Abstract: DNA Alkylation is thought to be the reason for the efficacy of lomustine while carbamylation has been implicated as the cause for the side effects seen with lomustine treatment such as hepatotoxicity. In the alkylation study we show that lomustine and its metabolites form similar levels of the DNA adducts N7 hydroxyethylguanine and O6 hydroxyethyldeoxyguanosine. In terms of carbamylation, lomustine showed greater extent of carbamylation in the canine hepatocytes and lymphoma cell lines. The DNA repair enzyme O6 methylguanine DNA methyltransferase (MGMT) causes resistance of tumor cells to bifunctional nitrosourea, like lomustine. There is no data available regarding MGMT expression/activity in canine cells or tissues. Our study shows that there is low MGMT activity in the canine lymphoid cell line 17–71 while the GL-1 cells did not show any detectable enzyme activity or mRNA expression. The MGMT enzyme activity measured in canine hepatocytes is about 250–350 fmol/mg protein as compared to about 90 fmol/mg protein in 17–71 cells. We also show that MGMT mRNA expression in 17–71 cells and canine hepatocytes positively correlates with its enzyme activity in these cells.
Abstract: The characteristics of canine IL-17-producing cells are incompletely understood. Expression of mRNA encoding orthologs of IL-17 and the IL-17 receptor has been documented in tissues from dogs with arthritis, inflammatory bowel disease, and lymphoma; however, no associations have been found between IL-17 gene expression and disease phenotype in these conditions. Robust assessment of the role of IL-17-producing cells in dogs will require measuring the frequency of these cells in health and disease in balance with other lymphocyte subsets. The aim of this study was to confirm that the T-cell IL-17 response in dogs is evolutionarily conserved. Canine peripheral blood mononuclear cells were stimulated with Concanavalin A with or without polarizing cytokines. We used a canine specific IL-17 ELISA and flow cytometry to identify IL-17-producing T cells. Accumulation of intracellular IL-17 was observed in stimulated CD4 and CD8 T cells. The addition of pro-inflammatory cytokines appeared to enhance polarization of canine CD4 T cells to the Th17 phenotype. Conversely, the addition of IL-2 in the presence of TGF-β resulted in expansion of Treg cells. We conclude that canine IL-17-producing cells behave similarly to those from humans and mice when stimulated with mitogens and polarized with pro-inflammatory or immune regulatory cytokines.
Abstract: The objective of this study was to develop and validate a Taqman real-time PCR assay for the detection of Mycoplasma bovis (M. bovis). Unique primers targeting the highly conserved house-keeping gene (uvrC) were designed and the probe sequence was derived from a previously published microarray study. There was 100% agreement in the outcome between our assay and the other two published assays for M. bovis detection. The analytical limit of detection of our assay is 83 copies of the uvrC gene. This assay was validated on a total of 214 bovine clinical specimens that were submitted to the Texas A&M Veterinary Medical Diagnostic Laboratory (TVMDL), Texas, USA. The specificity of the assay was assessed to be 100% since no cross-reactivity occurred with 22 other bacterial and other Mycoplasma species. We conclude that the uvrC gene serves as a good and reliable diagnostic marker for the accurate and rapid detection of M. bovis from a wider variety of specimen matrices.
Abstract: Colostrum contains substantially higher concentrations of immunoglobulins compared to serum, which may help to improve the utility of diagnostic tests. The aim of this study was to determine the diagnostic value of colostrum antibody concentrations in identifying Bovine Viral Diarrhoea Virus (BVDV) PI (persistently infected) calf carrying beef heifers following an experimental infection. Colostrum was collected within 12 hours of parturition and tested in undiluted, 1:5, 1:10, 1:100, 1:200, and 1:500 dilutions using an enzyme-linked immunosorbent assay (ELISA) for BVDV antibody. Cows were determined to be carrying a PI calf based on positive quantitative Real Time-Polymerase Chain Reaction and antigen ELISA result on pre-colostral serum and ear notch samples collected from their calf. The median ELISA sample-to-positive (S/P) ratio for colostrum collected from heifers that carried a PI calf were significantly higher than the median ELISA S/P ratio for colostrum collected from heifers that did not carry a PI calf at dilutions of 1:100, 1:200, and 1:500. This study provides further evidence for increased antigenic stimulation in utero by the BVDV viraemic PI calf, which can also be identified with 100% diagnostic sensitivity when using 1:500 dilution colostrum.
Abstract: One Health is one of the most important movements and emerging concepts in health today. The convergence of the fields of human and animal medicine has the potential to generate novel scientific hypotheses, create effective new therapies and potentially transform how physicians, veterinarians and their patients understand health and disease. Despite this potential, One Health has not yet gained significant awareness or traction in human medical communities. From its inception, One Health, sometimes also called One Medicine, has been piloted primarily by leaders from the world of veterinary medicine. Although the specific term was coined perhaps 10 years ago, comparative medicine has been quietly evident on university campuses with veterinary and medical schools for decades longer. Although a few physicians have played major leadership roles in One Health, in the United States, despite over ten years of the movement’s robust growth, many have still not heard of it. Furthermore, physicians with some awareness of One Health often believe it to be primarily and exclusively about zoonotic infections and global health. The much broader scope and potential of One Health as also including comparative physiology and medicine is not being communicated effectively. Consequently, the human medical community remains largely disengaged. This is problematic because without significant engagement from physicians, nurses and other human health care professionals, the potential of One Health cannot be realized. To advance One Health it is imperative that we first understand the roots of under-engagement of the human medical community. This, in turn, can guide the development of novel and engaging opportunities for physician which demonstrate the power relevance of One Health’s comparative, collaborative and cooperative approach.[...]
Abstract: Gingival overgrowth is an uncommon adverse effect of cyclosporine administration in veterinary species. In people, gingival overgrowth is a common complication of cyclosporine administration for immunosuppression, generally following transplant procedures. Azithromycin has been used successfully for managing gingival overgrowth in human transplant patients when cyclosporine administration cannot be reduced or discontinued. This case series describes six dogs being administered cyclosporine for various dermatologic diseases that developed gingival overgrowth. The dogs were prescribed systemic azithromycin, with or without concurrent dose reduction of cyclosporine. Oral administration of 6.6–10.8 mg/kg of azithromycin once daily for 4–14 weeks was effective for complete clinical resolution of gingival overgrowth. In most cases, gingival overgrowth did not recur even with continued cyclosporine administration long-term. Adverse events of long-term azithromycin administration did not occur in any of the dogs. This series highlights a potentially beneficial medical treatment option for gingival overgrowth even when cyclosporine dose reduction is not possible or elected, without the need for surgical resection of proliferative gingival tissue.