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Abstract CRISPR-based genome editing of pseudogene-associated disorders, such as p47 phox -deficient chronic granulomatous disease (p47 CGD), is challenged by chromosomal rearrangements due to presence of multiple targets. We report that interactions between highly homologous sequences that are localized on the same chromosome contribute substantially to post-editing chromosomal rearrangements. We successfully employed editing approaches at the NCF1 gene and its pseudogenes, NCF1B and NCF1C, in a human cell line model of p47 CGD and in patient-derived human hematopoietic stem and progenitor cells. Upon genetic engineering, a droplet digital PCR-based method identified cells with altered copy numbers, spanning megabases from the edited loci. We attributed the high aberration frequency to the interaction between repetitive sequences and their predisposition to recombination events. Our findings emphasize the need for careful evaluation of the target-specific genomic context, such as the presence of homologous regions, whose instability can constitute a risk factor for chromosomal rearrangements upon genome editing.

The paper highlights the significance of the drying process in post-harvest cucurbit seed treatment technology. To improve the effectiveness of drying high-moisture seeds, a plant construction design was introduced, incorporating the concept of providing varied heat distribution to a vigorously agitated seed layer. The paper validates the necessity of refining the dimensional, frictional, and aerodynamic properties of pumpkin seeds to determine the optimal operating modes for the proposed dryer. (Research purpose) To identify the physical and mechanical properties of pumpkin seeds by defining their dimensional, frictional and aerodynamic characteristics. (Materials and methods) The study employs one-factor experimental research methods with subsequent statistical data processing. An experimental study was conducted on the physical and mechanical properties of Volzhskaya Grey pumpkin seeds possessing a standard moisture content ranging between 9.3 and 9.7 percent. Well-known laboratory apparatuses, namely the «conical tank» and the «inclined plane», were used to determine the natural repose angle and to enhance the accuracy of friction coefficients determination. For analyzing the aerodynamic properties of the seeds, the K-293 Petkus air separator was employed. (Results and discussion) Research methods were developed and experimental plants were described. It was established that with a probability confidence level of 0.95, the static friction coefficients of pumpkin seeds on a solid steel sheet, a perforated steel sieve and on rubber are 0.473, 0.418, 0.481, respectively, and the dynamic friction coefficients under the identical conditions are measured as 0.331, 0.293, 0.337. The average angle of natural repose is found to be 22 degrees, the midsection area is 97.94 square millimeters, the soaring velocity is 7.083 meters per second, the windage coefficient stands at 0.196 and the aerodynamic drag coefficient is 0.136. (Conclusions). The assumption that enhancing the drying efficiency of vegetable seeds can be achieved by implementing a diverse thermal energy supply to continuously moving mass inside the apparatus is confirmed, provided that the seeds do not stick together in layers resulting in blockage of the coolant flow. An improved design of the dryer configurations has been developed and, to validate its optimum design and technological specifications, an in-depth analysis of the seeds' dimensional, frictional, and aerodynamic attributes has been conducted.

Aim. Xenobiotic metal mercury is one of the major crucial pollutants of global public health concerns according to the World Health Organization assessment. Carbon-containing nanoparticles (CNPs) are promising in nanotechnology. CNPs were obtained by the combustion of citric acid and urea. Methods. The cortex nerve terminals isolated from Wistar rats were used in the experiments. [14C]glutamate uptake and release in the nerve terminals were monitored using a radiolabeled assay. Results. It was shown that HgCl2 starting from 5 µM caused a concentration-dependent increase in the extracellular L-[14C]glutamate level in nerve terminals resulted from weak functioning of glutamate transporter, and so significantly decreased L-[14C] glutamate uptake. Combined effects of Hg2+ and CNPs obtained by heating of citric acid and urea were analysed. CNPs were able to mitigate in an acute manner excitotoxic Hg2+-induced increase in the extracellular L-[14C]glutamate level in nerve terminals by 37%, thereby being a provisional Hg2+ scavenger. Conclusion. Besides biotechnological implementation of data, developed approach can be applicable for monitoring capability of different particles and compounds to mitigate Hg2+ -mediated threat.

Relevance. Tense ecological situation in the impact zone of the large industrial complex of Usolye-Sibirskoe on the Angara river, search for sources and ways of pollutants inflow during different periods of technogenic load, necessary to assess and forecast the current changes in the aquatic ecosystem. Aim. To study chemical composition of wastewater from the industrial zone of Usolye-Sibirskoe during different periods of technogenic load and assess their impact on hydrochemical parameters of the Angara river water. Objects. Wastewater from the industrial zone of Usolye-Sibirskoe and Angara river waters in the impact area during the period of high technogenic load (2010) and the period of its reduction (2020). Methods. Chemical composition of wastewater and natural water was determined in the Center of Collective Use «Isotope-Geochemical Investigations» of Vinogradov Institute of Geochemistry SB RAS (Irkutsk, Russia). Concentrations of major ions in waters were measured using a set of methods generally accepted in hydrochemistry (atomic absorption and atomic emission flame spectrometry, spectrophotometry, potentiometry, titrimetry), Hg concentration was measured using atomic absorption spectrometry, concentrations of other trace elements – using mass spectrometry method (ISP MS). Results. The authors have revealed significant changes in chemical composition of wastewater of the industrial zone in different periods of technogenic load, reflecting the transformation of pollution sources during the liquidation of technogenic objects of the area. Over the ten-year period particularly important changes are expressed in a significant decrease of Hg, Al, Cs, Ti, Cr, Cl– and Na+ concentrations and increase of Mn concentrations in the wastewater of the industrial zone. This is reflected, in its turn, in the dynamics of hydrochemical composition of Angara river water in the impact area. In 2010 a regular consecutive decrease of most of elements concentrations was observed with distance of Angara river waters from technogenic sources. Where as in 2020 their spatial distribution is opposite: with a significant decrease in concentrations of most of the studied elements at the river site 50 m downstream of industrial discharges, their increase was detected at more distant monitoring stations (1.5 and 5 km) where Mn, Cu, Cd and Tl concentrations were the highest. Despite significant changes in wastewater chemical composition in different periods of technogenic load, the Angara river waters at all monitoring stations in the impact zone of the industrial complex of Usolye-Sibirskoe are polluted.

Relevance. The necessity to control the hydrochemical regime of water bodies significantly affected by anthropogenic impacts. Aim. To study the spatial distribution of major ions and trace element concentrations and identify natural and anthropogenic formation factors of the hydrochemical composition of the Vikhoreva river and Ust-Vikhorevsky bay of the Ust-Ilimsk reservoir. Objects. Waters of the Vikhoreva river before and after inflow of wastewater from the Bratsk industrial zone, waters of the Ust-Vikhorevsky bay (Vikhorevskaya and Burdoyskaya parts) of the Ust-Ilimsk reservoir. Methods. Determination of micronutrients concentrations in water was performed by inductive coupled plasma mass spectrometry, mercury – by atomic absorption method. Cations and anions were determined using standard methods accepted in hydrochemical studies, Sorg – by titrimetric method, oxygen – by iodometric method. Results. In the Vihoreva river water after wastewater inflow from the Bratsk industrial zone, significant transformations of its hydrochemical composition were detected. Along the water area of river decrease of O2 concentrations (up to 0.3 mg O2/l) and increase of Сorg (up to 131 mg/l), SO42– (up to 262 mg/l), K+ (up to 10.6 mg/l), Cl– (up to 220 mg/l), Na+ (up to 240 mg/l), Mn (up to 248.5 mg/l), Cr (up to 7.4 mg/l), to a lesser extent Al (up to 224.1 mg/l), Co (up to 0.26 mg/l), Cu (up to 5.4 mg/l), Zn (up to 43.2 mg/l), Cd (up to 0.126 mg/l), Pb (up to 0.40 mg/l), Hg (up to 0.0032 mg/l) are observed. In surface waters of the Ust-Vikhorevsky bay concentrations of most components of the main ionic composition and trace elements decrease, as compared to the Vikhoreva river water. Maximum concentrations of Mn (511.5 mg/l), Fe (1567.2 mg/l), Co (0.80 mg/l), Pb (0.77 mg/l) were detected in bottom waters. The main ionic composition and trace element concentrations in the Vihoreva river water are determined primarily by anthropogenic factor. At the same time, the hydrochemical regime of the river is effected by natural factors. Inflow of both waters of the highly polluted and forming waters of the channel part of the Ust-Ilimsk reservoir impacts the hydrochemistry of the Ust-Vikhorevsky bay.

Graphene materials are widely used in different technologies and certainly released into aquatic and air surroundings being environmental pollution components. Nitrogen‑doped graphene nanomaterials have great potential for application, in particular, in energy storage, as electrochemical sensors and waste water treatment. Aim. Evaluate neurotoxic risk of nitrogen-doped multilayer graphene. Methods. Here, nitrogen-doped multilayer graphene nanoparticles (N-MLG) were synthesized by means of electrochemical exfoliation of high-purity graphite rods in NaN3-based electrolyte and characterised using TEM, AFM and UV-vis spectroscopy. Neuroactive features of N-MLG were assessed in isolated cortex nerve terminals (synaptosomes) analysing the extracellular level of excitatory neurotransmitter L-[14C] glutamate and inhibitory one [3H]GABA. Results. It was revealed that N-MLG did not affect the extracellular synaptosomal levels of L-[14C] glutamate and [3H]GABA within the concentration range 0.01–0.5 mg/ml, and an increase in a concentration up to 1 mg/ml caused an insignificant increase (tendency to increase) in these levels for both neurotransmitters. To analyse a capability of interaction with heavy metals in biological system, N-MLG was investigated using model of acute Cd2+/Pb2+/Hg2+-induced neurotoxicity in nerve terminals. In was revealed that Cd2+/Pb2+/Hg2+-induced increase in the extracellular level of L-[14C] glutamate and [3H]GABA was not changed by N-MLG. Conclusions. N-MLG does not possess neurotoxic signs and is biocompatible within the concentration range 0.01–1 mg/ml. In biological system, N-MLG did not mitigate/aggravate Cd2+/Pb2+/Hg2+-induced neurotoxicity in nerve terminals.

Abstract Tremendous deposits of disposable medical facemask waste after the COVID-19 pandemic require improvement of waste management practice according to WHO report 2022, moreover facemasks are still in use around the world to protect against numerous airborne infections. Here, water-suspended smoke preparations from the combustion of disposable medical facemasks (polypropylene fibers) were collected; size, zeta potential, surface groups of smoke particulate matter were determined by dynamic light scattering, FTIR and Raman spectroscopy, and their optical properties were characterized. Neurochemical study using nerve terminals isolated from rat cortex revealed a significant decrease in the initial rate of the uptake/accumulation of excitatory and inhibitory neurotransmitters, L-[14C]glutamate and [3H]GABA, and exocytotic release, and also an increase in the extracellular level of these neurotransmitters. Fluorescent measurements revealed that ROS generation induced by hydrogen peroxide and glutamate receptor agonist kainate decreased in nerve terminals. A decrease in the membrane potential of nerve terminals and isolated neurons, the mitochondrial potential and synaptic vesicle acidification was also shown. Therefore, accidental or intentional utilization of disposable medical facemask waste by combustion results in the release of neuroactive ultrafine particulate matter to the environment, thereby contributing to plastic-associated pollution of air and water resources and neuropathology development and expansion.

Boron-enhanced proton therapy has recently appeared as a promising approach to increase the efficiency of proton therapy on tumor cells, and this modality can further be improved by the use of boron nanoparticles (B NPs) as local sensitizers to achieve enhanced and targeted therapeutic outcomes. However, the mechanisms of tumor cell elimination under boron-enhanced proton therapy still require clarification. Here, we explore possible molecular mechanisms responsible for the enhancement of therapeutic outcomes under boron NP-enhanced proton therapy. Spherical B NPs with a mode size of 25 nm were prepared by methods of pulsed laser ablation in water, followed by their coating by polyethylene glycol to improve their colloidal stability in buffers. Then, we assessed the efficiency of B NPs as sensitizers of cancer cell killing under irradiation with a 160.5 MeV proton beam. Our experiments showed that the combined effect of B NPs and proton irradiation induces an increased level of superoxide anion radical generation, which leads to the depolarization of mitochondria, a drop in their membrane mitochondrial potential, and the development of apoptosis. A comprehensive gene expression analysis (via RT-PCR) confirmed increased overexpression of 52 genes (out of 87 studied) involved in the cell redox status and oxidative stress, compared to 12 genes in the cells irradiated without B NPs. Other possible mechanisms responsible for the B NPs-induced radiosensitizing effect, including one related to the generation of alpha particles, are discussed. The obtained results give a better insight into the processes involved in the boron-induced enhancement of proton therapy and enable one to optimize parameters of proton therapy in order to maximize therapeutic outcomes.