Microtubule distribution in somatic cell nuclear transfer bovine embryos following control of nuclear remodeling type

This study was conducted to evaluate the microtubule distribution following control of nuclear remodeling by treatment of bovine somatic cell nuclear transfer (SCNT) embryos with caffeine or roscovitine. Bovine somatic cells were fused to enucleated oocytes treated with either 5 mM caffeine or 150 µM roscovitine to control the type of nuclear remodeling. The proportion of embryos that underwent premature chromosome condensation (PCC) was increased by caffeine treatment but was reduced by roscovitine treatment (p < 0.05). The microtubule organization was examined by immunostaining β- and γ-tubulins at 15 min, 3 h, and 20 h of fusion using laser scanning confocal microscopy. The γ-tubulin foci inherited from the donor centrosome were observed in most of the SCNT embryos at 15 min of fusion (91.3%) and most of them did not disappear until 3 h after fusion, regardless of treatment (82.9-87.2%). A significantly high proportion of embryos showing an abnormal chromosome or microtubule distribution was observed in the roscovitine-treated group (40.0%, p < 0.05) compared to the caffeine-treated group (22.1%). In conclusion, PCC is a favorable condition for the normal organization of microtubules, and inhibition of PCC can cause abnormal mitotic division of bovine SCNT embryos by causing microtubule dysfunction.     

64-Channel multi-detector row CT angiographic evaluation of the micropigs for potential living donor lung transplantation

Micropigs are the most likely source animals for xenotransplantation. However, an appropriate method for evaluating the lung of micropigs had not been established. Therefore, this study was performed to evaluate the feasibility of 64-channel multi-detector row computed tomography (MDCT) to measure the diameter of the pulmonary arteries and the lung volume in micropigs. The mean diameters of the trachea, and left and right bronchi were 1.6 ± 0.17, 1.18 ± 0.14, and 1.1 ± 0.11 cm, respectively. The mean diameters of the main, right, and left pulmonary arteries were 1.38 ± 0.09, 1.07 ± 0.26, and 0.98 ± 0.13 cm and the diameters of right, left, and common inferior pulmonary veins were 0.97 ± 0.20, 0.76 ± 0.20, and 1.99 ± 0.26 cm, respectively. The mean lung volume was 820.3 ± 77.11 mL. The data presented in this study suggest that the MDCT may be a noninvasive, rapid, and accurate investigational method for pulmonary anatomy in living lung donors.     

The relationship between the size of caudolateral curvilinear osteophyte of the canine femoral neck and the radiographic view

Caudolateral curvilinear osteophyte (CCO), an osteophyte at the site of joint capsule attachment on the caudal aspect of the femoral neck, has been advocated as a radiographic criterion for coxofemoral subluxation. The correlation between the presence of CCO on radiographs (radiographic-CCO), the size of the CCO (CCO index) on three-dimensional computed tomographic (CT) images, and hip evaluation using transverse CT images was assessed in 22 Border Collies. CCOs were detected on the radiographs and CT images of 32% and 100% femurs, respectively. The CCO index correlated significantly with radiographic-CCO, but a large CCO index did not necessarily imply that the CCO was visible on radiographs. Hence, radiographic-CCO findings should be used cautiously in hip evaluation of Border Collies.     

Molecular identification of the vaccine strain from the inactivated oil emulsion H9N2 low pathogenic avian influenza vaccine

In order to control the H9N2 subtype low pathogenic avian influenza (LPAI), an inactivated vaccine has been used in Korea since 2007. The Korean veterinary authority permitted the use of a single H9N2 LPAI vaccine strain to simplify the evolution of the circulating virus due to the immune pressure caused by the vaccine use. It is therefore important to determine the suitability of the vaccine strain in the final inactivated oil emulsion LPAI vaccine. In this study, we applied molecular rather than biological methods to verify the suitability of the vaccine strain used in commercial vaccines and successfully identified the strain by comparing the nucleotide sequences of the hemagglutinin and neuraminidase genes with that of the permitted Korean LPAI vaccine strain. It is thought that the method used in this study might be successfully applied to other viral genes of the LPAI vaccine strain and perhaps to other veterinary oil emulsion vaccines.     

Antigenic and immunogenic investigation of the virulence motif of the Newcastle disease virus fusion protein

Newcastle disease (ND) caused by virulent Newcastle disease virus (NDV) is a highly contagious viral disease of poultry. Virulent NDVs characteristically have a multibasic amino acid sequence (virulence motif) such as ¹¹²RRQKRF¹¹⁷ at the cleavage site of the precusor fusion (F0) protein. The antigenic and immunogenic characteristics of the virulence motif ¹¹²RRQKRF¹¹⁷ in the F0 protein of virulent NDVs were investigated. Epitope mapping analysis revealed that a RRQKRF-specific monoclonal antibody 4G2 recognized the KRF section of the motif. A synthetic peptide bearing the RRQKRF motif reacted strongly with sera from virulent NDV (with RRQKRF motif)-infected chickens. These sera also showed reactivity to peptides bearing other virulence motifs (¹¹²KRQKRF¹¹⁷, ¹¹²RRQRRF¹¹⁷ and ¹¹²RRRKRF¹¹⁷) but not an avirulence motif (¹¹²GRQGRL¹¹⁷) by ELISA. The synthetic bearing RRQKRF motif reacted with 60% to 91% of sera taken from surviving chickens on ND outbreak farms but not with sera from vaccinated birds, even though most of the sera had antibody to NDV due to vaccination. This indicates that the virulence motif has the potential to differentiate virulent NDV infected birds from vaccinated birds.     

Inhibitory effects of interleukin-10 plasmid DNA on the development of atopic dermatitis-like skin lesions in NC/Nga mice

Interleukin (IL)-10 exerts potent anti-inflammatory effects by suppression of both T-help (Th) 1 and Th2 cells. Previous studies have reported that IL-10 can ameliorate various inflammatory disorders. The present study was performed to examine whether IL-10 plasmid DNA could suppress development of atopic dermatitis (AD)-like skin lesions in NC/Nga mice, as an initial step towards the development of an appliance for use in dogs with AD. Intradermal injection of IL-10 plasmid DNA markedly inhibited the development of AD-like skin lesions, as evidenced by a marked decrease in skin symptoms and reduced inflammation within the skin lesions. Efficacy was confirmed by significant decreases in eosinophil ratio and serum IgE concentration, and a reduction in the number of Staphylococcus aureus recovered from the ear. Moreover, relative mRNA expression levels of IL-4 and interferon-γ in the skin lesions of mice injected with IL-10 plasmid DNA were also decreased compared with those of control mice. Of note, higher serum IL-10 levels in mice injected with IL-10 plasmid DNA were maintained compared with those in control mice. Taken together, the results indicate that IL-10 plasmid DNA can suppress the development of AD-like skin lesions by suppressing both Th1 and Th2 cell responses. Beneficial effects of IL-10 plasmid DNA may be expected in dogs with AD.     

Potential risks,prognostic indicators,and diagnostic and treatment modalities affecting survival in dogs with presumptive aspiration pneumonia: 125 cases (2005–2008)

Objective – To evaluate a clinical population of dogs diagnosed with presumptive aspiration pneumonia (AP) and determine diagnostic and treatment modalities contributing to survival. Design – Retrospective study. Setting – A university veterinary teaching hospital in an urban setting. Animals – One hundred and twenty‐five dogs with presumed AP treated from 2005 to 2008. Interventions – None. Measurements and Main Results – Dogs with presumptive AP identified by a review of medical records had an overall survival of 81.6% (102/125). Male large‐breed dogs (mean 24.9 kg; 82/125) were overrepresented and were more likely to develop AP in this study population. Recent anesthesia had been performed in 16% (20/125), and vomiting was reported in 64% (80/125). The most common radiographic findings were a predominantly alveolar pattern (187/272, [68.8%] total lung lobes) in the right middle lung lobe (80/115, [69.6%]). A mean of 2 lung lobes were involved radiographically, and the relationship between survival and the number of lung lobes affected was statistically significant (P=0.04). Neutrophilia with a left shift was common with no significant change on consecutive daily evaluations. The mean PaO₂ was 77.7 mm Hg (SD, 17.5 mm Hg) (range, 40.7–100 mm Hg) with a median alveolar‐arterial gradient of 41.1 mm Hg (range, 8.1–81.8 mm Hg). In this study population, 37.6% (47/125) of dogs had microbial cultures performed and of these, 76.6% (36/47) were positive for growth; Escherichia coli (38.8%), Mycoplasma spp. (21.3%), Pasturella spp. (19.1%), and Staphylococcus spp. (17%) were the most common isolates in either single or multiagent infections. No treatment modality was statistically associated with increased survival. Colloid therapy was a negative prognostic indicator. Conclusions – In this study the overall prognosis for AP was good. Patients with only 1 affected lung lobe appeared more likely to survive. Supportive treatment modalities are warranted for the hospitalized patient, although no individual treatment method was found to be clearly superior to others.     

Effect of silicon on growth and salinity stress of soybean plant grown under hydroponic system

Silicon (Si) is abundant in the soil, yet its role in plant biology has been poorly understood. The role of Si in soybean growth and its effectiveness in salt stress alleviation was investigated. Sodium metasilicate (Na₂SiO₃) was given as Si source to hydroponically grown soybean (Glycine max (L.) Merr.). The plant growth attributes, i.e. plant height, plant fresh and dry biomass, chlorophyll contents and endogenous gibberellins (GAs) level improved with 2.5 mM Si, while endogenous abscisic acid (ABA) and free proline contents were not affected as compared to control. Sodium chloride (NaCl) significantly decreased growth attributes and endogenous gibberellins levels but markedly enhanced ABA and proline contents of soybean leaves. An addition of Si to salt stressed plants substantially alleviated the adverse effects of NaCl on growth, as it enhanced endogenous gibberellins, while reduced the levels of ABA and proline. GAs analysis of soybean leaves also showed that both early C13 hydroxylation and non C13 hydroxylation pathways of gibberellin biosynthesis were operating in soybean. The major GA biosynthesis pathway was identified as non C13 hydroxylation, which led to the formation of bioactive GA₄. Current study suggests that Si application alleviates the detrimental effect of salinity stress on growth and development of soybean.