Detection of mutations in circulating cell‐free DNA in relation to disease stage in colorectal cancer

Enthusiasm has emerged for the potential of liquid biopsies to provide easily accessible genetic biomarkers for early diagnosis and mutational cancer characterization. We here systematically investigated the suitability of circulating cell-free DNA (cfDNA) analysis for mutation detection in colorectal cancer (CRC) patients with respect to clinicopathological disease stage. Droplet Digital PCR (ddPCR) was performed to detect common point mutations in the KRAS and BRAF oncogenes in cfDNA from 65 patients and compared to mutations in tumor tissue. Stage of disease was classified according to UICC (Union for International Cancer Control) criteria. In tumor tissue, KRAS or BRAF mutations were present in 35 of 65 cases (44% UICC stage I, 50% stage II, 47% stage III, and 62% stage IV). Although cfDNA was detected in 100% of patients, ddPCR displayed the tumor tissue mutation in only 1 of 6 (17%) stage II patients, whereas 10 of 18 (56%) reported variants were verified in cfDNA samples of the stage IV cohort. No BRAF or KRAS mutation was detected in cfDNA from patients with wild-type tumor tissue. In one case of mutant stage II colon cancer (KRAS-G12C), the G12D variant was detected in cfDNA instead. Further workup revealed that circulating tumor-derived DNA and liver metastases originated from a synchronous KRAS-mutated cancer of the pancreas. Our results demonstrate that ddPCR-based analysis is highly specific and useful for mutation monitoring, but the sensitivity limits its usefulness for early cancer detection

Improvement of the commercial quality of chilled Norway lobster (Nephrops norvegicus) stored in slurry ice: Effects of a preliminary treatment with an antimelanosic agent on enzymatic browning

8 páginas, 7 figuras, 2 tablasThe use of slurry ice is gaining increasing importance as an advanced method for the hygienic and efficient chilling and sub-zero storage of aquatic food products. In this work, this technology was applied as a novel technique for the chilling and storage of Norway lobster (Nephrops norvegicus) – a crustacean species of high-commercial value – under refrigeration conditions at 1.5 C. In addition, the effects of a preliminary treatment with 0.5% Na HSO3 on surface browning were evaluated and compared with the results obtained in control batches not subjected to such treatment. The processing of lobster in slurry ice significantly (p < 0.05) slowed down microbial spoilage, as determined by the counts of aerobes, psychrotrophs, proteolytic bacteria, and lactose-fermenting Enterobacteriaceae, and by the formation of volatile amines. Likewise, the autolytic breakdown mechanisms – as determined by the K value – were also significantly (p < 0.05) inhibited in the slurry ice batch. Remarkably, preliminary treatment with 0.5% sodium metabisulphite permitted better maintenance of the parameters involved in sensory quality – especially as regards the aspect of the carapace – as compared with nontreated batches, and allowed a shelf life of 9 days without surpassing the 150 mg/kg legal limit established for this food additive. On contrast, the non-treated batch stored in slurry ice exhibited a shelf life of 5 days. The combination of technological treatments proposed in this work – preliminary antimelanosic treatment and storage in slurry ice – may be successfully applied to other fresh and frozen shellfish species with a view to extending shelf life and to avoiding the legal and toxicological problems derived from current abuse of such antimelanosic agents to prevent shellfish browning.This work was supported by the Dirección Xeral de I + D + I from the Xunta de Galicia (Galicia, Spain) (Project PGIDTI02RMA18E).Peer reviewe

Interleukin-6 Gene Expression Changes after a 4-Week Intake of a Multispecies Probiotic in Major Depressive Disorder—Preliminary Results of the PROVIT Study

Major depressive disorder (MDD) is a prevalent disease, in which one third of sufferers do not respond to antidepressants. Probiotics have the potential to be well-tolerated and cost-efficient treatment options. However, the molecular pathways of their effects are not fully elucidated yet. Based on previous literature, we assume that probiotics can positively influence inflammatory mechanisms. We aimed at analyzing the effects of probiotics on gene expression of inflammation genes as part of the randomized, placebo-controlled, multispecies probiotics PROVIT study in Graz, Austria. Fasting blood of 61 inpatients with MDD was collected before and after four weeks of probiotic intake or placebo. We analyzed the effects on gene expression of tumor necrosis factor (TNF), nuclear factor kappa B subunit 1 (NFKB1) and interleukin-6 (IL-6). In IL-6 we found no significant main effects for group (F(1,44) = 1.33, p = ns) nor time (F(1,44) = 0.00, p = ns), but interaction was significant (F(1,44) = 5.67, p &lt; 0.05). The intervention group showed decreasing IL-6 gene expression levels while the placebo group showed increasing gene expression levels of IL-6. Probiotics could be a useful additional treatment in MDD, due to their anti-inflammatory effects. Results of the current study are promising, but further studies are required to investigate the beneficial effects of probiotic interventions in depressed individuals