Is caretta caretta a carrier of antibiotic resistance in the mediterranean sea?

Sea turtles can be considered a sentinel species for monitoring the health of marine ecosystems, acting, at the same time, as a carrier of microorganisms. Indeed, sea turtles can acquire the microbiota from their reproductive sites and feeding, contributing to the diffusion of antibiotic-resistant strains to uncontaminated environments. This study aims to unveil the presence of antibiotic-resistant bacteria in (i) loggerhead sea turtles stranded along the coast of Sicily (Mediterranean Sea), (ii) unhatched and/or hatched eggs, (iii) sand from the turtles’ nest and (iv) seawater. Forty-four bacterial strains were isolated and identified by conventional biochemical tests and 16S rDNA sequencing. The Gram-negative Aeromonas and Vibrio species were mainly found in sea turtles and seawater samples, respectively. Conversely, the Gram-positive Bacillus, Streptococcus, and Staphylococcus strains were mostly isolated from eggs and sand. The antimicrobial resistance profile of the isolates revealed that these strains were resistant to cefazolin (95.5%), streptomycin (43.2%), colistin and amoxicillin/clavulanic acid (34.1%). Moreover, metagenome analysis unveiled the presence of both antibiotic and heavy metal resistance genes, as well as the mobile element class 1 integron at an alarming percentage rate. Our results suggest that Caretta caretta could be considered a carrier of antibiotic-resistant genes

Analysis of HDL-microRNA panel in heterozygous familial hypercholesterolemia subjects with LDL receptor null or defective mutation

In the last years increasing attention has been given to the connection between genotype/phenotype and cardiovascular events in subjects with familial hypercholesterolemia (FH). MicroRNAs (miRs) bound to high-density lipoprotein (HDL) may contribute to better discriminate the cardiovascular risk of FH subjects. Our aim was to evaluate the HDL-miR panel in heterozygous FH (HeFH) patients with an LDLR null or defective mutation and its association with pulse wave velocity (PWV). We evaluated lipid panel, HDL-miR panel and PWV in 32 LDLR null mutation (LDLR-null group) and 35 LDLR defective variant (LDLR-defective group) HeFH patients. HDL-miR-486 and HDL-miR-92a levels were more expressed in the LDLR-null group than the LDLR-defective group. When we further stratified the study population into three groups according to both the LDLR genotype and history of ASCVD (LDLR-null/not-ASCVD, LDLR-defective/not-ASCVD and LDLR/ASCVD groups), both the LDLR/ASCVD and the LDLR-null/not-ASCVD groups had a higher expression of HDL-miR-486 and HDL-miR-92a than the LDLR-defective/not-ASCVD group. Finally, HDL-miR-486 and HDL-miR-92a were independently associated with PWV. In conclusion, the LDLR-null group exhibited HDL-miR-486 and HDL-miR-92a levels more expressed than the LDLR-defective group. Further studies are needed to evaluate these HDL-miRs as predictive biomarkers of cardiovascular events in FH

Serum coding and non-coding RNAs as biomarkers of NAFLD and fibrosis severity

BACKGROUND & AIMS: In patients with non-alcoholic fatty liver disease (NAFLD), liver biopsy is the gold standard to detect non-alcoholic steatohepatitis (NASH) and stage liver fibrosis. We aimed to identify differentially expressed mRNAs and non-coding RNAs in serum samples of biopsy-diagnosed mild and severe NAFLD patients with respect to controls and to each other. METHODS: We first performed a whole transcriptome analysis through microarray (n = 12: four Control: CTRL; four mild NAFLD: NAS 64 4 F0; four severe NAFLD NAS 65 5 F3), followed by validation of selected transcripts through real-time PCRs in an independent internal cohort of 88 subjects (63 NAFLD, 25 CTRL) and in an external cohort of 50 NAFLD patients. A similar analysis was also performed on liver biopsies and HepG2 cells exposed to oleate:palmitate or only palmitate (cellular model of NAFL/NASH) at intracellular/extracellular levels. Transcript correlation with histological/clinical data was also analysed. RESULTS: We identified several differentially expressed coding/non-coding RNAs in each group of the study cohort. We validated the up-regulation of UBE2V1, BNIP3L mRNAs, RP11-128N14.5 lncRNA, TGFB2/TGFB2-OT1 coding/lncRNA in patients with NAS 65 5 (vs NAS 64 4) and the up-regulation of HBA2 mRNA, TGFB2/TGFB2-OT1 coding/lncRNA in patients with Fibrosis stages = 3-4 (vs F = 0-2). In in vitro models: UBE2V1, RP11-128N14.5 and TGFB2/TGFB2-OT1 had an increasing expression trend ranging from CTRL to oleate:palmitate or only palmitate-treated cells both at intracellular and extracellular level, while BNIP3L was up-regulated only at extracellular level. UBE2V1, RP11-128N14.5, TGFB2/TGFB2-OT1 and HBA2 up-regulation was also observed at histological level. UBE2V1, RP11-128N14.5, BNIP3L and TGFB2/TGFB2-OT1 correlated with histological/biochemical data. Combinations of TGFB2/TGFB2-OT1 + Fibrosis Index based on the four factors (FIB-4) showed an Area Under the Curve (AUC) of 0.891 (P = 3.00E-06) or TGFB2/TGFB2-OT1 + Fibroscan (AUC = 0.892, P = 2.00E-06) improved the detection of F = 3-4 with respect to F = 0-2 fibrosis stages. CONCLUSIONS: We identified specific serum coding/non-coding RNA profiles in severe and mild NAFLD patients that possibly mirror the molecular mechanisms underlying NAFLD progression towards NASH/fibrosis. TGFB2/TGFB2-OT1 detection improves FIB-4/Fibroscan diagnostic performance for advanced fibrosis discrimination

Prospective memory performance in older people, adults and youth

The aims of this study were to verify the potential differences in prospective memory (PM) among young people, adults and the elderly; analyze the relationships between variables of comprehension and verbal fluency and PM; and finally, verify the existence of a relationship between self-reported health status and performance on PM. A cross- sectional design was used. The study involved 270 participants divided into three age groups: young people aged 18 to 28 years; adults 45 to 55, and seniors 60 to 80. Their comprehension and verbal fluency skills were assessed as well as their self-perceived health status. Subsequently, an experiment was carried out where participants were presented with paragraphs of three sentences on a computer screen and they had to recognize previously agreed words that would indicate their level of MP. The results established significant differences in prospective memory between adults and older people and between young people and the elderly. But no differences between youth and adults were found..The importance of verbal comprehension and verbal fluency in solving prospective memory experimental tasks was also significant. In addition, a better self-perception of well-being was linked to a higher performance in PMEste trabajo fue posible gracias a la financiación del grupo de investigación HUM634 de la Junta de Andalucía y al contrato OTRI de este grupo con la Diputación provincial de Cádiz. El primer autor fue becario AECID en el Departamento de Psicología de la UCA durante la realización del estu

Identification of nuclear retention domains in the alternative splicing regulator RBM20

RNA binding motif protein type 20 (RBM20) is a nuclear protein which regulates alternative splicing of expressed genes that have a key role in cardiac functions. Mutations in the RBM20 gene are linked to familial dilated cardiomyopathy and most of them alter residues in the RS domain of the protein. Functional motifs in the RBM20 protein have been poorly characterized. Our study provides functional annotations to structural domains within the RBM20 protein required for its nuclear localization. By cloning the human and mouse RBM20 genes, producing expressing vectors for truncated proteins and comparing their sub-cellular distribution in transfected cells, we have identified the sequences necessary for RBM20 full nuclear retention. The region overlaps both an RNA binding motif and a serine-arginine domain. The sequence is conserved in many species but belongs only to RBM20 protein orthologs. The RMB20 tissue specificity, together with the properties of its nuclear localization determinant, demonstrates a specific evolutionary selection for post-transcriptional regulation in the heart, highlighting RBM20 as a key factor for regulation of splicing events required for cardiac function

Molecular characterization of the RNA binding motif protein 20: determination of nuclear localization signals.

RNA splicing is a tightly regulated process that involves the spliceosome and additional RNA binding proteins that can repress or activate splice sites selection. Recent studies have indicated that mutations in RBM20, a gene encoding a novel ribonucleic acid - binding protein, are associated to human dilated cardiomyopathy (DCM). RBM20 regulates alternative splicing of expressed genes that have a key role in cardiac function, including ion homeostasis, sarcomere biology, and signal transduction. The functional motifs of the RBM20 protein have been poorly investigated.The focus of this study is to characterize the protein domains that contribute to the nuclear function of RBM20. Predictive in silico analysis of the translated RBM20 gene identifies an RNA recognition motif (RRM motif), a serine /arginine (RS) domain and Zn2+ finger domains. We have produced GFP-RBM20 fusion proteins in order to map the functional domains of the protein that contribute to subcellular distribution. We have produced truncated mutants of the RBM20 proteins and analyzed separately in immunofluorescence assays in transfected cells. We identified a region necessary and sufficient to nuclear localization of RBM20 protein that maps between the RRM and the RS domain. Actually we are producing RBM20 mutant proteins in order to characterize the nuclear localization signal (NLS). Further structural and functional characterization of RBM20 may contribute to understand the molecular pathogenesis of familiar DCM