Patterns of aquaporin expression in the canine eye

Aquaporins (AQPs) function as water channels in many types of cells involved in fluid transport. More than 10 isoforms have been identified, and these are differentially expressed in many types of mammalian cells in the body. Six AQPs (AQP0, AQP1, AQP3, AQP4, AQP5, and AQP9) have been identified in the eyes of humans and/or rodents. The unique permeability characteristics and distribution of AQPs indicate their diverse roles in the regulation of water homeostasis in the eye. The aim of this study was to investigate the localisation of AQPs in normal canine eyes, with AQP0 protein expressed in the crystalline lens and retina. Although AQP1 mRNA was detected in various areas of the canine eye, its protein expression was limited to the cornea, iris and ciliary body. AQP4 was identified in the iris, retina and optic nerve. AQP3 and AQP5 were found in the cornea and conjunctiva, and their expression was particularly high in the limbus. AQP3 and AQP5 were present in the nictitating membrane indicating that they play a role in water transport within the membrane. The observations suggested that several subtypes of the AQP family are involved in the regulation of water homeostasis in the canine eye.     

Purification of chicken carbonic anhydrase isozyme-III (CA-III) and its measurement in White Leghorn chickens

Background

The developmental profile of chicken carbonic anhydrase-III (CA-III) blood levels has not been previously determined or reported. We isolated CA-III from chicken muscle and investigated age-related changes in the levels of CA-III in blood.

Methods

CA-III was purified from chicken muscle. The levels of CA-III in plasma and erythrocytes from 278 female chickens (aged 1-93 weeks) and 68 male chickens (aged 3-59 weeks) were determined by ELISA.

Results

The mean level of CA-III in female chicken erythrocytes (1 week old) was 4.6 μg/g of Hb, and the CA-III level did not change until 16 weeks of age. The level then increased until 63 weeks of age (11.8 μg/g of Hb), decreased to 4.7 μg/g of Hb at 73 weeks of age, and increased again until 93 weeks of age (8.6 μg/g of Hb). The mean level of CA-III in erythrocytes from male chickens (3 weeks old) was 2.4 μg/g of Hb, and this level remained steady until 59 weeks of age. The mean plasma level of CA-III in 1-week-old female chickens was 60 ng/mL, and this level was increased at 3 weeks of age (141 ng/mL) and then remained steady until 80 weeks of age (122 ng/mL). The mean plasma level of CA-III in 3-week-old male chickens was 58 ng/mL, and this level remained steady until 59 weeks of age.

Conclusion

We observed both developmental changes and sex differences in CA-III concentrations in White Leghorn (WL) chicken erythrocytes and plasma. Simple linear regression analysis showed a significant association between the erythrocyte CA-III level and egg-laying rate in WL-chickens 16-63 weeks of age (p < 0.01).     

Acute myelomonocytic leukemia (AML-M4) in a dog with the extradural lesion

A two-year-old dog having presented with neurological signs showed marked leukocytosis and appearance of blast cells in the peripheral blood. Hematological and bone marrow examination showed an increase in blasts having both myeloid and monocytic cells characteristics. The dog was diagnosed with acute myelomonocytic leukemia (AML-M4) on the basis of bone marrow findings. Although the dog was treated with a multi-combination chemotherapy, the neurological abnormalities progressed and the dog was euthanized. Myelographic examination and necropsy revealed the extradural lesion formed by AML-M4 around the cervical spinal cord and this lesion was considered as a cause of the neurological signs.     

Non-CpG methylation occurs in the regulatory region of the Sry gene

The Sry (sex determining region on Y chromosome) gene is a master gene for sex determination. We previously reported that the Sry gene has tissue-dependent and differentially methylated regions (T-DMRs) by analyzing the DNA methylation states at CpG sites in the promoter regions. In this study, we found unique non-CpG methylation at the internal cytosine in the 5'-CCTGG-3' pentanucleotide sequence in the Sry T-DMR. This non-CpG methylation was detected in four mouse strains (ICR, BALB/c, DBA2 and C3H), but not in two strains (C57BL/6 and 129S1), suggesting that the CCTGG methylation is tentative and unstable. Interestingly, this CCTGG methylation was associated with demethylation of the CpG sites in the Sry T-DMR in the developmental process. A methylation-mediated promoter assay showed that the CCTGG methylation promotes gene expression. Our finding shows that non-CpG methylation has unique characteristic and is still conserved in mammals.     

Evaluation of the renin-angiotensin system in cardiac tissues of cats with pressure-overload cardiac hypertrophy

OBJECTIVE: To clarify regulation of the renin-angiotensin (RA) system in cardiac tissues by measuring angiotensin-converting enzyme (ACE) and chymase activities in cats with pressure-overload cardiac hypertrophy. ANIMALS: 13 adult cats. PROCEDURES: Pressure-overload cardiac hypertrophy was induced by coarctation of the base of the ascending aorta in 6 cats, and 7 cats served as untreated control animals. Cats were examined before and 3 months and 2 years after surgery. Two years after surgery, cardiac hypertrophy was confirmed by echocardiography, and the blood pressure gradient was measured at the site of constriction. Cats were euthanized, and ACE and chymase activities were measured in cardiac tissues. RESULTS: Mean +/- SD pressure gradient across the aortic constriction was 63 +/- 6 mm Hg. Chymase activity predominated (75% to 85%) in the RA system of the cardiac tissues of cats. Fibrosis in the wall of the left ventricle was detected in cats with hypertrophy, and fibrosis of the papillary muscle was particularly evident. CONCLUSIONS AND CLINICAL RELEVANCE: Chronic pressure overload on the heart of cats can activate the RA system in cardiac tissues. A local increase in angiotensin II was one of the factors that sustained myocardial remodeling.     

Plasma metabolite concentrations and hepatic enzyme activities in pregnant Romney ewes with restricted feeding

Plasma metabolite concentrations and activities of enzymes related to energy metabolism in plasma, peripheral leukocytes and liver of pregnant Romney ewes with restricted feeding were measured to assess those metabolites and enzymes as indicators for evaluating metabolic conditions in the ewes. The body weights and plasma lactate concentrations of the low-feeding ewes (about 1.0 times maintenance) were significantly lower than those of the high-feeding ewes (about 1.5 times maintenance). There were no significant differences in plasma protein, glucose, triglyceride, cholesterol, pyruvate and immunoreactive insulin concentrations and plasma and leukocyte enzyme activities between both groups. Hepatic malate dehydrogenase activities were significantly higher and hexokinase activities were significantly lower in the low-feeding ewes than in the high-feeding ewes. Restricted feeding could maintain pregnancy and some plasma metabolites and peripheral leukocyte enzymes may be useful indicators for evaluating metabolic changes in ewes.     

Contact dermatitis in dairy cattle caused by calcium cyanamide

Nine of 250 cows on a dairy farm initially developed severe dermatitis on parts of their bodies that touched the floor, and it then spread over their entire body. The cause was suspected to be calcium cyanamide, which had been added to the material spread on the floor to prevent environmental mastitis. Experimental exposure of the skin of a cow to calcium cyanamide induced the same type of contact dermatitis, and histopathological investigations showed that it caused irritant and allergic reactions. To identify the cause of the dermatitis, a patch test with calcium cyanamide and its breakdown products, cyanamide, urea and ammonium bicarbonate, was carried out on four cows. Three of them had a positive reaction to calcium cyanamide and cyanamide; delayed and amplified reactions suggesting an allergic response were observed.     

Changes in metabolite, energy metabolism related enzyme activities and peripheral blood mononuclear cell (PBMC) Populations in beef heifers with two differing liveweight change profiles in New Zealand

Metabolite and immunoreactive insulin (IRI) concentrations, energy metabolism related enzymes activities and peripheral blood mononuclear cell (PBMC) populations were measured in blood of pregnant Angus heifers with differing liveweight change profiles (gaining or losing), in New Zealand to investigate the meanings of those parameters in the restricted feeding beef heifers. Beef heifers losing liveweight (−412 g/day) showed significantly lower concentrations of plasma IRI, and higher concentrations of plasma free fatty acid (FFA) than heifers gaining liveweight (483 g/day). The cytosolic and mitochondrial malate dehydrogenase (MDH) activities and MDH/lactate dehydrogenase (M/L) ratio in leukocytes of the liveweight losing heifers were significantly higher than those the liveweight gaining heifers. Percentages of cluster of differentiation (CD) 3 positive cells and natural killer (NK) cells in PBMC decreased significantly in the liveweight losing heifers compared to those in the liveweight gaining heifers. Plasma IRI and FFA concentrations, leukocyte cytosolic and mitochondrial MDH activities and CD3 positive and NK cell populations may be useful markers to evaluate metabolic conditions and immunity in the restricted feeding beef heifers.     

Dose-dependent effects of isoflurane and dobutamine on cardiovascular function in dogs with experimental mitral regurgitation

Objective

To investigate the dose-dependent effects of isoflurane and dobutamine on haemodynamics in dogs with experimentally induced mitral valve insufficiency (MI).

Comparison of nutrient uptake and antioxidative response among four Labiatae herb species under salt stress condition

Herbs of the Labiatae have relatively low salt tolerance. They are widely grown in drylands, but salt stress there is a typical problem and may reduce yields. To examine their salt tolerance mechanisms, we grew basil, sage, thyme, and oregano in nutrient solution containing 50 mM NaCl and determined the biomass; contents of Na, K, and Mg in leaf blades, stems, and roots; contents of total chlorophyll, malondialdehyde (MDA), hydrogen peroxide in leaf blades; and activities of antioxidative enzymes in leaf blades. The salt tolerance decreased in the order of basil ≈ sage > thyme > oregano. The good salt tolerance of basil was explained by a significant increase in the activity of catalase, in addition to the low Na/K ratio of leaf blades due to the retention of Na in stems and roots and of K in leaf blades. The good salt tolerance of sage was explained by the low Na/K ratio in leaf blades and the prevention of lipid peroxidation by high antioxidative enzyme activities, despite its poorer management of nutrient uptake. In thyme, although catalase activity increased significantly to alleviate salt-induced oxidative stress caused by Na influx into all parts, low K and Mg allowed shoot weight in particular to decrease. In oregano, antioxidative responses appeared as significant increases in ascorbate peroxidase and glutathione reductase activity, and K was accumulated in leaf blades, but serious salt-induced oxidative stress caused by high Na influx into all parts reduced the growth of all parts. These results show that despite similar responses among species, salt tolerance is not necessarily the same. In this experiment, we revealed the salt tolerance mechanism of each of four Labiatae herbs by revealing their strengths and weaknesses in nutrient uptake and antioxidative responses.