116 research outputs found
Studio dell'espressione di microRNA, geni target e pathways indotti da NGF in cellule epiteliali di cornea: confronto tra differenti bioformulazioni.
No full textNerve Growth Factor (NGF) is a member of the neurotrophins family that plays a key role in the maintenance and functions of both the central and peripheral nervous systems, as well as in non-neuronal cells. It exerts its action through two classes of transmembrane receptors, one high affinity, TrkA, and one low affinity, p75NTR, regulating different and important biological functions. Besides several neuropathies, some studies showed that NGF revealed translational clinical application for ulcers of the eye’s anterior segment, and a NGF bioformulation has been EMA and FDA approved as first-in-class treatment for neurotrophic keratitis (NK), a rare degenerative corneal disease due to corneal trigeminal innervation defects causing spontaneous injuries and wounds. However, NGF-induced biomolecular mechanisms in corneal cells are still unknown.
MicroRNAs (or miRNAs) are a novel class of endogenous, small non-coding RNAs (~22 nucleotides), able to regulate target genes, at the post-transcriptional level, in plants and animals. MiRNAs are involved in maintaining cell homeostasis and tuning fundamental biological processes, such as growth, proliferation, apoptosis, cell metabolism; dysregulated expression of miRNAs’ is observed and strictly linked to relevant human diseases (e.g. viral, immune, neurodegenerative and cardiovascular diseases, cancer).
In this study we evaluated miRNAs’ expression modulation in corneal cells after treatment with different NGF bioformulations, by focusing the analysis on target genes and pathways as well.
Tens of miRNAs were significantly dysregulated, mainly hypo-expressed, after treatment. Among those, 3 were shared by all the bioformulations, 37 by 2 of them, whereas 12, 16 and 20 were correlated to each single bioformulation. A total number of 91 unique pathways were identified by DIANA experimentally-supported Tarbase analysis: 46, including the neurotrophin signalling pathway, were shared by all three bioformulations, 28 by 2 of them, whereas 9, 2 and 6 were related to each single bioformulation. More than 2.000 experimentally supported target genes were identified. After focusing the analysis on the neurotrophin signalling pathway, no relevant differences were globally detected among the NGF bioformulations, all appearing to drive through proliferative and survival signals. However, one of the three bioformulations here used appeared to be more specific than the remaining two, as, when proportionally compared to them, it involved less significant miRNAs which, however, can target higher numbers of genes involved in this pathway. The same result was achieved when analysis was restricted to miRNAs’ sub-groups specifically related to each single bioformulation. Genemania gene and functional enrichment analysis further confirmed data. Futhermore, the gene list can be in depth analysed in order to identify targets suitable for further molecular analyses and validations, especially for each single bioformulation, with the aim to possibly identify target genes and functions specifically related to them. For this reason, more experiments by immunoblotting to evaluate expression levels of interesting target genes are ongoing.
We examined the effects induced by different NGF bioformulations on epithelial corneal cells, by focusing the attention on miRNAs expression levels, and in silico target genes and pathways analysis. Although further functional studies to validate results are needed, original and novel insights about genes and epigenetics mechanisms induced by NGFs were provided. Our data may leave to hypothesize that NGF1 might be more specific and effective in inducing neurotrophin signalling pathway compared with NGF2 and NGF3. Importantly, given the putative role of miRNAs as biomarkers or therapeutic targets, this study makes available data potentially exploitable in clinical practice
Investigational neuroprotective compounds in clinical trials for retinal disease
No full textIntroduction: Retinal neurodegeneration causes irreversible vision loss, impairing quality of life. By targeting neurotoxic conditions, such as oxidative stress and ischemia, neuroprotectants can slow or stop sight loss resulting from eye disease. Despite limimted clinical use of neuroprotectants, there are several promising compounds in early clinical trials (pre-phase III) which may fulfil new therapeutic roles. Search terms relating to neuroprotection and eye disease were used on ClinicalTrials.gov to identify neuroprotective candidates. Areas covered: Research supporting neuroprotection in eye diseases is focused on, ranging from preclinical to phase II, according to the ClinicalTrials.gov database. The compounds discussed are explored in terms of future clinical applications. Expert opinion: The major challenge in neuroprotection research is translation from basic research to the clinic. A number of potential neuroprotectants have progressed to ophthalmology clinical trials in recent years, with defined mechanisms of action–saffron and CoQ10–targeting mitochondria, and both CNTF and NGF showing anti-apoptotic effects. Enhancements in trial design and patient cohorts in proof-of-concept trials with enriched patient populations and surrogate endpoints should accelerate drug development. A further important consideration is optimising drug delivery to improve individualised management and patient compliance. Progress in these areas means that neuroprotective strategies have a much improved chance of translational success
Mitotic phosphorylation of Tau/MAPT modulates cell cycle progression in prostate cancer cells
No full textPurposeTau/MAPT (microtubule associated protein tau) protein is actively studied for the pathologic consequences of its aberrant proteostasis in central nervous system leading to neurodegenerative diseases. Besides its ability to generate insoluble toxic oligomers, Tau homeostasis has attracted attention for its involvement in the formation of the mitotic spindle. This evidence, in association with the description of Tau expression in extra-neuronal tissues, and mainly in cancer tissues, constitutes the rationale for a more in-depth investigation of Tau role also in neoplastic diseases.MethodsIn our study, we investigated the expression of phosphorylated Tau in prostate cancer cell lines with particular focus on the residue Thr231 present in microtubule binding domain.ResultsThe analysis of prostate cancer cells synchronized with nocodazole demonstrated that the expression of Tau protein phosphorylated at residue Thr231 is restricted to G2/M cell cycle phase. The phosphorylated form was unable to bind tubulin and it does not localize on mitotic spindle. As demonstrated by the use of specific inhibitors, the phosphorylation status of Tau is under the direct control of cdk5 and PP2A, while cdk1 activation was able to exert an indirect control. These mechanisms were also active in cells treated with docetaxel, where counteracting the expression of the dephosphorylated form, by kinase inhibition or protein silencing, determined resistance to drug toxicity.ConclusionsWe hypothesize that phosphorylation status of Tau is a key marker for G2/M phase in prostate cancer cells and that the forced modulation of Tau phosphorylation can interfere with the capacity of cell to efficiently progress through G2/M phase
Innovative unattended SEM-EDS analysis for asbestos fiber quantification
No full textScanning electron microscopy with energy dispersive spectrometry (SEM-EDS) is the only affordable analytical
technique that can discriminate both morphology and elemental composition of inorganic fibers. SEM-EDS is
indeed required to quantify asbestos in confounding natural matrixes (e.g. ophiolites), but is also time-consuming,
operator dependent, and strongly relies on the stochastic distribution of the fibers on the filter surface.
The balance between analytical time/cost and the method sensibility allows only about 0.5% of the filter to be
analyzed, strongly affecting the statistical significance of results. To improve sensitivity and precision and enhance
productivity, an unattended quantitative measurement of the asbestos fibers by SEM-EDS is proposed. The
method identifies the particle shape first and determines their chemical composition later, saving EDS analytical
time. Our approach was tested on four asbestos standards and the relative error on replicated measurements
was< 10%. The proposed unattended method quantifies asbestos in natural confounding matrix, also with a
very low asbestos content
"Quer acender uma vela a Deus e outra ao diabo" : as (não) mudanças discursivas do integralista Luiz A. Compagnoni - Jornal Pioneiro 1948-1950
No full textA necessidade de aprofundamento nos estudos sobre os movimentos conservadores e autoritários nacionais se mostra cada vez mais presente, além de trazer respostas e outras tantas perguntas sobre o presente que vivenciamos no momento de escrita deste trabalho. A pesquisa analisa as transformações discursivas nos escritos de Luís Alexandre Compagnoni, publicadas no jornal O Pioneiro, entre os anos 1948 e 1950. O personagem em questão foi membro da Ação Integralista Brasileira na década de 1930 e, em 1948, foi um dos principais fundadores do jornal analisado no trabalho. Através da Análise de Conteúdo (BARDIN, 2011) e da Análise de Discurso Crítica (FAIRCLOUGH, 2001), atentamos para a constituição e para as possíveis mudanças no discurso empregado pelo autor nos primeiros anos de circulação do impresso. A análise das mudanças e permanências discursivas é feita através da comparação entre os escritos de Compagnoni na década de 1930 e o que escreveu em fins da década de 1940 e início da de 1950. [resumo fornecido pelo autor]The need for further studies on national conservative and authoritarian movements is increasingly present, in addition to providing answers and many other questions about the present that we experienced at the time of writing this work. The research analyzes the discursive transformations in the writings of Luís Alexandre Compagnoni, published in the newspaper O Pioneiro, between the years 1948 and 1950. The character in question was a member of the Brazilian Integralist Action in the 1930s and, in 1948, was one of the main founders of the newspaper analyzed at this study. Through Content Analysis (BARDIN, 2011) and Critical Discourse Analysis (FAIRCLOUGH, 2001), we pay attention to the constitution and the possible changes in the discourse used by the author in the first years of circulation of the print. The analysis of discursive changes and permanences is made through the comparison between Compagnoni's writings in the 1930s and what he wrote in the late 1940s and early 1950s. [resumo fornecido pelo autor
What Arthroscopic Skills Need to Be Trained Before Continuing Safe Training in the Operating Room?
No full textThe purpose of this study was to generate consensus among experienced surgeons on "what skills a resident should possess before continuing safe training in the operating room (OR)." An online survey of 65 questions was developed and distributed to surgeons in the European community. A total of 216 responded. The survey included 15 questions regarding generic and specific skills; 16 on patient and tissue manipulation, 11 on knowledge of pathology and 6 on inspection of e-anatomical structures; 5 methods to prepare residents; and 12 on specific skills exercises. The importance of each question (arthroscopic skill) was evaluated ranging from 1 (not important at all) to 6 (very important). Chi-square test, respondent agreement, and a qualitative ranking method were determined to identify the top ranked skills (p 134, p 0.85, and all " high priority" level). The top ranked 2 specific arthroscopic skills were " portal placement" and " triangulating the tip of the probe with a 30-degree scope" (chi-square test > 176, p <0.001, excellent agreement, and assigned high priority). The online survey identified consensus on skills that are considered important for a trainee to possess before continuing training in the OR. Compared with the Canadian colleagues, the European arthroscopy community demonstrated similar rankin
Il romanzo italiano 1816-26
No full textLa tesi indaga la produzione di romanzi negli stati italiani nel periodo dal 1816 al 1826 in relazione al dibattito, alla cultura della restaurazione, alla specificità letteraria italiana e ai modelli stranieri. Il corpus di opere identificato rivela, tra gli autori, alcuni dei poligrafi attivi anche nei periodici, da Giuseppe Compagnoni a Davide Bertolotti, da Defendente Sacchi a Giuseppe Bianchetti, e, tra le autrici, oltre alle già note Orintia Romagnoli Sacrati e Carolina Decio Cosenza, un'oscura figura di monaca toscana, Orsola Cozzi, che può considerarsi la prima romanziera italiana. Il lavoro dedica una particolare attenzione agli aspetti delle resistenze conservatrici nei confronti del romanzo, della ricerca di popolarità, che accomuna autori di diverse tendenze, e degli intenti educativi informati al progetto di costruire la nazione italiana. L'ultima parte è dedicata all'approfondimento testuale di alcuni dei romanzi ritenuti più significativi
MicroRNA profiling in women with migraine: effects of CGRP-targeting treatment
No full textAbstract Background Migraine lacks biomarkers that can trace the biological pathways of the disease and predict the effectiveness of treatments. Monoclonal antibodies targeting calcitonin gene-related peptide pathway – including erenumab – offer the opportunity of investigating potential migraine biomarkers due to their specific mechanism of action in preventing both episodic (EM) and chronic (CM) migraine. Our study aims at evaluating the expression levels of circulating microRNAs (miRNAs) according to migraine type, before and after treatment with erenumab and based on treatment response, in order to identify miRNAs with potential role as epigenetic biomarkers. Methods The study included women aged 25–50 years with EM or CM treated with erenumab according to clinical indications. MiRNAs expression levels were assessed before (baseline) and after a 16-week treatment with erenumab, 140 mg every four weeks (post-treatment). An extensive miRNAs profiling was performed by qRT-PCR in small, pooled groups of ≤ 8 women each, classified according to migraine frequency (EM and CM) and the degree of response to erenumab. The expression levels of selected miRNAs were also validated using single miRNA assays in each woman with EM and CM. Results During the study, 36 women with migraine (19 with EM and 17 with CM) out of 40 who were initially screened, performed the assessment of miRNA expression at baseline and post-treatment, Erenumab treatment significantly improved migraine burden in both EM and CM. MiRNA profiling revealed differential expression levels of a wide set of miRNAs (hsa-let-7d-3p, hsa-miR-106b-3p, hsa-miR-122-5p, hsa-miR-143-3p, hsa-miR-144-3p, hsa-miR-16-5p, hsa-miR-181a-5p, hsa-miR-221-3p, hsa-miR-25-3p, hsa-miR-29b-2-5p, hsa-miR-326, miR-363-3p, hsa-miR-424-5p, hsa-miR-485-3p, hsa-miR-532-5p, hsa-miR-543, hsa-miR-629-5p, hsa-miR-660-5p, hsa-miR-92a-3p) depending on treatment response. Among them, single miRNA assays confirmed the progressive decrease of hsa-miR-143-3p expression levels in relation to increasing response to erenumab in women with EM (7 with low, 6 with medium, and 6 with high response; p = 0.02). Additionally, single assays showed higher hsa-miR-34a-5p and hsa-miR-382-5p expression levels at baseline in women with CM compared with those with EM (p = 0.0002 and p = 0.0007, respectively), as well as their expression level decrease in women with CM from baseline to follow-up (p = 0.04 and p = 0.02, respectively). Conclusions Our study suggests that targeting the CGRP pathway in migraine changes the expression levels of certain miRNAs. These miRNA levels are linked to the levels of response to CGRP receptor blockage. Future research challenges include assigning specific functions to the modulated miRNAs to unravel pathways modulated by the disease and the treatment. Trial registration The study was registered in clinicaltrials.gov with code NCT04659226 and in the Novartis database with code CAMG334AIT05T
Applications of Next Generation Sequencing to the Analysis of Familial Breast/Ovarian Cancer
No full textNext generation sequencing (NGS) provides a powerful tool in the field of medical genetics, allowing one to perform multi-gene analysis and to sequence entire exomes (WES), transcriptomes or genomes (WGS). The generated high-throughput data are particularly suitable for enhancing the understanding of the genetic bases of complex, multi-gene diseases, such as cancer. Among the various types of tumors, those with a familial predisposition are of great interest for the isolation of novel genes or gene variants, detectable at the germline level and involved in cancer pathogenesis. The identification of novel genetic factors would have great translational value, helping clinicians in defining risk and prevention strategies. In this regard, it is known that the majority of breast/ovarian cases with familial predisposition, lacking variants in the highly penetrant BRCA1 and BRCA2 genes (non-BRCA), remains unexplained, although several less penetrant genes (e.g., ATM, PALB2) have been identified. In this scenario, NGS technologies offer a powerful tool for the discovery of novel factors involved in familial breast/ovarian cancer. In this review, we summarize and discuss the state of the art applications of NGS gene panels, WES and WGS in the context of familial breast/ovarian cancer
Circulating MicroRNAs as Prognostic and Therapeutic Biomarkers in Breast Cancer Molecular Subtypes
No full textBreast cancer (BC) is a common and heterogeneous disease, of which six molecular subtypes, characterized by different biological features and clinical outcomes, were described. The identification of additional biomarkers able to further connote and distinguish the different BC subtypes is essential to improve the diagnostic, prognostic and therapeutic strategies in BC patients. MicroRNAs (miRNAs) are short non-coding RNA involved in several physiological and pathological processes, including cancer development and progression. In particular, circulating miRNAs, which can be found in an adequately stable structure in serum/plasma of cancer patients, are emerging as very promising non-invasive biomarkers. Several studies have analyzed the potential role of circulating miRNAs as prognostic and therapeutic markers in BC. In the present review we describe circulating miRNAs, identified as putative biomarker in BC, with special reference to different BC molecular subtypes
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