The ESCRT-III machinery participates in the production of extracellular vesicles and protein export during Plasmodium falciparum infection

Infection with Plasmodium falciparum enhances extracellular vesicle (EV) production in parasitized red blood cells (pRBCs), an important mechanism for parasite-to-parasite communication during the asexual intraerythrocytic life cycle. The endosomal sorting complex required for transport (ESCRT), and in particular the ESCRT-III sub-complex, participates in the formation of EVs in higher eukaryotes. However, RBCs have lost the majority of their organelles through the maturation process, including an important reduction in their vesicular network. Therefore, the mechanism of EV production in P. falciparum-infected RBCs remains to be elucidated. Here we demonstrate that P. falciparum possesses a functional ESCRT-III machinery activated by an alternative recruitment pathway involving the action of PfBro1 and PfVps32/PfVps60 proteins. Additionally, multivesicular body formation and membrane shedding, both reported mechanisms of EV production, were reconstituted in the membrane model of giant unilamellar vesicles using the purified recombinant proteins. Moreover, the presence of PfVps32, PfVps60 and PfBro1 in EVs purified from a pRBC culture was confirmed by super-resolution microscopy and dot blot assays. Finally, disruption of the PfVps60 gene led to a reduction in the number of the produced EVs in the KO strain and affected the distribution of other ESCRT-III components. Overall, our results increase the knowledge on the underlying molecular mechanisms during malaria pathogenesis and demonstrate that ESCRT-III P. falciparum proteins participate in EV production

Hematopoietic reconstitution dynamics of mobilized- and bone marrow-derived human hematopoietic stem cells after gene therapy

Mobilized peripheral blood is increasingly used instead of bone marrow as a source of autologous hematopoietic stem/progenitor cells for ex vivo gene therapy. Here, we present an unplanned exploratory analysis evaluating the hematopoietic reconstitution kinetics, engraftment and clonality in 13 pediatric Wiskott-Aldrich syndrome patients treated with autologous lentiviral-vector transduced hematopoietic stem/progenitor cells derived from mobilized peripheral blood (n = 7), bone marrow (n = 5) or the combination of the two sources (n = 1). 8 out of 13 gene therapy patients were enrolled in an open-label, non-randomized, phase 1/2 clinical study (NCT01515462) and the remaining 5 patients were treated under expanded access programs. Although mobilized peripheral blood- and bone marrow- hematopoietic stem/progenitor cells display similar capability of being gene-corrected, maintaining the engineered grafts up to 3 years after gene therapy, mobilized peripheral blood-gene therapy group shows faster neutrophil and platelet recovery, higher number of engrafted clones and increased gene correction in the myeloid lineage which correlate with higher amount of primitive and myeloid progenitors contained in hematopoietic stem/progenitor cells derived from mobilized peripheral blood. In vitro differentiation and transplantation studies in mice confirm that primitive hematopoietic stem/progenitor cells from both sources have comparable engraftment and multilineage differentiation potential. Altogether, our analyses reveal that the differential behavior after gene therapy of hematopoietic stem/progenitor cells derived from either bone marrow or mobilized peripheral blood is mainly due to the distinct cell composition rather than functional differences of the infused cell products, providing new frames of references for clinical interpretation of hematopoietic stem/progenitor cell transplantation outcome.</p

Approaches to interim analysis of cancer randomised clinical trials with time to event endpoints: A survey from the Italian National Monitoring Centre for Clinical Trials

<p>Abstract</p> <p>Background</p> <p>Although interim analysis approaches in clinical trials are widely known, information on current practice of planned monitoring is still scarce. Reports of studies rarely include details on the strategies for both data monitoring and interim analysis. The aim of this project is to investigate the forms of monitoring used in cancer clinical trials and in particular to gather information on the role of interim analyses in the data monitoring process of a clinical trial. This study focused on the prevalence of different types of interim analyses and data monitoring in cancer clinical trials.</p> <p>Methods</p> <p>Source of investigation were the protocols of cancer clinical trials included in the Italian registry of clinical trials from 2000 to 2005. Evaluation was restricted to protocols of randomised studies with a time to event endpoint, such as overall survival (OS) or progression free survival (PFS). A template data extraction form was developed and tested in a pilot phase. Selection of relevant protocols and data extraction were performed independently by two evaluators, with differences in the data assessment resolved by consensus with a third reviewer, referring back to the original protocol. Information was obtained on a) general characteristics of the protocol b) disease localization and patient setting; c) study design d) interim analyses; e) DSMC.</p> <p>Results</p> <p>The analysis of the collected protocols reveals that 70.7% of the protocols incorporate statistical interim analysis plans, but only 56% have also a DSMC and be considered adequately planned. The most concerning cases are related to lack of any form of monitoring (20.0% of the protocols), and the planning of interim analysis, without DSMC (14.7%).</p> <p>Conclusion</p> <p>The results indicate that there is still insufficient attention paid to the implementation of interim analysis.</p

The impact of chest CT body composition parameters on clinical outcomes in COVID-19 patients

We assessed the impact of chest CT body composition parameters on outcomes and disease severity at hospital presentation of COVID-19 patients, focusing also on the possible mediation of body composition in the relationship between age and death in these patients. Chest CT scans performed at hospital presentation by consecutive COVID-19 patients (02/27/2020-03/13/2020) were retrospectively reviewed to obtain pectoralis muscle density and total, visceral, and intermuscular adipose tissue areas (TAT, VAT, IMAT) at the level of T7-T8 vertebrae. Primary outcomes were: hospitalization, mechanical ventilation (MV) and/or death, death alone. Secondary outcomes were: C-reactive protein (CRP), oxygen saturation (SO2), CT disease extension at hospital presentation. The mediation of body composition in the effect of age on death was explored. Of the 318 patients included in the study (median age 65.7 years, females 37.7%), 205 (64.5%) were hospitalized, 68 (21.4%) needed MV, and 58 (18.2%) died. Increased muscle density was a protective factor while increased TAT, VAT, and IMAT were risk factors for hospitalization and MV/death. All these parameters except TAT had borderline effects on death alone. All parameters were associated with SO2 and extension of lung parenchymal involvement at CT; VAT was associated with CRP. Approximately 3% of the effect of age on death was mediated by decreased muscle density. In conclusion, low muscle quality and ectopic fat accumulation were associated with COVID-19 outcomes, VAT was associated with baseline inflammation. Low muscle quality partly mediated the effect of age on mortality.We assessed the impact of chest CT body composition parameters on outcomes and disease severity at hospital presentation of COVID-19 patients, focusing also on the possible mediation of body composition in the relationship between age and death in these patients. Chest CT scans performed at hospital presentation by consecutive COVID-19 patients (02/ 27/2020-03/13/2020) were retrospectively reviewed to obtain pectoralis muscle density and total, visceral, and intermuscular adipose tissue areas (TAT, VAT, IMAT) at the level of T7-T8 vertebrae. Primary outcomes were: hospitalization, mechanical ventilation (MV) and/or death, death alone. Secondary outcomes were: C-reactive protein (CRP), oxygen saturation (SO2), CT disease extension at hospital presentation. The mediation of body composition in the effect of age on death was explored. Of the 318 patients included in the study (median age 65.7 years, females 37.7%), 205 (64.5%) were hospitalized, 68 (21.4%) needed MV, and 58 (18.2%) died. Increased muscle density was a protective factor while increased TAT, VAT, and IMAT were risk factors for hospitalization and MV/death. All these parameters except TAT had borderline effects on death alone. All parameters were associated with SO2 and extension of lung parenchymal involvement at CT; VAT was associated with CRP. Approximately 3% of the effect of age on death was mediated by decreased muscle density. In conclusion, low muscle quality and ectopic fat accumulation were associated with COVID-19 outcomes, VAT was associated with baseline inflammation. Low muscle quality partly mediated the effect of age on mortality

Ruolo dell'espressione dei geni mts1 e nm23 nelle variazioni della capacita' metastatica di tumori mammari umani

Dottorato di ricerca in scienze biologiche e molecolari applicate alla odontostomatologia. 9. ciclo. A.a. 1995-96. Coordinatore B. Venerando. Docente guida F. CajoneConsiglio Nazionale delle Ricerche - Biblioteca Centrale - P.le Aldo Moro, 7, Rome; Biblioteca Nazionale Centrale - P.za Cavalleggeri, 1, Florence / CNR - Consiglio Nazionale delle RichercheSIGLEITItal

Introduzione alla World Literature. Percorsi e prospettive

Nell’ultimo capitolo del saggio La letteratura postcoloniale. Dall’Impero alla World Literature (Carocci, 2013), Silvia Albertazzi ribadiva la necessità di superare il concetto di “letteratura postcoloniale” per arrivare a definire “una letteratura senza frontiere, che contamina stili, generi e tematiche, in cui si attua uno scambio fecondo tra periferia e centro, in un contagio positivo senza soluzione di continuità”. Questo volume, che nasce dal lavoro congiunto dei membri del CLOPEx (Centro studi sulle letterature omeoglotte dei paesi extraeuropei) dell’Università di Bologna, si propone di identificare la World Literature, esaminando le varie accezioni attribuite al termine nel corso del tempo e alle varie latitudini, le prese di posizione di studiosi e critici, nel passato e nel presente, i suoi rapporti con la globalizzazione, la storia, la memoria, l’oralità e le voci indigene, la traduzione, il mercato editoriale. Si articola in sette brevi capitoli e una introduzione: Introduzione metodologico/teorica 1) World Literature come super-canone/Letterature dal mondo 2) WL, etica e globalizzazione 3) World Poetry 4) WL, trauma, storia, memoria 5) World Literature e Sud Globale. Una questione aperta 6) Le riscritture globali 7) Transmedialità e WL A una puntuale introduzione metodologica sullo stato dell’arte, ad opera di Silvia Albertazzi, farà seguito un primo capitolo, sempre a mia firma, sulla World Literature intesa come “canone dei canoni”, ovvero scelta delle opere migliori da tutto il mondo. Si affrontano di conseguenza in questo capitolo anche problemi di canonizzazione e politiche traduttive/editoriali. Elena Lamberti indaga la WL rispetto alle grandi sfide del contemporaneo, che sono prima di tutto sociali e, poi, anche letterarie: neocolonialismi, omologazione culturale, migrazioni (forzate o meno), accessibilità alle risorse, sostenibilità (non solo ambientale), controllo e vulnerabilità, diversità, nuove identità transumane o postumane. Francesco Benozzo si concentra sulla poesia: rilevando come tutti gli studiosi di WL abbiano lavorato essenzialmente sul romanzo, Benozzo identifica le caratteristiche di una possibile World Poetry. Edoardo Balletta si occupa del rapporto della World Literature con i Sud del mondo, fino al rinvenimento di un ipotetico rinvenimento di un “Sud globale”. Maria Chiara Gnocchi si dedica alle riscritture "globali", che inneggiano ad un'infinita pratica di riscrittura, che non è più solo quella "uno contro uno" tra autori postcoloniali e capolavori del canone occidentali, segno di una nuova fase della letteratura che è diventata mondiale e che nasce in un momento storico che ha superato decolonizzazione e postcolonialismo. Francesco Vitucci, infine, offre un particolarissimo case study di transmedialità giapponese

Caring for health-care workers. Experience with a psychological support program for nurses working in Internal Medicine

IntroductionNurses working in an Internal Medicine ward must have very specific training and aptitude. Dealing with different types of patients with widely varying ages and different medical issues requires flexibility in managing emergencies and in choosing between various professional interventions, as well as strong communication skills. Because of this variety, the workload is perceived as being particularly heavy.Materials and methodsThe article describes the intervention of a psychologist in support of the nursing staff of an Internal Medicine ward. The intervention was prompted by findings of high staff turnover. The work began with an analysis of the group dynamics in the nursing team, and the psychologist's action was based on a group approach. In this way, specific problems of the group were solved through the instrument of the group itself, which became the true promoter of change.ResultsNurses worked to recognize their professional identity and to strengthen their self-esteem, and this changed their perception of their workload. The team also became more aware of its individual and group resources. These changes decreased staff turnover and reduced arguments between the nurses themselves and between the nurses and patients’ relatives.DiscussionThe nursing team become more solid and better organized. It dealt with emotional problems and has become more receptive to changes in the way the work is organized.</p

Caring for health-care workers. Experience with a psychological support program for nurses working in Internal Medicine

IntroductionNurses working in an Internal Medicine ward must have very specific training and aptitude. Dealing with different types of patients with widely varying ages and different medical issues requires flexibility in managing emergencies and in choosing between various professional interventions, as well as strong communication skills. Because of this variety, the workload is perceived as being particularly heavy.Materials and methodsThe article describes the intervention of a psychologist in support of the nursing staff of an Internal Medicine ward. The intervention was prompted by findings of high staff turnover. The work began with an analysis of the group dynamics in the nursing team, and the psychologist's action was based on a group approach. In this way, specific problems of the group were solved through the instrument of the group itself, which became the true promoter of change.ResultsNurses worked to recognize their professional identity and to strengthen their self-esteem, and this changed their perception of their workload. The team also became more aware of its individual and group resources. These changes decreased staff turnover and reduced arguments between the nurses themselves and between the nurses and patients’ relatives.DiscussionThe nursing team become more solid and better organized. It dealt with emotional problems and has become more receptive to changes in the way the work is organized.</p

2-D proteomic analysis of urine in diabetic patients with nephropathy

Mortality and morbidity associated with diabetes is diabetic nephropathy. We carried out a research project on the identification of specific proteomic profiles in patients with diabetic nephropathy. The rationale for this study is based on studies demonstrating that chronic inflammation and oxidative stress determine the complications observed in diabetic patients. Such a pathogenesis will determine post-translational protein modification, bringing to the identification of prognostic and predictive markers. Recent advances in proteomic profiling technologies have allowed preliminary profiling. Various specific proteomic profiles have been described for various pathologies, in particular tumors. Studies on proteomic profiling in diabetic nephropathy have not been performed yet. We analyzed easily available biological samples such as urine to evaluate the expression and function levels of urinary molecules that could become diagnostic, prognostic and predictive markers of disease