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See detailMEASUREMENT OF SERUM ALBUMIN BY PROTEIN ELECTROPHORESIS AND BROMOCRESOL GREEN METHODS IN CANINE AND EQUINE PATIENTS
Ramery, Eve ULg; Bureau, Fabrice ULg

in Veterinary Clinical Pathology (in press)

Background: Bromocresol green method (BCG) is routinely used in veterinary laboratories to measure albumin. A good agreement between BCG and electrophoresis (SPE) in healthy domestic animals is reported ... [more ▼]

Background: Bromocresol green method (BCG) is routinely used in veterinary laboratories to measure albumin. A good agreement between BCG and electrophoresis (SPE) in healthy domestic animals is reported. However, in human medicine, at low albumin and high globulin concentrations, BCG also binds to α- and β-globulins fractions. Surprisingly, there is no recent literature evaluating the BCG method in diseased domestic animals. Objective: The purpose of the present study was therefore to compare the serum albumin values obtained by SPE and BCG in canine and equine patients. Methods: Albumin measured by BCG and SPE was compared in the serum from 30 canine and 30 equine patients. Results: Good correlation was found (Spearman´s rho rs = 0.91 in dogs and 0.84 in horses). However, Altman and Bland analysis of results showed a positive bias of BCG with SPE (3.41 g/l in horses and 0.41 g/l in dogs) that increased when only hypoalbuminemic patients were considered (4.17 g/l in horses and 1.31 g/l in dogs). Conclusion: Results confirm that, despite good correlation, BCG method tends to overestimate albumin, especially in hypoalbuminemic patients. This is particularly evident in horses. [less ▲]

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See detailExercise and epinephrine affect equine leukocytes distribution and gene expression in a distinct but closely related pattern
Ramery, Eve ULg; Schoenecker, Julie; Art, Tatiana ULg et al

in Veterinary Clinical Pathology (2011, December), 40(4), 583-584

Background: In horses-athletes, exercise has been shown to be an important regulator of immune cells and their functions, and catecholamines to be essential components of the physiological response to ... [more ▼]

Background: In horses-athletes, exercise has been shown to be an important regulator of immune cells and their functions, and catecholamines to be essential components of the physiological response to exercise. Objectives: The purpose of the present study was to compare immunological reactions and molecular events during exercise and epinephrine infusion. Methods: Horses were submitted to exercise or epinephrine infusion for 10 min. Blood samples were collected at T0, T5, Tl0, T20 and T40 min for biochemistry and haematology at T0 and T20 min for RNA extraction and microarray analysis. Results: In both protocols, packed-cell-volume and neutrophils peaked at T5 and T10, and returned to baseline at T40. Lymphocytes increased at T5 with exercise and at T20 with epinephrine and then remained high until the end of the protocols. Between T0 and T20, 55 probes, coding for 25 genes, and 19 probes, coding for 11 genes, were differentially regulated with epinephrine and exercise respectively (P<0.001). Ten of the genes over-expressed with epinephrine were related to inflammatory response and leukocytes trafficking and activation. Among the genes with the highest significant fold changes following exercise were FOS, DUSP-1 and CXCL1. FOS was over-expressed in both conditions. Among genes differentially regulated, Ingenuity software identified 3 overlapping functional networks with epinephrine and 1 functional network with exercise that overlapped the 3 epinephrine networks. Conclusions: Circulating leukocytes distribution and gene expression are rapidly affected following exercise and epinephrine infusion. Distinct but closely-related sets of gene activities were shown to be altered with exercise and epinephrine, with lesser changes in the immunological pattern with physical stress than epinephrine alone, arguing for compensatory mechanisms in exercise. [less ▲]

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See detailEVALUATION OF ORGANIC DUST COMPONENTS CYTOTOXICITY ON THP1 MONOCYTES-DERIVED-MACROPHAGES USING HIGH CONTENT ANALYSIS
Ramery, Eve ULg; O'Brien, Peter James

in Veterinary Clinical Pathology (2011, December), 40(4), 583

Background: Organic dust contains pathogen associated molecular patterns (PAMPs) which can induce, following chronic exposure, significant airway diseases. Mononuclear phagocytes are key protecting cells ... [more ▼]

Background: Organic dust contains pathogen associated molecular patterns (PAMPs) which can induce, following chronic exposure, significant airway diseases. Mononuclear phagocytes are key protecting cells of the respiratory tract. Several studies have investigated the effects of PAMPs, and mainly endotoxins, on cytokine production. However the sub-lethal cytotoxicity of organic dust components on macrophages has not been tested yet. The novel technology of high content analysis (HCA) is already used to assess subclinical drug-induced toxicity. It combines the capabilities of flow cytometry, intracellular fluorescence probes, and image analysis and enables to perform rapid multiple analysis in large numbers of samples. Objectives: The purpose of the present study was, by using HCA, to investigate the cytotoxicity of the 3 major PAMPs contained in organic dust, ie. endotoxin (LPS), peptidoglycan (PGN) and β-glucans (zymosan) on THP-1 monocyte-derived macrophages. Methods: LPS was used at concentrations of 0.005, 0.01, 0.02, 0.05, 0.1 and 1 μg/ml; PGN and zymosan were used at concentrations of 1, 5, 10, 50, 100 and 500 μg/ml. Cells were exposed to PAMPs for 24 hours. In addition, the oxidative burst and the phagocytic capabilities of the cells were tested. Results: An overlap between PGN intrinsic fluorescence and red/far-red fluorescent dyes occurred, rendering the evaluation of some parameters impossible for PGN. LPS induced sub-lethal cytotoxicity at the lowest dose (from 10 ng/ml). However, the most spectacular changes occurred with zymosan. In addition, zymosan, but not LPS, induced phagosome maturation and oxidative burst. Conclusions: Given the fact that β-glucans can be up to 100 fold more concentrated in organic dust than LPS, these results suggest that β-glucans could play a major role in macrophages impairment following heavy dust exposure and will deserve further investigation in the near future. [less ▲]

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See detailRelationship between haptoglobin, serum amyloid A, and clinical status in a survey of dairy herds during a 6-month period
Humblet, Marie-France ULg; Guyot, Hugues ULg; Boudry, Benjamin et al

in Veterinary Clinical Pathology (2006), 35(2), 188-193

Haptoglobin and serum amyloid A are major acute phase proteins in cattle. Dairy cattle often develop pathologic conditions in the peripartum period; acute phase proteins may be useful in their diagnosis ... [more ▼]

Haptoglobin and serum amyloid A are major acute phase proteins in cattle. Dairy cattle often develop pathologic conditions in the peripartum period; acute phase proteins may be useful in their diagnosis. Objectives: The purpose of this study was to compare the accuracy of serum haptoglobin (Hp) and serum amyloid A (SAA) concentrations with clinical health status for diagnosing disease during the peripartum period in dairy cattle. Methods: Dairy cows from 4 herds were evaluated every 15 days over a 6-month period. Health status was determined by thorough clinical examination. Haptoglobin and SAA concentrations were measured in serum using validated methods and the results were classifed as positive or negative based on defined cutoff points. Disease prevalence, sensitivity, and specificity were compared using clinical examination as the gold standard. Results: A total of 1896 samples from 158 cows were analyzed. Significant increases in mean Hp and SAA concentrations were observed in the week following parturition in both primiparous and multiparous cows, although high interindividual variability was observed. Both Hp and SAA had low sensitivity but higher specificity in determining disease status compared with clinical examination. Increased concentrations of Hp and SAAwere found in ,10% of samples from clinically healthy cows, except in the week after parturition. Conclusions: Haptoglobin and serum amyloid A should be used with caution as markers of inflammation in the week after calving. Poor sensitivity in other postpartum periods could be related to the higher incidence of chronic (vs acute) inflammation. Haptoglobin may be appropriate for routine screening, but further work needs to be done to assess its value as an indicator of herd health. [less ▲]

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