Pro-Arrhythmic Potential of Accumulated Uremic Toxins Is Mediated via Vulnerability of Action Potential Repolarization.

Chronic kidney disease (CKD) is represented by a diminished filtration capacity of the kidneys. End-stage renal disease patients need dialysis treatment to remove waste and toxins from the circulation. However, endogenously produced uremic toxins (UTs) cannot always be filtered during dialysis. UTs are among the CKD-related factors that have been linked to maladaptive and pathophysiological remodeling of the heart. Importantly, 50% of the deaths in dialysis patients are cardiovascular related, with sudden cardiac death predominating. However, the mechanisms responsible remain poorly understood. The current study aimed to assess the vulnerability of action potential repolarization caused by exposure to pre-identified UTs at clinically relevant concentrations. We exposed human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) and HEK293 chronically (48 h) to the UTs indoxyl sulfate, kynurenine, or kynurenic acid. We used optical and manual electrophysiological techniques to assess action potential duration (APD) in the hiPSC-CMs and recorded IKr currents in stably transfected HEK293 cells (HEK-hERG). Molecular analysis of KV11.1, the ion channel responsible for IKr, was performed to further understand the potential mechanism underlying the effects of the UTs. Chronic exposure to the UTs resulted in significant APD prolongation. Subsequent assessment of the repolarization current IKr, often most sensitive and responsible for APD alterations, showed decreased current densities after chronic exposure to the UTs. This outcome was supported by lowered protein levels of KV11.1. Finally, treatment with an activator of the IKr current, LUF7244, could reverse the APD prolongation, indicating the potential modulation of electrophysiological effects caused by these UTs. This study highlights the pro-arrhythmogenic potential of UTs and reveals a mode of action by which they affect cardiac repolarization.

Fluoride-Ion-Responsive Sol-Gel Transition in an L-Cysteine/AgNO3 System: Self-Assembly Peculiarities and Anticancer Activity.

Supramolecular hydrogels based on low-molecular-weight compounds are a unique class of so-called "soft" materials, formed by weak non-covalent interactions between precursors at their millimolar concentrations. Due to the variety of structures that can be formed using different low-molecular-weight gelators, they are widely used in various fields of technology and medicine. In this study, we report for the first time an unusual self-assembly process of mixing a hydrosol obtained from L-cysteine and silver nitrate (cysteine-silver sol-CSS) with sodium halides. Modern instrumental techniques such as viscosimetry, UV spectroscopy, dynamic light scattering, zeta potential measurements, SEM and EDS identified that adding fluoride anions to CSS is able to form stable hydrogels of a thixotropic nature, while Cl-, Br- and I- lead to precipitation. The self-assembly process proceeds using a narrow concentration range of F-. An increase in the fluoride anion content in the system leads to a change in the gel network morphology from elongated structures to spherical ones. This fact is reflected in a decrease in the gel viscosity and a number of gel-sol-gel transition cycles. The mechanism of F-'s interaction with hydrosol includes the condensation of anions on the positive surface of the CSS nanoparticles, their binding via electrostatic forces and the formation of a resulting gel carcass. In vitro analysis showed that the hydrogels suppressed human squamous carcinoma cells at a micromolar sample concentration. The obtained soft gels could have potential applications against cutaneous malignancy and as carriers for fluoride anion and other bioactive substance delivery.

Preparation of Composite Hydrogels Based on Cysteine-Silver Sol and Methylene Blue as Promising Systems for Anticancer Photodynamic Therapy.

In this study, a novel supramolecular composite, "photogels", was synthesized by mixing of cysteine-silver sol (CSS) and methylene blue (MB). A complex of modern physico-chemical methods of analysis such as viscosimetry, UV spectroscopy, dynamic and electrophoretic light scattering, scanning electron microscopy and energy-dispersive X-ray spectroscopy showed that MB molecules are uniformly localized mainly in the space between fibers of the gel-network formed by CSS particles. Molecules of the dye also bind with the surface of CSS particles by non-covalent interactions. This fact is reflected in the appearance of a synergistic anticancer effect of gels against human squamous cell carcinoma even in the absence of light irradiation. A mild toxic influence of hydrogels was observed in normal keratinocyte cells. Photodynamic exposure significantly increased gel activity, and there remained a synergistic effect. The study of free-radical oxidation in cells has shown that gels are not only capable of generating reactive oxygen species, but also have other targets of action. Flow cytometric analysis allowed us to find out that obtained hydrogels caused cell cycle arrest both without irradiation and with light exposure. The obtained gels are of considerable interest both from the point of view of academics and applied science, for example, in the photodynamic therapy of superficial neoplasms.

Ca2+-dependent oxidation of exogenous NADH and NADPH by the mitochondria of spring wheat and its relation with AOX capacity and ROS content at high temperatures.

Mitochondria are sources of reactive oxygen species (ROS) in a plant cell under high temperature. Mitochondrial alternative NAD(P)H dehydrogenases (type II NAD(P)H DHs) and cyanide-resistant oxidase (AOX) can regulate ROS production, but their role at high temperatures is unknown. This study investigates the influence heat acclimation (37 °C) and heat shock (50 °C) temperatures on ROS content, activity and protein abundance of external Ca2+-dependent NAD(P)H DHs (NDB) and AOX in mitochondria of 4- and 8-day-old seedlings of spring wheat (Triticum aestivum L., var. Novosibirskya 29). The shoots of 4-day-old seedlings contained more carbohydrates, had a higher rate of total respiration and a high rate of oxidation of exogenous NADH, a greater AOX capacity and a lower of ROS content, as compared to leaves of 8-day-old seedlings, and were more resistant to heat shock. The activity of external NADH DH was higher than the one of NADPH DH in mitochondria of both shoots and leaves. At 37 °C, high NADH oxidation was associated with increased AOX capacity in mitochondria of both shoots and leaves, whereas NADPH oxidation with COX capacity. At 50 °C, the NADPH oxidation by shoots' mitochondria increased and the NADH oxidation stayed high. The content of NDB and AOX proteins depends on heat treatments and differs between mitochondria of shoots and leaves. Our data indicate that Ca2+-dependent type II NAD(P)H DHs can regulate the ROS content and together with AOX are involved in heat tolerance, depending on the development phase of spring wheat and is, probably, tissue-specific.

Cysteine-Silver-Polymer Systems for the Preparation of Hydrogels and Films with Potential Applications in Regenerative Medicine.

Herein, the problem concerning the poorer mechanical properties of gels based on low molecular weight gelators (LMWGs)-L-cysteine and silver nitrate-was solved by the addition of various polymers-polyvinyl alcohol (PVA), polyvinyl pyrrolidone (PVP) and polyethylene glycol (PEG)-to the initial cysteine-silver sol (CSS). The physicochemical methods of analysis-viscosimetry, UV spectroscopy, DLS, and SEM-identified that cysteine-silver hydrogels (CSG) based on PVA possess the best rheological properties and porous microstructure (the average pore size is 2-10 µm) compared to gels without the polymer or with PVP or PEG. Such gels are able to form cysteine-silver cryogels (CSC) and then porous cysteine-silver films (CSF) with an average pore size of 10-20 µm and good mechanical, swelling, and adhesion to skin characteristics as long as the structure of CSS particles remains stable. In vitro experiments have shown that hydrogels are non-toxic to normal human fibroblast cells. The obtained materials could potentially be applied to regenerative medicine.