This study was conducted to evaluate the ELISA kit for measuring the level of cortisol in the urine. The CV of within-run variation and day to day variation were 0.4-2.8 and 1.8-5.7, respectively. the minimum limitation of measurement was 1ng/ml. the cross reaction was high (CR50(%)=11.4-43.2) in prednisolone, 11-deoxycortisol, 21-deoxycortisol and predinosone. There was low and no cross reaction in other steroid. To develop the ELISA kit we measured the cortisol level in diluted urine with PBS (procedure I), extracted urine with methylene chloride (procedure II) and extracted methylene chloride-extracted urine from thin-layer chromatography (procedure III). The CV value of procedure I, II, III was 9.4-28.3%, 7.2-8.9% and 2.5-5.7%, respectively. There was significant difference between procedure I with II, and pro-cedure I with III(p0.01), but no difference between procedure II with III significantly(p0.01). The mean UCCR of urine collected through am 8 to 10 ws 9.5+_7.6(0.14-28.0) in 12-month-old dog(n

Since Robert Koch found tubercle bacilli in 1882, the studies on tubercle bacilli of human and animal had been carried out. Being old tuberculin(OT) introduced in 1890, the specificity of the diagnosis of tuberculosis has been improved by continual uses of heat concentrated synthetic medium(HCSM) and purified protein derivatives(PPD) tuberculin. Now, two types of tuberculin test are used worldwidly; the single intradermal test(SIT) using bovine tuberculin and the single intradermal comparative tuberculin test(SICTT) using avian and bovine tuberculins. In the SICTT, each countries have used with different combination of both avain and bovine tuberculins' titers. However, this kinds of studies have not reported in Korea. Therefore, the studies on the combination of their tuberculins' titers were performed through intradermal test of guinea pigs sensitized with either Mycobacterium bovis or M avium and were examined in 10 cattles of SIT positive reactors. Also, IFN-y assay, the latest diagnostic method of bovine tuberculosis, was experimentally applied to SIT positive reactors. For determinint the optimal titers, sensitized guinea pigs with M bovis and M avium were intradermally injected avain and bovine tuverculin. In guinea pigs sensitized with M bovis, bovine tuberculin 50 T.U. showed significant difference from all tested concentrations of avian tuberculin(p0.05). In guinea pigs sensitized with M avium, there is significantly different between bovine tuberculin and avian tuberculin by 25 T.U.(p0.01). Therefore, optimal titers of bovine and avian PPD tuberculins' titers for the SICTT in Korea were 5,000 and 2,500 tuberculin units, respectively, and the swelling differences between bobine and avian site in SIT positive reactors were above 3mm. Also, in IFN-y assay, the 9 SIT positive reactors were showed all the positive reactions.

We have studied, by a nonisotopic in situ end-laveling(ISEL) technique, frequency of apoptosis in the external granular layer(EGL) of the cerebellum of immature mice by Y-rays irradiation from 60Co or diagnostic ultrasound exposure. The total number of normal cells and cells showing morphological features of apoptosis were counted. The frequency of apoptotic cells was expressed as a percentage of the total number of cells in EGL. The extent of changes following 200 cGy(1090 cGy/min) was studied at 2, 4, 6, 8, 12, or 24 hours after exposure. The maximal frequency was found 6~8 hours after exposure. The immature mice that received 18, 36, 54, 108, 198, 396, cGY of y-rays or diagnostic ultrasound(7.5MHz, 4.2mW, Ispta=7.9mW/cm2, Ispta=114.3W/cm2) for 10 or 30 minutes were examined 6 hours after irradiation. Measurements performed after y-ray irradiation showed a dose-related increase in apoptotic cells in each of the mice studied. The dose-response curves were analyzed by a linear-quadratic model;frequency of apoptotic cell in the EGL was y=(0.1349+_0.01175)D+(-0.0001522+_0.0000334)D2+0.048(r2=0.981, D-dose in cGy). In the experiment of ultrasound exposure, the frequency of apoptotic cell was 0.106+_0.130(10 minutes exposure) and 0.167+_0.220(30 minutes exposure). We estimated the relative dose of the yield from the experiment with ultrasound by substituting the yield from ultrasound exposure into the curve from the y-irradiation. The relative dose of ultrasound exposure compared with y-irradiation were 0.432 cGY(10 minutes exposure) and 0.885 cGY(30 minutes exposure). We have found that there is no evidence to indicate that diagnostic ultrasound involves a significant risk.

This study was conducted to examine the outbreak rate and the causative agents of otitis externa in 26 dogs(49 ears; 23 dogs = bilateral, 3 dogs = unilateral), and the normal microfloras of external ear canal in 68 dogs(133 ears; 65 dogs = bilateral, 3 dogs = unilateral) in Taegu, 1997. The breed, living environment, sex, age and season distribution of otitic dogs were as follows: Dogs with erect and hairy ears(42.3%), pendulous and hairy ears(38.5%), indoor(92.3%), female(65.4%) and below one year old(38.5%) were more prevalent. According to season, otitis externa was mainly occurred between July and October. The major causative agents of canine otitis externa were Malassezia pachydermatis(32.7%), Staphylococcus aureus(26.5%) and S intermedius(16.3%). The major causative agents of canine otitis externa were Malasszia pachydermatis(32.7%), Staphylococcus aureus(26.5%) and S intermedius(16.3%). In the microorganism isolated 39 otitic ear canals, single infection was 53.8% and mixed infection was 46.2%. The normal microfloras of canine external ear canal were fungi including M pachydermaits, Aspergillus spp, Microsporum canis, Alternaria spp, Verticillium spp and Yeast, and bacteria including Staphylococcus spp(10 species including S xylosus), Bacillus spp, Corynebacterium spp, Listeria spp, Actinomyces pyogenes and Escherichia coli. No growth was 34.6%.

This study was designed to evaluate the effect of cyclophospaide(CY) on dietynitrosamine(DEN)-induced hepatic tumors in rats. Thirty five male of femal Sprague Dawley rats were continiously given water containing O.01% DEN for 10 weeks and then were give with CY 25mg/rat/day in water for 3, 5, 7 or 9 days. The livers of rats were removed and fixed in 10% buffered neutral formalin. The appearances of positive cells by immunohistochemical methods using proliferating cell nuclear antigen (PCNA) antibody, p53 antibody and apoptotic kit were investigated. The livers of rats given with CY were grossly brilliant, red-brown color, flexible, and thin border, and stainability of the liver cells were restored microscopically, and the vaccuolated and degenerated regions were differentiated from restored regions. These restored findings also were advanced in control group becouse of no DEN treatment but tended to be less avanced. In immunohistochemistry, positive cells to PCNA antibody appeared more numerous in control groups than that of CY treated groups. Appearance of positive cells in CY-treated group for 7 days and for 9 days were more numerous than those of CY-treated groups for 3 days and for 5 days, respectively. So these findings suggested that CY suppressed cell proliferations and effects of these action were decreased with CY-treated days. The numbers of positive cells to PCNA antibody were more prominent in hepatocelular carcinoma regions and cholangiocarcinoma regions, and then were ranked as order of large liver cell regions and normal liver cell regions. Also the numbers of the positive cells by apoptotic kit tended to be higher in hepatocelular carcinoma regions and cholangiocarcinoma regions but not uniformly in order in all regions nd were much less numbers than those of OCNA positive cells. So immunohistochemical methods using PCNA antibody together than using apoptotic kit alone when anti-carcinogen experiments. Rats with positive cells by p53 antibody were 11 of 15 rats(73.4%) in control groups and 12 of 18 rats(66.7%) in CY treated group, respectively. These positive cells appeared focally in early vacuole-occurring regions and were low in numbers.

Although it has been reported that hormones or chemicals, which increse in intracellular cAMP, produced Mg2+ release from the heart, it is not well characterized whether a specific Mg2+ exchanger is involved in cAMP-induced Mg2+ efflux in themammalian hearts In this work, we studied the relationship between the increase in intracellular cAMP and ion transport system on Mg2+ regulation in the perfused rat heart and isolated myocytes. The Mg2+ content in the perfusate and supernatant were measured by atomic absorption spectrophotometer. The addition of membrane permeable cAMP analogue to the perfused hearts and myocytes. cAMP-induced Mg2+ efflux was ingibited by H7, benzamil or imipramine in the perfused hearts and myocytes, but not by EIPA. We confirmed that a significant Mg2+ efflux was induced by an increase in intracellular cAMP in the hearts and myocytes. The cAMP-induced increase of Mg2+ efflux in the hearts may be involved in ion transport system(Na+-Ca2+ and Na+-Mg2+ exchanger)

The purpose of the present study was to investigate the effect of bovine amniotic membrane grafts on healing of full-thickness skin wound in dogs. Two 3cm*3cm area-matched full-thickness skin wounds were induced bilaterally on the dorsolateral aspect of the trunk of 15 dogs Chlorhexidine-treated amnion, dried amnion, silver sulfadiazine and 0.9% sterile saline solution were applied on the wound area and examined grossly and histopathologically. Begining 14 days after wounding, amnion applied group had appreciably less amount of inflammatory exudate and hemorrhage than sulfadiazine and saline treated groups. From 14 days after wounding, the degree of wound contraction in amnion groups, especially in the dried amnion group was greater than that of the sulfadiazine and saline treated groups. The percentages of wounds completely healed on 28 days after wounding in saline treated group, chlorhexidine-treated amnion group, dried amnion group and sulfadiazine treated group were 33%, 50%, 83% and 50%, respectively. Microscopically neovascularization and fibrosis were first noticed on 5 days after wounding in the dried amnion group and sulfadiazie treated group, on 7 days in the chlorhexidine-treated amnion group and on 14 days in the saline treated group. Epithlialization in thedried amion and sulfadizine treated groups ws first noticed on 9 days after wounding, which was faster than that in the other groups. The present study suggests that bovine amniotic membrane, especially dried bovine amnion is effective on healing of full-thickness skin wound in dogs through both wound contraction and epithelialization.

The study was designated to investigate the electron microscopic findings following diethylnitrosamine (DEN) treatment in rats. Forty four male (Srague Dawley) rats were continuously given water containing 0.01% DEN for 13 weeks and livers of five rats with more tumor lesions at 16 and 17 weeks after initial treatment were used as EM materials. In transmission electron microscopic findings, most small-sized hepatocytes were active cells containing large mount of organelles, but light (pale staining) hepatocyte among small-sized hepatocytes were injured cells containg disorganized organelles. Tumor cells among small-sized hepatocytes were irregularly arranged and have ;leomorphic nuclei containing electron dense chromatin but the organelles in cytoplasm were swelled. Large-sized hepatocytes were active cells with condensed chromatin but the cytoplasm of these cells were pale due to be injured and dilated organelles. Dark hepatocytes were apoptotic cells with homogenous pyknotic nuclei and cytoplasm, and the cytoplasm of these cells contained dilated smooth endoplasmic reticulum (sER) but these sER were non-vesiculated. Cholangiocarninoma cells were crowed and were pale by far less number of organelles in cytoplasm and nuclei. In scanning electron microscopic findings, the lumens of portal veins, bile canaliculi, bile ductules, bile ducts and sinusoids were dilated and have irregular folded inner surface by protruded parenchyma.

This investigation was carried out to study morphological and chronological aspects of the development of the Harderian gland in the Mongolian gerbil(Meriones unguiculatus). Male and female Mongolian gerbils were sacrificed on days 1, 3, 5, 10, 30 and 60 after birth and their Harderian glands were processed for light microscopic observation. The result obtained were summarized as follows; 1. In 1-day-old Mongolian gerbil, Harderian gland was well distinguished from other tissue structures. It was composed of several immature tubules, and these tubules were separated each other by undifferentiated mesenchymal connective tissues. 2. In 3-day and 5-day-old Mongolian gerbils, the arrangement of tubules in the gland was more condensed than that of 1-day-old Mongolian gerbil. The excretory ducts started to appear in the connective tissues located between lobes. 3. In-10-day-old Mongolian gerbil, small lipid vacuoles began to be found in the cytoplasm of the secretory cells of the Harderian gland. There were some mucus-secreting cells within the epithelium of the excretory duct found in the interlobar connective tissues. 4. In 30-day-old Mongolian gerbil, there was markdely increased number of the tubules in the glands. The epithelial cells of the tubules were typically columnar in shape. Most of the columnar epithelial cells contained many small lipid vacuoles, although a few cells contained large lipid vacuoles. 5. In 60-day-old Mongolian gerbil, the Harderian gland exhibited the typical structural characteristicsl of the adult gland. The mature glandular structrues were more distinct than those of 30-day-old animals.

A 5-month-old, 5kg, intact male mixed dog was admitted to the Veterinary Medical Teaching Hospital of the College of Veterinary Medicine Cornell University because of severe systolic murmur. On physical examination, the dog appeared to be thin and slightly small for his age. Radiographic studies with barium sulfate clearly showed the presence of loops of intestines in the pericardium. Based on these findings peritoneopericardial diaphragmatic hernia was diagnosed. Many abnormalities were found in dog's heart: ventricular septal defect, pulmonic stenosis with moderate pulmonic value insufficiency, and mild mitral and tricuspid valve insufficiency. Puppy's longterm prognosis was fair. His interstines were removed from his pericardium and the defect in his deaphragm was closed. The surgery went smoothly and puppy recovered very nicely from anesthesia.