Hermeneutic single case efficacy design: A systematic review of published research and current standards

open4siThis article systematically reviews the methodological characteristics of Hermeneutic Single Case Efficacy Design (HSCED) studies published in peer-reviewed journals. HSCED provides researchers with a flexible and viable alternative to both between-groups and within-subject experimental designs. This article includes a description of the evolution of the methodology distinctive to HSCED; a discussion of results of HSCED studies considered within a framework of contemporary standards and guidelines for systematic case study research; a presentation of recommendations for key characteristics (e.g., diagnosis, hermeneutic analysis, adjudication procedure). Overall, the aim is provide researchers and reviewers with a resource for conducting and evaluating HSCED research. The results of a systematic review of 13 studies suggests that published HSCED research meets contemporary criteria for systematic case study research. Hermeneutic analysis and adjudication emerged as areas of HSCED practice characterized by a diversity of procedures. Although consensus exists along key dimensions of HSCED, there remains a need for further evaluation of adjudication procedures and reporting standards.openBenelli, Enrico; De Carlo, Alessandro; Biffi, Diana; Mcleod, JohnBenelli, Enrico; De Carlo, Alessandro; Biffi, Diana; Mcleod, Joh

Tailored Exercise with an Innovative Mechanical Device: Effects on Cervical-Dorsal Rachis

3 volunteers for magnetic resonance imaging (MRI) data, collected before and after a training perio

Beyond Total Cost of Management (TCM) to Systemic Value Management (SVM): Transformational Trends and a Research Manifesto for an Evolving Discipline

Proposizione di un modello evolutivo della disciplina dell'ingegneria dei costi applicata ad opere a ciclo di vita intera, con ipotesi di ridefinizione dei criteri di valutazione del valore dell'opera da logiche strettamente economico-finanziare a logiche adottanti criteri di valore qualitativi.Total cost management (TCM) has developed as a systematic approach to managing resources, costs, profitability, and risks throughout the lifecycle of any enterprise, program, facility, project, product or service. However, a number of trends are today creating a new socio-technical scenario, characterized by increasing volatility, uncertainty, complexity, and ambiguity (VUCA), which is affecting the strategic scope and applicative dimensions of TCM. A logic of sustainability and multi-stakeholder value is increasingly required to account for the competing and multidimensional needs of customers, employees, partners, and large stakeholder ecosystems. This article presents a review of cross-disciplinary literature and the use of authors’ engagement and consolidated expertise in the field to drive a group model building process aimed to design a conceptual framework and a research manifesto for the evolving TCM discipline. The study provides a classification of nine major trends and evaluates the impact of those trends on a number of TCM dimensions. Next, a research agenda is showed, including nine trajectories for scholars and practitioners engaged to support the evolution of TCM towards a new idea of systemic value management (SVM). The study advances the current knowledge on value-based and sustainable approaches to management and offers to experts and practitioners a basis to implement innovative development projects in the field of TCM

Cardiac resynchronization therapy in persistent left superior vena cava: Can you do it two-leads-only?

Cardiac resynchronization therapy in persistent left superior vena cava: Can you do it two-leads-only

690. Permanent Epigenetic Silencing of Human Genes With Artificial Transcriptional Repressors

There are several diseases whereby the goal of gene therapy is to silence rather than replace a gene function. Paradigmatic examples are diseases caused by a dominant negative mutation or those in which silencing of a host gene confers resistance to a pathogen or compensates the function of the missing gene. Yet, gene silencing can be used to enhance efficacy of cell therapy and for biotechnological applications. Until now, two technologies have been used to silence gene expression, namely RNA interference with short harping RNAs (shRNA) and gene disruption with Artificial Nucleases (ANs). Although some promising pre-clinical and clinical data have been already obtained, the low efficiency of knock-down with shRNA and of biallelic disruption with ANs may limit efficacy of these treatments, especially when residual gene activity can exert a biological function. To overcome this issue, we have developed a novel modality of gene silencing that exploits endogenous epigenetic mechanisms to convey robust and heritable states of repression at the desired target gene. We have generated Artificial Transcriptional Repressors (ATRs), chimeric proteins containing a custom-made DNA binding domain fused to the effector domain of a chromatinmodifying enzyme involved in silencing of Endogenous RetroViruses (ERVs). By performing iterative rounds of selection in human cell lines and primary cells engineered to report for synergistic activity of candidate effector domains, we identified a combination of 3 domains that, when transiently co-assembled on the promoter of the reporter cassette, fully abrogated transgene expression in up to 90% of treated cells. Importantly, silencing was maintained for more than 250 days in cultured cell lines, was resistant to in vitro differentiation or metabolic activation of primary cells, and was confined to the reporter cassette. Silencing was associated with high levels of de novo DNA methylation at the targeted locus and was dependent on this epigenetic mark for its propagation. Finally, transient transfection of 3 ATRs targeted to the promoter region of the Beta-2-microglobulin (B2M) gene resulted in the loss of surface expression of B2M and, consequently, of the MHC-I molecules in up to 80% of treated cells. This phenotype was associated with a switch in the epigenetic and transcriptional state of the constitutively active B2M gene, which became highly decorated with DNA methylation and deprived of RNA PolII and of its transcript. Of note, silencing was resistant to IFN-γ treatment, a potent B2M inducer. Overall, these data provide the first demonstration of efficient and stable silencing of an endogenous gene upon transient delivery of ATRs. This result was made possible by repurposing the machinery involved in silencing of ERVs, which instructs self-sustaining repressive epigenetic states on the gene of interest. While silencing of B2M might be used to generate universally transplantable allogeneic cells, our hit-and-run strategy provides a powerful new alternative to conventional gene silencing for the treatment of several diseases. (LN & AL co-authorship

729 inheritable silencing of endogenous gene by hit and run targeted epigenetic editing

Gene silencing holds great promise for the treatment of several diseases and can be exploited to investigate gene function and activity of the regulatory genome. Here, we develop a novel modality of gene silencing that exploits epigenetics to achieve stable and highly efficient repression of target genes. To this end, we generated Artificial Transcriptional Repressors (ATRs), chimeric proteins containing a custom-made DNA binding domain fused to the effector domain of chromatin-modifying enzymes involved in silencing process of Endogenous RetroViruses (ERVs). By performing iterative rounds of selection in cells engineered to report for synergistic activity of candidate effector domains, we identified a combination of 3 domains (namely KRAB, DNMT3A and DNMT3L) that, when transiently co-assembled on the promoter of the reporter cassette, recreate a powerful embryonic-specific repressive complex capable of inducing full and long-term (>150 days) silencing of transgene expression in up to 90% of the cells. The ATR-induced silencing was cell type and locus independent, and resistant to metabolic activation of the cells. Importantly, these findings were holding true also for endogenous genes embedded in their natural chromatin context, as shown for the highly and ubiquitously expressed B2M gene. Here, transient co-delivery of TALE-based ATRs resulted in loss of surface expression of B2M and, consequently, of the MHC-I molecules in up to 80% of the cells. This phenotype was associated with a drastic switch in the epigenetic and transcriptional state of the constitutively active B2M promoter, which become highly decorated with de novo DNA methylation and deprived of RNAP II. Importantly, silencing was sharply confined to the targeted gene and resistant to INF-γ, a potent natural activator of B2M. We further extended these studies by showing that our silencing approach is portable to the CRISPR/dCas9 DNA binding technology. In this setting, comparable levels of B2M silencing (up to 80%) were achieved using either pools or even individual sgRNAs coupled to dCas9-based ATRs. Yet, adoption of this technology allowed performing simultaneous, highly efficient multiplex gene silencing within the same cell, as shown for B2M, IFNAR1 and VEGFA. Finally, we assessed resistance of the silenced gene to activity of potent artificial transcription activators and chromatin remodelers, and found that only targeted DNA demethylation was able to reawaken the silent gene. This allowed performing iterative cycles of silencing and reactivation of the same gene in the same cell population. Overall, these data provide the first demonstration of efficient and stable epigenetic silencing of endogenous genes upon transient delivery of ATRs. This was accomplished by repurposing the ERVs silencing machinery, which instructs self-sustaining repressive epigenetic states to the target gene. While silencing of B2M might be used to generate universally transplantable allogeneic cells, our hit-and-run strategy provides a powerful new alternative to conventional gene silencing for both basic and translational research

A new tool for investigation platelet activation in endometriosis patients

Objectives: Endometriosis (EM) is a gynecological disease characterized by chronic inflammation, due to the interaction of inflammatory cells with ectopic endometrium (1). Platelets (PLTs), recruited by procoagulant factors released from endometriotic stromal cells, secrete angiogenetic factors and induce overexpression of genes involved in pro-survival/ anti-apoptotic propensity, inflammationand extracellular matrix remodeling (2). We aimed to develop a tool to measure PLT activation (by small extracellular vesicles, s-EVs) in EM peritoneal fluids, as a potential predictive marker of EM severity. Materials & methods: S-EVs were isolated from EM peritoneal fluids and characterized with imaging (Atomic Force Microscopy; AFM) and protein expression analyses (Western blot, WB) (3). We explored gene expression in peritoneum and EM lesions using EndometDB (4). Results: We demonstrated the presence of s-EVs isolated from EM peritoneal fluids by liquid AFM, as showed by contact angle vs diameter scatterplot (Fig.1A-B), and by WB detecting the s-EV markers CD63, CD9, and TSG101 (Fig.1C). Using Endomet-DB, we highlighted the differentially expressed genes between control and EM peritoneum samples (Fig.1D). The protein expression of a panel of biomarkers of PTL in s-EVs was further confirmed by WB (Fig.1E). Conclusions: We propose applying s-EV research to EM investigation, generating a novel biochemical tool for PLT activation assessment and for the development of new diagnostics and therapies

The effects of gender on electrical therapies for the heart: physiology, epidemiology, and access to therapies: A report from the XII Congress of the Italian Association on Arrhythmology and Cardiostimulation (AIAC)

The difference between men and women is clear even just by looking at an electrocardiogram: females present higher resting heart rate, a shorter QRS complex length and greater corrected QT interval. The development of these differences from pubertal age onward suggests that sexual hormones play a key role, although their effect is far from being completely understood. Different incidences between sexes have been reported for many arrhythmias, both ventricular and supraventricular, and also for sudden cardiac death. Moreover, arrhythmias are an important issue during pregnancy, both for diagnosis and treatment. Interestingly, cardiovascular structural and electrophysiological remodelling promoted by exercise training enhances this 'gender effect'. Despite all these relevant issues, we lack gender specific recommendations in the current guidelines for electrical therapies for heart rhythm disorders and heart failure. Even more, we continue to see that fewer women are included in clinical trials and are less referred than men for these treatments

Cardiac Contractility Modulation in Patients with Heart Failure with Reduced Left Ventricular Ejection Fraction

Cardiac contractility modulation is an innovative therapy conceived for the treatment of heart failure. It is a device-based therapy, employing multiple electrodes to deliver relatively high-voltage (~7.5 V) biphasic signals to the endocardium of the right ventricular septum, in order to improve heart failure symptoms, exercise capacity and quality of life. Multiple clinical and mechanistic studies have been conducted to investigate the potential usefulness of this technology and, as of now, they suggest that it could have a place in therapy and meet a relevant medical need for a specific sub-category of underserved heart failure patients with reduced left ventricular ejection fraction. More studies are needed to further investigate its effect on outcomes such as mortality and rate of hospitalizations

Exercise Prescription in Patients With Arrhythmias

Epidemiological, clinical and laboratory studies have yielded definitive evidence that physical activity is able to reduce the morbidity and mortality of cardiovascular diseases and to improve physical fitness and quality of life. Moreover, physical activity seems to be capable of significantly reducing the risk of developing other chronic diseases, such as obesity, osteoporosis, diabetes, some neoplasms and depression. For these reasons, physical exercise has been proposed as an efficacious, low-cost, physiological means of disease prevention and therapy. However, although enormous amounts of scientific data have demonstrated the benefits of a physically active lifestyle, only a minority of people in Europe take regular physical exercise. One of the prime objectives of healthcare institutions is therefore to promote physical activity in the general population, as well as in patients with cardiovascular diseases.In the present review, we analyze the following points: some clinical aspects of arrhythmias and their relationships with physical exercise; the specific recommendations for exercise prescription in the various arrhythmias and in arhythmogenic heart diseases; the interactions between physical activity and antiarrhythmic therapies