RAS assessment is mandatory for therapy decision in metastatic colorectal cancer (mCRC) patients. This determination is based on tumor tissue, however, genotyping of circulating tumor (ct)DNA offers ...clear advantages as a minimally invasive method that represents tumor heterogeneity. Our study aims to evaluate the use of ctDNA as an alternative for determining baseline RAS status and subsequent monitoring of RAS mutations during therapy as a component of routine clinical practice.
RAS mutational status in plasma was evaluated in mCRC patients by OncoBEAM™ RAS CRC assay. Concordance of results in plasma and tissue was retrospectively evaluated.RAS mutations were also prospectively monitored in longitudinal plasma samples from selected patients.
Analysis of RAS in tissue and plasma samples from 115 mCRC patients showed a 93% overall agreement. Plasma/tissue RAS discrepancies were mainly explained by spatial and temporal tumor heterogeneity. Analysis of clinico-pathological features showed that the site of metastasis (i.e. peritoneal, lung), the histology of the tumor (i.e. mucinous) and administration of treatment previous to blood collection negatively impacted the detection of RAS in ctDNA. In patients with baseline mutant RAS tumors treated with chemotherapy/antiangiogenic, longitudinal analysis of RAS ctDNA mirrored response to treatment, being an early predictor of response. In patients RAS wt, longitudinal monitoring of RAS ctDNA revealed that OncoBEAM was useful to detect emergence of RAS mutations during anti-EGFR treatment.
The high overall agreement in RAS mutational assessment between plasma and tissue supports blood-based testing with OncoBEAM™ as a viable alternative for genotyping RAS of mCRC patients in routine clinical practice. Our study describes practical clinico-pathological specifications to optimize RAS ctDNA determination. Moreover, OncoBEAM™ is useful to monitor RAS in patients undergoing systemic therapy to detect resistance and evaluate the efficacy of particular treatments.
Aims/hypothesis Inflammation is associated with obesity and has been implicated in the development of diabetes and atherosclerosis. During gram-negative bacterial infection, lipopolysaccharide causes ...an inflammatory reaction via toll-like receptor 4 (TLR4), which has an essential function in the induction of innate and adaptative immunity. Our aim was to determine what role TLR4 plays in the development of metabolic phenotypes during high-fat feeding. Materials and methods We evaluated metabolic consequences of a high-fat diet in TLR4 mutant mice (C3H/HeJ) and their respective controls. Results TLR4 inactivation reduced food intake without significant modification of body weight, but with higher epididymal adipose tissue mass and adipocyte hypertrophy. It also attenuated the inflammatory response and increased glucose transport and the expression levels of adiponectin and lipogenic markers in white adipose tissue. In addition, TLR4 inactivation blunted insulin resistance induced by lipopolysaccharide in differentiated adipocytes. Increased feeding efficiency in TLR4 mutant mice was associated with lower mass and lower expression of uncoupling protein 1 gene in brown adipose tissue. Finally, TLR4 inactivation slowed the development of hepatic steatosis, reducing the liver triacylglycerol content and also expression levels of lipogenic and fibrosis markers. Conclusions/interpretation TLR4 influences white adipose tissue inflammation and insulin sensitivity, as well as liver fat storage, and is important in the regulation of metabolic phenotype during a fat-enriched diet.
Phase separation of mixtures of oppositely charged polymers provides a simple and direct route to compartmentalisation via complex coacervation, which may have been important for driving primitive ...reactions as part of the RNA world hypothesis. However, to date, RNA catalysis has not been reconciled with coacervation. Here we demonstrate that RNA catalysis is viable within coacervate microdroplets and further show that these membrane-free droplets can selectively retain longer length RNAs while permitting transfer of lower molecular weight oligonucleotides.
Self-assessment and peer assignment have clear advantages for the training of responsible, critical, and reflective professionals. In recent years, self and peer evaluation have also been shown to be ...even more effective than lecturer evaluation when we assure anonymity through online platforms learning tools. Therefore, self and peer assessments are to become a core aspect of student-centred evaluation processes in Higher Education. Besides, a high concordance with lecturer evaluation may allow lecturers to also benefit from self and peer evaluation without an increase in their workload. In the present work, we compare the formative evaluation from the lecturer with the self and peer assessments through a virtual learning environment. The subject of study if formed by assessments prepared by students in a first-year course in a Social Sciences degree at the Universidade de Vigo, Spain. We find a strong concordance between peer assessment and lecturer assignment, and a moderate agreement between self-assessment and lecturer assignment. These results show that students perform well as peer evaluators, with peer assignment being a procedure with high validity and reliability.
Method-dependent Etest epidemiological cutoff values (ECVs) are not available for susceptibility testing of either Candida or Aspergillus species with amphotericin B or echinocandins. In addition, ...reference caspofungin MICs for Candida spp. are unreliable. Candida and Aspergillus species wild-type (WT) Etest MIC distributions (microorganisms in a species-drug combination with no detectable phenotypic resistance) were established for 4,341 Candida albicans, 113 C. dubliniensis, 1,683 C. glabrata species complex (SC), 709 C. krusei, 767 C. parapsilosis SC, 796 C. tropicalis, 1,637 Aspergillus fumigatus SC, 238 A. flavus SC, 321 A. niger SC, and 247 A. terreus SC isolates. Etest MICs from 15 laboratories (in Argentina, Europe, Mexico, South Africa, and the United States) were pooled to establish Etest ECVs. Anidulafungin, caspofungin, micafungin, and amphotericin B ECVs (in micrograms per milliliter) encompassing ≥97.5% of the statistically modeled population were 0.016, 0.5, 0.03, and 1 for C. albicans; 0.03, 1, 0.03, and 2 for C. glabrata SC; 0.06, 1, 0.25, and 4 for C. krusei; 8, 4, 2, and 2 for C. parapsilosis SC; and 0.03, 1, 0.12, and 2 for C. tropicalis The amphotericin B ECV was 0.25 μg/ml for C. dubliniensis and 2, 8, 2, and 16 μg/ml for the complexes of A. fumigatus, A. flavus, A. niger, and A. terreus, respectively. While anidulafungin Etest ECVs classified 92% of the Candida fks mutants evaluated as non-WT, the performance was lower for caspofungin (75%) and micafungin (84%) cutoffs. Finally, although anidulafungin (as an echinocandin surrogate susceptibility marker) and amphotericin B ECVs should identify Candida and Aspergillus isolates with reduced susceptibility to these agents using the Etest, these ECVs will not categorize a fungal isolate as susceptible or resistant, as breakpoints do.
Recent findings suggest that use of colistin as a last resort antibiotic is seriously threatened by the rise of a new plasmid mediated mechanism of resistance (MCR-1). This work identifies, for the ...first time in Southern Europe, the gene mcr-1 in nine strains from farm animals (poultry and swine) corresponding to five Escherichia coli and four Salmonella enterica, among which three belong to serovar Typhimurium and one to Rissen. The MCR-1 was found encoded by a plasmid highly mobilizable by conjugation to the E. coli J53 strain. Two E. coli strains carried two determinants, mcr-1 plus pmrA or pmrB mutations, known to confer colistin resistance.
•MCR-1 is detected in E. coli and Salmonella from poultry and swine, in Spain.•The plasmid carrying mcr-1 has approximately 60kb.•Colistin resistance is efficiently mobilized by conjugation from MCR-1 positive strains.
Research has suggested that university students are at risk from certain unhealthy habits, such as poor diet or alcohol abuse. At the same time, anxiety levels appear to be higher among university ...students, which may lead to high levels of emotional eating. The aim of this study was to analyze the degree of adherence to the Mediterranean diet (AMD), emotional eating, alcohol intake, and anxiety among Spanish university students, and the interrelationship of these variables. A total of 252 university students filled out the Mediterranean Diet Quality Index (KIDMED) questionnaire for Mediterranean diet adherence, an Alcohol Use Disorders Identification Test, a State-Trait Anxiety Inventory and the Emotional Eater Questionnaire. We analyzed descriptive data, a
-test and analysis of variance (ANOVA) for differences, a Pearson correlation, and multiple regression tests. Results showed low levels of AMD among university students (15.5%) and considerable levels of emotional eating (29%) and anxiety (23.6%). However, levels of alcohol dependence were low (2.4%). State-anxiety was a predictor of the emotional eater score and its subscales, and sex also was predictive of subscale guilt and the total score. However, AMD was predicted only by trait-anxiety. These models accounted for between 1.9% and 19%. The results suggest the need for the implementation of educational programs to promote healthy habits among university students at risk.
Short-term oil exposure impacts on the harmful algal bloom (HAB) species Pseudonitzschia australis and the coccolithophore Emiliania huxleyi were studied by investigating physiological responses in ...growth, production of transparent exopolymer particles (TEP), P. australis domoic acid (DA) content, and E. huxleyi calcification, with the goal of better understanding the potential effects of the 2015 oil spill near Refugio State Beach in the Santa Barbara Channel, California (USA). While oil exposure negatively impacted growth of both species, P. australis appeared to be more sensitive to oil exposure compared to E. huxleyi. Increased cellular TEP (both species) and DA (P. australis), and abnormal calcification (E. huxleyi) were observed in the presence of oil. The physiological changes detected in these locally important phytoplankton species have implications for other ecosystem processes, human health, and the fate of the spilled oil. The results from this study enhance our understanding of the repercussions of oil pollution events, but further research is necessary to explore ecosystem-wide impacts and chronic or long-term effects of oil in marine environments.
Adenosine triphosphate (ATP) production and utilization is critically important for animal development. How these processes are regulated in space and time during tissue growth remains largely ...unclear. We used a FRET‐based sensor to dynamically monitor ATP levels across a growing tissue, using the Drosophila larval wing disc. Although steady‐state levels of ATP are spatially uniform across the wing pouch, inhibiting oxidative phosphorylation reveals spatial differences in metabolic behavior, whereby signaling centers at compartment boundaries produce more ATP from glycolysis than the rest of the tissue. Genetic perturbations indicate that the conserved Hedgehog signaling pathway can enhance ATP production by glycolysis. Collectively, our work suggests the existence of a homeostatic feedback loop between Hh signaling and glycolysis, advancing our understanding of the connection between conserved developmental patterning genes and ATP production during animal tissue development.
Synopsis
Morphogen signaling in the developing Drosophila wing can promote adenosine triphosphate (ATP) production from glycolysis upon loss of oxidative phosphorylation (OxPhos).
An ATP FRET sensor reveals regional differences in energy metabolism across the wing pouch upon treatment with inhibitors of OxPhos and glycolysis.
Regions of high morphogen signaling near compartment boundaries produce more ATP from glycolysis than the rest of the wing pouch, resulting in slower decline of ATP upon OxPhos inhibition.
Genetic perturbations indicate that Hh signaling promotes glycolytic ATP production under energy stress.
Enhancement of glycolytic ATP production by Hh signaling could preserve important morphogen signaling centers during brief periods of physiologically relevant energy stress, including hypoxia.
Morphogen signaling in the developing Drosophila wing can promote adenosine triphosphate production from glycolysis upon loss of oxidative phosphorylation (OxPhos).
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