Clinical use of Whole Genome Sequencing for Mycobacterium tuberculosis

Drug resistant tuberculosis (TB) remains a major challenge to global health and to healthcare in the UK. In 2014, England recorded 6520 cases of TB of which 1.4% were multi-drug resistant (MDR-TB). Extensively drug resistant TB (XDR-TB) occurs at a much lower rate, but the impact on the patient and hospital is severe. Current diagnostic methods such as drug susceptibility testing and targeted molecular tests are slow to return or examine only a limited number of target regions respectively. Faster, more comprehensive diagnostics will enable earlier use of the most appropriate drug regimen thus improving patient outcome and reducing overall healthcare costs. Whole genome sequencing has been shown to provide a rapid and comprehensive view of the genotype of the organism and thus enable reliable prediction of the drug susceptibility phenotype within a clinically relevant time frame. In addition it provides the highest resolution when investigating transmission events in possible outbreak scenarios. However, robust software and database tools need to be developed for the full potential to be realized in this specialized area of medicine

Global Chronic Total Occlusion Crossing Algorithm

The authors developed a global chronic total occlusion crossing algorithm following 10 steps: 1) dual angiography; 2) careful angiographic review focusing on proximal cap morphology, occlusion segment, distal vessel quality, and collateral circulation; 3) approaching proximal cap ambiguity using intravascular ultrasound, retrograde, and move-the-cap techniques; 4) approaching poor distal vessel quality using the retrograde approach and bifurcation at the distal cap by use of a dual-lumen catheter and intravascular ultrasound; 5) feasibility of retrograde crossing through grafts and septal and epicardial collateral vessels; 6) antegrade wiring strategies; 7) retrograde approach; 8) changing strategy when failing to achieve progress; 9) considering performing an investment procedure if crossing attempts fail; and 10) stopping when reaching high radiation or contrast dose or in case of long procedural time, occurrence of a serious complication, operator and patient fatigue, or lack of expertise or equipment. This algorithm can improve outcomes and expand discussion, research, and collaboration.info:eu-repo/semantics/publishedVersio

Atorvastatin reduces β-Adrenergic dysfunction in rats with diabetic cardiomyopathy.

In the diabetic heart the β-adrenergic response is altered partly by down-regulation of the β1-adrenoceptor, reducing its positive inotropic effect and up-regulation of the β3-adrenoceptor, increasing its negative inotropic effect. Statins have clinical benefits on morbidity and mortality in diabetic patients which are attributed to their "pleiotropic" effects. The objective of our study was to investigate the role of statin treatment on β-adrenergic dysfunction in diabetic rat cardiomyocytes.β-adrenergic responses were investigated in vivo (echocardiography) and ex vivo (left ventricular papillary muscles) in healthy and streptozotocin-induced diabetic rats, who were pre-treated or not by oral atorvastatin over 15 days (50 mg.kg-1.day-1). Micro-array analysis and immunoblotting were performed in left ventricular homogenates. Data are presented as mean percentage of baseline ± SD.Atorvastatin restored the impaired positive inotropic effect of β-adrenergic stimulation in diabetic hearts compared with healthy hearts both in vivo and ex vivo but did not suppress the diastolic dysfunction of diabetes. Atorvastatin changed the RNA expression of 9 genes in the β-adrenergic pathway and corrected the protein expression of β1-adrenoceptor and β1/β3-adrenoceptor ratio, and multidrug resistance protein 4 (MRP4). Nitric oxide synthase (NOS) inhibition abolished the beneficial effects of atorvastatin on the β-adrenoceptor response.Atorvastatin restored the positive inotropic effect of the β-adrenoceptor stimulation in diabetic cardiomyopathy. This effect is mediated by multiple modifications in expression of proteins in the β-adrenergic signaling pathway, particularly through the NOS pathway

General characteristics of healthy and diabetic treated (<i>atorvastatin</i>, 50 mg. kg^-1.day^-1) and control rats.

<p>General characteristics of healthy and diabetic treated (<i>atorvastatin</i>, 50 mg. kg^-1.day^-1) and control rats.</p

Atorvastatin reduces β-Adrenergic dysfunction in rats with diabetic cardiomyopathy - Fig 3

<p><b>Representative western blot and densitometric data reflecting protein expressions of β1-adrenoceptor (Panel A) and β3-adrenoceptor (Panel B) in left ventricles homogenates of healthy or diabetic rats, treated or not by atorvastatin (50 mg kg-1.day-1) during 15 days.</b> Western blot experiments were normalized using proteins using Ponceau S solution. Data are means ± SD (n = 4 to 9). *: <i>p</i><0.05 versus healthy untreated rats; †: <i>p</i><0.05 diabetic statin versus diabetic untreated rats.</p

Effects of atorvastatin on the transcriptome of left ventricles of healthy or diabetic rats.

<p>Panel A-B Heat Map of RNA expression profiles in diabetic versus healthy left ventricles (Panel A) or in statin diabetic versus untreated diabetic left ventricles (Panel B); Color scale indicate relative expression ratio for each gene in diabetic versus healthy left ventricle (Panel A) or in statin versus untreated diabetic left ventricle. Panel C-D Volcano Plot for the modification of genes expression by diabetes in heart ventricle (Panel C) and by atorvastatin in diabetic left ventricle (Panel D). The vertical axis represents the <i>p</i> value (-log¹⁰ <i>p</i> value) and the horizontal axis range the fold change (log² ratio) between diabetic and healthy left ventricles (Panel C) or statin diabetic versus untreated diabetic left ventricles (Panel D) (by t-test). Genes in the area delimited in red have a fold change greater than 1.5 with a <i>p</i> value < 0.05. Genes in the area delimited in green have a fold change greater than -1.5 (ratio <0.67) with a <i>p</i> value < 0.05. Panel E Venn diagram representing the differently expressed genes in diabetic versus healthy left ventricles in blue and in statin diabetic versus untreated diabetic left ventricles in red (<i>p</i><0.05). D is for down-regulation in diabetic versus healthy left ventricles, U for up-regulation. The overlapping part represents the genes modified by diabetes as well as statin, with up- or down-regulation for each comparison.</p

Active force (% of baseline value) variation of left ventricle papillary muscle exposed to isoproterenol in diabetic rats pretreated with statin (atorvastatin, 50 mg kg-1 day-1) during 15 days (8 rats per group) with or without L-NAME administration.

<p>Control refers to diabetic rats not receiving statin. AF/s = active force normalized per cross-sectional area during isometric contraction. Data are expressed as mean ± SD. NS: non-significant.</p

Representative Western Blot and densitometric data reflecting protein expressions of multidrug resistance protein 4 (MRP4) in left ventricles homogenates of healthy or diabetic rats, treated or not by atorvastatin (50 mg kg-1.day-1) during 15 days.

<p>Western blot experiments were normalized using GAPDH (37kDa). Data are means ± SD (n = 6). *: <i>p</i><0.05 versus healthy untreated rats; †: <i>p</i><0.05 diabetic statin versus diabetic untreated rats.</p