Fecal Calprotectin: A Comparison of Two Commercial Enzymoimmunoassays and Study of Fecal Extract Stability at Room Temperature.

BACKGROUND: The aims of the study were: 1) to compare the fecal calprotectin (fCal) assay results with Calprolab™ ELISA (HRP) (Calpro AS) versus our routine method, Elia™ fluoroenzymoimmunoassay (Thermo Fisher), and 2) to determine whether the fCal assay results do not vary following storage of the extract at room temperature for 4 days with the Calpro AS buffer, this being the estimated shipment time from the home of the patient, and an aspect little studied to date. METHODS: The fCal was determined in 198 patients divided into three groups: inflammatory bowel disease (IBD), organic intestinal disease, and functional intestinal disorders. Fecal extraction was carried out using the Roche Diagnostics kit with the corresponding specific buffers. RESULTS: The fCal assay with the Thermo Fisher method was found to be more sensitive but less specific than with the Calpro AS technique. The positive predictive value was low (just over 50%), though the negative predictive value was high (over 90%) with both methods. The likelihood ratios revealed small but occasionally important pre- versus post-test differences. When we compared the two methods, the Spearman correlation coefficient (ρ) was 0.819 (95% CI: 0.768 - 0.860) (P < 0.0001), reflecting a positive correlation. Similarly, when stratifying the fCal results into < 50 µg/g, 50 - 100 µg/g and > 100 µg/g, the resulting Cohen's kappa coefficient was 0.7766 (95% CI: 0.7025 - 0.8507), reflecting a substantial agreement between both methods. The stability of fCal was high in fecal extracts with the Calpro AS extraction buffer at room temperature for 4 days, which yielded a Spearman correlation coefficient of 0.951 (95% CI: 0.933 - 0.965), when the results were compared to those of the recent extracts (P < 0.0001). CONCLUSIONS: A positive correlation was observed between the two methods. In view of the high negative predictive value obtained with fCal, the presence of organic disease is highly unlikely in the presence of a normal concentration of this marker. We also confirmed the excellent stability of fCal in fecal extracts with the Calpro AS extraction buffer stored at room temperature. Thus, and for the sake of convenience and hygiene, it would be ideal for the patient to perform the extraction at home.

One-year follow-up of clinical, metabolic and oxidative stress profile of morbid obese patients after laparoscopic sleeve gastrectomy. 8-oxo-dG as a clinical marker.

Obesity has grown worldwide over the last few decades. In its different degrees, obesity is accompanied by many clinical and biochemical alterations reflecting the pathological condition of various body tissues. Among the mechanisms underlying the pathogenesis of obesity and associated complications, oxidative stress (OS) may be playing an important role. In the present study, we have characterized at systemic level the degree of OS status in a group of morbid obese patients (BMI>40kg/m2) at basal sate and its modulation during one year after bariatric surgery using the laparoscopic sleeve gastrectomy (LSG) technique. As compared with normal weight subjects matched in age, peripheral blood mononuclear cells (PBMc) of obese patients present a significant reduction of the antioxidant enzyme activities superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) as well as a significant increase of the oxidized/reduced glutathione ratio (GSSG/GSH) in these cells. Lipid peroxidation is significantly increased in the patient group as shown by the increased levels of malondialdehyde (MDA) in PBMc and the amount of F2-Isoprostanes (F2-IsoPs) released in urine. In addition, the DNA damage product 8-oxo-7,8-2'-deoxyguanosine (8-oxo-dG) was also observed to be increased in serum and urine of morbid obese patients as compared with the control group. After LSG, an improvement of their ponderal and metabolic profile was accompanied by a progressive recovery of antioxidant enzyme activities and the decline of oxidative byproducts both in PBMc and biological fluids. The observed changes of urinary 8-oxo-dG levels correlate positively with its serum concentration, the lipid peroxidation products MDA and F2-IsoPs, triglycerides, glucose, insulin, HOMA index and body weight and negatively with the percentage of weight and BMI loss and antioxidant activities. We conclude that the analysis of urinary 8-oxo-dG could be validated as a useful marker for the monitoring of ponderal and metabolic status of morbid obese patients.

Deregulated hepatic microRNAs underlie the association between non-alcoholic fatty liver disease and coronary artery disease.

BACKGROUND & AIMS: Non-alcoholic fatty liver disease (NAFLD) appears to be a new risk factor for the development of coronary artery disease (CAD). Members of a class of non-coding RNAs, termed microRNAs (miRNAs), have been identified as post-transcriptional regulators of cholesterol homoeostasis and can contribute to the development of NAFLD. The aims of this study were to (i) to assess the relationship between NAFLD and sudden cardiac death (SCD) from severe CAD in forensic autopsies and (ii) to quantify several hepatic miRNAs previously associated with lipid metabolism and NAFLD to correlate their expression with the presence of NAFLD, CAD, obesity parameters and postmortem lipid profile. METHODS: A total of 133 cases of autopsies with SCD and established CAD (patient group, CAD-SCD) and 106 cases of non-CAD sudden death (control group, non-CAD-SD) were included. miRNAs were quantified in frozen liver tissues. RESULTS: Males predominated in both groups. Patients more frequently exhibited NAFLD and necroinflammatory steatohepatitis (NASH) than controls (62% vs 26%, P = 0.001 and 42% vs 26%, P = 0.001 respectively). In both groups, the presence of NAFLD correlated with body mass index and abdominal circumference (P < 0.05). An increase in miR-34a-5p and a decrease in miR-122-5p and -29c-3p in patients with NASH vs controls without NAFLD were observed (P < 0.05). Finally, significant correlations between miR-122-5p and unfavourable lipid profile and also hs-CRP and miR-34a-5p were noted. CONCLUSIONS: CAD is associated with NAFLD and NASH. The hepatic miRNAs studied appear to be associated with NAFLD severity and may promote CAD through lipid metabolism alteration and/or promotion of the systemic inflammation.