RBM20 Mutations Induce an Arrhythmogenic Dilated Cardiomyopathy Related to Disturbed Calcium Handling

BACKGROUND: Mutations in RBM20 (RNA-binding motif protein 20) cause a clinically aggressive form of dilated cardiomyopathy, with an increased risk of malignant ventricular arrhythmias. RBM20 is a splicing factor that targets multiple pivotal cardiac genes, such as Titin (TTN) and CAMK2D (calcium/calmodulin-dependent kinase II delta). Aberrant TTN splicing is thought to be the main determinant of RBM20-induced dilated cardiomyopathy, but is not likely to explain the increased risk of arrhythmias. Here, we investigated the extent to which RBM20 mutation carriers have an increased risk of arrhythmias and explore the underlying molecular mechanism

The MoS2 Nanotubes with Defect-Controlled Electric Properties

We describe a two-step synthesis of pure multiwall MoS2 nanotubes with a high degree of homogeneity in size. The Mo6S4I6 nanowires grown directly from elements under temperature gradient conditions in hedgehog-like assemblies were used as precursor material. Transformation in argon-H2S/H2 mixture leads to the MoS2 nanotubes still grouped in hedgehog-like morphology. The described method enables a large-scale production of MoS2 nanotubes and their size control. X-ray diffraction, optical absorption and Raman spectroscopy, scanning electron microscopy with wave dispersive analysis, and transmission electron microscopy were used to characterize the starting Mo6S4I6 nanowires and the MoS2 nanotubes. The unit cell parameters of the Mo6S4I6 phase are proposed. Blue shift in optical absorbance and metallic behavior of MoS2 nanotubes in two-probe measurement are explained by a high defect concentration

Cardiovascular development: towards biomedical applicability: Regulation of cardiomyocyte differentiation of embryonic stem cells by extracellular signalling

Investigating the signalling pathways that regulate heart development is essential if stem cells are to become an effective source of cardiomyocytes that can be used for studying cardiac physiology and pharmacology and eventually developing cell-based therapies for heart repair. Here, we briefly describe current understanding of heart development in vertebrates and review the signalling pathways thought to be involved in cardiomyogenesis in multiple species. We discuss how this might be applied to stem cells currently thought to have cardiomyogenic potential by considering the factors relevant for each differentiation step from the undifferentiated cell to nascent mesoderm, cardiac progenitors and finally a fully determined cardiomyocyte. We focus particularly on how this is being applied to human embryonic stem cells and provide recent examples from both our own work and that of others

TECRL, a new life‐threatening inherited arrhythmia gene associated with overlapping clinical features of both LQTS and CPVT

Genetic causes of many familial arrhythmia syndromes remain elusive. In this study, whole-exome sequencing (WES) was carried out on patients from three different families that presented with life-threatening arrhythmias and high risk of sudden cardiac death (SCD). Two French Canadian probands carried identical homozygous rare variant in TECRL gene (p.Arg196Gln), which encodes the trans-2,3-enoyl-CoA reductase-like protein. Both patients had cardiac arrest, stress-induced atrial and ventricular tachycardia, and QT prolongation on adrenergic stimulation. A third patient from a consanguineous Sudanese family diagnosed with catecholaminergic polymorphic ventricular tachycardia (CPVT) had a homozygous splice site mutation (c.331+1G>A) in TECRL Analysis of intracellular calcium ([Ca(2+)]i) dynamics in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) generated from this individual (TECRLHom-hiPSCs), his heterozygous but clinically asymptomatic father (TECRLHet-hiPSCs), and a healthy individual (CTRL-hiPSCs) from the same Sudanese family, revealed smaller [Ca(2+)]i transient amplitudes as well as elevated diastolic [Ca(2+)]i in TECRLHom-hiPSC-CMs compared with CTRL-hiPSC-CMs. The [Ca(2+)]i transient also rose markedly slower and contained lower sarcoplasmic reticulum (SR) calcium stores, evidenced by the decreased magnitude of caffeine-induced [Ca(2+)]i transients. In addition, the decay phase of the [Ca(2+)]i transient was slower in TECRLHom-hiPSC-CMs due to decreased SERCA and NCX activities. Furthermore, TECRLHom-hiPSC-CMs showed prolonged action potentials (APs) compared with CTRL-hiPSC-CMs. TECRL knockdown in control human embryonic stem cell-derived CMs (hESC-CMs) also resulted in significantly longer APs. Moreover, stimulation by noradrenaline (NA) significantly increased the propensity for triggered activity based on delayed afterdepolarizations (DADs) in TECRLHom-hiPSC-CMs and treatment with flecainide, a class Ic antiarrhythmic drug, significantly reduced the triggered activity in these cells. In summary, we report that mutations in TECRL are associated with inherited arrhythmias characterized by clinical features of both LQTS and CPVT Patient-specific hiPSC-CMs recapitulated salient features of the clinical phenotype and provide a platform for drug screening evidenced by initial identification of flecainide as a potential therapeutic. These findings have implications for diagnosis and treatment of inherited cardiac arrhythmias

Present state and future perspectives of using pluripotent stem cells in toxicology research

The use of novel drugs and chemicals requires reliable data on their potential toxic effects on humans. Current test systems are mainly based on animals or in vitro–cultured animal-derived cells and do not or not sufficiently mirror the situation in humans. Therefore, in vitro models based on human pluripotent stem cells (hPSCs) have become an attractive alternative. The article summarizes the characteristics of pluripotent stem cells, including embryonic carcinoma and embryonic germ cells, and discusses the potential of pluripotent stem cells for safety pharmacology and toxicology. Special attention is directed to the potential application of embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) for the assessment of developmental toxicology as well as cardio- and hepatotoxicology. With respect to embryotoxicology, recent achievements of the embryonic stem cell test (EST) are described and current limitations as well as prospects of embryotoxicity studies using pluripotent stem cells are discussed. Furthermore, recent efforts to establish hPSC-based cell models for testing cardio- and hepatotoxicity are presented. In this context, methods for differentiation and selection of cardiac and hepatic cells from hPSCs are summarized, requirements and implications with respect to the use of these cells in safety pharmacology and toxicology are presented, and future challenges and perspectives of using hPSCs are discussed

MBaaS-based Authentication service for handicraft products traceability system

International audienceCounterfeiting is copying a product without having the right. It affects all manufacturing sectors, mainly, the craft sector. Authentication is a solution to prevent handicraft products from counterfeiting. According to ISO (international organization for standardization) 27001, authentication is defined as the process used to conform that a claimed characteristics or attributes that appears to be true is in fact true. With the aim of guaranteeing the authenticity of product, various methodological approaches have been devised over the past years and several authentication schemes have been proposed. Although it is a widely discussed and studied topic, authentication remains a crucial issue.More specifically, the main issue that the craft sector is facing is the lack of a structure for communicating a product information requested by the customer (e.g. tourist) due to the organization of the craft industry. As a result, several questions are emerging which authentication system for the craft industry. How to collect the product data and make it available to this system while guaranteeing its quality? How to guarantee a real time answer to the user (e.g. tourist)?Thereby, the project we are developing is oriented towards the management of data related to the product manufactured by Moroccan artisans to preserve its authenticity. Our objective is to conceive a traceability system to manage the information of handicraft product throughout its life cycle based on smartest authentication technologies and features.The Mobile Backend as a service (MBaaS) will help for sharing authentication information about goods in real time, as the request of the customer, in other way, this service will provide the answer about the product authenticity

Diffusion in phthalocyanine thin film layers: study by AES, EELS, EPES and electrical measurements

Diffusion studies are necessary to the feasibility of molecular pn junctions. Electrical and spectroscopic measurements have been done in two superimposed phthalocyanine thin layers: Zinc phthaiocyanine (PcZn) (p material) and Lutetium bisphthalocyanine (PC$_2$Lu) (n material). These measurements are in a good agreement. In spite of the very large dimensions of these molecules, diffusion and doping of PcZn by PC$_2$Lu occur. Moreover, elastic peak electron spectroscopy (EPES) seems more available than Auger electron spectroscopy (AES) to follow the diffusion in such samples.Les études de diffusion sont nécessaires pour utiliser pratiquement les semiconducteurs organiques – en particulier pour la réalisation de jonctions pn. Des mesures électriques et spectroscopiques ont été faites sur des couches minces de phtalocyanines superposées : phtalocyanine de Zinc(PcZn) (matériau de type p) et bisphtalocyanine de Lutetium (PC$_2$Lu) (matériau de type n). L'ensemble de ces mesures est en bon accord, la diffusion et le dopage de PcZn par PC$_2$Lu se produisent malgré la taille importante de ces molécules. De plus la spectroscopie de pic élastique (EPES) semble être plus adaptée à ces mesures que la spectroscopie d'électrons Auger (AES) pour suivre la diffusion dans ces échantillons

Microbioreactor arrays for full factorial screening of exogenous and paracrine factors in human embryonic stem cell differentiation

Timed exposure of pluripotent stem cell cultures to exogenous molecules is widely used to drive differentiation towards desired cell lineages. However, screening differentiation conditions in conventional static cultures can become impractical in large parameter spaces, and is intrinsically limited by poor spatiotemporal control of the microenvironment that also makes it impossible to determine whether exogenous factors act directly or through paracrine-dependent mechanisms. We detail here the development of a continuous flow microbioreactor array platform that combines full-factorial multiplexing of input factors with progressive accumulation of paracrine factors through serially-connected culture chambers, and further, the use of this system to explore the combinatorial parameter space of both exogenous and paracrine factors involved in human embryonic stem cell (hESC) differentiation to a MIXL1-GFP(+) primitive streak-like population. We show that well known inducers of primitive streak (BMP, Activin and Wnt signals) do not simply act directly on hESC to induce MIXL1 expression, but that this requires accumulation of surplus, endogenous factors; and, that conditioned medium or FGF-2 supplementation is able to offset this. Our approach further reveals the presence of a paracrine, negative feedback loop to the MIXL1-GFP(+) population, which can be overcome with GSK-3 beta inhibitors (BIO or CHIR99021), implicating secreted Wnt inhibitory signals such as DKKs and sFRPs as candidate effectors. Importantly, modulating paracrine effects identified in microbioreactor arrays by supplementing FGF-2 and CHIR in conventional static culture vessels resulted in improved differentiation outcomes. We therefore demonstrate that this microbioreactor array platform uniquely enables the identification and decoding of complex soluble factor signalling hierarchies, and that this not only challenges prevailing strategies for extrinsic control of hESC differentiation, but also is translatable to conventional culture systems