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Let-7c, an intronic microRNA (miRNA) embedded in the long non-coding gene LINC00478, can act as a tumor suppressor by targeting oncogenes. Previous studies indicated that in acute promyelocytic leukemia (APL), a subtype of acute myelogenous leukemia (AML) bearing the leukemia promoting PML/RARα fusion protein, let-7c expression seems to be controlled by the host gene promoter, in which canonical Retinoic Acid Responsive Elements (RAREs) are bound by PML/RARα in an all transretinoic acid (ATRA)–sensitive manner. Here, let-7c transcriptional regulation was further investigated and a novel intronic promoter upstream of the pre-miRNA was identified. This new promoter has transcriptional activity strongly indicating that at least two promoters need to be considered for let-7c transcription: the distal host gene and the proximal intronic promoter. Therefore, epigenetic modifying enzymes and histone acetylation and methylation status were analyzed on both let-7c promoters. It was demonstrated that ATRA treatment leads to let-7c upregulation inducing a more open chromatin conformation of the host gene promoter, with an enrichment of epigenetic marks that correlate with a more active transcriptional state. Conversely, the epigenetic marks on the intronic promoter are not significantly affected by ATRA treatment. Interestingly, in solid tumors such as prostate and lung adenocarcinoma it was found that both host and intronic promoters are functional. These data suggest that while the host gene promoter may control let-7c expression in AML, in a nonleukemic tumor context instead the intronic promoter contributes or preferentially regulates let-7c transcription.Implications: Alternative promoter usage represents a regulatory mechanism of let-7c expression in different tissues. Mol Cancer Res; 12(6); 878–89. ©2014 AACR.

PDF file - 725KB, S2. Expression of the primary transcript of miR-99a and miR-125b in cell lines from different solid tumors. S3. UCSC track.

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Purpose: We previously identified novel thiazole derivatives able to reduce histone acetylation and histone acetyltransferase (HAT) activity in yeast. Among these compounds, 3-methylcyclopentylidene-[4-(4′-chlorophenyl)thiazol-2-yl]hydrazone (CPTH6) has been selected and used throughout this study.Experimental Design: The effect of CPTH6 on histone acetylation, cell viability and differentiation, cell-cycle distribution, and apoptosis in a panel of acute myeloid leukemia and solid tumor cell lines has been evaluated.Results: Here, we showed that CPTH6 leads to an inhibition of Gcn5 and pCAF HAT activity. Moreover, it inhibits H3/H4 histones and α-tubulin acetylation of a panel of leukemia cell lines. Concentration- and time-dependent inhibition of cell viability, paralleled by accumulation of cells in the G0/G1 phase and depletion from the S/G2M phases, was observed. The role of mitochondrial pathway on CPTH6-induced apoptosis was shown, being a decrease of mitochondrial membrane potential and the release of cytochrome c, from mitochondria to cytosol, induced by CPTH6. Also the involvement of Bcl-2 and Bcl-xL on CPTH6-induced apoptosis was found after overexpression of the two proteins in leukemia cells. Solid tumor cell lines from several origins were shown to be differently sensitive to CPTH6 treatment in terms of cell viability, and a correlation between the inhibitory efficacy on H3/H4 histones acetylation and cytotoxicity was found. Differentiating effect on leukemia and neuroblastoma cell lines was also induced by CPTH6.Conclusions: These results make CPTH6 a suitable tool for discovery of molecular targets of HAT and, potentially, for the development of new anticancer therapies, which warrants further investigations. Clin Cancer Res; 18(2); 475–86. ©2011 AACR.

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Background Altered microRNA profiles have been observed not only in tumour tissues but also in biofluids, where they circulate in a stable form thus representing interesting biomarker candidates. This study aimed to identify a microRNA signature as a non-invasive biomarker and to investigate its impact on glioma biology. Methods MicroRNAs were selected using a global expression profile in preoperative serum samples from 37 glioma patients. Comparison between serum samples from age and gender-matched controls was performed by using the droplet digital PCR. The ROC curve and Kaplan-Meier survival analyses were used to evaluate the diagnostic/prognostic values. The functional role of the identified signature was assessed by gain/loss of function strategies in glioma cells. Results A three-microRNA signature (miR-1-3p/−26a-1-3p/−487b-3p) was differentially expressed in the serum of patients according to the isocitrate dehydrogenase (IDH) genes mutation status and correlated with both patient Overall and Progression Free Survival. The identified signature was also downregulated in the serum of patients compared to controls. Consistent with these results, the signature expression and release in the conditioned medium of glioma cells was lower in IDH-wild type cells compared to the mutated counterpart. Furthermore, in silico analysis of glioma datasets showed a consistent deregulation of the signature according to the IDH mutation status in glioma tumour tissues. Ectopic expression of the signature negatively affects several glioma functions. Notably, it impacts the glioma invasive phenotype by directly targeting the invadopodia-related proteins TKS4, TKS5 and EFHD2. Conclusions We identified a three microRNA signature as a promising complementary or even an independent non-invasive diagnostic/prognostic biomarker. The signature displays oncosuppressive functions in glioma cells and impacts on proteins crucial for migration and invasion, providing potential targets for therapeutic intervention.

Protecting the health of pregnant women from environmental stressors is crucial for reducing the burden of non-communicable diseases. In industrially contaminated sites, this action is particularly challenging due to the heterogeneous pollutant mixtures in environmental matrices. Aim of this study was to evaluate distribution patterns of mercury, hexacholobenzene and polychlorobyfenils in the serum of 161 pregnant women recruited in the framework of the NEHO cohort and living both inside and outside the National Priority Contaminated Site (NPCS) of Priolo. Food macro-categories were determined, and serum levels of contaminants were used to perform k-means cluster analysis and identify the role of food in pollutant transfer from the environment. Two groups of mothers with high and low measured pollutant levels were distinguished. Concentrations in mothers in the high-exposure cluster were at least twofold for all the evaluated pollutants (pHighlights- HCB, PCBs and Hg were measured in pregnant women from a highly contaminated area- Pregnant women’s exposure profiles were obtained through k-means cluster analysis- Distance of residence from emission sources only partially explains exposure levels- Maternal age and food consumption appear to primarily drive contaminant levelsAbstract Figure

During pregnancy, maternal nutrition and lifestyle play a critical role in influencing fetal development and newborn health outcomes. The aim of this study is to investigate the factors influencing the adherence to dietary patterns in pregnant women living in highly contaminated areas, and whether women with higher environmental risk perception manifest different nutritional behaviors during pregnancy. Food consumption data on 816 pregnant women from the Neonatal Environment and Health Outcomes (NEHO) residential birth cohort were analyzed. Dietary patterns were computed by principal component analysis. A multinomial logistic regression was also applied to identify sociodemographic, lifestyle, and pregnancy-related determinants of adherence to dietary patterns during pregnancy. Three patterns of food consumption—explaining 24.9% of the total variance—were identified as “prudent”, “high energy”, and “vegetarian” patterns. Results suggest that food choices during pregnancy follow a social gradient and align with other health behaviors during pregnancy: older, better educated, and physically active women with higher risk perception are more likely to follow healthier dietary patterns. Knowledge about what is eaten can contribute to dietary choices. Interventions to improve the prenatal nutrition knowledge of pregnant women are needed, especially concerning younger mothers and those with lower educational levels.

Il contributo intende approfondire il ruolo che il diritto penale internazionale può e potrà avere nel contrasto ai disastri ambientali atteso che, sempre più spesso, dinnanzi a danni che vanno ad incidere gravemente sull’ambiente e sulle sue risorse – sulla scia dei reclami dell’opinione pubblica mondiale che invoca il ricorso alla sanzione penale – stanno emergendo proposte volte ad ampliare l’ambito di operatività del diritto penale internazionale per permettere allo stesso di intervenire anche nel caso di crimini contro l’ambiente sia nei confronti delle persone fisiche sia nei riguardi delle persone giuridiche. Obbiettivo del presente scritto è quindi quello di valutare se sia possibile «gestire» la minaccia ambientale con gli attuali strumenti forniti dal diritto penale internazionale e se sia necessario – o meno – introdurre un nuovo specifico crimine internazionale (quello cd. di ecocidio).

Diffuse large B-cell lymphoma (DLBCL) is an aggressive, heterogeneous neoplasm where prognostication and therapeutic decision are challenging. The available prognostic tools are not able to identify all patients refractory to treatment. MicroRNAs, small RNAs frequently deregulated in cancer, stably circulate in biofluids, representing interesting candidates for non-invasive biomarkers. Here we validated serum miR-22, an evolutionarily conserved microRNA, as a prognostic/predictive biomarker in DLBCL. Moreover, we found that its expression and release from DLBCL cells are related to therapy response and adversely affect cell proliferation. These results suggest that miR-22 is a promising complementary or even independent non-invasive biomarker for DLBCL management.

Colorectal cancer (CRC), despite the advances in screening and surveillance, remains the second most common cause of cancer death worldwide. The biological inadequacy of pre-clinical models to fully recapitulate the multifactorial etiology and the complexity of tumor microenvironment and human CRC’s genetic heterogeneity has limited cancer treatment development. This has led to the development of Patient-derived models able to phenocopy as much as possible the original inter- and intra-tumor heterogeneity of CRC, reflecting the tumor microenvironment’s cellular interactions. Implantation of patient tissue into immunodeficient mice hosts and the culture of tumor organoids have allowed advances in cancer biology and metastasis. This review highlights the advantages and limits of Patient-derived models as innovative and valuable pre-clinical tools to study progression and metastasis of CRC, develop novel therapeutic strategies by creating a drug screening platform, and predict the efficacy of clinical response to therapy.

Perianal fistula represents one of the most disabling manifestations of Crohn's Disease (CD) due to complete destruction of the affected mucosa, which is replaced by granulation tissue and associated with changes in tissue organization. To date, the molecular mechanisms underlying perianal fistula formation are not well defined. Here, we dissected the tissue changes in the fistula area and addressed whether a dysregulation of extracellular matrix (ECM) homeostasis can support fistula formation.Surgical specimens from perianal fistula tissue and the surrounding region of fistulizing CD were analyzed histologically and by RNA sequencing (RNA-seq). Genes significantly modulated were validated by RT-PCR, WB, and immunofluorescence assays. The effect of the protein product of TNF-stimulated gene-6 (TSG-6) on cell morphology, phenotype and ECM organization was investigated with endogenous Lentivirus-induced over-expression of TSG-6 in Caco-2 cells and with exogenous addition of recombinant human TSG-6 protein (rhTSG-6) to primary fibroblasts from region surrounding fistula. Proliferative and migratory assays were performed.A markedly different organization of ECM was found across fistula and surrounding fistula regions with an increased expression of integrins and metalloproteinases (MMPs) and hyaluronan (HA) staining in the fistula, associated with increased newly synthesized collagen fibers and mechanosensitive proteins. Among dysregulated genes associated with ECM, TNFAI6 (gene encoding for TSG-6) was as significantly upregulated in the fistula compared to area surrounding fistula, where it promoted the pathological formation of complexes between heavy chains (HCs) from inter-alpha-inhibitor and HA responsible for the formation of a crosslinked ECM. There was a positive correlation between TSG-6 expression and expression of mechanosensitive genes in fistula tissue. The over-expression of TSG-6 in Caco-2 cells promoted migration, epithelial mesenchymal transition (EMT), transcription factor SNAIL and hyaluronan synthase (Has) levels; while in fibroblasts, isolated from the area surrounding the fistula, it promoted an activated phenotype. Moreover, the enrichment of an HA scaffold with rhTSG-6 protein promoted collagen release, increase of SNAIL, ITGA4, ITGA42B, and PTK2B genes, the latter being involved in the transduction of responses to mechanical stimuli.By mediating changes in the ECM organization, TSG-6 triggers the EMT transcription factor SNAIL through the activation of mechanosensitive proteins. These data point to regulators of ECM as new potential targets for the treatment of CD perianal fistula.

Background: Pomegranate (Punica granatum) can be used to prepare a bioactive extract exerting anti-inflammatory activities. Clinical studies demonstrated an improvement in clinical response in inflammatory bowel disease (IBD) patients when pomegranate extract (PG) was taken as a complement to standard medications. However, the molecular mechanisms underlying its beneficial effects are still scarcely investigated. This study investigates the effect of PG on bacterial biofilm formation and the promotion of mucosal wound healing. Methods: The acute colitis model was induced in C57BL/6N mice by 3% dextran sodium sulfate administration in drinking water for 5 days. During the recovery phase of colitis, mice received saline or PG (200 mg/kg body weight) by oral gavage for 11 days. Colitis was scored daily by evaluating body weight loss, bleeding, and stool consistency. In vivo intestinal permeability was evaluated by fluorescein isothiocyanate-conjugated dextran assay, bacterial translocation was assessed by fluorescence in situ hybridization on tissues, whereas epithelial and mucus integrity were monitored by immunostaining for JAM-A and MUC-2 markers. Bacterial biofilm formation was assessed using microfluidic devices for 24 or 48 h. Primary fibroblasts were isolated from healthy and inflamed areas of 8 IBD patients, and Caco-2 cells were stimulated with or without PG (5 μg/mL). Inflammatory mediators were measured at the mRNA and protein level by RT-PCR, WB, or Bio-plex multiplex immunoassay, respectively. Results: In vivo, PG boosted the recovery phase of colitis, promoting a complete restoration of the intestinal barrier with the regeneration of the mucus layer, as also demonstrated by the absence of bacterial spread into the mucosa and the enrichment of crypt-associated fibroblasts. Microfluidic experiments did not highlight a specific effect of PG on Enterobacterales biofilm formation, even though Citrobacter freundii biofilm was slightly impaired in the presence of PG. In vitro, inflamed fibroblasts responded to PG by downregulating the release of metalloproteinases, IL-6, and IL-8 and upregulating the levels of HGF. Caco-2 cells cultured in a medium supplemented with PG increased the expression of SOX-9 and CD44, whereas in the presence of HGF or plated with a fibroblast-conditioned medium, they displayed a decrease in SOX-9 and CD44 expression and an increase in AXIN2, a negative regulator of Wnt signaling. Conclusions: These data provide new insight into the manifold effects of PG on promoting mucosal homeostasis in IBD by affecting pathogen biofilm formation and favoring the regeneration of the intestinal barrier through the regulation of the crosstalk between epithelial and stromal cells.

The present article addresses the issue of the right to life from a double perspective. If – in fact – the protection of life must induce the legislator to introduce particularly effective sanctions to protect this right from unjustified aggression, on the other hand, the punitive claim of the State cannot exclusively consist, in any case, in the neutralization of the offender for life, since this would constitute an exploitation of the human being for contingent porpouses of criminal policy and would be in contrast with the re-educational function of the penalty, as it is provided by the Article 27, par. 3., of the italian Constitution and with the principle of human dignity. A similar prejudice would seem to be recognized in the case of life imprisonment, which – as a perpetual penalty – limits (and we will see later how) any possibility of liberation of the condemned and frustrates his expectations and hopes. In detail, therefore, the paper will examine the recent developments of the jurisprudence of the European Court of Human Rights and of the Constitutional Court on life imprisonment, trying to highlight the illegality not only of the so-called “life imprisonment impediment” (as it was recently affirmed by the ECHR and by the Constitutional Court) but – more generally – also to life imprisonment in all its forms and to propose – de lege ferenda – possible alternatives to the perpetual sanction, necessary to protect the right to life and hope of the offender.

The paper aim to examine the recent developments of the jurisprudence of the European Court of Human Rights and of the Constitutional Court on life imprisonment, trying to highlight the illegality not only of the so-called “life imprisonment impediment” (as it was recently affirmed by the ECHR and by the Constitutional Court) but – more generally – also to life imprisonment in all its forms and to propose – de lege ferenda – possible alternatives to the perpetual sanction, necessary to protect the right to life and hope of the offender. O presente artigo aborda o direito à vida mediante uma dupla perspectiva. Se de um lado a proteção à vida deve induzir o legislador a introduzir determinadas sanções para protegê-lo contra agressões injustificadas, de outro lado, a pretensão punitiva do Estado não pode exclusivamente consistir, em qualquer caso, na neutralização do ofensor eternamente. Isso pois, haveria uma exploração da condição humana para fins criminais bem como seria contraditório a pena como função reeducadora, conforme determinam a dignidade da pessoa humana e o artigo 27, §3° da Constituição Italiana. Um prejuízo similar deve ser reconhecido no caso da prisão pela vida inteira, como uma pena perpétua – que limita, como será visto, qualquer possibilidade de liberação do condenado e frustração das suas expectativas e esperanças. Particularmente, este trabalho examinará desenvolvimentos jurisprudenciais recentes da Corte Europeia de Direitos Humanos e da Corte Constitucional sobre prisão perpétua, buscando iluminar a ilegalidade não apenas em virtude do chamado “impedimento de prisão perpétua” (como recentemente afirmado pelos tribunais citados), mas, de forma mais geral.

Hsa-mir-143 and hsa-let-7c have been reported to be deregulated in multiple neoplasms. The main purpose of this study was to investigate the expression of these miRNAs in bladder cancer (BCa) and to analyze the association between their expression profiles and clinical and epidemiological parameters. Ninety BCa specimens were included. Expression patterns of miR-143 and let-7c were assessed by qRT-PCR using Taqman specific probes. Validated and predicted targets of these miRNA's were identified using CSmiRTar and DAVID tools, respectively. miR-143 was downregulated in tumors compared to controls (mean fold-change (FC) = 0.076). Its expression was significantly higher in MIBC compared to NMIBC (p = 0,001). Its value as a potential biomarker discriminating non invasive tumors from the invasive ones was confirmed by ROC curve (AUC = 0.768; p = 0.0001). Also, this down-regulation positively correlates with frequency of tobacco use (p = 0,04) and chronic alcohol consumption (p = 0,04). Let-7c was overexpressed in BCa samples (mean (FC = 9.92) compared to non tumoral ones but was not associated to clinical and epidemiological parameters. A comprehensive overview of miR-143 targets and pathways implicated in BCa initiation, diagnosis or prognosis using bioinformatical analysis, was conducted. While both deregulated miRNAs may contribute to urothelial tumorigenesis, the deregulation of miR-143 was significantly correlated to epidemiological and clinical parameters.

Nontypeable Haemophilus influenzae (NTHi) is commonly isolated from airways of patients suffering from chronic respiratory diseases, such as COPD or cystic fibrosis (CF). However, to what extent NTHi long-term infection contributes to the lung inflammatory burden during chronic airway disease is still controversial. Here, we exploited human respiratory samples from a small cohort of CF patients and found that patients chronically infected with NTHi had significantly higher levels of interleukin (IL)-8 and CXCL1 than those who were not infected. To better define the impact of chronic NTHi infection in fuelling inflammatory response in chronic lung diseases, we developed a new mouse model using both laboratory and clinical strains. Chronic NTHi infection was associated with chronic inflammation of the lung, characterised by recruitment of neutrophils and cytokine release keratinocyte-derived chemokine (KC), macrophage inflammatory protein 2 (MIP-2), granulocyte colony-stimulating factor (G-CFS), IL-6, IL-17A and IL-17F) at 2 and 14 days post-infection. An increased burden of T-cell-mediated response (CD4+ and γδ cells) and higher levels of pro-matrix metalloproteinase 9 (pro-MMP9), known to be associated with tissue remodelling, were observed at 14 days post-infection. Of note we found that both CD4+IL-17+ cells and levels of IL-17 cytokines were enriched in mice at advanced stages of NTHi chronic infection. Moreover, by immunohistochemistry we found CD3+, B220+ and CXCL-13+ cells localised in bronchus-associated lymphoid tissue-like structures at day 14. Our results demonstrate that chronic NTHi infection exerts a pro-inflammatory activity in the human and murine lung and could therefore contribute to the exaggerated burden of lung inflammation in patients at risk., The pathological impact of long-term infection by nontypeable Haemophilus influenzae (NTHi) is still debated. Chronic NTHi infection fuels lung inflammation in human samples and in a new mouse model of bacterial long-term persistence. https://bit.ly/3lvyvge

Per la gestione dell’emergenza sanitaria legata alla diffusione del nuovo coronavirus SARS-CoV-2, sono stati emanati diversi decreti volti a contenere il diffondersi del contagio mediante l’impiego di misure urgenti, preventive e cautelari, dirette a coniugare la necessaria prosecuzione delle attività produttive con le garanzie di sicurezza e salubrità degli ambienti lavorativi. Ne sono conseguiti nuovi e significativi obblighi per i datori di lavoro, tenuti a salvaguardare la salute dei propri lavoratori, sia per dovere etico sia perché la posizione di garanzia che rivestono impone – per evitare responsabilità penali – di impedire il verificarsi eventi lesivi. Il contributo intende quindi approfondire i risvolti penali dell’emergenza Covid-19, prendendo in esame i profili di responsabilità delle persone fisiche e giuridiche datrici di lavoro nel caso in cui il contagio avvenga in occasione del lavoro e sia eziologicamente riconducibile ad una loro azione od omissione.

Il contributo si propone di analizzare il reato di avvelenamento di acque, recentemente esaminato dalla giurisprudenza di legittimità nella sentenza n. 25547/2018 e nella pronuncia n. 48548/2018. Entrambe le decisioni prendono dettagliatamente in considerazione il delitto ex art. 439 c.p., contestato nelle due vicende nella forma colposa di cui all’art. 452 c.p. . Le sentenze delineano i tratti caratteristici del reato de quo, qualificandolo come reato di pericolo presunto e concordano circa la sua natura di reato istantaneo ad effetti permanenti, tuttavia divergono circa il momento in cui possa dirsi concluso l’iter criminis e quindi sul dies a quo per il decorso della prescrizione.

Con la sentenza che si annota la suprema Corte esamina il reato di realizzazione e gestione di discarica non autorizzata. L’elaborato si propone di approfondire i principali orientamenti dottri- nali e giurisprudenziali circa la responsabilità del terzo proprietario del fondo per la discarica ivi realizzata da altri, soffermandosi – soprattutto – su quello (condiviso dalla pronuncia in commen- to) che esclude l’esistenza di una posizione di garanzia e di un potere impeditivo in capo al proprietario dell’area interessata dalla discarica, che – al più – potrà essere ritenuto concorrente nel reato.

Il presente contributo – partendo dall’esame di una recente pronuncia della Corte di Cassazione, che ha affermato la penale responsabilità del gestore di un locale pubblico, per aver violato la legge quadro sulle aree protette (L. 349/1991) e provocato nell’esercizio della propria attività, all’interno di un parco naturale, emissioni sonore ad altissimo volume ed emissioni luminose di forte intensità, entrambe idonee a causare un rilevante disturbo all’habitat naturale e agli utenti del parco – si propone di svolgere una disamina del c.d. inquinamento luminoso, su cui però non è ancora intervenuto il legislatore nazionale né per regolare il fenomeno dal punto di vista amministrativo né per sanzionarlo penalmente, indugiando poi sulla rilevanza che potrebbe avere il diritto penale nel contrasto a tale nuova forma di inquinamento.

Highlights • Hyperinflammation in COVID-19 activates blood coagulation increasing thrombotic risk. • Tocilizumab blocks IL-6 receptor and may improve inflammatory-induced hypercoagulability. • Tocilizumab was associated with rapid and sustained coagulation improvement. • These benefits were consistent independently of thromboprophylaxis dose., Background Many COVID-19 patients develop a hyperinflammatory response which activates blood coagulation and may contribute to the occurrence of thromboembolic complications. Blockade of interleukin-6, a key cytokine in COVID-19 pathogenesis, may improve the hypercoagulable state induced by inflammation. The aim of this study was to evaluate the effects of subcutaneous tocilizumab, a recombinant humanized monoclonal antibody against the interleukin-6 receptor on coagulation parameters. Methods Hospitalized adult patients with laboratory-confirmed moderate to critical COVID-19 pneumonia and hyperinflammation, who received a single 324 mg subcutaneous dose of tocilizumab on top of standard of care were enrolled in this analysis. Coagulation parameters were measured before tocilizumab and at day 1, 3, and 7 after treatment. All patients were followed-up for 35 days after admission or until death. Results 70 patients (mean age 60 years, interquartile range 52-75) were included. Treatment with tocilizumab was associated with a reduction in D-dimer levels (-56%; 95% confidence interval [CI], -68% to -44%), fibrinogen (-48%; 95%CI, -60% to -35%), C-reactive protein (-93%; 95%CI, -99% to -87%), prothrombin time (-4%; 95%CI,-9% to 0.8%), and activated thromboplastin time (-4%; 95%CI,-8.7% to 0.8%), and an increase in platelet count (34%; 95%CI, 23% to 45%). These changes occurred already one day after treatment with sustained reductions throughout day 7. The improvement in coagulation was consistently observed in patients receiving prophylactic or therapeutic dose anticoagulants, and was paralleled by a rapid improvement in respiratory function. Conclusions Subcutaneous tocilizumab was associated with significant improvement of blood coagulation parameters independently of thromboprophylaxis dose.

Gliomas are diffusely growing tumours arising from progenitors within the central nervous system. They encompass a range of different molecular types and subtypes, many of which have a well-defined profile of driver mutations, copy number changes and DNA methylation patterns. A majority of gliomas will require surgical intervention to relieve raised intracranial pressure and reduce tumour burden. A proportion of tumours, however, are located in neurologically sensitive areas and a biopsy poses a significant risk of a deficit. A majority of gliomas recur after surgery, and monitoring tumour burden of the recurrence is currently achieved by imaging. However, most imaging modalities have limitations in assessing tumour burden and infiltration into adjacent brain, and sometimes imaging is unable to discriminate between tumour recurrence and pseudo-progression. Liquid biopsies, obtained from body fluids such as cerebrospinal fluid or blood, contain circulating nucleic acids or extracellular vesicles containing tumour-derived components. The studies for this systematic review were selected according to PRISMA criteria, and suggest that the detection of circulating tumour-derived nucleic acids holds great promises as biomarker to aid diagnosis and prognostication by monitoring tumour progression, and thus can be considered a pathway towards personalized medicine.

BackgroundSphingosine-1-phosphate receptor 2 (S1PR2) mediates pleiotropic functions encompassing cell proliferation, survival, and migration, which become collectively de-regulated in cancer. Information on whetherS1PR2participates in colorectal carcinogenesis/cancer is scanty, and we set out to fill the gap.MethodsWe screened expression changes of S1PR2 in human CRC and matched normal mucosa specimens [N = 76]. We compared CRC arising in inflammation-driven and genetically engineered models in wild-type (S1PR2+/+) and S1PR2 deficient (S1PR2−/−) mice. We reconstituted S1PR2 expression in RKO cells and assessed their growth in xenografts. Functionally, we mimicked the ablation of S1PR2 in normal mucosa by treating S1PR2+/+organoids with JTE013 and characterized intestinal epithelial stem cells isolated from S1PR2−/−Lgr5-EGFP- mice.ResultsS1PR2 expression was lost in 33% of CRC; in 55%, it was significantly decreased, only 12% retaining expression comparable to normal mucosa. Both colitis-induced and genetic Apc+/minmouse models of CRC showed a higher incidence in size and number of carcinomas and/or high-grade adenomas, with increased cell proliferation in S1PR2−/−mice compared to S1PR2+/+controls. Loss of S1PR2 impaired mucosal regeneration, ultimately promoting the expansion of intestinal stem cells. Whereas its overexpression attenuated cell cycle progression, it reduced the phosphorylation of AKT and augmented the levels of PTEN.ConclusionsIn normal colonic crypts, S1PR2 gains expression along with intestinal epithelial cells differentiation, but not in intestinal stem cells, and contrasts intestinal tumorigenesis by promoting epithelial differentiation, preventing the expansion of stem cells and braking their malignant transformation. Targeting of S1PR2 may be of therapeutic benefit for CRC expressing high Lgr5.Graphical Abstract. Schematic drawing of the role of S1PR2 in normal mucosa and colorectal cancer. In the normal mucosa, S1PR2 is highly expressed by differentiated cells at the upper region of both colon and intestinal crypts (S1PR2 ON), but not by the undifferentiated stem cell at the base of the crypts (S1PR2 OFF), in which acts as a negative proliferative regulator promoting epithelial differentiation. Its loss leads to the expansion of stem cells and reduced levels of PTEN and Axin-2, two negative regulators respectively of PI3K/AKT and Wnt signaling that control β-catenin signaling. The translocation of β-catenin into the nucleus promotes the transcription of target genes involved in the proliferation and malignant transformation. Thereby, S1PR2 works in the intestine as a tumor suppressor

Highlights • Early treatment with low-dose tocilizumab TCZ (324 mg, administered subcutaneously in two simultaneous 162 mg injections) was used to prevent progression of disease in a subgroup of patients with moderate COVID-19 pneumonia and hyperinflammation. • Low-dose subcutaneous tocilizumab TCZ (324 mg) was well-tolerated with no significant adverse effects. • Low-dose subcutaneous tocilizumab TCZ (324 mg) was associated with a rapid and significant reduction in C reactive protein (CRP) paralleled by a progressive improvement in oxygenation as reflected by the ratio of partial pressure of oxygen to fraction of inspired oxygen (P/F) ratio. • None of the TCZ-treated patients had disease progression, suggesting that IL-6 receptor blockade with subcutaneous TCZ may prevent COVID-19 progression when administered early in the course of disease., Aim The aim of the study was to evaluate the safety and efficacy profile of low-dose tocilizumab (TCZ) administered subcutaneously to patients with moderate COVID-19 pneumonia and hyperinflammation to prevent disease progression. Methods We retrospectively analyzed clinical characteristics and outcomes of patients with laboratory-confirmed bilateral COVID-19 pneumonia, hyperinflammation (C reactive protein ≥20 mg/dl), no hypoxemia (oxygen saturation >90%) and no contra-indications to TCZ, who were treated with subcutaneous TCZ (324 mg) administered within 48 hours from hospitalization on top of standard of care (SOC), and compared them with matched controls treated with SOC only before TCZ was available at our center. Clinical data were available for all patients until death or, for those discharged from hospital, until day 35. Findings Ten consecutive patients (6 males, median age 55 years) treated with TCZ on top of SOC, and ten patients (6 males, median age 56 years) treated with SOC only were included. TCZ was well-tolerated, with no clinically relevant adverse events. TCZ was associated with a reduction in CRP at day 1 (−50%, IQR −28 to −80) and day 3 (−89%, IQR −79 to −96; P = 0.005 for within-group), whereas there was no significant change in CRP values in the SOC group (P

•AbstractNontypeable Haemophilus influenzae (NTHi) is commonly isolated from airway of cystic fibrosis (CF) patients. However, to what extent NTHi persistence contributes to the lung inflammatory burden during CF chronic airway disease is controversial. Here, we aimed at determining the pathological role of NTHi persistence in a cohort of CF patients and in a newly generated mouse model of NTHi persistence.In our study cohort, we found that CF patients chronically colonized by NTHi had significantly higher levels of IL-8 and CXCL1 than those who were not colonized. To better define the impact of NTHi persistence in fuelling inflammatory response, we developed a new mouse model using both laboratory and CF clinical strains. NTHi persistence was associated with chronic inflammation of the lung, characterized by recruitment of neutrophils and cytokine release (KC, G-CFS, IL-6 and IL-17A) at 2 and 14 days postinfection. An increased burden of T cell mediated response (CD4+ and γδ cells) and higher levels of matrix metalloproteinase 9, known to be associated with tissue remodelling, were observed at 14 days post-infection. Of note we found that both CD4+IL-17+ cells and levels of IL-17 cytokine were enriched in mice at advanced stage of NTHi chronic infection. Moreover, by immunohistochemistry we found CD3+, B220+ and CXCL-13+ cells localized in bronchus-associated lymphoid tissue-like structures at day 14.Our results demonstrate that NTHi persistence exerts a pro-inflammatory activity in the human and murine lung, and could therefore contribute to the exaggerated burden of lung inflammation in CF patients.

Chronic diseases such as hypertension and rheumatoid arthritis are persistent ailments that require personalized lifelong therapeutic management. However, the difficulty of adherence to strict dosing schedule compromises therapeutic efficacy and safety. Moreover, the conventional one-size-fits-all treatment approach is increasingly challenged due to the intricacies of inter- and intra-individual variabilities. While accelerated technological advances have led to sophisticated implantable drug delivery devices, flexibility in dosage and timing modulation to tailor precise treatment to individual needs remains an elusive goal. Here we describe the development of a subcutaneously implantable remote-controlled nanofluidic device capable of sustained drug release with adjustable dosing and timing. By leveraging a low intensity electric field to modify the concentration driven diffusion across a nanofluidic membrane, the rate of drug administration can be increased, decreased or stopped via Bluetooth remote command. We demonstrate in vitro the release modulation of enalapril and methotrexate, first-line therapeutics for treatment of hypertension and rheumatoid arthritis, respectively. Further, we show reliable remote communication and device biocompatibility via in vivo studies. Unlike a pulsatile release regimen typical of some conventional controlled delivery systems, our implant offers a continuous drug administration that avoids abrupt fluctuations, which could affect response and tolerability. Our system could set the foundation for an on-demand delivery platform technology for long term management of chronic diseases.

Background High grade non-muscle-invasive bladder cancer (HG-NMIBC) is a heterogeneous disease with variable risk of progression. Urinary microRNAs are promising biomarkers for BC detection and surveillance. Let-7c-5p miRNA, clustered with miR-99a-5p and -125b-5p, is deregulated in cancer, including BC. The aim of this study is to evaluate urinary let-7c cluster expression in Ta/T1 HG-NMIBC patients and its impact on progression-free survival (PFS). Methods Quantitative Real-Time-Polymerase-Chain-Reaction (qRT-PCR) was used to analyze the let-7c cluster expression in 57 urine and 49 neoplastic paired tissue samples prospectively collected from transurethral resection (TUR) HG-NMIBC patients. Twenty urine and 10 bladder tissue samples were collected and analyzed as normal controls. QRT-PCR was also used to detect intra−/extra-cellular let-7c cluster in BC cells. Receiver Operating Characteristic (ROC) curves were used to identify urinary miRNAs cut-off values predicting T-stage and PFS. Uni/multivariable Cox regression was performed to identify predictors of PFS. A nomogram predicting progression risk and a decision curve analysis (DCA) were performed. Results Urinary let-7c was significantly up-regulated in patients compared with controls, while the whole cluster was down-regulated in tumor tissues. Supporting these findings, in vitro comparison of extra−/intra-cellular ratios of cluster levels between BC cells, showed a higher ratio for let-7c in HG-NMIBC versus low-grade cells. Urinary let-7c cluster expression was increased in higher T-stage and was an independent predictor of progression. Lower EORTC-score and downregulation of urinary cluster were predictors of higher PFS on univariable Cox regression, while on multivariable analysis only cluster expression was an independent progression predictor. On DCA, a benefit was evident for patients with a PFS probability > 20%. Conclusions Urinary let-7c cluster evaluation may improve prognosis, identifying patients at risk of progression and addressing early radical treatment.

O presente artigo aborda o direito à vida mediante uma dupla perspectiva. Se de um lado a proteção à vida deve induzir o legislador a introduzir determinadas sanções para protegê-lo contra agressões injustificadas, de outro lado, a pretensão punitiva do Estado não pode exclusivamente consistir, em qualquer caso, na neutralização do ofensor eternamente. Isso pois, haveria uma exploração da condição humana para fins criminais bem como seria contraditório a pena como função reeducadora, conforme determinam a dignidade da pessoa humana e o artigo 27, §3° da Constituição Italiana. Um prejuízo similar deve ser reconhecido no caso da prisão pela vida inteira, como uma pena perpétua – que limita, como será visto, qualquer possibilidade de liberação do condenado e frustração das suas expectativas e esperanças. Particularmente, este trabalho examinará desenvolvimentos jurisprudenciais recentes da Corte Europeia de Direitos Humanos e da Corte Constitucional sobre prisão perpétua, buscando iluminar a ilegalidade não apenas em virtude do chamado “impedimento de prisão perpétua” (como recentemente afirmado pelos tribunais citados), mas, de forma mais geral.

L'elaborato si propone l’esame della disciplina concernente la gestione dei rifiuti da parte delle strutture sanitarie, siano queste pubbliche o private, e le – conseguenti – responsabilità pena- li che possono sorgere in capo alle persone fisiche e giuridiche laddove si ravvisi una scorretta am- ministrazione dei rifiuti prodotti ovvero nel caso in cui si verifichino pregiudizi per la salute pub- blica e l’ambiente. A tal fine si rende necessario un esame delle principali disposizioni normative di riferimento, una menzione degli illeciti che possono essere commessi nel suddetto ambito ed, ancora, un esame del- la giurisprudenza di legittimità.

Electric power can be wirelessly transferred to power supply medical devices, in order to reduce the size of the implants and increase their lifetime. The inductive power transmission, through the interaction between two coils, is substitute by a novel magnetoelectric (ME) technology. In this case, the external coil delivers the energy to a ME receiver, made with an electroactive (e.g piezoelectric) material, bonded between two magnetostrictive layers. The maximum power density obtained is 175 mW/cm3, that means 25 times more than current state of the art. This new technology can increase the power density and overpass the issues of directionality, present in the inductive transmission technique, and open new prospective for wireless powering medical implants.