Idiopathic thrombocytopenia in Japanese black cattle

A female Japanese Black cow showed hemorrhage under the mucous membrane of the eye conjunctiva and the vagina, and thrombocytopenia in peripheral blood. The clinical manifestations repeated spontaneously during long-term observation. The clinical findings were stable except for severe periodic bleeding. Prothrombin time, activated partial thromboplastin time and fibrinogen levels were normal. The anti-platelet antibody in the serum was analyzed by indirect platelet coagulation tests. Because the platelets in the micro-plate wells of the subject series showed coagulation up to 1:128, it was judged that anti-platelet antibody was present in the serum. A number of megakaryocytes were recognized in smear samples obtained from bone marrow of the scapula immediately after euthanasia. Therefore, this case was diagnosed as idiopathic thrombocytopenia.     

Clinical availability of urinary N-acetyl-beta-D-glucosaminidase index in dogs with urinary diseases

Urinary excretion of N-acetyl-beta-D-glucosaminidase (NAG) was examined in healthy dogs and dogs with urinary diseases, and its clinical usefulness as an indicator of urinary diseases was discussed. Twenty-eight healthy dogs and 20 dogs with urinary diseases were used. Urinary NAG activity was measured using p-nitrophenyl N-acetyl-beta-D-glucosaminide as substrate, and expressed as units per gram of urinary creatinine (NAG index). Urinary NAG index in urine of healthy dogs was 3.2+/-2.4 U/g, and NAG index in the dogs with chronic renal failure or lower urinary tract infection accompanied by pyelonephritis was higher than that in healthy dogs. However, the dogs with lower urinary tract infection without pyelonephritis showed normal values of NAG index. Some dogs with diabetic mellitus showed elevated values of NAG index when control of blood sugar was not successful. Increase of NAG index was observed in some dogs with pyometra before increases of BUN and serum creatinine concentration. Therefore, NAG index in urine seems to be a good indicator for urinary diseases in dogs.     

Urinary excretion of N-acetyl-beta-D-glucosaminidase and its isoenzymes in cats with urinary disease

The present study was designed to assess the clinical usefulness of measurement of urinary N-acetyl-beta-D-glucosaminidase (NAG) activity and its isoenzymes in cats with urinary disease. Thirty-five healthy cats and 9 cats with renal disease were used. Furthermore, a 5-year-old female cat was administered a large amount of sulfonamide in order to induce acute renal failure, and urine samples were collected for the assay of NAG activity and its isoenzymes. Urinary NAG activity was measured using p-nitrophenyl N-acetyl-beta-D-glucosaminide, and expressed as units per gram of urinary creatinine (NAG index). Urinary NAG isoenzymes were assayed by use of the mini-column method and electrophoresis. The overall mean value of urinary NAG index in healthy cats was 1.6+/-1.5 U/g. Urinary NAG index varied from 6.2 to 35.5 U/g in cats with chronic renal failure. There was no significant correlation between BUN, serum creatinine concentration and urinary NAG index. In cats with feline lower urinary tract disease, normal values of urinary NAG index were observed. In the urine samples of healthy cats, the proportions of NAG isoenzyme A (NAG-A) and isoenzyme B (NAG-B) were 79.1+/-4.4% and 21.0+/-4.4%, respectively, as assayed by the mini-column method. In the assay of NAG isoenzymes by electrophoresis, the proportions of NAG-A and NAG-B in healthy cats were 66.6+/-5.8% and 33.4+/-5.8%, respectively. The proportion of NAG-B as measured by electrophoresis was significantly larger (p<0.05) than that obtained with the mini column method. A feline case of acute renal failure experimentally-induced by sulfonamide showed elevation of urinary NAG index, NAG-A and NAG-B after injection of sulfonamide. The increase in NAG-B was larger than that of NAG-A. From the results reported here, measurement of urinary NAG and its isoenzymes seems to yield information about tubular damage at an early stage in cats with urinary disease.     

Strain differences in the spontaneous incidence of sperm morphological abnormalities in Hatano rats

Two inbred strains of Hatano rats, which had been bred selectively on the basis for shuttlebox avoidance responses (high- and low-avoidance animals; HAA and LAA), were different in sperm motility. We have found that there are strain differences in the spontaneous incidence of sperm morphological abnormalities. The percentages of abnormalities were extraordinarily higher in the LAA rats, whose sperm motility was lesser. Since the high sperm abnormality was an exceptional case in rats, these rats may be useful models for sperm quality.     

Bile acid extraction rate in the liver of cows fed high-fat diet and lipid profiles in the portal and hepatic veins

The purposes of this study are to assess the responses of increased supplemental dietary fat in the cow, without upsetting rumen fermentation, on the bile acid (BA) extraction rate in the liver and to determine whether this diet would affect the postprandial lipid profiles in the portal and hepatic venous blood. Six Holstein cows were equipped with catheters fitted in the portal and hepatic veins. Two cows each were assigned randomly to a sequence of three dietary treatments of 21-day period. The methodology of this study was based on the supplementation of the basal concentrate diet with 0 (control), 5, or 10% calcium salts of fatty acids (CSFA). The total bile acids were significantly increased in the portal and hepatic veins with the 5% CSFA diet, whereas no increase occurred with the 10% CSFA diet. Data obtained in this study showed that 10% CSFA diet failed to stimulate BA secretion to exceed the values obtained with 5% CSFA-diet. Moreover, there was no change in the hepatic extraction rate of BA in animals fed either the 0 or 5% CSFA diets which ranged from 2.4 to 6.5-fold and 3.1 to 7.3-fold, respectively. However, the extraction rate increased sharply with the 10% CSFA diet (27-fold). The median portal and hepatic concentrations of total lipids, triglycerides, total cholesterol, phospholipids and non-esterified fatty acids did not show any significant increase during feeding of the control diet. Moreover, feeding either the 5 or 10% CSFA diet did not significantly increase these values in either vein.     

Nicotinic acetylcholine receptor binding of imidacloprid-related diaza compounds with various ring sizes and their insecticidal activity against Musca domestica

Fifteen 5-substituted 1-(6-chloro-3-pyridylmethyl)-2-nitromethylene-1,3- diazacyclohexanes and three other related compounds having a five- or seven-membered ring were synthesized and their biological activities were measured in vivo and in vitro. The insecticidal (in vivo) activity was evaluated against houseflies Musca domestica L under synergistic conditions with propargyl propyl phenyl phosphonate and piperonyl butoxide. The binding activity of each compound to nicotinic acetylcholine receptor in vitro was measured using [125I] alpha-bungarotoxin. The insecticidal activities of the unsubstituted diazacyclohexane analogues were slightly higher than those of the imidazolidine analogues, but the enlargement of ring size to diazacycloheptane lowered the activity. Substitution of 1,3-diazacyclohexane or imidazolidine rings was not generally favourable for the activity, but the unsubstituted 1,3-diazacyclohexane analogue showed the highest binding activity. Ring substitutions and ring enlargement decreased the activity 100-30,000-fold.     

Identification of genes for two major sialoglycoproteins, glycophorin A and glycophorin C in canine red cell membranes

Glycophorins are the major sialoglycoproteins in red blood cell membranes, possessing various physiological and pathological roles. We examined membrane glycoproteins in canine red cells and cloned cDNAs for two major glycophorins, glycophorins A (GPA) and C (GPC) from bone marrow cells. Periodic acid-Schiff staining and immunoblotting analyses showed that canine red cell membranes contained several glycoproteins immunoreactive to an anti-bovine GPC antibody, whereas the most abundant sialoglycoproteins, the candidates for GPA, did not react with an anti-human GPA antibody. The amino acid sequences of the extracellular domains of GPA and GPC had no significant homology to those from other mammalian species, including humans, and had O-linked and/or N-linked glycosylation sites. On the other hand, the C-terminal cytoplasmic domain and/or the transmembrane helices of GPA and GPC were conserved among species, indicating some functional significance of those regions in red cell membranes that include dimerization of GPA in the membrane-spanning region, and association of GPC with membrane skeletal proteins through binding with protein 4.1 and p55 in the cytoplasmic domain. These findings provide insights for clinical studies to evaluate the involvement of GPA and GPC in the pathogenesis of red cell diseases.     

A BOLD-fMRI study of cerebral activation induced by injection of algesic chemical substances into the anesthetized rat forepaw

This study was performed to examine whether the brain activities induced by noxious algesic chemical substances in anesthetized animals could be detected by blood oxygen-level-dependent functional magnetic resonance imaging (BOLD-fMRI). Multislice gradient echo images of the primary somatosensory cortex were obtained using a 7.05 T superconducting system and a one-turned surface coil centered over the primary somatosensory cortex of the 1.0%-isoflurane-anesthetized rat. The Z-score t-map of BOLD signals and its time-course analysis revealed that subcutaneous injection of formalin into the left forepaw immediately induced an early response in the contralateral primary sensory cortex lasting for a few minutes, followed by a late response until 20 min after stimulation. In contrast, injection of capsaicin into the left forepaw evoked only the early response. Furthermore, pretreatment with morphine completely abolished these responses induced by the chemical algesic substances. Thus BOLD-fMRI is a useful method to analyze the brain activities of painful stimulation in anesthetized animals.     

Expression of Bcl-2 in feline lymphoma cell lines

BACKGROUND: The Bcl-2 gene is a member of the rapidly expanding Bcl-2 family of genes that regulate apoptosis. Bcl-2 has been shown to repress cell death triggered by a diverse array of stimuli, including chemotherapy and gamma irradiation. OBJECTIVE: The purpose of this study was to determine feline Bcl-2 expression level in feline lymphoma cells using an immunoblot assay with anti-human and anti-canine Bcl-2 monoclonal antibodies. METHODS: About 708 base pairs containing the coding sequence of the feline Bcl-2 gene were transformed into Escherichia coli. The recombinant Bcl-2 was used as a positive control for an immunoblot assay using mouse monoclonal antibodies against human and canine Bcl-2. An immunoblot assay using the monoclonal antibodies was carried out to determine the level of feline Bcl-2 expression in lymphoma and lymphocytic leukemia cell lines. RESULTS: The recombinant feline Bcl-2 protein produced in E. coli had a molecular weight of about 26 kDa and was detected by immunoblot assay by using anti-human Bcl-2 mouse monoclonal antibody. Feline Bcl-2 expression was high in lymphoma cell lines (FL-74-UDC-1 and FT-1) and low in the cell line from peripheral blood mononuclear cells from a healthy cat (FeTJ-1) but not low in freshly isolated peripheral blood mononuclear cells from a healthy cat. The anti-human Bcl-2 mouse monoclonal antibody was found to cross-react with feline Bcl-2. CONCLUSIONS: These results confirm the expression of Bcl-2 in T-cell lymphoma cell lines and indicate that it is suitable to detect feline Bcl-2 using an immunoblot assay. Pending further evaluation, Bcl-2 expression might be useful in the differential diagnosis of feline tumors.