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. J Clin Med Res. 2018;10(5):396-404 doi: https://doi.org/10.14740/jocmr3275e

Oxidative stress (OS) and inflammation have been related to colorectal cancer (CRC), but the influence of the Mediterranean diet (MD) on these parameters is unknown. Therefore, the aim of this study was to determine the association between adherence to the MD and markers of OS and DNA damage in CRC patients and to study the influence of adherence to the MD on metabolic and tumor-related factors. This prospective observational study included a total of 80 patients diagnosed with CRC. Adherence to the MD was estimated by the 14-item Mediterranean Diet Adherence Screener (MEDAS) questionnaire. The levels of OS markers (catalase, glutathione peroxidase, and glutathione system in serum; 8-oxo-7′8-dihydro-2′-deoxyguanosine and F2-isoprotanes in urine) and tumor and metabolic factors were determined. A total of 51.2% of our CRC patients showed a high adherence to the MD. These patients presented decreased levels of 8-oxodG, increased GPX and HDL–cholesterol levels, and a downward trend in the GSSG/GSH ratio with respect to patients with low adherence to the MD. In addition, a high adherence to the MD was associated with a lower histological grade of the tumor and a lower presence of synchronous adenomas. We conclude that a high adherence to the MD has a protective role against metabolic and oxidative DNA damage and improves antioxidant systems in CRC patients.

Ewing sarcoma (ES) is an aggressive bone and soft-tissue pediatric cancer. High vitronectin (VN) expression has been associated with poor prognosis in other cancers, and we aimed to determine the utility of this extracellular matrix glycoprotein as a biomarker of aggressiveness in ES. Silk fibroin plus gelatin–tyramine hydrogels (HGs) were fabricated with and without cross-linked VN and cultivated with A673 and PDX73 ES cell lines for two and three weeks. VN secretion to culture media was assessed using ELISA. Morphometric analysis was applied for phenotypic characterization. VN release to culture media was higher in 3D models than in monolayer cultures, and intracellular, intercellular, and pericluster presence was also observed. A673-HGs showed lower density of clusters but a proportion of larger clusters than PDX73-HGs, which presented low cluster circularity. The cluster density of A673-HGs without added VN was higher than with added VN and slightly lower in the case of PDX73-HGs. Furthermore, a culture time of three weeks provided no benefits in cluster growth compared to two weeks, especially in A673-HGs. These advances in 3D modeling and digital quantification pave the way for future studies in ES and other cancers to deepen understanding about intra- and intercellular heterogeneity and anti-adhesion VN therapies.