A simplified genomic profiling approach predicts outcome in metastatic colorectal cancer

The response of metastatic colorectal cancer (mCRC) to the first-line conventional combination therapy is highly variable, reflecting the elevated heterogeneity of the disease. The genetic alterations underlying this heterogeneity have been thoroughly characterized through omic approaches requiring elevated efforts and costs. In order to translate the knowledge of CRC molecular heterogeneity into a practical clinical approach, we utilized a simplified Next Generation Sequencing (NGS) based platform to screen a cohort of 77 patients treated with first-line conventional therapy. Samples were sequenced using a panel of hotspots and targeted regions of 22 genes commonly involved in CRC. This revealed 51 patients carrying actionable gene mutations, 22 of which carried druggable alterations. These mutations were frequently associated with additional genetic alterations. To take into account this molecular complexity and assisted by an unbiased bioinformatic analysis, we defined three subgroups of patients carrying distinct molecular patterns. We demonstrated these three molecular subgroups are associated with a different response to first-line conventional combination therapies. The best outcome was achieved in patients exclusively carrying mutations on TP53 and/or RAS genes. By contrast, in patients carrying mutations in any of the other genes, alone or associated with mutations of TP53/RAS, the expected response is much worse compared to patients with exclusive TP53/RAS mutations. Additionally, our data indicate that the standard approach has limited efficacy in patients without any mutations in the genes included in the panel. In conclusion, we identified a reliable and easy-to-use approach for a simplified molecular-based stratification of mCRC patients that predicts the efficacy of the first-line conventional combination therapy

Developing a participatory process for soil fertility:A case study in an urban area of Italy

Approaches that are transdisciplinary and participatory can help to address complex socio-ecological issues by integrating multiple disciplinary perspectives while taking into account the different needs and experiences of community members and other stakeholders. Despite this promise, such approaches are rarely applied within the scientific community, as researchers and public actors often lack the training, practice and reference cases required to handle the working relationships and translations of terminology, ideas and values across multiple bodies of knowledge. A case study described in this manuscript depicts a group of researchers, artists and citizens consciously engaged in the construction of a transdisciplinary process as part of a 40-day ‘citizen science’ experiment focussed on assessing soil fertility in the urban area of Milan, Italy. The group drew from recognised scientific approaches, applied agronomic methodologies, artistic practices and technological tools, integrating them into a hybrid process of collective and participatory inquiry. As a quantitative outcome of the experiment, a dataset of bio-chemical parameters was generated, which was enriched by agronomic interpretations but also by artistic and reflective materials. Importantly, the process developed transdisciplinary and participatory skills, as it created a potentially replicable procedure of engagement, analysis and presentation for use in other citizen science settings. This article presents the context, the multiple objectives of the research and the applied approach and its timeline. Described in detail are the process of designing and conducting the experiment by involving an extended research community—including both junior and senior researchers—in progressive steps. Quantitative and qualitative results are provided. The findings are meant to contribute case material and methods to inform the advancement of transdisciplinary research approaches within the scientific community as well as examples of ways to transcend the boundaries of science to include artists and community stakeholders. The aspiration is to inform and inspire concrete application of transdisciplinary and participatory methods in concert to address complex socio-environmental challenges

Methylation-associated down-regulation of RASSF1A and up-regulation of RASSF1C in pancreatic endocrine tumors

<p>Abstract</p> <p>Background</p> <p><it>RASSF1A </it>gene silencing by DNA methylation has been suggested as a major event in pancreatic endocrine tumor (PET) but <it>RASSF1A </it>expression has never been studied. The <it>RASSF1 </it>locus contains two CpG islands (<it>A </it>and <it>C</it>) and generates seven transcripts (<it>RASSF1A</it>-<it>RASSF1G</it>) by differential promoter usage and alternative splicing.</p> <p>Methods</p> <p>We studied 20 primary PETs, their matched normal pancreas and three PET cell lines for the (i) methylation status of the <it>RASSF1 </it>CpG islands using methylation-specific PCR and pyrosequencing and (ii) expression of <it>RASSF1 </it>isoforms by quantitative RT-PCR in 13 cases. CpG island A methylation was evaluated by methylation-specific PCR (MSP) and by quantitative methylation-specific PCR (qMSP); pyrosequencing was applied to quantify the methylation of 51 CpGs also encompassing those explored by MSP and qMSP approaches.</p> <p>Results</p> <p>MSP detected methylation in 16/20 (80%) PETs and 13/20 (65%) normal pancreas. At qMSP, 11/20 PETs (55%) and 9/20 (45%) normals were methylated in at least 20% of <it>RASSF1A </it>alleles.</p> <p>Pyrosequencing showed variable distribution and levels of methylation within and among samples, with PETs having average methylation higher than normals in 15/20 (75%) cases (<it>P </it>= 0.01). The evaluation of mRNA expression of <it>RASSF1 </it>variants showed that: i) <it>RASSF1A </it>was always expressed in PET and normal tissues, but it was, on average, expressed 6.8 times less in PET (<it>P </it>= 0.003); ii) <it>RASSF1A </it>methylation inversely correlated with its expression; iii) <it>RASSF1 </it>isoforms were rarely found, except for <it>RASSF1B </it>that was always expressed and <it>RASSF1C </it>whose expression was 11.4 times higher in PET than in normal tissue (<it>P </it>= 0.001). A correlation between <it>RASSF1A </it>expression and gene methylation was found in two of the three PET cell lines, which also showed a significant increase in <it>RASSF1A </it>expression upon demethylating treatment.</p> <p>Conclusions</p> <p><it>RASSF1A </it>gene methylation in PET is higher than normal pancreas in no more than 75% of cases and as such it cannot be considered a marker for this neoplasm. <it>RASSF1A </it>is always expressed in PET and normal pancreas and its levels are inversely correlated with gene methylation. Isoform <it>RASSF1C </it>is overexpressed in PET and the recent demonstration of its involvement in the regulation of the Wnt pathway points to a potential pathogenetic role in tumor development.</p

Clinical Features, Cardiovascular Risk Profile, and Therapeutic Trajectories of Patients with Type 2 Diabetes Candidate for Oral Semaglutide Therapy in the Italian Specialist Care

Introduction: This study aimed to address therapeutic inertia in the management of type 2 diabetes (T2D) by investigating the potential of early treatment with oral semaglutide. Methods: A cross-sectional survey was conducted between October 2021 and April 2022 among specialists treating individuals with T2D. A scientific committee designed a data collection form covering demographics, cardiovascular risk, glucose control metrics, ongoing therapies, and physician judgments on treatment appropriateness. Participants completed anonymous patient questionnaires reflecting routine clinical encounters. The preferred therapeutic regimen for each patient was also identified. Results: The analysis was conducted on 4449 patients initiating oral semaglutide. The population had a relatively short disease duration (42%  60% of patients, and more often than sitagliptin or empagliflozin. Conclusion: The study supports the potential of early implementation of oral semaglutide as a strategy to overcome therapeutic inertia and enhance T2D management

Azimuthal anisotropy of charged jet production in root s(NN)=2.76 TeV Pb-Pb collisions

We present measurements of the azimuthal dependence of charged jet production in central and semi-central root s(NN) = 2.76 TeV Pb-Pb collisions with respect to the second harmonic event plane, quantified as nu(ch)(2) (jet). Jet finding is performed employing the anti-k(T) algorithm with a resolution parameter R = 0.2 using charged tracks from the ALICE tracking system. The contribution of the azimuthal anisotropy of the underlying event is taken into account event-by-event. The remaining (statistical) region-to-region fluctuations are removed on an ensemble basis by unfolding the jet spectra for different event plane orientations independently. Significant non-zero nu(ch)(2) (jet) is observed in semi-central collisions (30-50% centrality) for 20 <p(T)(ch) (jet) <90 GeV/c. The azimuthal dependence of the charged jet production is similar to the dependence observed for jets comprising both charged and neutral fragments, and compatible with measurements of the nu(2) of single charged particles at high p(T). Good agreement between the data and predictions from JEWEL, an event generator simulating parton shower evolution in the presence of a dense QCD medium, is found in semi-central collisions. (C) 2015 CERN for the benefit of the ALICE Collaboration. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).Peer reviewe

Production of He-4 and (4) in Pb-Pb collisions at root(NN)-N-S=2.76 TeV at the LHC

Results on the production of He-4 and (4) nuclei in Pb-Pb collisions at root(NN)-N-S = 2.76 TeV in the rapidity range vertical bar y vertical bar <1, using the ALICE detector, are presented in this paper. The rapidity densities corresponding to 0-10% central events are found to be dN/dy4(He) = (0.8 +/- 0.4 (stat) +/- 0.3 (syst)) x 10(-6) and dN/dy4 = (1.1 +/- 0.4 (stat) +/- 0.2 (syst)) x 10(-6), respectively. This is in agreement with the statistical thermal model expectation assuming the same chemical freeze-out temperature (T-chem = 156 MeV) as for light hadrons. The measured ratio of (4)/He-4 is 1.4 +/- 0.8 (stat) +/- 0.5 (syst). (C) 2018 Published by Elsevier B.V.Peer reviewe

Pseudorapidity and transverse-momentum distributions of charged particles in proton-proton collisions at root s=13 TeV

The pseudorapidity (eta) and transverse-momentum (p(T)) distributions of charged particles produced in proton-proton collisions are measured at the centre-of-mass energy root s = 13 TeV. The pseudorapidity distribution in vertical bar eta vertical bar <1.8 is reported for inelastic events and for events with at least one charged particle in vertical bar eta vertical bar <1. The pseudorapidity density of charged particles produced in the pseudorapidity region vertical bar eta vertical bar <0.5 is 5.31 +/- 0.18 and 6.46 +/- 0.19 for the two event classes, respectively. The transverse-momentum distribution of charged particles is measured in the range 0.15 <p(T) <20 GeV/c and vertical bar eta vertical bar <0.8 for events with at least one charged particle in vertical bar eta vertical bar <1. The evolution of the transverse momentum spectra of charged particles is also investigated as a function of event multiplicity. The results are compared with calculations from PYTHIA and EPOS Monte Carlo generators. (C) 2015 CERN for the benefit of the ALICE Collaboration. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).Peer reviewe

Centrality evolution of the charged-particle pseudorapidity density over a broad pseudorapidity range in Pb-Pb collisions at root s(NN)=2.76TeV

Peer reviewe

Strategie cardioprotettive nella senescenza: modulazione farmacologica del canale mitoBK da parte del flavonoide naringenina

Le patologie cardiovascolari, in particolare l’infarto del miocardio, risultano incrementate con l’avanzare dell’età; in più è stato dimostrato che i mitocondri ed in particolare le variazioni delle funzioni mitocondriali legate all’invecchiamento sono strettamente coinvolte in questo processo. Recentemente è stato dimostrato come alcune sostanze provenienti dal regno vegetale tra cui i flavonoidi, svolgono un’azione cardioprotettiva. Tra questi la Naringenina (NAR), flavonoide presente in vari frutti appartenenti al genere Citrus, possiede interessanti attività cardioprotettive dimostrate attraverso modelli sperimentali in vivo ed ex vivo su animali giovani-adulti. Inoltre è stato caratterizzato il target d’azione di NAR, responsabile almeno in parte, delle sue proprietà benefiche nei modelli di ischemia/riperfusione (I/R) ed è stato individuato il bersaglio nel canale del potassio calcio attivato localizzato a livello mitocondriale (mitoBK). Con questo lavoro di tesi si è valutata l’azione cardioprotettiva di NAR su un modello in vivo di infarto del miocardio (più vicino alla clinica) utilizzando animali senescenti di 10-12 mesi di età, più sensibili al danno da I/R. Inoltre, visto il meccanismo d’azione di NAR sui canali mitoBK, si è investigata l’espressione di tali canali sui mitocondri isolati da tessuto cardiaco senescente e su cellule di cardiomioblasti sottoposte a processo di invecchiamento per esposizione alla doxorubicina. I risultati ottenuti in questa tesi di Laurea dimostrano l’efficacia antiischemica di NAR anche sugli animali senescenti. Infatti, quando somministrata intraperitonealmente alla dose di 100mg/Kg, NAR era capace di ridurre significativamente, in ratti senescenti sottoposti ad infarto acuto del miocardio, l’estensione delle aree ischemiche rispetto all’ estensione totale del ventricolo sinistro; tale effetto è risultato ancora una volta mediato dall’ attivazione dei canali mitoBK, visto che il bloccante selettivo, paxillina, ne aboliva gli effetti benefici. Il secondo obiettivo di questa tesi di Laurea è stato la valutazione dell’espressione delle subunità alfa e beta che compongono il canale mitoBK. Studi di western blot hanno permesso di dimostrare che in condizioni di senescenza, l’ espressione del canale mitoBK risultava essere ridotta ma comunque presente, sia su mitocondri isolati da tessuto cardiaco senescente di ratto sia su mitocondri ottenuti da cellule invecchiate con doxorubicina. Questo rappresenta un dato interessante che getta le basi per successivi studi e ci porta a fare speculazioni su possibili usi nutraceutici di NAR sulle persone più predisposte al danno da I/R, ossia i più anziani

Glycemic control in the coronary care unit: Prognostic value and new therapeutic strategies

Type 2 diabetes and acute coronary syndromes are widely interconnected. Individuals with type 2 diabetes are more likely than non-diabetic subjects to experience silent or manifest episodes of myocardial ischemia as the first presentation of coronary artery disease. Insulin resistance, inflammation, microvascular disease and a tendency to thrombosis are common in these patients. Intensive blood glucose control with intravenous insulin infusion has been demonstrated to significantly reduce morbidity and mortality in critically ill hyperglycemic patients admitted to an intensive care unit. Direct glucose toxicity likely plays a crucial role in explaining the clinical benefits of intensive insulin therapy in such critical patients. However, the difficult implementation of nurse-driven protocols for insulin infusion, able to achieve more rapid and effective blood glucose control without significant episodes of hypoglycemia, has led physicians to consider alternative drugs for this purpose. New intravenous or oral agents include the incretin glucagon-like peptide-1, its analogs, and dipeptidyl peptidase-4 inhibitors, which potentiate the activity of glucagon-like peptide-1 and thus enhance glucose-dependent insulin secretion. Improved glycemic control with protective effects on myocardial and vascular tissue, with lesser side effects and a better therapeutic compliance may represent an important therapeutic potential for this class of drugs in acutely ill patients in general, and in patients with acute coronary syndromes in particular. Such drugs should be known by practicing cardiologists for their possible use in intensive care units in the years to come. © 2008 AIM Publishing Srl