Tibial plateau symmetry and the effect of osteophytosis on tibial plateau angle measurements

A novel technique was developed to estimate the caudal medial tibial plateau landmark in the face of osteophytosis to improve accuracy in tibial plateau angle measurements. Using this technique, tibial plateau angles were evaluated in 31 normal dogs before and 8 months after right cranial cruciate ligament transection. There was no significant difference in mean tibial plateau angle before or after induction of osteophytosis. Additionally, it was determined that 90% of dogs had a difference of =2 degrees between right and left tibial plateau angles, which was considered symmetrical.     

Molecular cloning and expression analysis of pig CD81

CD81, also known as TAPA-1 (target of antiproliferative antibody 1), is a member of the tetraspanin family of proteins and a component of the B cell co-receptor complex. Several studies have shown that CD81 plays significant roles in a variety of immune responses, including activation of B cells and T cells. In this study, we cloned pig Cd81 cDNA using RT-PCR coupled with rapid amplification of cDNA ends (RACE)-PCR and determined the complete cDNA sequence of pig Cd81. Pig Cd81 cDNA contains an open reading frame (711 bp) encoding 236 amino acids. The identity of pig CD81 with those of human, cattle, rat, and mouse are 90.30%, 92.26%, 86.22%, and 86.22%, respectively. Alignment of the CD81 amino acid sequence with those of mammalian species showed that the large extracellular loop (LEL) is the most divergent, whereas other domains are largely conserved. Pig Cd81 mRNA was detected by RT-PCR in a broad range of tissues, including lymphoid tissues as well as nonlymphoid tissues, indicated variety of cellular functions of CD81 in most pig tissues. Flow cytometry analyses demonstrated that human CD81 antibody recognizes a pig CD81 on the cell surface. Further, immunohistochemistry analysis using human CD81 antibody on pig spleen was revealed that CD81 expression is widely diffused in spleen tissue. Future study will be focused on defining the functional role of CD81 during the course of pig infectious diseases.     

Changes in glial fibrillary acidic protein immunoreactivity in the dentate gyrus and hippocampus proper of adult and aged dogs

Astrocytes perform neuron-supportive tasks, repair and scarring process in the central nervous system. In this study, we observed glial fibrillary acidic protein (GFAP), a marker for astrocytes, immunoreactivity in the dentate gyrus and hippocampus proper (CA1-3 region) of adult (2-3 years of age) and aged (10-12 years of age) dogs. In the adult group, GFAP immunoreactive astrocytes were distributed in all layers of the dentate gyrus and CA1-3 region, except in the stratum pyramidale of the CA1-3 region. In the aged group, GFAP immunoreactivity decreased markedly in the molecular layer of the dentate gyrus. However, GFAP immunoreactivity in the CA1-3 region increased in all layers, and the cytoplasm of GFAP immunoreactive astrocytes was hypertrophied. GFAP protein levels in the aged dentate gyrus decreased; however, GFAP levels in the CA1-3 region increased. These results suggest that the morphology of astrocytes and GFAP protein levels in the hippocampal dentate gyrus and CA1 region are changed, respectively, with age.     

Getting cosy in freshwater: Assumed to be brackish pike are not so brackish after all

Pike (Esox lucius) occupy coastal streams and rivers of the Baltic Sea, where they attain large sizes (>5 kg). These large sizes are perhaps due to the fact that they can tolerate relatively high salinities and can thus forage in the nearby more productive brackish environments. In an attempt to quantify the extent to which pike utilise brackish environments, and to provide some insight into the underlying causes for brackish water migrations, we tagged 30 pike from a western Baltic river with acoustic transmitters and were able to track 21 individuals for 1 year. Based on experienced from local anglers, this population was assumed to be brackish in nature, where individuals underwent freshwater migrations to spawn. Our findings however suggest that the smallest and most active individuals make short exits into brackish waters and do so on rare occasions. Our results further indicate that neither sex nor size is related to activity level. We suggest that these patterns reflect two distinct behaviours—active and passive—and that large pike can be supported by the food availability in the river, without the need to venture into coastal zones, thus defying the conventional view that Baltic pike are all brackish in nature.     

Watermelon lycopene β-cyclase: promoter characterization leads to the development of a PCR marker for allelic selection

In the carotenoid biosynthetic pathway, lycopene β-cyclase (LCYB) catalyzes the cyclization that converts lycopene into β-carotene. Only a single copy of LCYB was identified and was suggested to encode a chromoplast-specific LCYB (CYCB type) in watermelon [Citrullus lanatus (Thunb.), Matsum & Nakai]. Splicing variants in the 5′-untranslated region were identified, but alternative splicing did not provide an explanation of the regulation of carotenoid accumulation in watermelon flesh. A quantitative assay using real time-PCR showed that differential expression was not detected between red- and canary yellow-fleshed watermelon cultivars. LCYB promoter regions were isolated and characterized, and a sequence difference was identified in the promoter region between red and canary yellow LCYB alleles. This polymorphism did not change the expression of LCYB, but does provide a reliable marker for discriminating LCYB alleles for red and canary yellow flesh. To develop a PCR-based marker to distinguish between the two LCYB alleles, we designed primers flanking the polymorphic region. The newly developed marker, designated Clcyb.600, co-segregated perfectly with flesh color phenotypes and single nucleotide polymorphism (SNP) markers developed in our previous study. Moreover, the Clcyb.600 marker offers easier discrimination of LCYB alleles than SNP or cleaved amplified polymorphic sequence markers, as it does not require restriction enzyme digestion for genotyping. Genotyping of LCYB promoter alleles in various commercial cultivars and plant introductions indicated that watermelon cultivars can be classified into two groups, those carrying a red LCYB allele or a canary yellow LCYB allele.     

Three AFLP markers tightly linked to the genic male sterility <Emphasis Type="Italic">ms</Emphasis><Subscript><Emphasis Type="Italic">3</Emphasis></Subscript> gene in chili pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis> L.) and conversion to a CAPS marker

Genic male sterility (GMS) has long been used as a tool for hybrid seed production in chili pepper (Capsicum annuum L.). We developed DNA markers linked to the GMS ms ₃ gene in a segregating population using bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) techniques. The segregating population was subjected to BSA-AFLP with 512 primer combinations. Three AFLP markers (Eagg/Mccc₂₇₆, Eagc/Mctt₁₇₈, and Ecag/Mtgc₂₀₄) were identified as tightly linked to the ms ₃ locus. Among them, we converted the AFLP marker Ecag/Mtgc₂₀₄ to the cleavage amplified polymorphic sequence (CAPS) marker, named GMS3-CAPS, based on sequencing analysis of internal and flanking regions for the markers between male-fertile and sterile plants. This marker will be useful for pepper breeding using the GMS system.     

Genetic and antigenic characterization of a poxvirus isolate from ostriches

Avian poxvirus was isolated from nodules on the heads and conjunctiva of two 3-to-4-wk-old ostrich chicks. The ostriches from which poxvirus was isolated had been placed on premises where turkeys that had shown evidence of poxvirus infection had been raised earlier. Microscopically, the nodules from the ostriches were composed of proliferating and hypertrophic epithelial cells that formed large fronds. Most of the hypertrophic epithelial cells contained large eosinophilic intracytoplasmic inclusion bodies characteristic of poxvirus. Characterization of the avian poxvirus isolated from the cutaneous lesions in ostriches was based on western blotting of virus antigen, restriction fragment length polymorphism of genomic DNA, pathogenesis, and cross-protection studies in chickens. Antigenic and genetic studies did not reveal any significant difference between the poxvirus isolated from ostriches (PVO) and fowl poxvirus (FPV). Further, susceptible chickens immunized with the PVO were protected when challenged with a virulent strain of FPV. Thus, the poxvirus isolated from ostriches had similar antigenic, genetic, and biological properties to FPV.     

Metabolism of flupyrazofos in the isolated perfused rat liver

To investigate the hepatic metabolism of the new insecticide flupyrazofos [O,O-diethyl O-(1-phenyl-3-trifluoromethylpyrazol-5-yl) phosphorothioate], isolated rat liver was perfused with flupyrazofos under single-pass conditions. In outflow perfusate and bile, 1-phenyl-3-trifluoromethyl-5-hydroxyprazole (PTMHP), PTMHP-sulfate and PTMHP-glucuronide conjugates were identified as the metabolites of flupyrazofos. However, O,O-diethyl O-(1-phenyl-3-trifluoromethylpyrazol-5-yl) phosphate (flupyrazofos oxon) was not detected. A HPLC method with UV detection was used to investigate the hepatic disposition of flupyrazofos and its metabolite PTMHP. The concentrations of flupyrazofos, PTMHP and PTMHP conjugates in outflow perfusate reached steady-state levels within 20 min after commencing perfusion of 7.3 microM flupyrazofos. At steady state, the mean extraction ratio of flupyrazofos was 0.93 (+/- 0.01) and clearance was 26.1 (+/- 0.2) ml min-1 which nearly approached perfusate flow rate (28 ml min-1). PTMHP accounted for 55.7 (+/- 5.8)% of eliminated flupyrazofos and was recovered as unchanged PTMHP, PTMHP-sulfate and PTMHP-glucuronide in the bile as well as the outflow perfusate.     

Effect of catheter size and injection rate of contrast agent on enhancement and image quality for triple‐phase helical computed tomography of the liver in small dogs

Rapid contrast injection is recommended for triple‐phase helical computed tomography (CT) of the liver. However, a large‐gauge catheter is needed for faster contrast injection and this is not practical for small breed dogs or cats. The purpose of this crossover group study was to evaluate applicability of a lower injection rate with a small‐gauge (G) catheter for triple‐phase hepatic CT in small dogs. Triple‐phase CT images were acquired for six beagle dogs using three protocols: an injection rate of 1.5 ml/s with a 24 G catheter, 3.0 ml/s with a 22 G catheter, and 4.5 ml/s with a 20 G catheter. Enhancement of the aorta, portal vein, and hepatic parenchyma was measured in each phase (arterial, portal, and delayed) and image quality was scored subjectively by two observers. Injection duration, time to scan delay, and time to peak enhancement were also recorded. Contrast injection duration decreased with a higher injection rate (n = 6, P ≤ 0.01), but time to peak enhancement and time to scan delay were not significantly affected by injection rates and catheter sizes. Contrast injection rate did not significantly affect aortic, portal, and hepatic enhancement. In addition, separation between each phase and quality of images was subjectively scored as good regardless of injection rate. Findings from the current study supported using an injection rate of 1.5 ml/s with a catheter size of 24 G for triple‐phase hepatic CT in small dogs (weight < 12 kg).     

Effect of heat treatment on the antioxidant activity of extracts from citrus peels

The effect of heat treatment on the antioxidant activity of extracts from Citrus unshiu peels was evaluated. Citrus peels (CP) (5 g) were placed in Pyrex Petri dishes (8.0 cm diameter) and heat-treated at 50, 100, or 150 degrees C for 10, 20, 30, 40, 50, and 60 min in an electric muffle furnace. After heat treatment, 70% ethanol extract (EE) and water extract (WE) (0.1 g/10 mL) of CP were prepared, and total phenol contents (TPC), radical scavenging activity (RSA), and reducing power of the extracts were determined. The antioxidant activities of CP extracts increased as heating temperature increased. For example, heat treatment of CP at 150 degrees C for 60 min increased the TPC, RSA, and reducing power of EE from 71.8 to 171.0 microM, from 29.64 to 64.25%, and from 0.45 to 0.82, respectively, compared to non-heat-treated control. In the case of WE from CP heat-treated at the same conditions (150 degrees C for 60 min), the TPC, RSA, and reducing power also increased from 84.4 to 204.9 microM, from 15.81 to 58.26%, and from 0.27 to 0.96, respectively. Several low molecular weight phenolic compounds such as 2,3-diacetyl-1-phenylnaphthalene, ferulic acid, p-hydroxybenzaldoxime, 5-hydroxyvaleric acid, 2,3-diacetyl-1-phenylnaphthalene, and vanillic acid were newly formed in the CP heated at 150 degrees C for 30 min. These results indicated that the antioxidant activity of CP extracts was significantly affected by heating temperature and duration of treatment on CP and that the heating process can be used as a tool for increasing the antioxidant activity of CP.