Holistic Health Model Of Sustainable Development

Health can be interpreted not only as an important element of sustainable development. It is connected to its all the three pillars namely its environmental, economic and social dimension as well. Health can be found directly or indirectly in every goal related to sustainable development. Considering the importance of health, the aim of our research was to elaborate a holistic health conscious model encouraging sustainable development. Holistic health conscious model means a model including the physical, psychical and mental health of the individual. Furthermore, besides the responsibility of the individual environmental effects are also taken into account as well as the intervening factors. In addition, it integrates the aspect of sustainable development. In our research, first of all we were looking for the answer to the following (1) what kind of stakeholders play a role in developing health consciousness? (2) What is the role of individuals, the state, the companies and NGOs in developing health consciousness? The article presents the holistic health model which was created as a result of the empirical research

Mixed-effects modeling for concentration effect profiling in cardiomyocyte contractility assays

Présentation PosterInternational audienceBackground. With the advent of new realtime technologies such as impedance assays, extracellular field potential measurement and optical sensing for in vitro cardiac safety screening studies, researchers have now to frequently deal with analyzing voluminous amounts of complex time responses. In this context, main issues are to speed up the data analysis process and to extract accurate information for cardiotoxicity profiling. Objectives. A first objective is the development of an innovative computational method able to globally process a large set of in vitro cardiac signals (provided by 96, 384 and 1536-well microplates) instead of analyzing them once at a time. Such a statistical population approach has the advantage the account for the common characteristics between the individual responses. A second objective is to handle qualitative factors (type of cardiomyocytes, compounds and media, etc.) in the computational process. Methods. The proposed estimation method relies on the combination of a dynamic system identification method and a mixed-effect modeling technique. An output-error polynomial model structure is used for the system identification step and a stochastic approximation expectation maximization is implemented for the estimation of the hyperparameters. Input signals to be analyzed are the contractility amplitudes of cardiomyocytes submitted to compounds to be tested. Impedance signals and contractility amplitude were provided by a CardioExcyte96 system (Nanion Technologies). human iPSC-derived cardiomyocytes were provided by Cellartis Takara with 30,000 cells per well. Results. Our data-driven profiling method extracted four parameters that completely fit the contractility time variations and fully characterize the effect of compound concentration on the contractility amplitude. The proposed method not only estimates the values of the model parameters but also their uncertainty distribution. The latter allows to compute p-values associated with each effect.Conclusion. We show that the population-based estimation method developed in this study is suited to the fully characterize dynamic effects in cardiomyocyte contractility assays. Each parameter becomes a profiling characteristics of the concentration effect. It can be applied to estimate concentration and compounds effects with an optimal accuracy and could be extended directly to multielectrode array and optical sensing responses

Cross - site comparison of excitation-contraction coupling using impedance and field potential recordings in hiPSC cardiomyocytes

Introduction: Since 2005 the S7B and E14 guidances from ICH and FDA have been in place to assess a potential drug candidate’s ability to cause long QT syndrome. To refine these guidelines, the FDA proposed the Comprehensive in vitro Proarrhythmia Assay (CiPA) initiative, where the assessment of drug effects on cardiac repolarization was one subject of investigation. Within the myocyte phase II study, effects of pharmaceutical compounds on human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) were assessed and this article will focus on an evaluation of the proarrhythmic potential of 23 drugs in four hiPSC-CM cell lines. Methods: Experiments were performed on the CardioExcyte 96 at different sites. A combined readout of contractility (via impedance) and electrophysiology endpoints (field potentials) was performed. Results: Our data demonstrates that hERG blockers such as Dofetilide and further high risk categorized compounds prolong the field potential duration. Arrhythmic events were detected in both impedance as well as EFP recordings. Intermediate risk compounds induced arrhythmia in almost all cases at the highest dose. In the case of low risk compounds, either a decrease in FPDmax was observed, or not a significant change. Discussion: All sources of hiPSC-CMs are sensitive enough to detect delayed or shortened repolarization and arrhythmia after drug application and can provide predictive cardiac electrophysiology data. However, the baseline electrophysiological parameters vary between iPS cells from different sources

Frequency-Dependent Multi-Well Cardiotoxicity Screening Enabled by Optogenetic Stimulation

Side effects on cardiac ion channels causing lethal arrhythmias are one major reason for drug withdrawals from the market. Field potential (FP) recording from cardiomyocytes, is a well-suited tool to assess such cardiotoxic effects of drug candidates in preclinical drug development, but it is currently limited to the spontaneous beating of the cardiomyocytes and manual analysis. Herein, we present a novel optogenetic cardiotoxicity screening system suited for the parallel automated frequency-dependent analysis of drug effects on FP recorded from human-induced pluripotent stem cell-derived cardiomyocytes. For the expression of the light-sensitive cation channel Channelrhodopsin-2, we optimised protocols using virus transduction or transient mRNA transfection. Optical stimulation was performed with a new light-emitting diode lid for a 96-well FP recording system. This enabled reliable pacing at physiologically relevant heart rates and robust recording of FP. Thereby we detected rate-dependent effects of drugs on Na+, Ca2+ and K+ channel function indicated by FP prolongation, FP shortening and the slowing of the FP downstroke component, as well as generation of afterdepolarisations. Taken together, we present a scalable approach for preclinical frequency-dependent screening of drug effects on cardiac electrophysiology. Importantly, we show that the recording and analysis can be fully automated and the technology is readily available using commercial products

Advantages and Limitations of Different p62-Based Assays for Estimating Autophagic Activity in Drosophila

<div><p>Levels of the selective autophagy substrate p62 have been established in recent years as a specific readout for basal autophagic activity. Here we compared different experimental approaches for using this assay in Drosophila larvae. Similar to the more commonly used western blots, quantifying p62 dots in immunostained fat body cells of L3 stage larvae detected a strong accumulation of endogenous p62 aggregates in null mutants for Atg genes and S6K. Importantly, genes whose mutation or silencing results in early stage lethality can only be analyzed by microscopy using clonal analysis. The loss of numerous general housekeeping genes show a phenotype in large-scale screens including autophagy, and the p62 assay was potentially suitable for distinguishing bona fide autophagy regulators from silencing of a DNA polymerase subunit or a ribosomal gene that likely has a non-specific effect on autophagy. p62 accumulation upon RNAi silencing of known autophagy regulators was dependent on the duration of the knockdown effect, unlike in the case of starvation-induced autophagy. The endogenous p62 assay was more sensitive than a constitutively overexpressed p62-GFP reporter, which showed self-aggregation and large-scale accumulation even in control cells. We recommend western blots for following the conversion of overexpressed p62-GFP reporters to estimate autophagic activity if sample collection from mutant larvae or adults is possible. In addition, we also showed that overexpressed p62 or Atg8 reporters can strongly influence the phenotypes of each other, potentially giving rise to false or contradicting results. Overexpressed p62 aggregates also incorporated Atg8 reporter molecules that might lead to a wrong conclusion of strongly enhanced autophagy, whereas expression of an Atg8 reporter transgene rescued the inhibitory effect of a dominant-negative Atg4 mutant on basal and starvation-induced autophagy.</p> </div