Technical Validation and Clinical Implications of Ultrasensitive PCR Approaches for EGFR-Thr790Met Mutation Detection in Pretreatment FFPE Samples and in Liquid Biopsies from Non-Small Cell Lung Cancer Patients

In pretreatment tumor samples of EGFR-mutated non-small cell lung cancer (NSCLC) patients, EGFR-Thr790Met mutation has been detected in a variable prevalence by different ultrasensitive assays with controversial prognostic value. Furthermore, its detection in liquid biopsy (LB) samples remains challenging, being hampered by the shortage of circulating tumor DNA (ctDNA). Here, we describe the technical validation and clinical implications of a real-time PCR with peptide nucleic acid (PNA-Clamp) and digital droplet PCR (ddPCR) for EGFR-Thr790Met detection in diagnosis FFPE samples and in LB. Limit of blank (LOB) and limit of detection (LOD) were established by analyzing negative and low variant allele frequency (VAF) FFPE and LB specimens. In a cohort of 78 FFPE samples, both techniques showed an overall agreement (OA) of 94.20%. EGFR-Thr790Met was detected in 26.47% of cases and was associated with better progression-free survival (PFS) (16.83 ± 7.76 vs. 11.47 ± 1.83 months; p = 0.047). In LB, ddPCR was implemented in routine diagnostics under UNE-EN ISO 15189:2013 accreditation, increasing the detection rate of 32.43% by conventional methods up to 45.95%. During follow-up, ddPCR detected EGFR-Thr790Met up to 7 months before radiological progression. Extensively validated ultrasensitive assays might decipher the utility of pretreatment EGFR-Thr790Met and improve its detection rate in LB studies, even anticipating radiological progression.     

From latex specifications to parallel codes

The advent of multicore systems, joined to the potential acceleration of the graphics processing units, has given us a low cost computation capability unprecedented. The new systems alleviate some well-known important architectural problems at the expense of a considerable increment of the programmability wall. Heterogeneity, at both the architectural and programming levels, poses a great challenge to programmers. As a contribution, we propose a development methodology for the automatic source-to-source transformation on specific domains. This methodology is successfully instantiated as a framework to solve Dynamic Programming problems. As a result of applying our framework, the end user (a physicist, a mathematician or a biologist) can express her problem through a latex equation and automatically derive efficient parallel codes for current homogeneous or heterogeneous architectures. The approach allows an easy portability to new emergent architectures.     

Observation and formation mechanism of individual graphene foils inside multi-walled carbon nanotubes

The formation of individual graphene foils (GF) has been evidenced in the present study inside multi-walled carbon nanotubes. The graphene foils are attached on one side to a catalyst nanoparticle and on the other side to the internal walls of the nanotube. Moreover, results suggest that a necking process occurs in which internal carbon walls are deformed until formation of the graphene foil. A possible mechanism for the GF formation is then proposed.     

Solution‐Processed GaSe Nanoflake‐Based Films for Photoelectrochemical Water Splitting and Photoelectrochemical‐Type Photodetectors

Gallium selenide (GaSe) is a layered compound, which has been exploited in nonlinear optical applications and photodetectors due to its anisotropic structure and pseudodirect optical gap. Theoretical studies predict that its 2D form is a potential photocatalyst for water splitting reactions. Herein, the photoelectrochemical (PEC) characterization of GaSe nanoflakes (single‐/few‐layer flakes), produced via liquid phase exfoliation, for hydrogen evolution reaction (HER) and oxygen evolution reaction (OER) in both acidic and alkaline media is reported. In 0.5 m H₂SO₄, the GaSe photoelectrodes display the best PEC performance, corresponding to a ratiometric power‐saved metric for HER (Φ_saved,HER) of 0.09% and a ratiometric power‐saved metric for OER (Φ_saved,OER) of 0.25%. When used as PEC‐type photodetectors, GaSe photoelectrodes show a responsivity of ≈0.16 A W^?1 upon 455 nm illumination at a light intensity of 63.5 µW cm^?2 and applied potential of ?0.3 V versus reversible hydrogen electrode (RHE). Stability tests of GaSe photodetectors demonstrated a durable operation over tens of cathodic linear sweep voltammetry scans in 0.5 m H₂SO₄ for HER. In contrast, degradation of photoelectrodes occurred in both alkaline and anodic operation due to the highly oxidizing environment and O₂‐induced (photo)oxidation effects. The results provide new insight into the PEC properties of GaSe nanoflakes for their exploitation in photoelectrocatalysis, PEC‐type photodetectors, and (bio)sensors.     

Experimentation with a water tank including a PCM module

Storage of heat is seen as a major issue for the development of solar energy for house heating and cooling under all climates. Most of the storage systems available on the market use water as the storage medium. The idea studied here was to add a phase change material (PCM) module at the top of a hot-water storage tank with stratification. An experimental solar pilot plant was constructed to test the PCM behaviour in real conditions. The PCM module geometry adopted was to use several cylinders. A granular PCM–graphite compound was chosen as the PCM for the experiments presented here.     

β-hydroxyacyl-coa-dehydrogenase (HADH) activity of unfrozen and frozen—thawed frog (Rana esculenta) legs

The β-hydroxyacyl-CoA-dehydrogenase (HADH) activity of unfrozen and thawed frog legs was investigated. The enzyme was extracted by either immersing frog legs in phosphate buffer 0.1 M, pH 6.0 at 25°C for 15 min or pressing them between trichinoscopy glasses. The enzyme activity was assayed using acetoacetyl-CoA as substrate and measured spectrophotometrically at 340 nm. It was possible by both extraction methods to distinguish between thawed and unfrozen samples although when the juice was obtained by pressing the HADH activity of the dilution was ～ 1.5 times higher than that obtained by immersion. The HADH activity was significantly higher (P≤0·001) in frozen-thawed frogs than in unfrozen legs because during freezing there is a release of HADH. No significative differences were found in the HADH activity in samples frozen in the temperature range -10 to -196°C. HADH activity was not affected by the storage time in crushed ice up to 6 days.     

A comparison of methods for linear prediction of apple flavour from gas chromatographic measurements

A comparative study of linear methods for prediction of sensory profiles from gas chromatography (GC) measurements was performed. The data used came from an experiment on the effect of storing apples at various oxygen concentrations. Partial least-squares regression and continuum regression showed the best performance, measured by a two-step cross-validation principle. The traditional prediction error sum of squares (PRESS) overestimated the predictive ability of a multiple linear regression approach. The quality of the predictions of sensory properties from GC analyses was measured in terms of a ‘panel size equivalent’. Thus, the predictions obtained in the present study were as accurate as predictions from an assessor panel consisting of 4–9 assessors, depending on the sensory property in question.     

Effect of fat-enriched diets on rabbit meat fatty acid composition

The effects of beef tallow-, soya and sunflower olein- and soya bean oil-enriched (3%) diets on the chemical and fatty acid composition of total, a polar and polar lipids of rabbit meat have been studied. No significant differences (P > 0.05) in the meat chemical composition were observed between groups. However, a large effect of the diet on the total and apolar lipid fatty acids was found, with C18:1 and C18:2 the most affected fatty acids. The influence of the diets on the fatty acids of the polar fraction was less marked. It is concluded that the enrichment of the rabbit diet with either soya and sunflower oleins or soya bean oil allows the production of rabbit meat with a higher unsaturation degree than is obtained by using conventional diets, which constitutes an important nutritional benefit to the human being.     

Solid-Phase Microextraction (SPME) of Pyrazines in Model Reaction Systems

Headspace solid-phase microextraction (HS-SPME) combined with gas chromatography (GC) was used for isolating and analysing pyrazines formed in a model system. This system consisted of an aqueous mixture of glucose, glycine and sodium hydroxide. Pyrazine formation was accelerated by heating in a microwave oven. Results were compared with our earlier ones using conventional techniques for isolation of pyrazines. HS-SPME-GC showed several advantages over traditional methods. The method is rapid, sensitive, easy to use and reproducible. Headspace and liquid of the model reaction were extracted and the results obtained using headspace sampling showed good agreement with the methylene chloride extraction method. In all cases, coefficients of variation of 5% or lower were obtained when the SPME extraction time was consistent.     

Keeping quality of vacuum-packed smoked sardine fillets: microbiological aspects

The effect of mixed smoking (2 h at 30 degrees C and 45 min at 75 degrees C was studied on the keeping quality of lean and fatty sardine fillets from fish caught in the Mediterranean Sea in March and June, respectively. Raw material for smoking included both fresh and frozen sardine fillets. Microbiological and sensory analyses were performed over the storage period, which lasted for 120 days at 0 +/- 1 degrees C. Smoking reduced the viable microbial flora values from 3-5 x 10(4) cfu/g to 2 x 10(2) cfu/g. The antimicrobial action of smoking, combined with that of vacuum packaging, had the effect of producing a gram-positive flora made up mainly of lactic acid bacteria. A practical consequence of this was to extend the shelf life of the smoked sardines for a period of 4 months. Frozen storage of samples at -18 degrees C for 6 months prior to smoking affected the behaviour of the microbial flora during cold storage at 0 degrees C, but did not affect the behaviour of the flora over a period of 3 months frozen storage, since the initial numbers of bacteria decreased and microbial growth was retarded. Furthermore, at the end of the storage period, the number of bacteria in the samples of frozen fatty sardines were higher than in the samples of frozen lean sardine. Physical conditions, therefore appear to exert an influence on bacterial growth. Sulphite-reducing Clostridia were not detected during the storage period.