Diagnostic Efficacy of a Single Progesterone Determination to Assess Full-Term Pregnancy in the Bitch.

In clinical settings, when the reproductive history of a near-term bitch is limited to mating dates, the possibility to accurately assess whether pregnancy is at term could be very useful in order to be able to plan a correct management of parturition or to safely perform an elective Caesarean section. The aim of this study was to assess the diagnostic efficacy of a single progesterone determination, measured by chemiluminescent immunoassay (CLIA), in predicting the occurrence of parturition on the following day. At least one blood sample was collected from 51 pre-partum bitches during the 3 days before parturition and on day of parturition. The efficacy of progesterone as a marker of the end of pregnancy was tested using a receiver operating characteristic (ROC) analysis. Youden's index was calculated to select the optimal cut-off value (with 95% confidence interval), aiming at maximizing the correct identification of negative events, so not to risk to diagnose as full term a bitch which is not. Progesterone concentration lower than 3.4 ng/ml correctly identified the bitches whelping the following day; however, because of the obliged prudential approach, sensitivity was low (46.88%), and 17 of 32 full-term bitches were missed. Due to a very large individual variation, a single progesterone determination has low diagnostic efficacy, although it can represent a useful first screening.

Oxytocin precursor gene expression in bovine skeletal muscle is regulated by 17ß-oestradiol and dexamethasone.

Growth promoter administration, in livestock, potentially poses a major threat to public health, due to the potential endocrine and carcinogenic activity of residues, accumulating in edible tissues, such as skeletal muscle. Therefore, development of new screening tests and methods for the detection of illicit treatments of food animals would be useful. In this study the serum concentrations of oxytocin peptide were measured in beef cattle receiving 17ß oestradiol, dexamethasone or placebo over a period of 40 days. Changes in gene expression of oxytocin precursor in skeletal muscle were also examined in these animals. Serum analysis using an oxytocin EIA kit indicated a significant up-regulation of the biosynthesis of this nonapeptide only in cattle after 17ß oestradiol, but not after dexamethasone or placebo treatment. Quantitative PCR (qPCR) analysis showed a significant overexpression of the oxytocin precursor gene by 33.5 and 13.3-fold in cattle treated with 17ß oestradiol and dexamethasone, respectively, in comparison to placebo treated animals. Regulation of gene expression by some myogenic regulatory factors in skeletal muscle was also evaluated in these animal groups, confirming the activity of both growth promoters on this gene. To investigate the use of the oxytocin precursor gene as biomarker for 17ß oestradiol and dexamethasone treatment in beef cattle, an absolute quantification of this gene by qPCR was developed. A standard curve was generated and developed with TaqMan® technology and optimal criterion value, sensitivity and specificity of this screening method were established through ROC analysis. This analysis suggested that the up-regulation of oxytocin precursor gene expression in skeletal muscle tissue is a valid marker for detection of illicit 17ß oestradiol and/or dexamethasone use in beef cattle. This method may serve as a novel diagnostic tool in the screening phase, and, if introduced in routine testing, may significantly improve overall efficacy and success of the food screening process ordered by state authorities.

E-cadherin and ß-catenin expression in canine colorectal adenocarcinoma.

E-cadherin and its associated cytoplasmic proteins, including ß-catenin, have been examined as potential oncogenic markers due to the significant correlation between tumour dedifferentiation and the invasive capacity of epithelial tumours. The purpose of this study was to evaluate the expression of E-cadherin and ß-catenin in canine colorectal cancer using immunohistochemistry and to examine the relationship between this expression and various clinicopathological variables. The expression pattern of E-cadherin and ß-catenin was investigated in 44 colorectal canine carcinomas. In the intestinal mucosa of noncancerous areas, epithelial cells demonstrated equally strong membranous expression of E-cadherin and ß-catenin localised to the cell-cell junctions. Reduced expression of E-cadherin and ß-catenin was demonstrated in 75% and 81.8% of the colorectal carcinoma cases, respectively. The down-regulation of both E-cadherin and ß-catenin was correlated with decreased differentiation and increased tumour grade. In addition, the expression of ß-catenin was correlated with tumour size. These results suggest that dysfunction of the E-cadherin-catenin complex starts in the early stages of carcinogenesis and that the disruption of the tissue architecture is progressively associated with the invasion of the tumour.

Health status of a population of nutria (Myocastor coypus) living in a protected area in Italy.

Ninety trapped nutria (Myocastor coypus) from a protected area of Piedmont (Italy), including the Po river, were examined for the prevalence for lesions in major viscera, selected serum antibodies and enteric bacteria. Coccidial lesions in the liver included cholangitis, calcification and necrosis. Renal lesions were nonsuppurative interstitial nephritis and a single case of renal adenocarcinoma. The lungs had a 41.1% prevalence of nonsuppurative interstitial pneumonia. Ten of 87 sera (11.5%) had antibodies against Leptospira bratislava, 3 of 87 (3.4%) against Leptospira ichterohaemorrhagiae, 15 of 41 (36.6%) against Toxoplasma gondii, and antibodies against encephalomyocarditis virus were detected in 5 of 78 sera (6.4%). All fecal samples were negative for Salmonella, Shigella, and Pseudomonas, and growth of enterobacteriaceae was in the normal range.

p53 expression in cultured blood human monocytes infected with mycobacterial strains.

BACKGROUND: An upregulation of the cell-cycle associated proteins p53 and p21/Waf1/Cip1 induced by mycobacteria was previously reported. We aimed to evaluate the expression of such proteins in peripheral blood human monocyte cultures infected with strains of different mycobacterial pathogens. METHODS: The study relied on the immunocytochemical determination of p53, p21/Waf1/Cipl, bcl-2 and on the Tunel detection of apoptosis in monocytes populations cultured on four-welled chamber slides (10(6) cells/well) infected with Mycobacterium tuberculosis, M. bovis and M. avium for four consecutive days (mycobacterium/monocyte ratio 10:1). The results were expressed as mean values and SD of the percentages of stainings recorded in five fields per slide. RESULTS: The statistical analysis with Fischer test demostrated that at most sampling times the p53 and p21/Waf1/Cip1 expression and the apoptosis index were significantly higher in M. tuberculosis infected cultures than in controls (p<0.05). The M. bovis related picture diverged from the previous one for a lower p53 expression (p<0.05) at all sampling times. The M. avium infected culture values did not diverge significantly from the controls. CONCLUSIONS: The p53 and p21/Wafl/Cipl upregulation is compatible with both host defense strategies and pathogen strategies (safeguard of intracellular sanctuaries). The discrepancies among different cultures suggest a direct relationship between p53 activation and mycobacterial ability to enter host cells.