Differential integrity of TALE nuclease genes following adenoviral and lentiviral vector gene transfer into human cells

The array of genome editing strategies based on targeted double-stranded DNA break formation have recently been enriched through the introduction of transcription activator-like type III effector (TALE) nucleases (TALENs). To advance the testing of TALE-based approaches, it will be crucial to deliver these custom-designed proteins not only into transformed cell types but also into more relevant, chromosomally stable, primary cells. Viral vectors are among the most effective gene transfer vehicles. Here, we investigated the capacity of human immunodeficiency virus type 1- and adenovirus-based vectors to package and deliver functional TALEN genes into various human cell types. To this end, we attempted to assemble particles of these two vector classes, each encoding a monomer of a TALEN pair targeted to a bipartite sequence within the AAVS1 'safe harbor' locus. Vector DNA analyses revealed that adenoviral vectors transferred intact TALEN genes, whereas lentiviral vectors failed to do so, as shown by their heterogeneously sized proviruses in target cells. Importantly, adenoviral vector-mediated TALEN gene delivery resulted in site-specific double-stranded DNA break formation at the intended AAVS1 target site at similarly high levels in both transformed and non-transformed cells. In conclusion, we demonstrate that adenoviral, but not lentiviral, vectors constitute a valuable TALEN gene delivery platfor

Systematic review and meta-analysis of the diagnostic accuracy of ultrasonography for deep vein thrombosis

Background Ultrasound (US) has largely replaced contrast venography as the definitive diagnostic test for deep vein thrombosis (DVT). We aimed to derive a definitive estimate of the diagnostic accuracy of US for clinically suspected DVT and identify study-level factors that might predict accuracy. Methods We undertook a systematic review, meta-analysis and meta-regression of diagnostic cohort studies that compared US to contrast venography in patients with suspected DVT. We searched Medline, EMBASE, CINAHL, Web of Science, Cochrane Database of Systematic Reviews, Cochrane Controlled Trials Register, Database of Reviews of Effectiveness, the ACP Journal Club, and citation lists (1966 to April 2004). Random effects meta-analysis was used to derive pooled estimates of sensitivity and specificity. Random effects meta-regression was used to identify study-level covariates that predicted diagnostic performance. Results We identified 100 cohorts comparing US to venography in patients with suspected DVT. Overall sensitivity for proximal DVT (95% confidence interval) was 94.2% (93.2 to 95.0), for distal DVT was 63.5% (59.8 to 67.0), and specificity was 93.8% (93.1 to 94.4). Duplex US had pooled sensitivity of 96.5% (95.1 to 97.6) for proximal DVT, 71.2% (64.6 to 77.2) for distal DVT and specificity of 94.0% (92.8 to 95.1). Triplex US had pooled sensitivity of 96.4% (94.4 to 97.1%) for proximal DVT, 75.2% (67.7 to 81.6) for distal DVT and specificity of 94.3% (92.5 to 95.8). Compression US alone had pooled sensitivity of 93.8 % (92.0 to 95.3%) for proximal DVT, 56.8% (49.0 to 66.4) for distal DVT and specificity of 97.8% (97.0 to 98.4). Sensitivity was higher in more recently published studies and in cohorts with higher prevalence of DVT and more proximal DVT, and was lower in cohorts that reported interpretation by a radiologist. Specificity was higher in cohorts that excluded patients with previous DVT. No studies were identified that compared repeat US to venography in all patients. Repeat US appears to have a positive yield of 1.3%, with 89% of these being confirmed by venography. Conclusion Combined colour-doppler US techniques have optimal sensitivity, while compression US has optimal specificity for DVT. However, all estimates are subject to substantial unexplained heterogeneity. The role of repeat scanning is very uncertain and based upon limited data

Gene expression profiling of gluteal adipose tissue after prolonged bedrest

Adipose tissue is a complex and highly active metabolic and endocrine organ, with an important role in immune and inflammatory processes. Its dysfunction has been linked to development of insulin resistance, type 2 diabetes and cardiovascular disease. A sedentary lifestyle is associated to a quantitative and qualitative alteration of adipose tissue mass and function. Bed rest has been utilized as an experimental model to simulate physical inactivity. The aim of this study was to investigate the impact of physical inactivity on adipose tissue gene expression in healthy subjects. A total of 7 healthy subjects underwent gluteal adipose tissue biopsies before and after 14 days of bed rest; RNA was isolated and hybridized on RNA microarray chips in order to detect gene expression differences. A total of 308 genes were differently expressed after bed rest intervention; these genes were analyzed with bioinformatics database support (DAVID gene functional classification tool), to find potential functional-related gene groups and gene-disease associations. In silico analysis showed that 14 genes in our list have been previously involved in cardiovascular diseases, 8 genes in metabolic disorders and 12 genes in both. Among these, UCP3 (Uncoupling Protein 3) and BMP7 (bone morphogenetic protein 7) have been shown to be involved in energy expenditure and adipose tissue “browning”; TTR (transthyretin) is a known transport protein that carries the thyroid hormones (T4) and retinol-binding protein bound to retinol. Moreover several genes involved in inflammatory response such as CRP (CReactive Protein), IL1-beta and IL18 are differentially expressed after bedres

Aspirin In Primary Cardiovascular Disease Prevention In Type-2 Diabetic Patients: A Longitudinal Observational Study.

Aspirin In Primary Cardiovascular Disease Prevention In Type-2 Diabetic Patients: A Longitudinal Observational Study

Gene expression profiling of gluteal adipose tissue after prolonged bedrest.

Gene expression profiling of gluteal adipose tissue after prolonged bedrest

Study of the potential influence of obesity on the activities of serum Paraoxonase-1 and lipoprotein-associated phospholipase A2

INTRODUCTION: Obesity is independently associated with disturbances in lipid and lipoprotein metabolism, oxidative stress and is a well-established independent risk factor cardiovascular diseases (CVD) [1]. Human paraoxonase 1 (PON1) is a pleotropic high density lipoprotein (HDL)-associated enzyme with antioxidant and anti-inflammatory proprieties which have been suggested to contribute to the athero-protective function of the lipoprotein [2]. In contrast, lipoprotein-associated phospholipase A2 (Lp-PLA2) is prevalently bound to low density lipoprotein (LDL) and might act as a pro-atherogenic player. This study aimed to explore whether and in which extent PON1 and Lp-PLA2 are associated with obesity and, thus, could act as mediators of the adverse effects of this condition on cardiovascular health [3]. METHODS: The serum activities of these two enzymes were spectrophotometrically measured in 101 obese and severely obese, 101 overweight and 129 normal-weight women (controls) women (mean age: 54, intequartile range: 48-61). Markers of inflammation (high-sensitivity-C-reactive protein, hs-CRP) and oxidative stress (hydroperoxides), and lipid profile were also assessed. RESULTS: PON1 activity showed significant differences across the BMI groups (ANOVA, p 40kg/m2 compared to controls (p<0.001). PON1 and lp-PLA2 were both significantly associated to BMI but with opposite directions (negative, r= -0.280, p<0.01 and positive, r=201, p<0.05, respectively). At the multivariate analysis, PON1 emerged to be correlated with BMI in a way independent of age, inflammation and oxidative stress, thus suggesting that other factors may mediate the link between the enzyme expression/activity and obesity. CONCLUSIONS: our data suggest that lower PON1 and/or higher lp-PLA2 activities might contribute to increase the risk of cardio-metabolic disorders in obese women. 1. Keaney JF, et al. Arterioscler Thromb Vasc Biol. 2003;23(3):434–9. 2. Cervellati C, et al. 2016;64(1):21–32. 3. Tellis CC, Tselepis AD. Biochim Biophys Acta. 2009 May;1791(5):327–38

Effect Of Acute Immobilization On Plasma Irisin Levels And On Gene Expression Of Fndc5 And Related Genes In Gluteal Subcutaneous Adipose Tissue.

Effect Of Acute Immobilization On Plasma Irisin Levels And On Gene Expression Of Fndc5 And Related Genes In Gluteal Subcutaneous Adipose Tissue