Using the immunohistochemical technique, we attempted to identify the source of secretion of steroid hormones between the mid- and late-terms of gestation in dogs by investigating steroid converting enzymes such as cholesterol side-chain cleavage enzyme (SCC), 3 beta-hydroxysteroid dehydrogenase/isomerase (3 beta-HSD), 17 alpha-hydroxylase/C17, 20lyase (c17), and aromatase in the ovaries and placenta. Aromatase positive cells were slightly confirmed in luteal cells in the mid-term of gestation (day 40), whereas, in the late-stage (day 50 and 60), the number of aromatase positive cells had increased. However, the oestrogen precursor (c-17 positive cells), could barely be identified in the marginal regions of the corpora lutea (CL) and completely disappeared in the late-stage of gestation. The androgen precursors, convertase SCC and 3 beta-HSD, were confirmed in all regions of the CL during the mid-stage of gestation (day 40), showing particularly strong cell reactions in the marginal region of the CL. Yet, these positive reactions of SCC and 3 beta-HSD in the marginal region of the CL disappeared in the late-stage of gestation. Moreover, it was discovered that the number of SCC and 3 beta-HSD positive cells had decreased in all regions of the CL. None of the enzymes were detected in the placenta. The above results indicated that the source of oestrogen secretion in pregnant dogs is considered to be the CL, and that, compared with the mid-stage of gestation, there was an increased number of oestrogen synthesizing cells within the CL in the late-stage. However, the biosynthetic site of oestrogen precursors from the luteal cells during the late-stage of gestation is still unknown.

Sphingosine 1-phosphate (S1P) is a platelet-derived bioactive sphingolipid that evokes a variety of biological responses. To understand the role of S1P in the central nervous system, we have examined the effect of S1P on the production of glial cell line-derived neurotrophic factor (GDNF) and growth regulation of cortical astrocytes from rat embryo. Moreover, we examined the possibility that the expression of GDNF is regulated differently in cultured astrocytes from the stroke-prone spontaneously hypertensive rat (SHRSP) than in those from Wistar kyoto rats (WKY). The mRNA expression was quantitated by RT-PCR based on the fluorescent TaqMan methodology. A new instrument capable of measuring fluorescence in real time was used to quantify gene amplification in astrocytes. GDNF protein was investigated by enzyme-linked immunosorbent assay. S1P induced the expression of GDNF mRNA and the production of GDNF protein in a dose-dependent manner in WKY astrocytes. Moreover, S1P increased cell numbers and induced the proliferation of astrocytes. In addition, the level of mRNA expression and protein production of GDNF was significantly lower in SHRSP than WKY astrocytes following exposure to S1P. These findings revealed that S1P augments GDNF protein production and cellular growth in astrocytes. Also, our results indicate that production in SHRSP astrocytes was attenuated in response to S1P compared with that observed in WKY. We conclude that S1P specifically triggers a cascade of events that regulate the production of GDNF and cell growth in astrocytes. Our results also suggest that the reduced expression of GDNF caused by S1P is a factor in the stroke proneness of SHRSP.

ABSTRACT. Increase in circulating vascular endothelial growth factor (VEGF) is suggested as a prognostic indicator in human patients with malignant tumors. The purpose of this study was to evaluate the clinical significance of circulating VEGF in dogs with mammary gland tumors (MGT). Both plasma and serum VEGF were significantly higher in dogs with MGT when compared with those in the healthy dogs. In dogs with MGT, the plasma and serum VEGF of the malignant group increased significantly compared with those of the benign group. Additionally, there was a significant difference between the plasma and serum VEGF in the groups with postoperative metastasis and no metastasis. Circulating VEGF is expected to be clinically available for the determination of prognosis in canine MGT. KEY WORDS: canine, mammary gland tumor, vascular endothelial growth factor.

Glial cell line-derived neurotrophic factor (GDNF) plays several important roles in the survival and recovery of mature neurons during ischemia. We examined the possibility that the expression of GDNF mRNA and the release of GDNF protein are regulated differentially in cultured astrocytes from the stroke-prone spontaneously hypertensive rat (SHRSP) compared with those from Wistar Kyoto rats (WKY) during hypoxia and reoxygenation (H/R) and after exposure to glutamate and hydrogen peroxide (H(2)O(2)). The mRNA expression was quantitated by reverse transcription-polymerase chain reaction (RT-PCR) based on the fluorescent TaqMan methodology. A new instrument capable of measuring fluorescence in real-time was used to quantify gene amplification in astrocytes. GDNF protein was investigated by enzyme-linked immunosorbent assay (ELISA). GDNF mRNA expression and GDNF protein release at normoxia were greater in SHRSP than in WKY astrocytes. During H/R, however, the mRNA expression and protein release tended to be reduced in SHRSP compared with WKY. Glutamate and H(2)O(2) induced the expression of GDNF mRNA and the release of GDNF protein in both WKY and SHRSP in a dose-dependent manner. Levels of GDNF mRNA and protein in SHRSP were significantly lower than in WKY. These findings indicate that GDNF production in SHRSP astrocytes was low in response to H/R, glutamate, and H(2)O(2), compared with that observed in WKY. We conclude that the attenuated production of GDNF in astrocytes is involved in neuronal vulnerability in SHRSP during H/R, as GDNF production, which is stimulated by glutamate and H(2)O(2), is closely related to the protective effect against H/R-mediated neurotoxicity.

ABSTRACT. To establish an accurate method for parentage testing in dogs, microsatellite DNA repeat length polymorphisms were examined. We selected twenty microsatellite markers reported previously and examined their application for parentage testing in Beagles and Labrador Retrievers. Heterozygosity (He), Polymorphism Information Content (PIC), the probabilities of Paternity Exclusion (PE) and the combined PE were calculated from allelic frequencies of the markers. All markers amplified by polymerase chain reactions were polymorphic and many markers showed high He and PIC in the both breeds. The final combined PEs in Beagles and Labrador Retrievers were 0.999994 and 0.999920, respectively. The results suggest that the twenty markers can be applied for routine parentage testing in dogs.

ABSTRACT. After intravenous (IV) infusion of various volumes of 1.35%-isotonic sodium bicarbonate solution (ISB), acid-base equilibrium, blood pressure, plasma volume and biochemical parameters in healthy Holstein calves were studied. Four calves each were randomly assigned to the low-dose (LD; IV infusion of 5 ml/kg ISB), middle-dose (MD; IV infusion of 10 ml/kg ISB) and the high-dose groups (HD; IV infusion of 15 ml/kg ISB). Administration volumes of ISB in the LD, MD and HD groups were decided based on the first half volumes of 5, 10 and 15 mEq of base requirement by the acceptable equation. Systemic, pulmonary artery and central venous pressures, cardiac output and plasma osmotic pressure were not changed by ISB infusion and remained constant throughout the experiment for all groups. There was good correlation ( r 2 = 0.950) between relative changes in base excess and infused volume of bicarbonate (y=2.491x). The coefficient of distribution for bicarbonate ions was calculated to be 0.401 (=1/2.491). Therefore, it is suggested that a value of 0.4 would be most appropriate when calculating the base requirements in calves. Therefore, the first half volumes of ISB correcting base deficits of 5, 10 and 15 mEq in calves were estimated to be 6, 12 and 18 ml/kg, respectively. On the basis of the findings in this study, ISB may be used to correct metabolic acidosis without altering the plasma osmotic pressure, hemodynamic status and respiratory function in the calves. KEY WORDS: acid-base equilibrium, calf, coefficient of distribution, metabolic acidosis, sodium bicarbonate.

Two peaks of aromatase activities were detected in bovine cotyledons, 2.76 +/- 0.96 pmol min-1 mg-1 protein at month five of gestation and 3.62 +/- 0.78 pmol min-1 mg-1 protein immediately after parturition, but aromatase activity in caruncles was very low throughout gestation. Oestrone concentrations in the cotyledons and caruncles were 0.93 +/- 0.50 and 1.51 +/- 0.69 ng mg-1 protein at month five of gestation, but 4.31 +/- 1.66 and 3.36 +/- 0.98 ng mg-1 protein immediately after parturition, indicating a biphasic pattern. Our findings suggest that oestrogen synthesis in the bovine placenta increased to a maximum at least twice during the period from gestation to parturition; at month five of gestation and at parturition the changes in aromatase activity may only be partially explained by changes in oestrogen production.

ABSTRACT. A 38-month-old female Golden retriever was presented with dysuria and dyschezia. It was difficult to visualize the vagina by vaginoscopy due to a cystic polyp on the hymen. The polyp was 2 × 3 cm in diameter, round, and pink in color. From clinical and imaging evaluations the original diagnosis was mucometra or pyometra. From endoscopic examination of the vagina an imperforate hymen was finally diagnosed. The ovaries, uterus, and half of the vagina were removed through a median abdominal incision. The vagina contained about 1.5 liters of fluid, but the uterus and ovaries appeared normal. This is a rare case with imperforate hymen and hydrocolpos with a polyp on the hymenal membrane in bitch. KEY WORDS: bitch, hydrocolpos, imperforate hymen.J. Vet. Med. Sci. 63(4): 475-477, 2001 Congenital abnormalities of the canine vagina and vulva are classified into six categories, including four vestibulovaginal ring strictures and two strictures in the vestibulovulvar area [12]. The female genital tract is formed from the Mullerian ducts and urogenital sinus [9]. The hymen is formed by fusion of the Mullerian ducts and urogenital sinus, and in domestic animals disappears during the fetal period or after birth [4,5,9]. An imperforate hymen causes hydrocolpos, hydrometra and hydorosalpinx (also hemato-, respectively) [10]. A similar accumulation of vaginal fluid is also caused from vaginal atresia and vaginal aplasia [10]. In the bitch and queen, there are a few reports of imperforate hymen [7,11] and vaginal atresia [2,7,13]. We report a Golden retriever bitch with imperforate hymen and hydrocolpos due to a polyp on the hymen, diagnosed by endoscopic examination.A 38-month-old female Golden retriever weighing 30 kg was presented to the Animal Medical Center, Nihon University, with dysuria and dyschezia. The dog had been treated successfully for vaginitis and cystic calculus at 3 months and 2 years of age, respectively. During estrus, 3 months prior to presentation, the owner noticed vulvar swelling (a normal estrous sign), but without proestrous bleeding.Physical examination revealed a normal bitch except for the cystic polyp in the vagina. Urinalysis revealed a pH 6, slight protein and occult blood, 20/hpf red blood cells and 8/ hpf white blood cells consisting mainly of neutrophils were observed in the urine sediment, and the urine specific gravity was 1.030. The size of the cystic polyp prevented full digital exploration of the cranial vagina, but no vulvar discharge was noted. Left lateral (Fig. 1-A) and ventrodorsal ( Fig. 1-B) radiographs of the abdomen revealed an extremely enlarged vagina (15 × 27 cm), initially diagnosed as the uterus, located above the urinary bladder and reaching to the posterior margin of the liver. On the ventrodorsal view, the cranial pole of the urinary bladder extended to the last rib, displaced toward the right side of the abdomen, with a very long proximal urethra. The vagina, which was thought to be a uterus simplex, displaced to the left side, occupied two-thirds of the abdom...