The development of sensing coatings, as important sensor elements that integrate functionality, simplicity, chemical stability, and physical stability, has been shown to play a major role in electrochemical sensing system development trends. Simple and versatile assembling procedures and scalability make polyelectrolytes highly convenient for use in electrochemical sensing applications. Polyelectrolytes are mainly used in electrochemical sensor architectures for entrapping (incorporation, immobilization, etc.) various materials into sensing layers. These materials can often increase sensitivity, selectivity, and electronic communications with the electrode substrate, and they can mediate electron transfer between an analyte and transducer. Analytical performance can be significantly improved by the synergistic effect of materials (sensing material, transducer, and mediator) present in these composites. As most reported methods for the preparation of polyelectrolyte-based sensing layers are layer-by-layer and casting/coating methods, this review focuses on the use of the latter methods in the development of electrochemical sensors within the last decade. In contrast to many reviews related to electrochemical sensors that feature polyelectrolytes, this review is focused on architectures of sensing layers and the role of polyelectrolytes in the development of sensing systems. Additionally, the role of polyelectrolytes in the preparation and modification of various nanoparticles, nanoprobes, reporter probes, nanobeads, etc. that are used in electrochemical sensing systems is also reviewed.

Azobenzene-based photochromic lipids are valuable probes for the analysis of ion channel–lipid interactions. Rapid photoisomerization of these molecules enables the analysis of lipid gating kinetics and provides information on lipid sensing. Thermal relaxation of the metastable cis conformation to the trans conformation of azobenzene photolipids is rather slow in the dark and may be modified by ligand–protein interactions. Cis photolipid-induced changes in pure lipid membranes as visualized from the morphological response of giant unilamellar vesicles indicated that thermal cis–trans isomerization of both PhoDAG-1 and OptoDArG is essentially slow in the lipid bilayer environment. While the currents activated by cis PhoDAG remained stable upon termination of UV light exposure (dark, UV-OFF), cis OptoDArG-induced TRPC3/6/7 activity displayed a striking isoform-dependent exponential decay. The deactivation kinetics of cis OptoDArG-induced currents in the dark was sensitive to mutations in the L2 lipid coordination site of TRPC channels. We conclude that the binding of cis OptoDArG to TRPC channels promotes transition of cis OptoDArG to the trans conformation. This process is suggested to provide valuable information on DAG–ion channel interactions and may enable highly selective photopharmacological interventions.

In this paper, the electrochemical behavior of cysteine is described, using carbon paste electrodes (CPEs) modified with ternary silver-copper sulfide containing intrinsic silver at two pH values (pH 3 and 5). Experiments have revealed that presence of cysteine has a large impact on the electrochemical behavior of modified CPEs. Observed phenomena take place in solution, as well as at the surface of the modified CPEs, and can be applied for electroanalytical purposes. Based on the electrochemical behavior observed in the examined system, differential pulse voltammetry (DPV) was selected as an electroanalytical method for determination of cysteine. The effects of the various parameters on the electroanalytical signal, such as the amount of electroactive material, electroanalytical parameters, pH etc., were investigated using differential pulse voltammograms. The results indicated that electrochemical signal characterized with well-defined cathodic peak at 0.055 V vs. Ag/AgCl (3 M) in acetic buffer solution at pH 5 can be used for indirect electrochemical determination of cysteine. The optimization procedure revealed that the most sensitive and stabile electrode was that containing 5% modifier. The DPV response of the electrode, in the presence of cysteine, showed two different linear concentration ranges of 0.1 to 2.5 μM, and 5.6 to 28 μM. The explanation of the origin of two linear ranges is proposed. The lower concentration range was characterized by remarkable sensitivity of the 11.78 μA μM⁻¹, owing to the chosen indirect method of determination. The calculated limit of detection (LOD), as well as limit of quantification (LOQ) were 0.032 and 0.081 μM, respectively. The influence of interfering agents on the electroanalytical response was examined, and low or no interference on the DPVs was observed. The proposed method was validated and applied for the determination of cysteine in pharmaceutical preparations with satisfactory recoveries in the range of 97 to 101.7%.

The interplay of SK3, a Ca²⁺ sensitive K⁺ ion channel, with Orai1, a Ca²⁺ ion channel, has been reported to increase cytosolic Ca²⁺ levels, thereby triggering proliferation of breast and colon cancer cells, although a molecular mechanism has remained elusive to date. We show in the current study, via heterologous protein expression, that Orai1 can enhance SK3 K⁺ currents, in addition to constitutively bound calmodulin (CaM). At low cytosolic Ca²⁺ levels that decrease SK3 K⁺ permeation, co-expressed Orai1 potentiates SK3 currents. This positive feedback mechanism of SK3 and Orai1 is enabled by their close co-localization. Remarkably, we discovered that loss of SK3 channel activity due to overexpressed CaM mutants could be restored by Orai1, likely via its interplay with the SK3–CaM binding site. Mapping for interaction sites within Orai1, we identified that the cytosolic strands and pore residues are critical for a functional communication with SK3. Moreover, STIM1 has a bimodal role in SK3–Orai1 regulation. Under physiological ionic conditions, STIM1 is able to impede SK3–Orai1 interplay by significantly decreasing their co-localization. Forced STIM1–Orai1 activity and associated Ca²⁺ influx promote SK3 K⁺ currents. The dynamic regulation of Orai1 to boost endogenous SK3 channels was also determined in the human prostate cancer cell line LNCaP.

Canonical transient receptor potential (TRPC) channels were identified as key players in maladaptive remodeling, with nuclear factor of activated T-cells (NFAT) transcription factors serving as downstream targets of TRPC-triggered Ca²⁺ entry in these pathological processes. Strikingly, the reconstitution of TRPC-NFAT signaling by heterologous expression yielded controversial results. Specifically, nuclear translocation of NFAT1 was found barely responsive to recombinant TRPC3, presumably based on the requirement of certain spatiotemporal signaling features. Here, we report efficient control of NFAT1 nuclear translocation in human embryonic kidney 293 (HEK293) cells by light, using a new photochromic TRPC benzimidazole activator (OptoBI-1) and a TRPC3 mutant with modified activator sensitivity. NFAT1 nuclear translocation was measured along with an all-optical protocol to record local and global Ca²⁺ pattern generated during light-mediated activation/deactivation cycling of TRPC3. Our results unveil the ability of wild-type TRPC3 to produce constitutive NFAT nuclear translocation. Moreover, we demonstrate that TRPC3 mutant that lacks basal activity enables spatiotemporally precise control over NFAT1 activity by photopharmacology. Our results suggest tight linkage between TRPC3 activity and NFAT1 nuclear translocation based on global cellular Ca²⁺ signals.

Various species of the Brassicaceae family are known to hyperaccumulate metals. Lunaria annua L., a plant from the Brassicaceae family, is an oilseed crop known for its pharmaceutical and nutraceutical applications. In this work, Lunaria annua L. was investigated for its accumulation potential in copper and lead-contaminated soil. Concentrations of copper and lead were measured before planting (in seeds and soils) and after the plant was harvested (in soils and plant). Two types of soils were used: a soil sample collected from the Botanical Garden of the Faculty of Science, University of Split (soil 1, S1) and a commercially available organic mineral substrate (soil 2, S2). Measured pH values showed that the S1 (pH = 8.58) was moderately alkaline soil. On the other hand, the purchased organic soil, S2 (pH = 6.35), was poorly acidic to neutral. For the determination of copper (Cu) and lead (Pb), square wave anodic stripping voltammetry (SWASV), using a glassy carbon electrode modified with mercury film, was applied. The concentrations of Pb and Cu were determined and calculated in the sample using the standard addition method. Obtained results have shown that Lunaria annua L. is a lead hyperaccumulator (4116.2 mg/kg in S1 and 3314.7 mg/kg in S2) and a potential copper accumulator (624.2 mg/kg in S1 and 498.9 mg/kg in S2). Likewise, the results have shown that the higher the pH is, the lower the possibility that metal accumulation exists.