samples were taken from 10 groups(10 days of incubation to hatching) and studied by immunogistochemical methods. The findings were as follows. Somatostatin-immunoreactive cells were observed from 12 days of incubation in the proventricular glands and after that increased with incubation periods. The first observation time of these cells in the epithelium were at 15 days of incubation in the basal portion but in 16 and 17 days of incubation, no immunoreactive cells were observed in the epithelium but after that a few immunoreactive cells were observed in the basal portion and gastric gland regions. The shpaes of these cells were spherical to spindle in the proventricular glands and spherical to round in the epithlium and gastric gland.

Two-day old house fly adults were exposed to six insect growth regulators, flufenoxuron, teflubenzuron, triflumuron, diflubenzuron, methoxyfenozide, tebufenozide, as a feed additive (milk + 5% sugar + chemical) in the laboratory for 6 days. The number of eggs deposited by the exposed-adults, viability of the eggs, and F1 larval development were checked. All the IGRs tested were found to have no adverse effect on the reproduction of house fly, except methoxyfenozide (210ppm). The most effective inhibitor to egg hatch was flufenoxuron, followed by teflubenzuron, triflumuron, and diflubenzuron. Exposure to flufenoxuron (over 5ppm), teflubenzuron (over 25ppm), triflumuron (over 125ppm), and diflubenzuron (over 125ppm) reduced egg hatchability to 0 to 1.3%, but lower concentrations of these IGRs were less effective (6.3 to 46.3% egg hatchability). Almost all the larvae emerged from eggs deposited by the adults exposed to diflubenzuron (62.5ppm) and teflubenzuron (12.5ppm) failed to develop into pupae, causing total mortalities of 98% and 100%, respectively. However, two IGRs, methoxyfenozide and tebufenozide, did not inhibit egg hatch and F1 larval development, except methoxyfenozide (210ppm) treatment. These results suggest that these 4 IGRs may be used in the development of autosterilization system for house fly control. However, further work is required to develop delivery systems capable of transferring an effective dose to the fly under field conditions.

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

This study was carried out to nvestigate the NPY-immunohistochemical characteristics of the olfactory bulb in the striped field mouse(Apodemus agrarius). The animals were anesthesized with thiopental sodium and perfused with 4% paraformaldehyde through left ventricle and aorta. Brains were removed and tranfered 10%, 20% and 30% sucrose. Sections were then cut on a cryostat into 40 micro meter-thick. The tissue immunostained with avidin-biotinylated complex method. The main olfactory bulb consisted of seven circumferential laminae: and olfactory nerve fiber layer, a glomerular layer with glomeruli surrounding by periglomerular cells, an external plexiform layer having granule and tufted cells, a mitral cell layer, a narrow internal plexiform layer, a granule cell layer forming several cell rows and alayer of white matter. The accessory olfactory bulb had four layers: an olfactory or vomeronasal nerve fiber layer, a glomerular layer consisting of small glomeruli, a mixed layer not distinguishing the external plexiform/mitral cell/granule cell layers and a granule cell layer. Most of NPY-immunoreactive(NPY-IR) neurons in main olfactory bulb were localized in the deeper portion of granule cell layer, white matter and anterior olfactory nucleus. In addition, some NPY-IR neurons were identified in the external plexiform layer. The shape of NPY-IR neurons of all olfactory bulb were predominant round or oval, sometime multipolar in shape. And most NPY-IR processes were parallel to long axis of white matter. In accessory olfactory bulb, NPY-IR neurons were not found in all region.

Ischemic preconditioning (IPC), i.e., a preliminary brief episode of ischemia and reperfusion, has been shown to reduce the cell damage induced by long ischemia and reperfusion. Superoxide radical which is produced during reperfusion after ischemia was recongnized as a factor of the ischemic injury and it is dismutated into H2O2 and O2 by two types of intracellular superoxide dismutase (SOD), Cu,Zn-SOD in cytoplasm and Mn-SOD in mitochondria. Recently oxygen free radicals are suggested to induce the apoptosis, however mechanism of the reduced apoptosis by ischemic precondition was unknown, while many studies performed in mammalian heart indicated that ATP-sensitive K+(K atp) channel activation related with the protective effects. The aim of present study is toinvestigate 1)whether IP upregulate the Cu,Zn-SOD and Mn-SOD activities, and 2)whether ischemic preconditioning decreases apoptosis via K atp channel activation in timely reperfused skeletal muscle after long ishemia. The experimental animals, Sprague-Dawley rats weighing 250~300g, were divided into 8 group; 1)control group, 2)ischemic preconditioning only groups, 3)pinacidil, a K atp channel opener, treatment only groups, 4)glibenclamide, a K atp channel blocker, treatment only groups, 5)ischemia groups, 6)ischemia after IPC groups, 7)ischemia and pinacidil treatment groups, and 8)IP and ischemia after glibenclamide pretreatment groups. Animals of the control group were administered with the vehicle (DMSO) alone. Pinacidil (1mg/kg) was administred intravenously 5 minutes after initiation of ischemia, and glibenclamide(0.5mg/kg) was injected intravenously 20 minutes before IPC. In rats that were ischemic preconditioned, the left common iliac artery was occluded for 5 minutes followed by 5 minutes of reperfusion by three times usign vascular clamp. Ischemia was done by occlusion of the same artery for 4 hours. The specimens of left rectus femoris muscle were obtained immediately (0 hours), 12 hours, 24 horus after drug administrations, IP or ischemia and reperfusion. The immunoreactivities of SOD and its alterations were observed by use of sheep antihuman Cu,Zn-SOD and Mn-SOD antibodies onthe 10 micro meter cryosections. The incidencies of apoptosis were observed by TUNEL methods with in situ apoptosis detection kit on 6 micro meter paraffine section. The results obtained were as follows: 1. After IPC, immunoreactivities of Cu,Zn-SOD mainly in the small-sized fibers were increased by 24 hours, that of Mn-SOD at 0 hour and 24 hours. 2. No significant changes in immunoreactivities of SDO was observed in the pinacidil and in the glibenclamide treatment only groups, and in the ischemia only groups. 3. The immunoreactivities of the Cu,Zn-SOD were incresed in the ischemia after IPC groups and the ischemia and pinacidil treatment groups. 4. The immunoreactivities of the Cu,Zn-SOD in the IPC and ischemia after glibenclamide pretreantment groups were not increased except for the 12 hours reperfusion group. But, Mn-SOD immunoreactivities were incresed in to 0 hours, 12 hours and 24 hours after reperfusion. 5. In the control group, the IPC only groups, and the pinacidil treatment only groups, negative or trace apoptotic reactions were observed, but the positive apoptotic reaction occured in the glibenclamide treatment groups. 6. Moderate or many number of apoptosis were revealed in the ischemia groups, and also the IPC and ischemia after glibenclamide pretreatment group except for 12 hours and 24 hours after reperfusion. However, the incidence of apoptosis was decreased in the ischemia after IPC groups and in theischemia and pinacidil treatment groups. 7. There is a coincidence between the increase of Cu,Zn-SOD immunoreactivities and the decrease of apoptosis in thepresence of ischemia and reperfusion. These results suggest that the protective effects of ishemic preconditioing may related to the SOD activation, and the ischemic preconditioning decreases the apoptosis partially via K atp channel activation in timely reperfused rat skeletal muscle. It is also suggested that inhibition of apoptosis by IPC may related with the SOD activation.