High eEF1A1 Protein Levels Mark Aggressive Prostate Cancers and the In Vitro Targeting of eEF1A1 Reveals the eEF1A1–actin Complex as a New Potential Target for Therapy

Although the eukaryotic elongation factor eEF1A1 plays a role in various tumours, there is little information on its prognosis/therapeutic value in prostate carcinoma. In high-grade and castration-resistant prostate carcinoma (CRPC), the identification of novel therapeutic markers/targets remains a priority. The expression of eEF1A1 protein was determined in formalin-fixed, paraffin-embedded prostate cancer and hyperplasia tissue by IHC. The role of eEF1A1 was investigated in a cellular model using a DNA aptamer (GT75) we previously developed. We used the aggressive CRPC cancer PC-3 and non-tumourigenic PZHPV-7 lines. Cytotoxicity was measured by the MTS assay and eEF1A1 protein levels by in-cell Western assays. The mRNA levels of eEF1A1 were measured by qPCR and ddPCR. Higher expression of eEF1A1 was found in Gleason 7–8 compared with 4–6 tissues (Gleason ≥ 7, 87% versus Gleason ≤ 6, 54%; p = 0.033). Patients with a high expression of eEF1A1 had a worse clinical outcome. In PC-3, but not in PZHPV-7, GT75 decreased cell viability and increased autophagy and cell detachment. In PC-3 cells, but not in PZHPV-7, GT75 mainly co-localised with the fraction of eEF1A1 bound to actin. Overexpression of the eEF1A1 protein can identify aggressive forms of prostate cancer. The targeting of eEF1A1 by GT75 impaired cell viability in PC-3 cancer cells but not in PZHPV-7 non-tumourigenic cells, indicating a specific role for the protein in cancer survival. The eEF1A1–actin complexes appear to be critical for the viability of PC-3 cancer cells, suggesting that eEF1A1 may be an attractive target for therapeutic strategies in advanced forms of prostate cancer.     

Root Exposure to 5-Aminolevulinic Acid (ALA) Affects Leaf Element Accumulation,Isoprene Emission,Phytohormonal Balance,and Photosynthesis of Salt-Stressed Arundo donax

Arundo donax has been recognized as a promising crop for biomass production on marginal lands due to its superior productivity and stress tolerance. However, salt stress negatively impacts A. donax growth and photosynthesis. In this study, we tested whether the tolerance of A. donax to salinity stress can be enhanced by the addition of 5-aminolevulinic acid (ALA), a known promoter of plant growth and abiotic stress tolerance. Our results indicated that root exposure to ALA increased the ALA levels in leaves along the A. donax plant profile. ALA enhanced Na⁺ accumulation in the roots of salt-stressed plants and, at the same time, lowered Na⁺ concentration in leaves, while a reduced callose amount was found in the root tissue. ALA also improved the photosynthetic performance of salt-stressed apical leaves by stimulating stomatal opening and preventing an increase in the ratio between abscisic acid (ABA) and indol-3-acetic acid (IAA), without affecting leaf methanol emission and plant growth. Supply of ALA to the roots reduced isoprene fluxes from leaves of non-stressed plants, while it sustained isoprene fluxes along the profile of salt-stressed A. donax. Thus, ALA likely interacted with the methylerythritol 4-phosphate (MEP) pathway and modulate the synthesis of either ABA or isoprene under stressful conditions. Overall, our study highlights the effectiveness of ALA supply through soil fertirrigation in preserving the young apical developing leaves from the detrimental effects of salt stress, thus helping of A. donax to cope with salinity and favoring the recovery of the whole plant once the stress is removed.     

Enzymes Action on Wheat–Soy Dough Properties and Bread Quality

High levels of soy flour added to wheat bread produce negative effects on gluten network formation, dough properties, and on bread final quality. The objective of this study was to assess the influence of three enzymes, transglutaminase (TG), glucose oxidase (GOX), and endoxylanase (XYL), on dough properties and final quality of high-protein breads. The addition of TG and GOX increased the mixing stability and maximum resistance of dough, decreased its extensibility, and produced stronger and more consistent dough samples. XYL incorporation produced opposite results. XYL addition and the lowest GOX dose increased bread volume significantly and decreased initial crumb firmness, while high doses of TG (0.3%) produced detrimental effects on bread volume and crumb firmness. In conclusion, XYL and GOX 0.001% addition improved the final quality of soy-fortified breads, but XYL was the best additive to improve dough properties, bread volume, and quality.     

Quantitative detection method for Roundup Ready soybean in food using duplex real‐time PCR MGB chemistry

BACKGROUND: Methodologies that enable the detection of genetically modified organisms (GMOs) (authorized and non‐authorized) in food and feed strongly influence the potential for adequate updating and implementation of legislation together with labeling requirements. Quantitative polymerase chain reaction (qPCR) systems were designed to boost the sensitivity and specificity on the identification of GMOs in highly degraded DNA samples; however, such testing will become economically difficult to cope with due to increasing numbers of approved genetically modified (GM) lines. Multiplexing approaches are therefore in development to provide cost‐efficient solution. RESULTS: Construct‐specific primers and probe were developed for quantitative analysis of Roundup Ready ^® soybean (RRS) event glyphosate‐tolerant soybean (GTS) 40‐3‐2. The lectin gene (Le1) was used as a reference gene, and its specificity was verified. RRS‐ and Le1‐specific quantitative real‐time PCR (qRTPCR) were optimized in a duplex platform that has been validated with respect to limit of detection (LOD) and limit of quantification (LOQ), as well as accuracy. The analysis of model processed food samples showed that the degradation of DNA has no adverse or little effects on the performance of quantification assay. CONCLUSION: In this study, a duplex qRTPCR using TaqMan minor groove binder‐non‐fluorescent quencher (MGB‐NFQ) chemistry was developed for specific detection and quantification of RRS event GTS 40‐3‐2 that can be used for practical monitoring in processed food products. Copyright © 2010 Society of Chemical Industry     

Germin-like protein Cit s 1 and profilin Cit s 2 are major allergens in orange (Citrus sinensis) fruits

Oranges are clinically relevant allergenic foods. To date, orange allergens have not been characterized in detail. The study is aimed at analyzing the sensitization profile in orange-sensitized subjects with and without clinical allergy, and to identify orange allergens. Fifty-six sensitized subjects with self-reported reactions to orange were grouped into reactors (anaphylaxis or multiple episodes of immediate reactions and/or positive challenge tests) and non-reactors (negative open food challenge tests). Allergens were characterized by IgE immunoblotting, N-terminal sequencing, IgE-inhibition assays, and mediator release assays were performed to determine the allergenic potency of orange profilin. Of 56 subjects, 23 were classified as orange allergic showing mainly an oral allergy syndrome. Of 23 subjects classified as orange allergic, 22 were sensitized to profilin, Cit s 2. In patients with mono-sensitization to profilin in vitro histamine releases up to 75% from basophils were induced using orange extract and purified plant profilins. Of the allergic patients 78% were sensitized to germin-like protein, Cit s 1. Both allergens showed retained IgE reactivity in heat-processed orange juice. Interestingly, subjects with and without clinical allergy showed a comparable sensitization profile. Profilin and germin-like proteins are major orange allergens. The potential clinical relevance of orange profilin was indicated by its strong capacity to release histamine from basophils. However, a predominant sensitization to both allergens in subjects without symptoms also indicates a high frequency of clinically insignificant sensitization.     

Multifunctional TiO2 coatings for Cultural Heritage

Environmental pollution arising from industrial implants and urban factors is constantly increasing, causing aesthetical and durability concerns to urban structures exposed to the atmosphere.     

Pig sire type and sex effects on carcass traits,meat quality and physicochemical and sensory characteristics of Serrano dry‐cured ham

This study demonstrates that improvements in animal line selection by breeding enterprises exert a strong effect on carcass traits, meat quality and sensory characteristics of Serrano dry‐cured ham. A total of 461 pigs from the offspring of a Duroc (DU) × Landrace (LD) sow mated with two DU boars and a DU × Large White (LW) boar from three breeding enterprises were evaluated. The two DU terminal sires were significantly different (P < 0.05) in carcass conformation, backfat thickness, ham and loin yields, raw ham traits, myoglobin concentration and total pigments formed during the curing process; in addition, the two lines provided different percentages of hams (54 vs 91%) with sufficient subcutaneous fat and weight to manufacture dry‐cured Serrano hams using a slow ripening process (11 months). The DU × LW sire had the best carcass and ham traits from an economic standpoint and obtained highest scores for sensory characteristics of Serrano ham evaluated by a trained panel test; furthermore, this line provided 84% of total hams suitable for manufacturing Serrano hams by a slow process. When the sex effect was analysed, carcass and ham traits of females were more favourable, but females presented a higher incidence of pale, soft and exudative (PSE) meat and a lower percentage of hams with sufficient subcutaneous fat and weight to produce Serrano hams using a slow ripening process (61% for females and 91% for castrates). On the other hand, castrates provided Serrano hams cured by a slow procedure with better organoleptic characteristics than females. Right and left hams were similar. Copyright © 2005 Society of Chemical Industry     

Effect of ultraviolet processing on selected properties of egg white

The effect of ultraviolet processing (10.6 and 63.7 kJ m(-2)) on selected properties of egg white (absorbance, particle size, protein fractions, free sulfhydryl content, immunoreactivity, viscosity, gelling and foaming properties) was investigated. Ultraviolet exposure induced the development of browning, the formation of large protein aggregates by disulfide exchange, and protein backbone cleavage. However, egg white proteins were differently sensitive to UV radiation. No changes in immunoreactivity, gelling temperature and gel firmness were observed. Independently on the UV dose, light treated egg white produced foams with higher stability. This effect was attributed to protein aggregates jamming in the fluid interstices between bubbles and/or to the higher viscosity of the aqueous phase. The latter was also associated to higher foam volume.     

Coordination Environment of Cu(II) Ions Bound to N-Terminal Peptide Fragments of Angiogenin Protein

Angiogenin (Ang) is a potent angiogenic factor, strongly overexpressed in patients affected by different types of cancers. The specific Ang cellular receptors have not been identified, but it is known that Ang–actin interaction induces changes both in the cell cytoskeleton and in the extracellular matrix. Most in vitro studies use the recombinant form (r-Ang) instead of the form that is normally present in vivo (“wild-type”, wt-Ang). The first residue of r-Ang is a methionine, with a free amino group, whereas wt-Ang has a glutamic acid, whose amino group spontaneously cyclizes in the pyro-glutamate form. The Ang biological activity is influenced by copper ions. To elucidate the role of such a free amino group on the protein–copper binding, we scrutinized the copper(II) complexes with the peptide fragments Ang(1–17) and AcAng(1–17), which encompass the sequence 1–17 of angiogenin (QDNSRYTHFLTQHYDAK-NH₂), with free amino and acetylated N-terminus, respectively. Potentiometric, ultraviolet-visible (UV-vis), nuclear magnetic resonance (NMR) and circular dichroism (CD) studies demonstrate that the two peptides show a different metal coordination environment. Confocal microscopy imaging of neuroblastoma cells with the actin staining supports the spectroscopic results, with the finding of different responses in the cytoskeleton organization upon the interaction, in the presence or not of copper ions, with the free amino and the acetylated N-terminus peptides.