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Detecting MLH1 promoter methylation is highly relevant to differentiate between possible Lynch syndrome patients or patients with sporadic causes of MLH1/PMS2 deficiency in colorectal (CRC) and endometrial cancers. Here, we aimed to develop a test for assessing MLH1 promoter methylation based in next generation sequencing (NGS), and to evaluate the concordance of MLH1 methylation and BRAF -V600 mutation status in CRC. For that, we performed a series of experiments with DNA from tumor, saliva and commercial control samples and our in house developed amplicon-based NGS test. In patients' samples, MLH1 methylation above 10% was only observed in tumors with MLH1/PMS2 loss. We confirmed the reproducibility and accuracy of MLH1 promoter analysis performing a serial dilution experiment with completely methylated and unmethylated control DNAs and a comparison between two NGS platforms (Ion Proton and Illumina). In MLH1/PMS2 deficient tumors, the MLH1 methylation status was concordant with the BRAF mutation status in 90% (18/20) of the cases. Our amplicon-based NGS test showed a great sensitivity and specificity for detecting MLH1 methylation in CRC samples, with a high agreement with the evaluation of BRAF mutation. This simple and affordable test could be used as a reflex test to identify patients with sporadic causes of MLH1/PMS2 deficiency in CRC, aiding to genetic test referral and identification of Lynch syndrome patients. • Detecting MLH1 promoter methylation is highly relevant to identify Lynch syndrome patients. • Amplicon based NGS is a simple and low-cost method for detecting MLH1 methylation. • The test showed high agreement with BRAF mutation status in colorectal cancer. [ABSTRACT FROM AUTHOR]

Sardine cooking effluents contain a high level of organic matter, such as proteins and lipids, which allows them to be forward into a chain exploiting high added-value compounds attained from these effluents, increasing their economic value while reducing their environmental effect. Thus, the purpose of this work was to develop an innovative pilot-scale integrated membrane process, with or without enzymatic hydrolysis, to obtain fractions with high protein/peptide and low NaCl contents, as well as optimized bioactive properties. The research strategy followed involved the use of ultrafiltration (UF) and nanofiltration (NF) technologies of the pretreated sardine cooking effluent followed by reverse osmosis (RO) at a pilot scale levels. Moreover, it allowed for the attainment of fractions rich in protein/peptides that might be used in the food, pharmaceutical, or cosmetic industries, particularly after RO, as they present a lower NaCl content. The RO retentate (hydrolyzed sample) coupled with UF and NF resulted in the fractions with the best bioactive properties (higher antioxidant capacity and antimicrobial activity) of all the analyzed samples. Overall, the current work demonstrated the feasibility of exploiting liquid by-products as a source of functional components as well as reinforcing this strategy's potential relevance in future effective management strategies for this type of effluents. [Display omitted] • Protein/peptide recovery from fish effluents using membrane technology. • Bioactive properties of proteins/peptides recovered from fishery effluents. • Efficiently valorizing fishery effluents. • Removing the mono-valent salts from sardine cooking effluents by NF membrane. [ABSTRACT FROM AUTHOR]