Nomogramma clinico-patologico predittivo dello stato mutazionale di BRAF nel tumore del colon-retto metastatico

Negli ultimi anni i progressi ottenuti nel trattamento del carcinoma del colon-retto metastatico (mCRC) hanno prolungato notevolmente l’aspettativa di vita dei pazienti, con valori mediani di sopravvivenza che oggi sono vicini ai 30 mesi. Ha certamente contribuito a questo risultato l’introduzione di alcuni farmaci a bersaglio molecolare, tra cui gli anticorpi monoclonali anti-epidermal growth factor receptor (EGFR). Poiché non tutti i pazienti ottengono beneficio da tali farmaci, è oggi necessario selezionare la popolazione da trattare attraverso l’analisi dello stato mutazionale dei geni RAS (KRAS e NRAS). I pazienti che presentano una mutazione a carico di questi geni non traggono beneficio dal ricevere i farmaci anti-EGFR e devono quindi essere esclusi dal trattamento. Il ruolo della mutazione V600E del gene BRAF, mutuamente esclusiva con le mutazioni di RAS, nel predire la risposta ai farmaci anti-EGFR è tutt’ora dibattuto e gli studi a riguardo non risultano conclusivi, anche a causa della bassa incidenza di questa mutazione nel mCRC (circa 10%). Tutti gli studi concordano invece sul valore prognostico negativo di questa mutazione che si associa a una sopravvivenza mediana di circa dodici mesi. Conoscere lo stato mutazionale di BRAF è un valore aggiunto nella scelta del miglior iter terapeutico per ogni paziente, spingendo a scegliere un trattamento di partenza più aggressivo quando possibile e a ponderare con molta attenzione interventi di resezione secondaria, spesso non risolutivi. Tuttavia, il test di BRAF non è disponibile né comunemente eseguito in tutte le realtà cliniche, con sensibili differenze geografiche. In letteratura sono riportate delle caratteristiche clinico-patologiche che si osservano più frequentemente nei mCRC BRAF mutati. Tuttavia, la forza dell’associazione tra ogni singola caratteristica e la mutazione di BRAF in un modello multivariato non è mai stata valutata. L’obiettivo di questa tesi è quello di costruire un nomogramma predittivo dello stato mutazionale di BRAF nei pazienti con tumore del colon-retto RAS wild-type, partendo dalle caratteristiche clinico-patologiche associate alla mutazione. Sono state analizzate due ampie popolazioni di pazienti con mCRC, una popolazione training (TP) e una popolazione di validazione (VP) di cui erano disponibili dati clinici e patologici inclusi i risultati delle analisi mutazionali dei geni RAS e BRAF. E’ stato creato un database specifico selezionando le caratteristiche già presenti in letteratura associate alla mutazione V600E del gene BRAF: sesso, età, performance status (secondo ECOG), sede del tumore primitivo, precedente resezione del tumore primitivo, istologia mucinosa, grado di differenziazione, tempo all’insorgenza delle metastasi (sincrono o metacrono), numero di siti metastatici alla diagnosi, metastasi peritoneali, metastasi ai linfonodi a distanza, metastasi polmonari. Nei 596 pazienti della TP 281 erano RAS wild-type (47%); 54 BRAFV600E mutati (9.1%) e le due mutazioni erano sempre mutuamente esclusive. Nella popolazione RAS wild-type i fattori predittivi della mutazione di BRAF erano la sede destra del tumore primitivo (OR: 7.80, 95% CI 3.05-19.92); il sesso femminile (OR: 2.90, 95% CI 1.14-7.37) e l’istologia mucinosa (OR: 4.95, 95% CI 1.90-12.90). Questi fattori si replicavano alla cross-validazione interna con tassi rispettivamente al 100%, 93% e 98%. Sulla base di queste evidenze è stato quindi costruito il nomogramma predittivo: i pazienti con sede destra del tumore primitivo, sesso femminile e istologia mucinosa avevano l’81% di probabilità di avere la mutazione BRAF. All’analisi ROC l’accuratezza predittiva del nomogramma era alta (AUC: 0.812, SE: 0.034) con una sensibilità dell’81.2% e una specificità del 72.1%. Nei 508 pazienti della VP 262 erano RAS wild-type (51.6%) e 49 BRAFV600E mutati (9.6%). La sede destra del tumore primitivo, il sesso femminile e l’istologia mucinosa si sono confermati fattori predittivi indipendenti della mutazione di BRAF. Il nomogramma predittivo derivato nella TP è stato quindi applicato alla VP. Anche nella VP, all’analisi ROC, l’accuratezza predittiva del nomogramma è stata alta (AUC=0.811, SE: 0.041), con una sensibilità del 73.5% e una specificità dell’80.3%, senza nessuna differenza significativa tra le prestazioni predittive del nomogramma nelle due popolazioni (p=1.0). In conclusione, tre semplici caratteristiche ampiamente disponibili consentono di predire lo stato mutazionale di BRAF con alta sensibilità e specificità. Sebbene il test molecolare resti chiaramente il gold standard per stabilire se un mCRC è BRAF mutato o meno, questo strumento può essere estremamente utile laddove il test non sia effettivamente disponibile o non venga adottato su larga scala

Strontium substituted hydroxyapatite with β-lactam integrin agonists to enhance mesenchymal cells adhesion and to promote bone regeneration

Multi-functionalization of calcium phosphates to get delivery systems of therapeutic agents is gaining increasing relevance for the development of functional biomaterials aimed to solve problems related to disorders of the muscolo-skeletal system. In this regard, we functionalized Strontium substituted hydroxyapatite (SrHA) with some β-lactam integrin agonists to develop materials with enhanced properties in promoting cell adhesion and activation of intracellular signaling as well as in counteracting abnormal bone resorption. For this purpose, we selected two monocyclic β-lactams on the basis of their activities towards specific integrins on promoting cell adhesion and signalling. The amount of β-lactams loaded on SrHA could be modulated on changing the polarity of the loading solution, from 3.5–24 wt% for compound 1 and from 3.2–8.4 wt% for compound 2. Studies on the release of the β-lactams from the functionalized SrHA in aqueous medium showed an initial burst followed by a steady-release that ensures a small but constant amount of the compounds over time. The new composites were fully characterized. Co-culture of human primary mesenchymal stem cells (hMSC) and human primary osteoclast (OC) demonstrated that the presence of β-lactams on SrHA favors hMSC adhesion and viability, as well as differentiation towards osteoblastic lineage. Moreover, the β-lactams were found to enhance the inhibitory role of Strontium on osteoclast viability and differentiation

Higher protein intake is associated with improved muscle strength in elite senior athletes

OBJECTIVE: The optimal protein intake for elderly individuals who exercise regularly has not yet been clearly defined. The aim of this study was to test the hypothesis that protein intake level is associated with muscle strength in elderly elite athletes. METHODS: We evaluated 50 elite senior athletes (38 men and 12 women) participating in the European Master Games 2011 in an observational cross-sectional study. Participants were divided into two groups-lower (LPI) or higher (HPI) protein intake-according to the median value of their ratio of urinary urea nitrogen to urinary creatinine (i.e., 8.8 g/L), as a marker of protein intake. A dietary interview confirmed differences in protein consumption between the LPI and HPI groups. We also evaluated body composition (bioimpedance), muscle strength, and hematochemical indices. RESULTS: LPI and HPI groups were homogeneous for age (72 [68-74] and 71 [68-74] y, respectively), fat-free mass index (18.4 [17-19.4] and 18.2 [17-19.1] kg/m2), body fat (18.3% [12.3-20.7%] and 16.6% [13.6-21.2%]), and glomerular filtration rate (57.7 [53.8-64.9] and 62.7 [56.1-69.3] mL/min/1.73 m2). The HPI group showed greater leg and trunk muscle strength (N) compared with the LPI group (left leg extension, 339 [238-369] versus 454 [273-561], respectively, P < 0.05; right leg extension, 319 [249-417] versus 432 [334-635], P 64 0.05; trunk extension, 435 [370-467] versus 464 [390-568], P 64 0.05). CONCLUSIONS: Higher protein intake in elite senior athletes is associated with a greater muscle strength

Higher risk for influenza-associated pulmonary aspergillosis (IAPA) in asthmatic patients: A Swiss multicenter cohort study on IAPA in critically ill influenza patients

Background: Influenza-associated pulmonary aspergillosis (IAPA) is an important complication of severe influenza with high morbidity and mortality. Methods: We conducted a retrospective multicenter study in tertiary hospitals in Switzerland during 2017/2018 and 2019/2020 influenza seasons. All adults with PCR-confirmed influenza infection and treatment on intensive-care unit (ICU) for >24 h were included. IAPA was diagnosed according to previously published clinical, radiological, and microbiological criteria. We assessed risk factors for IAPA and predictors for poor outcome, which was a composite of in-hospital mortality, ICU length of stay ≥7 days, mechanical ventilation ≥7 days, or extracorporeal membrane oxygenation. Results: One hundred fifty-eight patients (median age 64 years, 45% females) with influenza were included, of which 17 (10.8%) had IAPA. Asthma was more common in IAPA patients (17% vs. 4% in non-IAPA, P = 0.05). Asthma (OR 12.0 [95% CI 2.1-67.2]) and days of mechanical ventilation (OR 1.1 [1.1-1.2]) were associated with IAPA. IAPA patients frequently required organ supportive therapies including mechanical ventilation (88% in IAPA vs. 53% in non-IAPA, P = 0.001) and vasoactive support (75% vs. 45%, P = 0.03) and had more complications including ARDS (53% vs. 26%, P = 0.04), respiratory bacterial infections (65% vs. 37%, P = 0.04), and higher ICU-mortality (35% vs. 16.4%, P = 0.05). IAPA (OR 28.8 [3.3-253.4]), influenza A (OR 3.3 [1.4-7.8]), and higher SAPS II score (OR 1.07 [1.05-1.10]) were independent predictors of poor outcome. Interpretation: High clinical suspicion, early diagnostics, and therapy are indicated in IAPA because of high morbidity and mortality. Asthma is likely an underappreciated risk factor for IAPA. Keywords: asthma; influenza; influenza-associated aspergillosis; intensive care medicine; invasive aspergillosis

Higher risk for influenza-associated pulmonary aspergillosis (IAPA) in asthmatic patients: A Swiss multicenter cohort study on IAPA in critically ill influenza patients.

BACKGROUND Influenza-associated pulmonary aspergillosis (IAPA) is an important complication of severe influenza with high morbidity and mortality. METHODS We conducted a retrospective multicenter study in tertiary hospitals in Switzerland during 2017/2018 and 2019/2020 influenza seasons. All adults with PCR-confirmed influenza infection and treatment on intensive-care unit (ICU) for >24 h were included. IAPA was diagnosed according to previously published clinical, radiological, and microbiological criteria. We assessed risk factors for IAPA and predictors for poor outcome, which was a composite of in-hospital mortality, ICU length of stay ≥7 days, mechanical ventilation ≥7 days, or extracorporeal membrane oxygenation. RESULTS One hundred fifty-eight patients (median age 64 years, 45% females) with influenza were included, of which 17 (10.8%) had IAPA. Asthma was more common in IAPA patients (17% vs. 4% in non-IAPA, P = 0.05). Asthma (OR 12.0 [95% CI 2.1-67.2]) and days of mechanical ventilation (OR 1.1 [1.1-1.2]) were associated with IAPA. IAPA patients frequently required organ supportive therapies including mechanical ventilation (88% in IAPA vs. 53% in non-IAPA, P = 0.001) and vasoactive support (75% vs. 45%, P = 0.03) and had more complications including ARDS (53% vs. 26%, P = 0.04), respiratory bacterial infections (65% vs. 37%, P = 0.04), and higher ICU-mortality (35% vs. 16.4%, P = 0.05). IAPA (OR 28.8 [3.3-253.4]), influenza A (OR 3.3 [1.4-7.8]), and higher SAPS II score (OR 1.07 [1.05-1.10]) were independent predictors of poor outcome. INTERPRETATION High clinical suspicion, early diagnostics, and therapy are indicated in IAPA because of high morbidity and mortality. Asthma is likely an underappreciated risk factor for IAPA

The role of the WISE Consortium in the European DEDIPAC-KH project

WISE (Wellness, nutrItion, Sport and Exercise prevention) is a research consortium including five Italian research teams (University of Turin, University of Milan, University of Trieste, Universiy of Rome “Foro Italico”, University of Bari), operating within the broader framework of the DEDIPAC-KH joint action (Determinants of Diet and Physical Activity Knowledge Hub). Research actions within the WISE consortium, funded by the Italian Ministry of Higher Education & Research, are in line with the main objective of the DEDIPAC-KH of developing an international and interdisciplinary network of researchers on dietary, physical activity and sedentary behaviours, related determinant research and policy interventions. More specifically, the WISE consortium research aimed to contribute to the following task (1.2.4 - Task Leader: Prof. Alan Donnelly): perform SLRs to identify state-of the art methods for physical activity and sedentary behaviour measurements. The focus of task 1.2.4 was to examine the methodological effectiveness (validity, reliability and sensitivity/responsiveness) of measures of physical activity and sedentary behaviours. The approach taken with this task was to examine the methodological effectiveness of measures of physical activity and sedentary behaviours in two populations; i) child/adolescence and ii) adults. Findings on methodological effectiveness of measures of physical activity and sedentary behaviours constitute the basis for a variety of publication and reports, and conference communications. The DEDIPAC-KH project created an unique opportunity for developing a comprehensive analysis on the determinants of diet and physical activity in Italy, and fostered successful collaboration with leading international groups. The findings of the WISE project created valuable information for the implementation of successful policies in Italy.This work was supported by MIUR

Admixture into and within sub-Saharan Africa

Similarity between two individuals in the combination of genetic markers along their chromosomes indicates shared ancestry and can be used to identify historical connections between different population groups due to admixture. We use a genome-wide, haplotype-based, analysis to characterise the structure of genetic diversity and gene-flow in a collection of 48 sub-Saharan African groups. We show that coastal populations experienced an influx of Eurasian haplotypes over the last 7000 years, and that Eastern and Southern Niger-Congo speaking groups share ancestry with Central West Africans as a result of recent population expansions. In fact, most sub-Saharan populations share ancestry with groups from outside of their current geographic region as a result of gene-flow within the last 4000 years. Our in-depth analysis provides insight into haplotype sharing across different ethno-linguistic groups and the recent movement of alleles into new environments, both of which are relevant to studies of genetic epidemiology