The skull adaptation was functional-morphologically examined in 14 species of the tree shrews. From the data of the proportion indices, the similarities were confirmed between T. minor and T. gracilis, T. tana and T. dorsalis, and T. longipes and T. glis. We demonstrated that the splanchnocranium was elongated in terrestrial T. tana and T. dorsalis and shortened in arboreal T. minor and T. gracilis from the proportion data. In both dendrogram from the matrix of the Q-mode correlation coefficients and scattergram from the canonical discriminant analysis, the morphological similarities in the skull shape suggested the terrestrial-insectivorous adaptation of T. tana and T. dorsalis, and the arboreal adaptation of T. minor and T. gracilis. Since the osteometrical skull similarities were indicated among the three species of Tupaia by cluster and canonical discriminant analyses, the arbo-terrestrial behavior and its functional-morphological adaptation may be commonly established in T. montana, T. longipes and T. glis.
The olfactory system is indispensable to the survival of animals in finding foods and for the reproductive process. Odorant signals are conveyed through olfactory sensory neurons to the olfactory bulb, which modifies the signals and relays them to the neocortex. In the present study, a "full-length" cDNA library was constructed from the main and accessory olfactory bulbs of 5-week-old male pigs, in order to elucidate the expressed genes. The average insert size of the library was estimated to be 1.7 kb based on 54 randomly-selected clones. One thousand randomly selected clones were subjected to sequencing, and the resulting 883 sequences were then clustered into 753 sequences based on similarity. Since 723 of the 753 sequences had sufficient sequence information for homology analysis, the 723 sequences were subjected to BLAST analysis against GenBank/EMBL/DDBJ; 655 out of the 723 sequences showed similarities with known genes, and the remaining 68 were indicated to be novel sequences. The full-length rate of the library was estimated to be ca. 80%, using 70 sequences corresponding to human full-length cDNAs. The full-length cDNA sequences of a single gene appearing more than 6 times in the analysis were aligned to determine major transcription initiation sites for SLC25A, CKB, TUBB4, TUBB, YWHAH, TUBB2, and CNP genes.
To obtain a basal concentration of serum Haptoglobin (Hp) in cattle in Taiwan, Hp concentrations were measured from serum samples collected from 10 healthy heifers, every week for one year. The values were also compared with those collected from 15 cows diagnosed with postpartum metritis. The heifers were successfully impregnated by artificial insemination six months after the tests. Hp concentrations were also measured in the serum collected from 11 other cows within 3 weeks after parturition. The Hp assay developed in this study gave a good correlation (r=0.893)with Western blotting. The Hp concentration of 454 serum samples from the 10 heifers had a mean value of 83.6 ± 34.1 mg/l, and there was no significant difference among individual heifers. The basal value of Hp in heifers was calculated as less than 73.6 mg/l. No significant difference in Hp concentration was observed among the 10 heifers during cold and warm seasons (19.8 ± 2.2 °C vs 27.3 ± 1.4°C), or before and after pregnancy. The mean serum Hp concentration from cows suffering from postpartum reproductive disorders was 1133.5 ± 627.1 mg/l, which was significantly greater than the serum of healthy heifers and postpartum cows (104.6 ± 61.0 mg/l) (P<0.05). These results demonstrate that Hp concentration may be a useful indicator for cows with postpartum reproductive disorders.
Hepatocyte growth factor (HGF) is a pleiotropic cytokine that stimulates a wide array of cellular targets, including hepatocytes and other epithelial cells, melanocytes, endothelial and hematopoietic cells. We have cloned a different form of cDNA, with a deletion of 15 base pairs predicted to result in the loss of 5 amino acids from the first kringle domain. To investigate the biological activity, original and deleted variant of feline HGF cDNAs were transiently expressed in COS-7 cells. Both recombinant feline HGFs showed almost the same dose-response curves in the stimulation of the growth of BNL CL.2 cells (a mouse hepatocyte cell line) and scatter activity of Madin-Darby canine kidney (MDCK) cells. The findings reported here suggest that the deleted variant of feline HGF has almost the same biological activity as the original in terms of the proliferation and scatter activity.
Three dogs clinically diagnosed with allergic rhinitis (AR) were examined for their immunological findings. House dust mites (HDM) such as Dermatophagoides farinae (DF) and D. pteronyssinus (DP) were identified as positive allergens in the 3 dogs with both intradermal skin test and serum antigen-specific IgE test. Lymphocyte blastogenic response of peripheral blood mononuclear cells (PBMCs) under stimulation with DF antigen in dogs with AR was higher than that in 4 healthy control dogs. Expression level of IL-4 mRNA in PBMCs obtained from the 3 AR dogs was higher than that in PBMCs obtained from 4 healthy control dogs before and after stimulation with DF antigen. Expression level of IFN-γ mRNA in PBMCs was not different between the AR and control dogs before and after stimulation with DF antigen. These results suggested that allergic reaction to HDM antigen and TH2-type immune response were associated with the development of AR in 3 dogs examined in this study.
A 6-month-old female Holstein calf was referred with a history of respiratory distress, anorexia and dehydration. When first examined, a loud harsh murmur was clearly audible from both sides of the thorax. Two-dimentional echocardiography demonstrated a markedly dilated right ventricle and right atrium, hypertrophied myocardium, prominent papillary muscles and a thickened interventricular septum (IVS). A large-sized ventricular septal defect was imaged at the top of the IVS. The aorta was dextrally located and overriding both ventricles. The diagnosis was confirmed as tetralogy of fallot on postmortem examination.
To elucidate possible relationships between triglyceride (TG) levels and fatty acid composition in bovine liver, hepatic TG and seven individual fatty acids were measured in 23 Holstein dairy cows, of them 6 are healthy. Liver TG level was greater than 3 % in 12 cows which were ruled fatty liver. Palmitic and oleic acid proportions were significantly higher in fatty liver cows than in the healthy cows, while stearic acid was lower in fatty liver cows. With increased liver TG, stearic acid proportions decreased dramatically. Results indicate that hepatic lipidosis markedly alters the proportions of the various fatty acids in the liver of dairy cows.
To investigate the usefulness of ascites as a material for viral tests in cats with effusive feline infectious peritonitis (FIP), we attempted to detect anti-feline coronavirus antibody, anti-feline immunodeficiency virus antibody, and feline leukemia virus antigen in ascites from 88 cats clinically suspected with effusive FIP. In each of these three viral tests, all cats positive for serum antibody/antigen were also positive for ascitic antibody/antigen, while cats negative for serum antibody/antigen were also negative for ascitic antibody/antigen. This finding indicates that ascites is useful for these viral tests.
Polymorphonuclear neutrophil (PMN) apoptosis was examined in three dogs with pyometra by TUNEL assay in a 24-hr incubation period and compared with that in healthy control dogs (n=5). The incidence of apoptotic PMNs in dogs with pyometra was 26.4 ± 5% and that in healthy dogs was 54.3 ± 7%. The results indicated that apoptotic PMN rates in dogs with pyometra were significantly lower than those in control dogs (p<0.05), suggesting the prolongation of PMN survival.
Mutations of Fas (CD95/Apo-1) gene have been reported in various malignancies and therefore the Fas gene has been considered to be a tumor suppressor gene. To examine an involvement of Fas gene as a tumor suppressor gene in radiation lymphomagenesis, we examined the loss of heterozygosity (LOH) in lymphomas from (MSM/Ms x MRL-MpJ/Fas lpr) F1 and (BALB/cHeA × MRL-MpJ/Fas lpr) F1 hybrid mice. Lymphoma development by X-irradiation was efficiently observed in both F1 hybrids. Frequent LOH was found on chromosomes 12 and 4 in the tumors from both F1 mice, but no allelic loss on chromosome 19 containing Fas locus was found, and no wild-type allele of the Fas gene was lost in 51 lymphomas. Therefore, the putative tumor-suppressor gene regions responsible for lymphomagenesis might not considerably differ due to the Fas gene status.
A 12-year-old female miniature poodle showed a 3-month history of neurological signs. Magnetic resonance imaging disclosed a high intensity tumor mass in the right cerebral hemisphere with compression of the lateral ventricle. At necropsy, a 2 × 3 cm white, friable mass was found in the right ventral pyriform lobe. Microscopically, the tumor cells were large, polygonal to round cells supported by a sparse fibrovascular stroma. The tumor cells typically possesed finely granular, pale eosinophilic cytoplasm with strongly positive periodic acid-Schiff (PAS) reaction. The tumor cells were immunopositive for vimentin, NSE and S-100. Ultrastructurally, the tumor cells showed large amounts of granules in the cytoplasm, and absence of basement membrane. Based on the above-mentioned findings, the intracranial granular cell tumor was diagnosed.
A 5-year-old spayed male ferret showed a subcutaneous mass in the right lateral thoracic wall. Microscopic examination revealed that the neoplasm had proliferated in the subcutis with infiltration into the surrounding tissues. A packed bundle of large polymorphic neoplastic cells, containing abundant eosinophilic cytoplasm and a round to ovoid, occasionally bizarre nucleus, were arranged interwoven. The neoplasm had metastasized to the right axillary lymph node. The neoplastic cells were intensively positive for vimentin, desmin and myoglobin. Skeletal muscle type creatine phosphokinase-positive granules were detected in the cytoplasm. Ultrastructurally, various amounts of disorganized myofibrils with focal density resembling the Z-band were shown in the cytoplasm of the neoplastic cells. The neoplasia was diagnosed as rhabdomyosarcoma.
To clarify the role of nitric oxide (NO) in hemodialysis (HD)-related hypotension, the relationship between plasma NO metabolites (NOx) and blood pressure changes, and the effect of NG-monomethyl-L-arginine (L-NMMA), a NO synthase inhibitor, on changes in blood pressure were evaluated in an experimental renal dysfunctional dog model. In order to create a renal dysfunction model, gentamicin was administered to male beagles in which 7 of 8 renal artery branches had been ligated. Normal renal functional and dysfunctional dogs underwent 3 hr of HD per day for 3 days. HD induced a transient decrease in mean blood pressure in the normal renal functional dogs. In renal dysfunctional dogs, a continuous hypotension occurred with a gradual increase in the plasma NOx concentration during HD. Although L-NMMA prevented the fall in blood pressure, it did not significantly change the plasma NOx concentration during HD. These results suggest that NO contributes to HD-related hypotension in renal dysfunctional dogs but the plasma NOx concentration does not reflect the change in blood pressure.
Effects of physiological variables (age, body weight and sex) on lipemia (total lipids, phospholipids, triglycerides and NEFA), fecal fat and chymotrypsin excretion were examined in dogs. On comparing various ages and body weights, they found statistically significant differences, while only total lipids showed a statistically significant difference between the two sexes. Moreover, there was a significant correlation between body weight and fecal fat and chymotrypsin excretion. The results obtained contribute to the evaluation of small intestine functionality, especially in relation to some diseases, as malabsorption, maldigestion and steatorrhea in the dog.
To characterize spinal cord effects of needle placement using lumbar puncture myelography technique, lumbar puncture was performed in 5 dogs and computed tomography images of the spinal column were acquired in the transverse plane at the level of the puncture site after contrast injection and both before and after needle removal. The spinal cords were punctured during needle placement and parenchymal contrast enhancement was present in 4 of 5 dogs. Although no dogs exhibited overt neurological abnormalities following computed tomographic imaging, hemorrhage, gliosis and axonal degeneration were confirmed microscopically in all subjects. These results suggest that spinal cord morbidity is induced when lumbar myelography is performed using currently accepted technique.
In search of a safer myelographic technique, we performed myelography via the lumbosacral intervertebral space. Eight dogs, in which the backflow of cerebrospinal fluid was observed, received contrast media via the lumbosacral intervertebral space. The subarachnoid contrast columns were successfully observed in 6 dogs. During and after examination, no physiological changes or neurological signs were observed. We recommend that the lumbosacral intervertebral space be selected first before implementing the conventional lumbar myelography.
A 10-year-old, female poodle was presented with left paresis and vestibular signs, following left enophthalmus and atrophy of the cheek. Magnetic resonance imaging revealed a mass along the base of the brain and extending from the left cerebellopontine angle to the sella turcica. The mass showed isointensity on the T1-weighted image and T2-weighted image, and was enhanced by contrast medium (Gd-DTPA). Although occipital craniotomy was carried out and the mass removed, the dog died without recovering from the anesthesia. The tumor was diagnosed pathologically as fibroblastic meningioma.
Two pony mares were immunized against recombinant porcine inhibin α subunit three times with 39 day intervals. Clinical findings and endocrinological changes before immunization were taken as the control. The first significant rise in the anti-inhibin titre (P<0.05) in the circulation was found 27 days after the first injection. Maximum binding activity was reached by the 12th day after the second booster dose. The number of small, medium and large sized follicles had increased significantly compared to before immunization (11.75 ± 4.30, 2.75 ± 0.69 and 2.51 ± 0.63 vs 6.50 ± 1.43, 1.83 ± 0.44 and 1.33 ± 0.38, respectively), but the ovulation rate remained unchanged after immunization. The average plasma concentration of FSH and estradiol-17β during the estrous cycle increased significantly (P<0.05) after immunization. These results suggest that immunization against inhibin is a useful tool to increase the number of ovarian follicles during the estrous cycle of pony mares. Moreover, the present study supported the concept that inhibin plays a major role in the control of follicular growth through its inhibitory effect on FSH secretion synergistically with steroid hormones.
Freeze-storage of epididymal sperm is an important technique for the preservation of gametes in animals, including those becoming extinct. We froze canine sperm recovered from the cauda epididymis and investigated the fertility. The qualities of sperm from the cauda epididymis before freezing were: mean sperm motility, 89.4 ± 1.6 (SE) %; sperm viability, 89.1 ± 1.1%; and these were significantly higher than those of sperm from the caput-corpus epididymis (P<0.01, P<0.05). The number of sperm recovered from both cauda epididymides varied among animals: 6.3-122.3 × 107, mean 61.5 ± 10.0 × 107. Freezing was used only for sperm recovered from the cauda epididymis. The sperm motility and viability after thawing were 19.5 ± 2.5% and 53.1 ± 3.3%, respectively. These were slightly lower than those of frozen-thawed ejaculated sperm, but the differences were not significant. When 2 × 108, 3 × 108, or 4 × 10 8 sperm were inseminated in the unilateral uterus, only one animal inseminated with 3 × 108 sperm was fertilized (1/16, 6.3%). When 1 × 108 sperm were inseminated in the bilateral uterine tubes, one of six animals (16.7%) was fertilized. Therefore, although the qualities of epididymal sperm after thawing were similar to those of ejaculated sperm, the conception rate obtained with frozen-thawed epididymal sperm was low in beagle dogs. It is necessary to investigate the differences in damage between epididymal sperm after thawing and ejaculated sperm and to develop a method for improving the conception rate.
Aims: To determine the turnover of the first follicular wave in Japanese black cows and quantitative immunohistological cha-racteristics of the previously in vivo identified dominant follicle (DF) and largest subordinate follicle (SF) derived from ovariectomy on Day 7 (3 cows) and Day 10 (3 cows) (Day 0=estrus). Six cases of first follicular wave in cows were observed twice daily by ultrasound scanning. The number of follicles, diameter of DF and SF, and prevalence of apoptotic granulosa cells (GC) and theca cells (TC) were studied by TUNEL methods. At follicular wave emergence, 13.5 ± 9.5 Class I (2-5 mm in diameter follicles) were found 12 hr after ovulation, and increased its number until Day 1 pm. Future DF and SF observed retrospectively were 4.9 ± 0.8 mm and 4.9 ± 0.9 mm at wave emergence. Deviation of DF and SF occurred on Day 3 pm with mean diameters of 8.9 ± 1.3 mm and 6.8 ± 0.9 mm, respectively. DF developed until Day 8 am with a maximum diameter of 14.4 ± 1.8 mm (n=3) and then regressed. The follicular wall of the DF had a characteristic image of a healthy follicle on Day 7 and slightly atretic DF on Day 10, whereas SF showed heavy atresia on both Day 7 and Day 10 under HE staining. In the prevalence of apoptotic cells, DF were 4.4 ± 1.0% and 17.9 ± 4.9% on Day 7 and on Day 10 in GC, respectively, and 2.4 ± 0.7% and 8.0 ± 1.4% on Day 7 and on Day 10 in TC, respectively. These results showed that, 1) the first follicular wave in cows is characterized by 24 hr recruitment of small follicles and a gradual divergence of growth rates in future DF and SF, and 2) early regression of DF on Day 10 was preceded by severe apoptosis.
This study was conducted to evaluate whether refining mineral oil and the addition of hemoglobin and/or glucose to a serum-free medium could improve in vitro-development of embryos cultured in a chemicallysemi-defined microdroplet culture system. Block strain, outbred (ICR) mouse 1- or 2-cell embryos were cultured in 5 μl droplets of Chatot, Ziomek and Bavister medium overlaid with mineral oil of different types, and preimplantation development to the blastocyst stage was subsequently monitored. In the experiment 1, either Sigma (M-8410) or BDH (GPR TM) mineral oil with or without washing was used for embryo culture and, distilled water (DW) or culture medium was used as a washing agent. As results, better (P<0.0001) development of 1-cell embryos was found in the Sigma than in the BDH; more blastocysts developed in Sigma oil washed with culture medium than in the others (37% vs. 0%). Subsequently, 1- (experiment 2) or 2-cell (experiment 3) embryos were cultured in the droplets overlaid with medium-washed Sigma oil, to which 0.001 mg/ml hemoglobin and/or 5.6 mM glucose were supplemented at the 1-cell and the 4-cell stages, respectively. Regardless of embryo stages, blastocyst formation was significantly improved by the addition of hemoglobin (54 to 48% vs. 42 to 31% in 1-cell and 83 to 78% vs. 65 to 68% in 2-cell embryos) and this effect was independent of glucose addition. In conclusion, the selection and washing of mineral oil, and the addition of hemoglobin is beneficial for improving the efficacy of a drop embryo culture system using a serum-free medium.
Eight clones of monoclonal antibodies (Mabs) to Nipah virus (NV) were produced against formalin-inactivated NV antigens. They reacted positive by indirect immunofluorescent antibody test, and one of them also demonstrated virus neutralizing activity. They were classified into six different types based on their biological properties. These Mabs will be useful for immunodiagnosis of NV infections in animals and further research studies involving the genomes and proteins of NV.