Premature Termination Codon in 5′ Region of Desmoplakin and Plakoglobin Genes May Escape Nonsense-Mediated Decay through the Reinitiation of Translation

Arrhythmogenic cardiomyopathy is a heritable heart disease associated with desmosomal mutations, especially premature termination codon (PTC) variants. It is known that PTC triggers the nonsense-mediated decay (NMD) mechanism. It is also accepted that PTC in the last exon escapes NMD; however, the mechanisms involving NMD escaping in 5′-PTC, such as reinitiation of translation, are less known. The main objective of the present study is to evaluate the likelihood that desmosomal genes carrying 5′-PTC will trigger reinitiation. HL1 cell lines were edited by CRISPR/Cas9 to generate isogenic clones carrying 5′-PTC for each of the five desmosomal genes. The genomic context of the ATG in-frame in the 5′ region of desmosomal genes was evaluated by in silico predictions. The expression levels of the edited genes were assessed by Western blot and real-time PCR. Our results indicate that the 5′-PTC in PKP2, DSG2 and DSC2 acts as a null allele with no expression, whereas in the DSP and JUP gene, N-truncated protein is expressed. In concordance with this, the genomic context of the 5′-region of DSP and JUP presents an ATG in-frame with an optimal context for the reinitiation of translation. Thus, 5′-PTC triggers NMD in the PKP2, DSG2* and DSC2 genes, whereas it may escape NMD through the reinitiation of the translation in DSP and JUP genes, with no major effects on ACM-related gene expression.     

Effectiveness of combined preservation methods to extend the shelf life of Morcilla de Burgos

Morcilla de Burgos is the most famous blood sausage in Spain. However, while producers are interested in extending its shelf life, the consumer is increasingly demanding more natural food. This situation has led to the current search for new and mild preservation technologies. Two batches of four different products: control without any treatment, control with organic acid salts (CnOAS; a 3% mixture of potassium/sodium l-lactate), control with high hydrostatic pressure processing (CnHPP; 600MPa-10min), and a combination of both treatments (OAS+HPP), were carried out to evaluate any synergistic effect that occurs when combining OAS and HPP, and the influence of different preservative treatments on the spoilage bacterial population and their evolution. HPP (with or without addition of OAS) can be considered the most suitable method for preserving morcilla de Burgos as it does not produce negative changes in sensory attributes. No clear selective effect of different treatments on the composition of the spoilage bacteria was seen and similar spoilage patterns were observed independently of the preservation treatment used.     

Embryonic fatty acid composition as a function of yolk fatty acid composition in eggs of the lesser spotted dogfish (Scyliorhinus canicula L.)

Yolk and embryonic total lipids were extracted from spotted dogfish eggs at two developmental stages. Total lipids were fractionated into neutral lipids (NL) and polar lipids (PL), and the fatty acid composition of each group was determined. Yolk lipid composition was found to be quantitatively different (NL/PL≊1) from embryo lipid composition (NL/PL≊0.5), for both stages of development. However, individual fatty acid composition did not differ from younger to older eggs for either yolk or embryo. There were significant differences (p<0.05) in major fatty acid groups from yolk and embryonic PL for saturated fatty acids, monounsaturated fatty acids (MUFA) and n−3 polyunsaturated fatty acids (PUFA) for younger eggs, and for MUFA and n−3 PUFA for older eggs. For NL, only MUFA composition from the oldest eggs showed differences between yolk and embryo. Results are discussed in terms of embryonic needs for highly unsaturated fatty acid (HUFA) biosynthesis, as well as to provide some explanations for the unusually high levels of 20∶4n−6 in both yolk and embryonic neutral lipids and polar lipids.     

Enhancement of Ca3Co4O9 thermoelectric properties by Cr for Co substitution

Ca₃Co_4−xCr_xO₉ polycristalline thermoelectric ceramics with small amounts of Cr have been synthesized by the classical solid state method. Microstructural characterizations have shown that all the Cr has been incorporated into the Ca₃Co₄O₉ structure and no Cr-containing secondary phases have been produced for Cr contents≤0.05. Apparent density measurements have shown that all samples are very similar, with densities around 75% of the theoretical one. Electrical resistivity decreases and Seebeck coefficient slightly raises when Cr content increases until 0.05 Cr addition. The improvement in both parameters leads to higher power factor values than the usually obtained by conventional solid state routes.     

Prediction and identification of the evolution of solar-heated hot water temperature in a stratified tank in static mode using artificial neural networks

The results of analyses of thermal stratification using different initial water temperature profiles are reported and discussed. Stratification proved to be the main factor in the cooling process, since although ambient temperature is also a determining factor, in this case it played a lesser role because the tank’s location inside the laboratory meant that it remained almost constant.     

Role of Serum Cholesterol and Statin Use in the Risk of Prostate Cancer Detection and Tumor Aggressiveness

The aim of this study was to analyze the relationship between statin use along with serum cholesterol levels and prostate cancer (PCa) detection and aggressiveness. Statin users of three years or more and serum cholesterol levels (SC) were assessed in 2408 men scheduled for prostate biopsy. SC was classified as normal (NSC: <200 mg/dL) or high (HSC: >200 mg/dL). High-grade PCa (HGPCa) was considered if the Gleason score was greater than 7. Statin users comprised 30.9% of those studied. The PCa detection rate was 31.2% of men on statins and 37% of non-statin users (p < 0.006). The PCa detection rate was 26.3% in men with NSC and 40.6% in those with HSC (p < 0.001). In the subset of NSC men, the PCa rate was 26.5% for statin users and 26.2% for non-users (p = 0.939), while in men with HSC, the PCa rate was 36.4% for statin users and 42.0% for non-statin users (p = 0.063). The HGPCa rate was 41.8% for statin users and 32.5% for non-users (p = 0.012). NSC men had a 53.8% rate of HGPCa, while the rate was only 27.6% in HSC men (p < 0.001). NSC men on statins had an HGPCa rate of 70.2%, while non-statin users had a rate of 41.2% (p < 0.001). The HGPCa rate for HSC men on statins was 18.8%, while the rate was 30.0% (p = 0.011) for non-users. Logistic regression analysis suggested that serum cholesterol levels could serve as an independent predictor of PCa risk, OR 1.87 (95% CI 1.56–2.24) and HGPCa risk, OR 0.31 (95% CI 0.23–0.44), while statin usage could not. Statin treatment may prevent PCa detection through serum cholesterol-mediated mechanisms. A disturbing increase in the HGPCa rate was observed in statin users who normalized their serum cholesterol.     

Characterization by culture-dependent and culture-independent methods of the bacterial population of suckling-lamb packaged in different atmospheres

This study offers insight into the dynamics of bacterial populations in fresh cuts of suckling lamb under four different atmospheric conditions: air (A), and three Modified Atmosphere Packaging (MAP) environments, 15%O₂/30%CO₂/55%N₂ (C, commercial), 70%O₂/30%CO₂ (O), and 15%O₂/85%CO₂ (H) for 18 days. Microbial analyses by both conventional methods and PCR-DGGE were performed. Controversial and surprising results emerged from comparing both methods in relation to the genus Pseudomonas. Thus, conventional methods detected the presence of high numbers of Pseudomonas colonies, although PCR-DGGE only detected this genus in air-packaged samples. PCR-DGGE detected higher microbial diversity in the control samples (A) than in the modified atmospheres (C, O, H), having atmosphere H the fewest number of species. Brochothrix thermosphacta, LAB (Carnobacterium divergens and Lactobacillus sakei), and Escherichia spp. were detected in all the atmospheres throughout storage. Moreover, previously undescribed bacteria from lamb meat such as Enterobacter hormaechei, Staphylococcus equorum and Jeotgalicoccus spp. were also isolated in this study by DGGE. Additionally, qPCR analysis was used to detect and characterize strains of Escherichia coli. Virulence genes (stx₁, stx₂ and eae) were detected throughout storage in 97% of the samples. A high CO₂ atmosphere was the most effective packaging combination doubling storage time in comparison with commercial atmosphere.     

Effect of raspberry pomace extracts isolated by high pressure extraction on the quality and shelf‐life of beef burgers

Raspberry pomace extracts isolated with supercritical carbon dioxide (SCE) and pressurised ethanol/water (ETE) were tested in beef burgers. Only ETE additives effectively inhibited lipid oxidation and the growth of microorganisms, as it was observed by measuring the changes of thiobarbituric acid reactive substances, bacterial counts and the content of O₂ and CO₂ during storage in the modified atmosphere package. ETE additives also demonstrated some meat colour preservation effects, which were assessed by the intensity of hamburger colour and metmyoglobin concentration. However, ETE additives did not alter burger's taste at the applied concentrations (up to 1%). It may be concluded that the most effective extracts possessing strong antioxidant and antimicrobial activity may be isolated from raspberry pomace by the pressurised liquid extraction with a hydroethanolic solvent; such extracts may be considered as promising additives in meat products for improving their stability and enriching with beneficial to health phytochemicals.     

Thick unevenness compensation in a hot rolling mill havingautomatic gage control

Hot rolling products with improved quality features such as thickness profile, strip width, flatness and material mechanical properties, are in increasing demand. The project goal is to improve the production quality in an existing hot rolling mill at Aceralia Steel Company, Aviles, Spain. This paper describes the design and implementation of a supervisory system for real-time compensation of uneven thickness on both sides of a rolled strip. The design is based on a multivariable process model, whose parameters are calculated online using measurement data from the mill. As a result, a computer system was developed that corrects the automatic gage control output from online acquired mill operation data. Previously, this task was done manually and periodically by a human operator. Manual correction is tedious and error prone, since it is based on visual inspection. Moreover, the proposed method leads to notorious improvement in the output strip quality, as the correcting signal may be continuously applied before the uneven thickness effect is visible