Investigating LMNA-Related Dilated Cardiomyopathy Using Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

LMNA-related dilated cardiomyopathy is an inherited heart disease caused by mutations in the LMNA gene encoding for lamin A/C. The disease is characterized by left ventricular enlargement and impaired systolic function associated with conduction defects and ventricular arrhythmias. We hypothesized that LMNA-mutated patients’ induced Pluripotent Stem Cell-derived cardiomyocytes (iPSC-CMs) display electrophysiological abnormalities, thus constituting a suitable tool for deciphering the arrhythmogenic mechanisms of the disease, and possibly for developing novel therapeutic modalities. iPSC-CMs were generated from two related patients (father and son) carrying the same E342K mutation in the LMNA gene. Compared to control iPSC-CMs, LMNA-mutated iPSC-CMs exhibited the following electrophysiological abnormalities: (1) decreased spontaneous action potential beat rate and decreased pacemaker current (I_f) density; (2) prolonged action potential duration and increased L-type Ca²⁺ current (I_Ca,L) density; (3) delayed afterdepolarizations (DADs), arrhythmias and increased beat rate variability; (4) DADs, arrhythmias and cessation of spontaneous firing in response to β-adrenergic stimulation and rapid pacing. Additionally, compared to healthy control, LMNA-mutated iPSC-CMs displayed nuclear morphological irregularities and gene expression alterations. Notably, KB-R7943, a selective inhibitor of the reverse-mode of the Na⁺/Ca²⁺ exchanger, blocked the DADs in LMNA-mutated iPSC-CMs. Our findings demonstrate cellular electrophysiological mechanisms underlying the arrhythmias in LMNA-related dilated cardiomyopathy.     

cd1 Mutation in Drosophila Affects Phenoxazinone Synthase Catalytic Site and Impairs Long-Term Memory

Being involved in development of Huntington’s, Parkinson’s and Alzheimer’s diseases, kynurenine pathway (KP) of tryptophan metabolism plays a significant role in modulation of neuropathology. Accumulation of a prooxidant 3-hydroxykynurenine (3-HOK) leads to oxidative stress and neuronal cell apoptosis. Drosophila mutant cardinal (cd¹) with 3-HOK excess shows age-dependent neurodegeneration and short-term memory impairments, thereby presenting a model for senile dementia. Although cd gene for phenoxazinone synthase (PHS) catalyzing 3-HOK dimerization has been presumed to harbor the cd¹ mutation, its molecular nature remained obscure. Using next generation sequencing, we have shown that the cd gene in cd¹ carries a long deletion leading to PHS active site destruction. Contrary to the wild type Canton-S (CS), cd¹ males showed defective long-term memory (LTM) in conditioned courtship suppression paradigm (CCSP) at days 5–29 after eclosion. The number of dopaminergic neurons (DAN) regulating fly locomotor activity showed an age-dependent tendency to decrease in cd¹ relative to CS. Thus, in accordance with the concept “from the gene to behavior” proclaimed by S. Benzer, we have shown that the aberrant PHS sequence in cd¹ provokes drastic LTM impairments and DAN alterations.     

Copolymerization of Propylene with 1-Hexene and 4-Methyl-1-Pentene in Liquid Propylene Medium. Synthesis and Characterization of Random Metallocene Copolymers with Isotactic Propylene Sequences

Summary Propylene copolymerization with 1-hexene and 4-methyl-1-pentene in liquid propylene medium in presence of MAO-activated C₂-symmetry ansa-zirconocene rac-Me₂Si(4-Ph-2-MeInd)₂ZrCl₂ was studied. Random copolymers of propylene with 1-hexene and 4-methyl-1-pentene content up to 7 mol % were obtained at 60 °C. General kinetic characteristics of propylene/higher α-olefin copolymerization were evaluated. The distinct feature of propylene copolymerization with 1-hexene and 4-methyl-1-pentene in liquid propylene medium – the proximity of comonomer relative reactivity ratios (r₁∼r₂∼1) that indicates azeotropic nature of copolymerization processes in studied conditions. Synthesized copolymers were characterized with the use of IR, ¹³C NMR, GPC, WAXD, DSC techniques, and uniaxial tensile testing.     

Electron microscopic visualization of complementary labeled DNA with platinum‐containing guanine derivative

The object of the present report is to provide a method for a visualization of DNA in TEM by complementary labeling of cytosine with guanine derivative, which contains platinum as contrast‐enhanced heavy element. The stretched single‐chain DNA was obtained by modifying double‐stranded DNA. The labeling method comprises the following steps: (i) stretching and adsorption of DNA on the support film of an electron microscope grid (the hydrophobic carbon film holding negative charged DNA); (ii) complementary labeling of the cytosine bases from the stretched single‐stranded DNA pieces on the support film with platinum containing guanine derivative to form base‐specific hydrogen bond; and (iii) producing a magnified image of the base‐specific labeled DNA. Stretched single‐stranded DNA on a support film is obtained by a rapid elongation of DNA pieces on the surface between air and aqueous buffer solution. The attached platinum‐containing guanine derivative serves as a high‐dense marker and it can be discriminated from the surrounding background of support carbon film and visualized by use of conventional TEM observation at 100 kV accelerated voltage. This method allows examination of specific nucleic macromolecules through atom‐by‐atom analysis and it is promising way toward future DNA‐sequencing or molecular diagnostics of nucleic acids by electron microscopic observation. Microsc. Res. Tech. 79:280–284, 2016. © 2016 Wiley Periodicals, Inc.     

Effect of comonomer composition on the controlled free-radical copolymerization of styrene and maleic anhydride by reversible addition-fragmentation chain transfer (RAFT)

Controlled free-radical copolymerization of styrene and maleic anhydride was performed in 1,4-dioxane or tetrahydrofurane solution at 60 °C using the RAFT technique. The effect of monomer feed ratio on copolymerization kinetics and on control over molar mass distribution was examined. It was shown that polymerization was faster and quality of control was poorer when the proportion of maleic anhydride in the monomer feed was larger. These features were assigned to a decrease in the chain transfer constant of the polymeric RAFT agent, most probably due to an increase in the apparent rate constant of propagation with the proportion of maleic anhydride.     

Experimental and direct numerical analysis of hard-disk drive

The purpose of this paper is to study methods for enhancing the reliability and performance of hard-disk drives (HDD) because it is essential for improving recording density, speed of data access, and output signal. This study also investigates various techniques that can be used for head/disk contact detection. The acoustic emission (AE) and friction signal characteristics were observed with respect to the durability of the head/disk interface (HDI) under various operating conditions using a contact start-stop (CSS) test. In addition, to study the influence of surface topography on the stiction performance of the HDI, a modified and polished laser pump was proposed and CSS investigations were accomplished. Moreover, the static and dynamic properties of an HDD air slider were studied using a finite element method (FEM).     

Quantity-based buffer-constrained two-machine flowshop problem: active and passive prefetch models for multimedia applications

Conventional studies on buffer-constrained flowshop scheduling problems have considered applications with a limitation on the number of jobs that are allowed in the intermediate storage buffer before flowing to the next machine. The study in Lin et al. (Comput. Oper. Res. 36(4):1158?C1175, 2008a) considered a two-machine flowshop problem with ??processing time-dependent?? buffer constraints for multimedia applications. A???passive?? prefetch model (the PP-problem), in which the download process is suspended unless the buffer is sufficient for keeping an incoming media object, was applied in Lin et al. (Comput. Oper. Res. 36(4):1158?C1175, 2008a). This study further considers an ??active?? prefetch model (the AP-problem) that exploits the unoccupied buffer space by advancing the download of the incoming object by a computed maximal duration that possibly does not cause a buffer overflow. We obtain new complexity results for both problems. This study also proposes a new lower bound which improves the branch and bound algorithm presented in Lin et al. (Comput. Oper. Res. 36(4):1158?C1175, 2008a). For the PP-problem, compared to the lower bounds developed in Lin et al. (Comput. Oper. Res. 36(4):1158?C1175, 2008a), on average, the results of the simulation experiments show that the proposed new lower bound cuts about 38% of the nodes and 32% of the execution time for searching the optimal solutions.     

Cyclization dynamics of polymers: 7. Applications of the pyrene excimer technique to the internal dynamics of poly(dimethylsiloxane) chains

Poly(dimethylsiloxane) (PDMS) polymers end-capped on both ends with pyrene chromophores have been synthesized. Rate constants for end-to-end cyclization 〈 k₁ 〉 have been determined for dilute solutions of these polymers in toluene solution using a combination of fluorescence decay and steady-state fluorescence measurements. While precise values of the critical exponent for the chain length (N¯) dependence of 〈 k₁ 〉 are not yet available, these results are consistent with the N¯^{? 3/2} dependence predicted by Wilemski-Fixman theory. PDMS chains cyclize somewhat more than two times faster than polystyrene chains of the same length in solvents of similar solvating power and viscosity. These results provide strong support for similar predictions made several years ago by Perico and Cuniberti, who used intrinsic viscosity data to parametrize the Rouse-Zimm model for analysis of polymer cyclization dynamics.     

Branch‐and‐bound approach for optima localization in scheduling multiprocessor jobs

We consider multiprocessor task scheduling problems with dedicated processors. We determine the tight optima localization intervals for different subproblems of the basic problem. Based on the ideas of a computer‐aided technique developed by Sevastianov and Tchernykh for shop scheduling problems, we elaborate a similar method for the multiprocessor task scheduling problem. Our method allows us to find an upper bound for the length of the optimal schedule in terms of natural lower bound. As a byproduct of our results, a family of linear‐time approximation algorithms with a constant ratio performance guarantee is designed for the NP‐hard subproblems of the basic problem, and new polynomially solvable classes of problems are found.