Despite clear evidence that exosomal microRNAs (miRNAs) are able to modulate the cellular microenvironment and that exosomal RNA cargo selection is deregulated in pathological conditions, the ...mechanisms controlling specific RNA sorting into extracellular vesicles are still poorly understood. Here, we identified the RNA binding protein SYNCRIP (synaptotagmin-binding cytoplasmic RNA-interacting protein; also known as hnRNP-Q or NSAP1) as a component of the hepatocyte exosomal miRNA sorting machinery. SYNCRIP knockdown impairs sorting of miRNAs in exosomes. Furthermore, SYNCRIP directly binds to specific miRNAs enriched in exosomes sharing a common extra-seed sequence (hEXO motif). The hEXO motif has a role in the regulation of miRNA localization, since embedment of this motif into a poorly exported miRNA enhances its loading into exosomes. This evidence provides insights into the mechanisms of miRNA exosomal sorting process. Moreover, these findings open the way for the possible selective modification of the miRNAs exosomal cargo.
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•SYNCRIP interacts specifically with selected miRNAs•SYNCRIP has a functional role in exosomal sorting of specific miRNAs•A hEXO miRNA extra-seed motif determines exosomal loading
Santangelo et al. shed light on the mechanism that allows a cell to load exosomes with a specific repertoire of miRNAs, showing a functional role of a specific miRNA motif and a protein that specifically interacts with it, the RNA-binding protein SYNCRIP.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
is a lncRNA overexpressed in several epithelial cancers and strongly correlated with invasion. This lncRNA was proven a pivotal element of the epithelial-to-mesenchymal transition (EMT), a ...transdifferentiation process triggering metastasis. Snail, master inducer of EMT, requires
to recruit EZH2 on specific epithelial target genes (i.e.,
, and
) and cause their repression. Here, we designed a
deletion mutant form, named
-
, including the putative Snail-binding domain but depleted of the EZH2-binding domain.
-
acted as a dominant negative of the endogenous
. In both murine and human tumor cells,
-
impaired the ability of
to bind Snail and, in turn, trigger H3K27me3/EZH2-mediated repression of Snail epithelial target genes. Notably,
-
expression was proven to reduce cellular motility, invasiveness, anchorage-independent growth, and responsiveness to TGFβ-induced EMT. These data provide evidence on a lncRNA-based strategy to effectively impair the function of a master EMT-transcriptional factor. SIGNIFICANCE: This study defines an innovative RNA-based strategy to interfere with a pivotal function of the tumor-related lncRNA
, comprising a dominant negative mutant that was computationally designed and that impairs epithelial-to-mesenchymal transition.
Treatment of multi-drug resistant Tuberculosis (MDR-TB) is challenging because it mostly relies on drugs with lower efficacy and greater toxicity than those used for drug-susceptible TB.
Aim of the ...study was to describe the frequency and type of adverse drug reactions in a cohort of MDR-TB patients and their potential impact on treatment outcome.
We conducted a retrospective study in a cohort of MDR-TB patients enrolled at a tertiary referral hospital in Italy from January 2008 to December 2016. The records of patients were reviewed for epidemiological, clinical, microbiological and adverse drug reactions data.
Seventy-four MDR-TB patients (mean age 32 years, 58.1% males, 2 XDR, 12 pre-XDR TB) were extracted from the Institute data base and included in the retrospective study cohort in the evaluation period (January 2008-December 2016). Median length of treatment duration was 20 months (IQR 14-24). Treatment outcome was successful in 57 patients (77%; 51 cured, 6 treatment completed); one patient died and one failed (2.7% overall); 15 patients were lost to follow-up (20.3%). Sixty-six (89.2%) presented adverse drug reactions during the whole treatment period. Total number of adverse drug reactions registered was 409. Three hundred forty-six (84.6%) were classified as adverse events (AEs) and 63 (15.4%) were serious AEs (SAEs). One third of the total adverse drug reactions (134/409; 32.8%) was of gastrointestinal origin, followed by 47/409 (11.5%) ototoxic drug reactions, thirty-five (8.6%) regarded central nervous system and 33 (8.1%) affected the liver. All 63 SAEs required treatment suspension with 61 SAEs out of 63 (96.8%) occurring during the first six months of treatment. Factors associated with unsuccessful treatment outcome were smoking (p = 0.039), alcohol abuse (p = 0.005) and homeless condition (p = 0.044). Neither the number of antitubercular drugs used in different combinations nor the number of AEs showed significant impact on outcome. Patients who completed the treatment experienced a greater number of AEs and SAEs (p < 0.001) if compared to lost to follow-up patients.
Our data demonstrate that, despite the high frequency of adverse drug reactions and long term therapy, the clinical management of MDR-TB patients in a referral center could reach successful treatment according to WHO target, by implementing active and systematic clinical and laboratory assessment to detect, report and manage suspected and confirmed adverse drug reactions.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Abstract Introduction School-age children report much stress in their daily lives, which may lead to psychological and physical problems. Mindfulness-based Stress Reduction is a program of ...awareness-based practices effective with adults. The purpose of this study was to investigate the efficacy of mindfulness training through yoga with school-age girls to reduce perceived stress, enhance coping abilities, self-esteem, and self-regulation, and explore the relationship between the dose of the intervention and outcomes. Method Fourth- and fifth-grade girls were recruited from two public schools and randomly assigned to intervention and wait-list control groups. The intervention group met 1 hour a week for 8 weeks and completed 10 minutes of daily homework. Results Self-esteem and self-regulation increased in both groups. The intervention group was more likely to report greater appraisal of stress ( p < .01) and greater frequency of coping ( p < .05). Homework accounted for 7% of the variance in reported stress. Discussion Consistent with reports of mindfulness training, greater awareness of the feelings associated with stress may enhance coping abilities. However, it is possible that the increasing awareness of stressors in itself increased stress, possibly as part of the process of developing mindfulness or related to cognitive, emotional, or social development. Mindfulness in children may differ from mindfulness in adults and warrants further investigation.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK, ZRSKP
Marine Recreational Fishing (MRF) is a highly attended complex activity, extremely evident along the coastlines, and mainly practiced among riparian communities. For that reason, this activity plays ...an important role to effectively contribute to the collective well-being, both from the social and economic points of view. However, it may negatively affect the fish stocks and the marine environment in general, mainly due to the removal of biological resources. The growing need to evaluate the magnitude of marine recreational fishing is recognized worldwide, especially in the last decade, when inclusive fishing programs began to focus their attention on this fishing activity. Based on its unexpected evidence and its wider repercussion on social behavior, the COVID-19 pandemic is considered by the scientific community as one of the most unique opportunities to better understand the social phenomenon and their repercussion on the environment. In this work will be reported very preliminary results on the consistency of marine recreational fishing in the case study of the Marche region (Italy). Number of recreational fishers and fishing effort were estimated through a telephone survey conducted in the Italian side of the Northern Adriatic Sea (FAO GFCM Geographical Sub Area 17) by interviewing 580 households. The sampling strategy also included a recall survey, which was carried out every month on a list of recruited fishers. In this manner, additional information was collected, such as detailed fishing effort, catches, and expenditures. In addition, biological data of catches were estimated through several on-site surveys. The information collected from January and December 2020 was affected by the COVID-19 pandemic in terms of social restrictions and access to marine places, inevitably impacting on marine recreational fishing features, including the biological resources and the related economic aspects.
The ErbB-4 receptors are unique in the EGFR/ErbB family for the ability to associate with WW domain-containing proteins. To identify new ligands of the cytoplasmic tail of ErbB-4, we panned a brain ...cDNA phage library with ErbB-4 peptides containing sequence motifs corresponding to putative docking sites for class-I WW domains. This approach led to identification of AIP4/Itch, a member of the Nedd4-like family of E3 ubiquitin protein ligases, as a protein that specifically interacts with and ubiquitinates ErbB-4 in vivo. Interaction with the ErbB-4 receptors occurs via the WW domains of AIP4/Itch. Functional analyses demonstrate that AIP4/Itch is recruited to the ErbB-4 receptor to promote its polyubiquitination and degradation, thereby regulating stability of the receptor and access of receptor intracellular domains to the nuclear compartment. These findings expand our understanding of the mechanisms contributing to the integrity of the ErbB signaling network and mechanistically link the cellular ubiquitination pathway of AIP4/Itch to the ErbB-4 receptor.--Omerovic, J., Santangelo, L., Puggioni, E. M-R., Marrocco, J., Dall'Armi, C., Palumbo, C., Belleudi, F., Di Marcotullio, L., Frati, L., Torrisi, M-R., Cesareni, G., Gulino, A., Alimandi, M. The E3 ligase Aip4/Itch ubiquitinates and targets ErbB-4 for degradation.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
Although conducting research within schools poses challenges, the knowledge gained from studying children within the context of their lives and the availability of participants makes the school site ...a valuable site for research with children. The process of scheduling the research program, identifying an appropriate school site, recruiting and retaining participants, and knowing the optimal times and methods of contact is paramount to enhancing the success of a school based study.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK, ZRSKP
The expression of the long noncoding RNA HOTAIR (HOX Transcript Antisense Intergenic RNA) is largely deregulated in epithelial cancers and positively correlates with poor prognosis and progression of ...hepatocellular carcinoma and gastrointestinal cancers. Furthermore, functional studies revealed a pivotal role for HOTAIR in the epithelial-to-mesenchymal transition, as this RNA is causal for the repressive activity of the master factor SNAIL on epithelial genes. Despite the proven oncogenic role of HOTAIR, its transcriptional regulation is still poorly understood. Here hepatocyte nuclear factor 4-α (HNF4α), as inducer of epithelial differentiation, was demonstrated to directly repress HOTAIR transcription in the mesenchymal-to epithelial transition. Mechanistically, HNF4α was found to cause the release of a chromatin loop on HOTAIR regulatory elements thus exerting an enhancer-blocking activity.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
Background & Aims In each hepatocyte, the specific repertoire of gene expression is influenced by its exact location along the portocentrovenular axis of the hepatic lobule and provides a reason for ...the liver functions compartmentalization defined “metabolic zonation.” So far, few molecular players controlling genetic programs of periportal (PP) and perivenular (PV) hepatocytes have been identified; the elucidation of zonation mechanisms remains a challenge for experimental hepatology. Recently, a key role in induction and maintenance of the hepatocyte heterogeneity has been ascribed to Wnt/β-catenin pathway. We sought to clarify how this wide-ranging stimulus integrates with hepatocyte specificity. Methods Reverse transcriptase polymerase chain reaction (RT-PCR) allowed the transcriptional profiling of hepatocytes derived from in vitro differentiation of liver stem cells. The GSK3β inhibitor 6-bromoindirubin-3′-oxime (BIO) was used for β-catenin stabilization. Co-immunoprecipitations were used to study biochemical protein interactions while ChIP assays allowed the in vivo inspection of PV and PP genes regulatory regions. Results We found that spontaneous differentiation of liver stem cells gives rise to PP hepatocytes that, after Wnt pathway activation, switch into PV hepatocytes. Next, we showed that the Wnt downstream player LEF1 interacts with the liver-enriched transcriptional factor HNF4α. Finally, we unveiled that the BIO induced activation of PV genes correlates with LEF1 binding to both its own and HNF4α consensus, and the repression of PP genes correlates with HNF4α displacement from its own consensus. Conclusion Our data show a direct and hitherto unknown convergence of the canonical Wnt signaling on the HNF4α-driven transcription providing evidences of a mechanism controlling liver zonated gene expression.
Background & Aims
Hepatitis C virus (HCV) infection is known to cause major alterations in the cross‐talk between hepatic and immune cells thus contributing to the liver disease pathogenesis. ...Extracellular vesicles have been proved to act as major players in cell‐cell communication, and their cargo changes in relation to pathophysiological states. The aim of this study was to evaluate the effects of chronic HCV infection and direct‐acting antivirals (DAA) on exosome‐delivered microRNAs and on their ability to modulate the innate immune response.
Methods
Exosomes isolated from the plasma of healthy donors and naïve, viremic HCV patients before and after DAA treatment have been compared for their microRNAs cargo by quantitative polymerase chain reaction. Functional assays with peripheral blood cells from healthy donors were performed to assess exosome‐mediated immune responses.
Results
MicroRNAs associated with HCV‐related immunopathogenesis which were found to be enriched in exosomes of HCV viremic patients (in particular, miR‐122‐5p, miR‐222‐3p, miR‐146a, miR‐150‐5p, miR‐30c, miR‐378a‐3p and miR‐20a‐5p) were markedly reduced by DAA therapy. This exosome‐microRNA cargo modulation parallels changes in their immunomodulatory properties in ex vivo experiments. Exosomes from HCV patients inhibit NK degranulation activity and this effect correlates with miR‐122‐5p or miR‐222‐3p levels.
Conclusions
Enrichment of immunomodulatory microRNAs in exosomes of HCV patients was correlated with their inhibitory activity on innate immune cells function. Direct‐acting antivirals (DAA) treatment was observed to revert both microRNA content and functional profiles of systemic exosomes towards those of healthy donors. Exosome‐associated microRNAs may provide valuable biomarkers to monitor immune response recovery.
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BFBNIB, DOBA, FZAB, GIS, IJS, IZUM, KILJ, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBMB, UILJ, UKNU, UL, UM, UPUK
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