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Background Determination of ceruloplasmin (Cp) activity in plasma can provide an objective measure of the health of dairy cows as well as it can be used for various diagnostic purposes. The current study was designed to perform an analytical validation of a method for the determination of plasma Cp activity in dairy cows and to evaluate the influences of plasma storage times and temperatures as well as freeze–thaw cycles on the activity of this enzyme. This cohort was carried out on ten cows. For each cow, 24 aliquots of plasma, which were stored at different temperature regimes, were prepared. Both intra- and interassay coefficients of variation (CVs) were determined. The linearity was evaluated using bovine plasma Cp standard. Results The mean values of intra- and interassay CVs were 1.08 and 2.12%, respectively. Results of linearity testing showed a high correlation coefficient (r = 0.998, P

Wilson's disease (WD) is an autosomal recessive disorder of copper metabolism with copper accumulation in the liver as well as in the central nervous system. Treatment of WD includes oral chelating agents and diet and it is effective. However, once irreversible damage has occurred, the effect of treatment is diminished and the patient's quality of life is compromised. It is estimated that at least half of the patients with WD remain undiagnosed and die of untreated disease. Early detection of patients presymptomatically has been hampered by the lack of effective methods for mass screening. Recently, a sandwich ELISA method for ceruloplasmin measurement in blood spots was developed. We have used this method to analyze blood specimens collected on filter paper from 3667 children aged 3 months–15 years. The mean value of ceruloplasmin was 30.5±9.5 mg/dL. Among these children, we identified one WD case, a 32-month-old boy with markedly reduced ceruloplasmin concentration (2.3 mg/dL). Measurement of CP level in dried blood spot sample is proposed as a reliable method for population screening of WD.

Ceruloplasmin (Cp) determination could provide an objective measure of the health status of an animal and could be used as marker of animal health and welfare (Skinner, 2001) (Skinner, J. G., 2001. Special report. International standardization of acute phase proteins. Veterinary Clinical Pathology 30, 2-7.) but only manual methods have been reported to determine Cp concentrations in pigs (McCosker, 1961; Toussaint et al., 1995; Eckersall et al., 1996) (McCosker, P. J., 1961. Paraphenylenediamine oxidase activity and copper-levels in mammalian plasmas. Nature. 190, 887-889; Toussaint, M. J. M., Van Ederen, A. M., Gruys, E., 1995. Implication of clinical pathology in assessment of animal health and in animal production and meat inspection. Comparative Haematology International 5, 149-157; Eckersall, P. D., Saini, P. K., McComb, C., 1996. The acute phase response of acid soluble glycoprotein, alpha(1)-acid glycoprotein, ceruloplasmin, haptoglobin and C-reactive protein, in the pig. Veterinary Immunology and Immunopathology 51, 377-385). In the present study two automated methods based on the use of two different substrates for the determination of serum ceruloplasmin in pigs were developed, evaluated and compared. Both methods showed a good precision and detection limits, with no signs of inaccuracy and could be applied to biochemical autoanalyzers usually found in clinical laboratories using only minimal amounts of serum. Additionally the behaviour of Cp in experimental and clinical situations was studied showing an increase of around two fold after turpentine administration and significantly higher values in cases of pigs with inflammatory conditions when compared with healthy pigs.

INTRODUCTION AND AIMS:There are a limited number of studies devoted to the association between an antioxidant status and peritoneal dialysis (PD) adequacy and technical survival. The present study was undertaken to investigate some blood antioxidant biomarkers and their potential effects on PD technical survival. METHODS:This single-center, cross-sectional cohort study was conducted for between January 2010 and May 2015. 44 (31 males, 13 females) stable ambulatory non-diabetic patients with ESRD on PD were included in the study. All the patients had been undergoing continuous ambulatory PD (CAPD) for more than 3 months. They were observed during a period of 36 months to determine the impact of antioxidant status on the dialysis technical survival. The mean age in the patient population was 48.9 ± 13.2 yrs. The diabetics and the patients with a history of peritonitis or significant illness / hospitalization in the previous 3 months were excluded. The average duration of PD therapy at this study entry (at baseline) was 29 [18.5-37] months. The control group consisted of 30 healthy individuals. Survival curves were constructed according to the Kaplan-Meier method and compared using the log-rank test. A technical failure was defined as all causes of discontinuation of PD. The durations of technique survival were calculated from the date of inclusion in the study. For this analysis, in January 2010, the patients were categorized into two groups according to the baseline CP level. RESULTS:During the 3-yrs follow-up, 9 of 44 (20.5 %) patients dropped out from PD; 7 patients were considered to be ‘technique failures’, but, they continued PD treatment for various reasons: the inability to formation of an adequate vascular access 3 (6.8 %), the patient's refusal to transfer to hemodialysis 2 (4.5 %) and non-compliance 2 (4.5 %). The baseline CP level in the PD patients was significantly decreased compared to the control group (0.09 [0.07 - 0.1] g/L vs 0.22 [0.19 - 0.23] g/L; P < 0.0001). But, the average serum CP level in the 28 patients of the ‘technique survival’ group was significantly higher compared with the 16 patients of the ‘technique failure’ group (0.26 [0.22 - 0.32] g/L vs 0.09 [0.07 - 0.1] g/L, p < 0.0001). There were no significant differences in the demographic and baseline PD characteristics of the patients in the two groups stratified according to CP levels, excluding PD technique survival (P = 0.001). The results of the Kaplan-Meier analysis and log-rank test also demonstrated that there was a significant difference in the cumulative technical survival rate between the patients with CP level < 0.19 g/L and CP level ≥ 0.2 g/L (log-rank test: χ2= 11.3, P = 0.0008, Figure 1). The hazard ratio for the technique failures was significantly higher in the lowest CP tertile category compared with the highest CP level (HR, 4.8; 95% confidence interval, 1.2 to 18.7; P=0.002). CONCLUSIONS:We have found out a decrease in the serum ceruloplasmin level in the PD patients. Baseline serum ceruloplasmin level is an independent predictive factor for the PD technical survival. Further studies are needed to identify the opportunities for antioxidant therapy to improve the PD technique survival.