Genomic characterisation of bovine papillomavirus types 1 and 2 identified in equine sarcoids in Japan.

BACKGROUND: Bovine papillomavirus types 1 and 2 (BPV1/2) infection in horses has been associated with the development of equine sarcoids. Previous findings revealed the presence of sarcoid-associated BPV sequence variants that have been proposed as a key factor of cross-species infection in horses. To verify this hypothesis, sarcoid-associated BPV variants should be identified regardless of geographic location. OBJECTIVES: Sequence analyses of BPV1/2 derived from both horses and cattle were conducted to clarify the sarcoid-associated sequence variants. The aim of this study was to clarify the correlation between BPV phylogeny and the geographic origin/host species. STUDY DESIGN: Cross-sectional study. METHODS: Conventional PCR to detect BPV1/2 was performed with genomic DNA extracted from equine sarcoid (n = 10) and bovine papilloma (n = 10) samples collected in Japan. Direct sequencing results were compared between equine and bovine (equine/bovine)-derived BPV to identify sarcoid-associated variants of two early regions (E2, E5), one late region (L1) and the long control region (LCR). Phylogenetic and phylogeny-trait correlation were analysed using Bayesian Markov chain Monte Carlo (MCMC) method and Bayesian tip-association significance testing (BaTS). RESULTS: Seven BPV1 and three BPV2 were identified from equine sarcoids using PCR and direct sequencing. Sequence analysis of equine/bovine-derived samples showed no sarcoid-associated variants in four regions (E2, E5, L1 and LCR) of either BPV1 or BPV2. The phylogenetic tree of BPV1 E2, L1 and LCR tended to cluster within its geographic origins. BaTS analysis demonstrated that BPV1 sequence variability may be due to the geographic origin rather than host species difference. MAIN LIMITATIONS: There was a limitation in sample numbers. CONCLUSIONS: This study supports the geographic-specific hypothesis of sequence variability, suggesting that BPV1 is shared between local equids and bovids. However, more extensively collected sequences worldwide and functional evaluations are needed to verify the geographic-specific sequence variability of BPV1/2 between equine- and bovine-derived sequence.

The silicon concentration in cat urine and its relationship with other elements.

To understand the effects of silicon (Si) in the urine with respect to the formation of urinary stones, the distribution of Si in urine was observed. Urine samples from cats with urolithiasis (n=10) and healthy cats (n=15) were used. The concentration of Si in the cats with urolithiasis was significantly higher (P<0.001). A significant correlation (P<0.05) was observed between the concentration of Si and those of other elements, such as calcium, magnesium, phosphorus, potassium and iron, only in the urine of the healthy cats. The distribution of elements in the urine differed between the cats with urolithiasis and the healthy cats. The Si concentration and its relationship with other elements were suggested to be useful biomarkers for urolithiasis in cats.

Is experience on a farm an effective approach to understanding animal products and the management of dairy farming?

The understanding of animal products and dairy farming is important for the promotion of dairy farming. Thus, to examine the effects of farm experience on the understanding of animal products and the management of dairy farming, the interaction between students and dairy cows was investigated in groups of first-year veterinary nursing students in 2011 and 2012 (n = 201). These students included 181 women and 20 men. Nine items about dairy cows were presented in a questionnaire. The survey was performed before and after praxis on the educational farm attached to the authors' university. After praxis on the farm, increases occurred in the number of positive responses to the items involving the price of milk, dairy farming and the taste of milk. For these items, a significant difference (P < 0.05) was found between the scores obtained before and after training. The results of the study suggested that farm experience is useful for improving the understanding of animal products and dairy farming.

The distribution of several elements in cat urine and the relation between the content of elements and urolithiasis.

The concentrations of elements in urine obtained from cats with urolithiasis were compared with those of healthy cats. The concentration of several elements, such as sodium (Na), phosphorus (P), sulfur (S), and potassium (K), in urine obtained from cats with urolithiasis was significantly higher than that of healthy cats. A significant correlation (p<0.01) was found between the concentration of magnesium (Mg) and that of other elements, such as P (r=0.8913), S (r=0.6817), and K (r=0.8391), in the urine obtained from healthy cats. A significant correlation (r=0.7422, p<0.05) was also obtained between the concentration of K and that of P in urine collected from cats with urolithiasis, but the slope of regression line was significantly different from that of the urine obtained from healthy cats. Other correlations observed in healthy cats were not obtained from cats with urolithiasis. However, a significant correlation between the concentration of magnesium (Mg) and that of calcium was obtained only from cats with urolithiasis. The results of the present study suggest that urinary concentrations of various elements in cats with urolithiasis are higher than those of healthy cats. Furthermore, the balance of elements in the urine of cats with urolithiasis was altered.

Identification of annexin 1 as a novel autoantigen in acute exacerbation of idiopathic pulmonary fibrosis.

Consistent with the hypothesis that pulmonary epithelial apoptosis is the key to the acute exacerbation of idiopathic pulmonary fibrosis (IPF), we conducted serological identification of Ags by recombinant expression cloning (SEREX) analysis using type II alveolar cell carcinoma (A549) cell lines to identify disease-related Abs. In a survey of Abs to the recombinant autoantigens identified by SEREX analysis, five Abs were identified as novel candidates for the acute exacerbation of IPF. Abs to annexin 1 were detected in 47 and 53% of the sera and bronchoalveolar lavage materials from patients with acute exacerbation of IPF. Some identical TCR Vbeta genes were identified in sequential materials obtained at 1-3 mo in all 10 acute exacerbation IPF cases, suggesting that some infiltrating CD4-positive T cells sharing limited epitopes expand by Ag-driven stimulation during disease extension. The CDR3 region of these identical TCR Vbeta genes showed high homology with the N-terminal portion of annexin 1, including in the HLA-DR ligand epitopes predicted by TEPITOPE analysis. By Western blotting analysis and observation of the CD4-positive T cell responses in bronchoalveolar lavage samples, the N-terminal portion of annexin 1 was cleaved and found to induce marked proliferative responses of CD4-positive T cells in three patients. Our study demonstrates that annexin 1 is an autoantigen that raises both Ab production and T cell response in patients with acute exacerbation of IPF, and that the N-terminal portion of annexin 1 plays some role in the pathogenesis of acute exacerbation in IPF patients.

BCL-6 mutations in pulmonary lymphoproliferative disorders: demonstration of an aberrant immunological reaction in HIV-related lymphoid interstitial pneumonia.

We used a PCR and sequence procedure to analyze the Ig V(H) gene and the mutations in the 5' regulatory regions of BCL-6 genes in pulmonary lymphoproliferative disorders (mucosa-associated lymphoid tissue (MALT) lymphoma, HIV-related, EBV-related, and virus-negative lymphocytic interstitial pneumonia (LIP)). Eight of 20 (40%) pulmonary MALT lymphoma and 10 of 20 LIP (5 of 5 (100%) HIV-related, 2 of 5 (40%) EBV-related, and 3 of 10 (30%) virus-negative LIP) cases showed BCL-6 gene mutations. Intraclonal heterogeneity of the BCL-6 mutations was observed only in pulmonary MALT lymphoma cases whose Ig V(H) genes also showed intraclonal heterogeneity. Ongoing BCL-6 mutations might reflect re-entry into a germinal center pathway to further mutations. BCL-6 mutations in pulmonary MALT lymphoma and HIV-negative LIP showed some features (high transition to transversion ratio, standard polarity, and RGYW/WRCY bias) of Ig V(H) gene hypermutation, leading to the view that pulmonary MALT lymphomas and HIV-negative LIP are under the influence of germinal center hypermutation mechanisms. Because BCL-6 mutations in HIV-related LIP cases did not demonstrate features of Ig V(H) gene hypermutation, immunological reactions in HIV-related LIP are the result of a process different from that found in HIV-negative pulmonary lymphoproliferative disorders.

Oligoclonal T cell expansions in pulmonary lymphoproliferative disorders: demonstration of the frequent occurrence of oligoclonal T cells in human immunodeficiency virus-related lymphoid interstitial pneumonia.

We used a denaturing gradient gel electrophoresis (DGGE) procedure with 40-nucleotide guanine- and cytosine-rich sequences in the polymerase chain reaction (PCR) and sequencing analysis to analyze the T cell antigen receptor (TCR)-Vgamma gene repertoire of infiltrating T lymphocytes in pulmonary lymphoproliferative disorders. Six of 15 low-grade mucosa-associated lymphoid tissue (MALT) lymphomas and 8 of 15 cases of lymphocytic interstitial pneumonia (LIP) showed some oligoclonal bands for TCR-Vgamma genes on DGGE. Sequencing analysis demonstrated plural oligoclonal TCR-Vgamma clones among the oligoclonal PCR products on DGGE, leading to the conclusion that conventional antigen-specific oligoclonal expansions may play some role in the pathogenesis of pulmonary lymphoproliferative disorders. The frequency of oligoclonal infiltrating T cell expansions in human immunodeficiency virus (HIV)-related LIP (100%) was significantly higher than in low-grade pulmonary MALT lymphomas (40%) or in HIV-negative LIP (30%). Because recent evidence demonstrates that the V3 loop in the proviral amino acid sequences of mononuclear cells from bronchoalveolar lavage is more homogeneous than those from peripheral blood, this homogeneity might result in oligoclonal expansions of infiltrating T lymphocytes as a consequence of ongoing reactions against lung-specific viral strains.