Effects of in vitro digestion on the antioxidant activity of three phenolic extracts from olive mill wastewaters

The aim of this study was to assess the impact of in vitro digestion on the antioxidant activity of three extracts rich in phenols (two purified organic extracts (A20, A21) and one powdered extract stabilized with maltodextrins (SP)) obtained from olive mill wastewaters (OMWW). The content and composition of phenols and antioxidant activity was determined before and after in vitro digestion. The phenol content of the A20 and A21 samples were higher (>75%) than that of the SP sample before in vitro digestion. After the entire in vitro digestion, 89.3, 76.9, and 50% loss of phenols was found in A20, A21 and SP, respectively. ABTS•+ and ORAC values decreased during in vitro digestion of A20 and A21 samples, while they remained almost constant in SP. IC50 increased during digestion of A20 and A21, evidencing a loss of antioxidant capacity after the intestinal phase; an opposite IC50 trend was noted in SP, confirming the protective role of maltodextrins. For these reasons, SP represents a promising formulation to be used in the food field

Restoration of Correct Splicing of Thalassemic β-Globin Pre-mRNA by Modified U1 snRNAs

The T-->G mutation at nucleotide 705 in the second intron of the beta-globin gene creates an aberrant 5' splice site and activates a 3' cryptic splice site upstream from the mutation. As a result, the IVS2-705 pre-mRNA is spliced via the aberrant splice sites leading to a deficiency of beta-globin mRNA and protein and to the genetic blood disorder thalassemia. We have shown previously that in cell culture models of thalassemia, aberrant splicing of beta-thalassemic IVS2-705 pre-mRNA was permanently corrected by a modified murine U7 snRNA that incorporated sequences antisense to the splice sites activated by the mutation. To explore the possibility of using other snRNAs as vectors for antisense sequences, U1 snRNA was modified in a similar manner. Replacement of the U1 9-nucleotide 5' splice site recognition sequence with nucleotides complementary to the aberrant 5' splice site failed to correct splicing of IVS2-705 pre-mRNA. In contrast, U1 snRNA targeted to the cryptic 3' splice site was effective. A hybrid with a modified U7 snRNA gene under the control of the U1 promoter and terminator sequences resulted in the highest levels of correction (up to 70%) in transiently and stably transfected target cells

Effects of dietary supplementation of soybean lecithin on growth performance, nutrients digestibility and serum profiles of broilers fed fried soybean oil

The objective of this study was to evaluate the effects of soybean lecithin in broiler diets formulated with either fried or non-fried soybean oils (as an energy source), on growth performance, digestibility, and some serum parameters. A total of 600 broilers (50% male) were assigned to 4 experimental groups (with 10 replicates of 15 birds each): (i) group (S) fed with soybean oil only, (ii) group (SL) fed with soybean oil + lecithin (0.035% in feed), (iii) group (F) fed with fried soybean oil only, and iv) group (FL) fed with fried soybean oil + lecithin (0.035% in feed). The results showed that fried soybean oil in broilers diets had the same effects on the analysed parameters as the non-fried oil. Broilers that received lecithin with fried soybean oil (group FL) exhibited significantly higher body weights (1.228 vs. 1.210, p<0.05 and 1.935 vs. 1.917 kg, p<0.05) than group F in the last two experimental periods (22-28 and 29-35 d), respectively. On the contrary, the addition of soybean lecithin to diets formulated with either fried or non-fried soybean oil resulted in a significant increase in weight of some carcass cuts (thigh, breast) and a significant decrease in some visceral organs (intestine, liver, viscera), without affecting the digestibility parameters here assessed. Our findings revealed that the addition of soybean lecithin to standard soybean oil (group SL) significantly decreased serum TG (77.16 vs. 83.46, p<0.05) compared to group S. In conclusion, it was possible to use recovered frying soybean oil in broiler diets without a negative impact on growth performance and serum triglyceride level, while these parameters were improved by the addition of soybean lecithin

Modification of Alternative Splicing of Bcl-x Pre-mRNA in Prostate and Breast Cancer Cells: ANALYSIS OF APOPTOSIS AND CELL DEATH

There is ample evidence that deregulation of apoptosis results in the development, progression, and/or maintenance of cancer. Since many apoptotic regulatory genes (e.g. bcl-x) code for alternatively spliced protein variants with opposing functions, the manipulation of alternative splicing presents a unique way of regulating the apoptotic response. Here we have targeted oligonucleotides antisense to the 5'-splice site of bcl-x(L), an anti-apoptotic gene that is overexpressed in various cancers, and shifted the splicing pattern of Bcl-x pre-mRNA from Bcl-x(L) to Bcl-x(S), a pro-apoptotic splice variant. This approach induced significant apoptosis in PC-3 prostate cancer cells. In contrast, the same oligonucleotide treatment elicited a much weaker apoptotic response in MCF-7 breast cancer cells. Moreover, although the shift in Bcl-x pre-mRNA splicing inhibited colony formation in both cell lines, this effect was much less pronounced in MCF-7 cells. These differences in responses to oligonucleotide treatment were analyzed in the context of expression of Bcl-x(L), Bcl-x(S), and Bcl-2 proteins. The results indicate that despite the presence of Bcl-x pre-mRNA in a number of cell types, the effects of modification of its splicing by antisense oligonucleotides vary depending on the expression profile of the treated cells

The PRMT1 gene expression pattern in colon cancer

The methylation of arginine has been implicated in many cellular processes, such as regulation of transcription, mRNA splicing, RNA metabolism and transport. The enzymes responsible for this modification are the protein arginine methyltransferases. The most abundant methyltransferase in human cells is protein arginine methyltransferase 1. Methylation processes appear to interfere in the emergence of several diseases, including cancer. During our study, we examined the expression pattern of protein arginine methyltransferase 1 gene in colon cancer patients. The emerging results showed that the expression of one of the gene variants is associated with statistical significant probability to clinical and histological parameters, such as nodal status and stage. This is a first attempt to acquire an insight on the possible relation of the expression pattern of protein arginine methyltransferase 1 and colon cancer progression

Large-Scale Evidence for Conservation of NMD Candidature Across Mammals

BACKGROUND: Alternatively-spliced (AS) forms can vary protein function, intracellular localization and post-translational modifications. AS coupled with mRNA nonsense-mediated decay (NMD) can also control the transcript abundance. Here, we have investigated the genome-scale conservation of alternatively-spliced NMD candidates (AS-NMD candidates), in mammals. METHODOLOGY/PRINCIPAL FINDINGS: We mapped>12 million cDNA/EST library transcripts, comprising pooled data from both older and next-generation sequencing techniques, against genomic sequences to annotate AS-NMD candidates generated by in-frame premature termination codons (PTCs), in the human, mouse, rat and cow genomes. In these genomes, we found populations of genes that harbour AS-NMD candidates, varying in number from approximately 149 to 2,051 genes. We discovered that a highly-significant proportion (27%-35%) of AS-NMD candidate genes in mouse, rat and cow, also have human orthologs targeted for NMD. Intron retention was the most abundant type of AS-NMD, ranging from 43% to 67% of genes harbouring an AS-NMD candidate. Groupings of AS-NMD candidate genes either with or without intron retentions also have highly significant AS-NMD conservation, indicating that the trend is not due primarily to conservation of intron retentions. As a subset, the AS-NMD intron retentions are distinguished from non-retained introns by higher GC content, and codon usage similar to the usage in protein-coding sequences. This indicates that most of these alternatively spliced sequences have coded for proteins in the recent evolutionary past. In general, the AS-NMD candidate genes showed a similar pattern of Gene Ontology functional category enrichments in all four species. Genes linked to nucleic-acid interaction and apoptosis, and involved in pathways linked with cancer, were the most common. Finally, we mapped the AS-NMD candidates to mass spectrometry-derived proteomics data, and gathered evidence of truncated polypeptides for at least 10% of all human AS-NMD candidate transcripts. CONCLUSIONS/SIGNIFICANCE: In summary, our analysis provides strong statistical evidence for conservation of functional AS-NMD candidature across Mammalia for a large subset of genes. However, because codon usage of AS-NMD intron retentions is similar to the usage in exons, it is difficult to de-couple conservation of AS-NMD-based regulation from conservation for protein-coding ability, for intron retentions

Dynamics of Co-Transcriptional Pre-mRNA Folding Influences the Induction of Dystrophin Exon Skipping by Antisense Oligonucleotides

Antisense oligonucleotides (AONs) mediated exon skipping offers potential therapy for Duchenne muscular dystrophy. However, the identification of effective AON target sites remains unsatisfactory for lack of a precise method to predict their binding accessibility. This study demonstrates the importance of co-transcriptional pre-mRNA folding in determining the accessibility of AON target sites for AON induction of selective exon skipping in DMD. Because transcription and splicing occur in tandem, AONs must bind to their target sites before splicing factors. Furthermore, co-transcriptional pre-mRNA folding forms transient secondary structures, which redistributes accessible binding sites. In our analysis, to approximate transcription elongation, a “window of analysis” that included the entire targeted exon was shifted one nucleotide at a time along the pre-mRNA. Possible co-transcriptional secondary structures were predicted using the sequence in each step of transcriptional analysis. A nucleotide was considered “engaged” if it formed a complementary base pairing in all predicted secondary structures of a particular step. Correlation of frequency and localisation of engaged nucleotides in AON target sites accounted for the performance (efficacy and efficiency) of 94% of 176 previously reported AONs. Four novel insights are inferred: (1) the lowest frequencies of engaged nucleotides are associated with the most efficient AONs; (2) engaged nucleotides at 3′ or 5′ ends of the target site attenuate AON performance more than at other sites; (3) the performance of longer AONs is less attenuated by engaged nucleotides at 3′ or 5′ ends of the target site compared to shorter AONs; (4) engaged nucleotides at 3′ end of a short target site attenuates AON efficiency more than at 5′ end

Desarrollo de un queso vegano sin alérgenos y mejorado nutricionalmente

The agri-food technology center FUDin has developed a fermented vegan cheese prototype with unique attributes to meet consumers’ needs in terms of allergens, nutritional density, price and taste. Ingredients were chosen to ensure a low saturated fat product without allergens, flavourings and gelling agents, following a clean labelling approac

Honey discrimination by Volatile Organic Compounds analysis: comparison between GC-IMS and GC-E-Nose

The intrinsic characteristics of 54 honey samples were assessed by qualitative analysis of their volatile fraction through GC-E-NOSE and GC-IMS. Both systems detected differences among samples, due to botanical origin, breeding techniques and production processes. The application of IMS allowed to identify the discriminating molecules among honey samples