Ion Current Research Articles

Optimization of ion source structure for enhancing beam current of multiply charged ions

Optimization of ion source structure for enhancing beam current of multiply charged ions

Atypical antipsychotic drug olanzapine inhibits 5-HT3 receptor-mediated currents by allosteric and non-competitive mechanisms.

Olanzapine, an atypical antipsychotic, is widely used in the treatment of schizophrenia and bipolar disorder due to its modulation of dopamine and serotonin receptor systems. While its primary action involves antagonism of dopamine D2 and serotonin 5-HT (5-hydroxytryptamine)2A receptors, recent evidence suggests that olanzapine also inhibits 5-HT3 receptors, which are ligand-gated ion channels involved in synaptic transmission in central and peripheral nervous systems. The present study aimed to investigate the action of olanzapine on 5-HT3 receptor-mediated currents using whole-cell voltage-clamp recordings in NCB-20 neuroblastoma cells. Results of this study indicated that olanzapine could act as a non-competitive antagonist of the 5-HT3 receptor, exhibiting concentration-dependent inhibition of ion currents. Moreover, olanzapine facilitated both deactivation and desensitization kinetics, accelerating decay of 5-HT3 receptor-mediated currents. Recovery from desensitization was significantly delayed by olanzapine, whereas recovery from deactivation was largely unaffected by it. Current-voltage relationship analysis revealed that olanzapine reduced the amplitude of 5-HT3 receptor-mediated currents across all holding potentials without altering reversal potential, suggesting a voltage-independent inhibition. Furthermore, olanzapine exhibited use-dependent inhibition, with a greater reduction in current observed during more frequent 5-HT application. These findings provide novel insights into a non-competitive and allosteric inhibition of 5-HT3 receptors by olanzapine, contributing to a deeper understanding of its pharmacological profile in neuropsychiatric and gastrointestinal conditions where serotonergic neurotransmission is implicated.

Ankyrin-B is required for the establishment and maintenance of lens cytoarchitecture, mechanics and clarity.

The transparent ocular lens is essential for vision because it focuses light onto the retina. Despite recognition of the importance of its unique cellular architecture and mechanical properties, the molecular mechanisms governing these attributes remain elusive. This study aims to elucidate the role of ankyrin-B (AnkB, encoded by ANK2), a membrane scaffolding protein, in lens cytoarchitecture, growth and function using a conditional knockout (cKO) mouse model. The AnkB cKO mouse has no defects in lens morphogenesis but exhibited changes that supported a global role for AnkB in maintenance of lens clarity, size, cytoarchitecture, membrane organization and stiffness. Notably, absence of AnkB led to nuclear cataract formation, which was evident from postnatal day 16. AnkB cKO lens fibers exhibit progressive disruption in membrane organization of the spectrin-actin cytoskeleton, cell adhesion proteins and channel proteins; loss and degradation of several membrane proteins [such as NrCAM. N-cadherin (CDH2) and aquaporin-0 (also known as MIP)]; along with a disorganized plasma membrane and impaired membrane interdigitations. Furthermore, absence of AnkB led to decreased lens stiffness. Collectively, these results illustrate the essential role for AnkB in lens architecture, growth and function through its involvement in membrane skeletal and protein organization and stability.

Light-activated channelrhodopsins: a revolutionary toolkit for the remote control of plant signalling.

Channelrhodopsins (CHRs), originating within algae and protists, are membrane-spanning ion channel proteins that are directly activated and/or deactivated by specific wavelengths of light. Since 2005, CHRs have been deployed as genetically encoded optogenetic tools to rapidly advance understanding of neuronal networks. CHRs provide the opportunity to finely tune ion transport across membranes and regulate membrane potential. These are fundamental biochemical signals, which in plants can be translated into physiological and developmental responses such as changes in photosynthesis, growth, turgor, vascular hydraulics, phosphorylation or reactive oxygen species (ROS) status, gene expression, or even cell death. Exploration of CHR family diversity and structure-function engineering has led to the expansion of the CHR optogenetic toolbox, offering unparalleled opportunities to precisely control and understand electrical and secondary messenger signalling in higher plants. In this Tansley Insight, we provide an overview of the recent progress in the application of CHR optogenetics in higher plants and discuss their possible uses in the remote control of plant biology, illuminating a new future domain for plant research enabled through synthetic biology.

GLP-1 receptor signaling restores aquaporin 4 subcellular polarization in reactive astrocytes and promotes amyloid β clearance in a mouse model of Alzheimer's disease.

The physiological actions of a gut hormone, glucagon-like peptide-1 (GLP-1), in Alzheimer's disease (AD) brain remain poorly understood, although GLP-1 receptor (GLP-1R) expression in this organ has been shown in several experimental studies. Therefore, we explored whether the GLP-1R signaling promotes the clearance of amyloid β (Aβ) (1-42) which is a core pathological hallmark of AD, focusing on the water channel protein aquaporin 4 (AQP4) localized to astrocyte endfeet perivascular membranes in intact brain. First, we confirmed that Glp1r mRNA is predominantly expressed at perivascular site of astrocytes in normal mouse cerebral cortex through in situ hybridization analysis. Next, we observed that 20-week subcutaneous administration of a GLP-1R agonist (GLP-1RA) liraglutide significantly reduced Aβ (1-42) accumulation in the cerebral cortex and improved spatial working memory in an AD mouse model, AppNL-G-F/NL-G-F mice. Furthermore, our current data revealed that the 4-week liraglutide treatment relocalized subcellular AQP4 in morphologically injured reactive astrocytes of AppNL-G-F/NL-G-F mice to the cell surface perivascular site through PKA-mediated AQP4 phosphorylation. Such translocation of phosphorylated AQP4 to astrocyte cell surface following incubation with liraglutide was observed also in the present in vitro study using the cell line in which AQP4 cDNA was introduced into immortalized human astrocyte. These results suggest that enhanced intracerebral GLP-1R signaling following peripheral administration of GLP-1RA restores AQP4 subcellular polarization in reactive astrocytes and would promote Aβ excretion possibly through increasing AQP4-mediated intracerebral water flux in the brain in AD.

A deep learning approach to real-time Markov modeling of ion channel gating

The patch-clamp technique allows us to eavesdrop the gating behavior of individual ion channels with unprecedented temporal resolution. The signals arise from conformational changes of the channel protein as it makes rapid transitions between conducting and non-conducting states. However, unambiguous analysis of single-channel datasets is challenging given the inadvertently low signal-to-noise ratio as well as signal distortions caused by low-pass filtering. Ion channel kinetics are typically described using hidden Markov models (HMM), which allow conclusions on the inner workings of the protein. In this study, we present a Deep Learning approach for extracting models from single-channel recordings. Two-dimensional dwell-time histograms are computed from the idealized time series and are subsequently analyzed by two neural networks, that have been trained on simulated datasets, to determine the topology and the transition rates of the HMM. We show that this method is robust regarding noise and gating events beyond the corner frequency of the low-pass filter. In addition, we propose a method to evaluate the goodness of a predicted model by re-simulating the prediction. Finally, we tested the algorithm with data recorded on a patch-clamp setup. In principle, it meets the requirements for model extraction during an ongoing recording session in real-time.

Reduced 17β-estradiol following ovariectomy induces mitochondrial dysfunction and degradation of synaptic proteins in the entorhinal cortex

Reductions in circulating estrogens can contribute to cognitive decline, in part by impairing mitochondrial function within the hippocampal region. The entorhinal cortex provides the hippocampus with its main cortical input and, to assess the impact of estrogen deficiency on mitochondrial respiration and synaptic proteins in the entorhinal cortex, wildtype rats received either sham surgery, bilateral ovariectomy, or ovariectomy with implantation of a subdermal capsule to maintain low levels of 17β-estradiol (E2). Mitochondrial respiration in the entorhinal cortex was not significantly affected two weeks following ovariectomy, but there was a reduction in oxygen consumption four weeks after ovariectomy that was prevented by E2 supplementation. The expression of mitochondrial membrane integrity element voltage-dependent anion channel protein (VDAC1) was also reduced four weeks after ovariectomy, suggesting that respiration was reduced due to a decline in mitochondrial density. Ovariectomy also increased mitochondrial and cytoplasmic cytochrome c and upregulated superoxide dismutase 2 (SOD2) both two and four weeks after ovariectomy, reflecting mitochondrial electron leakage and oxidative redox imbalance. Further, the ovariectomy-induced changes in mitochondrial proteins were associated with reductions in postsynaptic density protein 95 (PSD95) and the presynaptic protein synaptophysin. There were no changes in mitochondrial or synaptic proteins in ovariectomized animals that received E2 supplementation. Our findings indicate that reductions in circulating 17β-estradiol induced by ovariectomy disrupt mitochondrial functions in the entorhinal cortex, and suggest that a resulting increase in oxidative stress contributes to the degradation in synaptic proteins that may affect cognitive functions mediated by the hippocampal region.

Seasonal remodelling of the fish heart alters sensitivity to petrochemical pollutant, 3-methylphenanthrene

Exploitation of offshore oil reserves, heightened traffic in marine transportation routes, and the release of petrochemicals from the thawing of permafrost and glaciers is increasing the bioavailability of polycyclic aromatic hydrocarbons (PAHs) to aquatic organisms. This availability may also change with the seasons as temperature changes accessibility of Arctic transport routes and the degree of land- and ice-melt and thus run-off into coastal ecosystems. Seasonal temperature change also remodels the ion channels in the heart of fish to facilitated preserved cardiac function across a range of temperatures. How this seasonal cardiac remodelling impacts vulnerability to pollutants is currently unknown. In this study we accessed the electrical activity of navaga cod (Eleginus nawaga) ventricular cardiomyocytes under the dual influence of seasonal change and varying concentrations of a pervasive PAH pollutant, 3-methylphenanthrene (3-MP). We used whole-cell patch-clamp to elucidate the effect of various doses of 3-MP on action potential (AP) parameters and the main ion currents (IKr, IK1, INa, ICa) in ventricular cardiomyocytes isolated from navaga cod in winter and summer at the White Sea, close to the Russian Arctic circle. Navaga cod ventricular cardiomyocytes were particularly vulnerable to 3-MP during the winter season. Exposure to 300 nM 3-MP resulted in significant changes in AP duration in winter-acclimatized fish, whereas no such changes were observed in summer-acclimatized fish. The IKr current was the most sensitive to 3-MP, with a winter IC50 of 49.7 nM and a summer IC50 of 53 μM. The INa current also exhibited seasonal shifts in sensitivity to 3-MP, with IC50 values of 2.39 μM in winter-acclimatized fish and 7.73 μM in summer-acclimatized fish. No significant differences were observed in the effect of 3-MP on the peak ICa current, although 3 μM of 3-MP caused a pronounced decrease in charge transferred by ICa (e.g. QCa) in both seasons. The IK1 current was insensitive to 3-MP in both winter and summer fish. These findings highlight how remodelling of the fish heart with changing season alters the potency of PAH pollution. This paper lays the groundwork for future research on the molecular mechanisms that drive the altered seasonal potency of pollutants in navaga cod and other species.

Model of the HVC neural network as a song motor in zebra finch.

The nucleus HVC within the avian song system produces crystalized instructions which lead to precise, learned vocalization in zebra finches (Taeniopygia guttata). This paper proposes a model of the HVC neural network based on the physiological properties of individual HVC neurons, their synaptic interactions calibrated by experimental measurements, as well as the synaptic signal into this region which triggers song production. This neural network model comprises of two major neural populations in this area: neurons projecting to the nucleus RA and interneurons. Each single neuron model of HVCRA is constructed with conductance-based ion currents of fast Na+ and K+ and a leak channel, while the interneuron model includes extra transient Ca2+ current and hyperpolarization-activated inward current. The synaptic dynamics is formed with simulated delivered neurotransmitter pulses from presynaptic cells and neurotransmitter receptor opening rates of postsynaptic neurons. We show that this network model qualitatively exhibits observed electrophysiological behaviors of neurons independent or in the network, as well as the importance of bidirectional interactions between the HVCRA neuron and the HVCI neuron. We also simulate the pulse input from A11 neuron group to HVC. This signal successfully suppresses the interneuron, which leads to sequential firing of projection neurons that matches measured burst onset, duration, and spike quantities during the zebra finch motif. The result provides a biophysically based model characterizing the dynamics and functions of the HVC neural network as a song motor, and offers a reference for synaptic coupling strength in the avian brain.

Transmembrane Ion Channels: From Natural to Artificial Systems.

Natural channel proteins allow the selective permeation of ions, water or other nutritious entities across bilayer membranes, facilitating various essential physiological functions in living systems. Inspired by nature, chemists endeavor to simulate the structural features and transport behaviors of channel proteins through biomimetic strategies. In this review, we start from introducing the inherent traits of channel proteins such as their crystal structures, functions and mechanisms. Subsequently, different kind of synthetic ion channels including their design principles, dynamic regulations and therapeutic applications were carefully reviewed. Finally, the potential challenges and opportunities in this research field were also carefully discussed. It is anticipated that this review could provide some inspiring ideas and future directions towards the construction of novel bionic ion channels with higher-level structures, properties, functions and practical applications.

Transmembrane Ion Channels: From Natural to Artificial Systems

Natural channel proteins allow the selective permeation of ions, water or other nutritious entities across bilayer membranes, facilitating various essential physiological functions in living systems. Inspired by nature, chemists endeavor to simulate the structural features and transport behaviors of channel proteins through biomimetic strategies. In this review, we start from introducing the inherent traits of channel proteins such as their crystal structures, functions and mechanisms. Subsequently, different kind of synthetic ion channels including their design principles, dynamic regulations and therapeutic applications were carefully reviewed. Finally, the potential challenges and opportunities in this research field were also carefully discussed. It is anticipated that this review could provide some inspiring ideas and future directions towards the construction of novel bionic ion channels with higher‐level structures, properties, functions and practical applications.

Time-resolved electromechanical and conductive behavior of nanostructured bilayers tethered to the surface of the electrode with incorporated channel proteins and peptides

The influence of incorporation of mitochondrial inner membrane potassium channel, and channel-forming peptide – Gramicidin on the ion transport and electromechanical properties of model lipid membranes tethered to gold electrode was electrochemically investigated by chronoamperometric and impedance spectroscopy techniques. In the case of the potassium channel the ion transport properties were modulated with channel-specific inhibitor – ATP-Mg2+ complex, whereas in the case of gramicidin peptide – by replacing potassium with sodium ions. The observed two exponential current–time responses of the systems studied were interpreted in terms of ion penetration and electrostriction of tethered lipid bilayer membrane, and conclusions supported with the experiments on alkanethiol self-assembled monolayers of different alkanethiol chain lengths deposited on gold.

Astroglial Dysfunction, Demyelination and Nodular inflammation in Necrotizing Meningoencephalitis.

Necrotizing Meningoencephalitis (NME), a form of Meningoencephalitis of Unknown Origin (MUO), is a progressive neuroinflammatory disease that primarily affects young, small-breed dogs. Due to limited understanding of its pathophysiology, early detection and the development of targeted therapies remain challenging. Definitive ante-mortem diagnosis is often unfeasible, and dogs with NME are frequently grouped under the broader MUO category. Our long-term objective is to identify distinct disease mechanisms within each MUO subtype to improve diagnostic accuracy, therapeutic approaches, and prognostic outcomes. To establish unique inflammatory patterns as they relate to neuropathologic changes in NME, we studied we studied the degree of immune cell infiltration, astrogliosis, demyelination, and microglial activation, comparing these factors with granulomatous meningoencephalomyelitis (GME), a closely related MUO subtype. We found that in the leptomeninges, NME is characterized by mild immune cell infiltration, in contrast to the prominent, B cell-rich aggregates seen in GME. In the neuroparenchyma, both diseases exhibit a comparable degree of lymphocyte infiltration; however, demyelination is more pronounced in NME, particularly within the subcortical white matter. Notably, areas of the brain affected by NME display a reduction in astrogliosis, which is associated with a marked decrease in the expression of the water channel protein aquaporin-4 (AQP4), a reduction not observed in GME. Additionally, we found that AQP4 expression levels correlate with the extent of microglial and macrophage activation. These findings suggest that astrocyte dysfunction in regions of microglial inflammation is a driver of NME and with adaptive immune responses likely playing a supportive role.

Abstract 4136901: Mechanosensitive channel protein Piezo1 synergizes with TLR4 to enhance osteogenic responses in human aortic valve interstitial cells

Background: Calcific aortic valve disease (CAVD), a common heart valve disease in the elderly, has a complex pathogenesis. Abnormal blood flow shear can be regarded as an important factor in the development of CAVD, and a key molecule that responds to mechanical stress is the mechanosensitive ion channel protein Piezo1; however, the role of Piezo1 in CAVD has not been clarified. Methods and results: We collected CAVD patients and their age- and sex-matched controls, and compared the differences in the protein expression levels of Piezo1 and TLR4 between the two using Western blot and other techniques, and found that the expression of Piezo1 and TLR4 proteins was elevated in the CAVD group. We gave LPS or Piezo1 agonist Yoda1 to stimulate aortic valve mesenchymal stromal cells, respectively, and the Western blot results showed that ALP and Runx2 expression were elevated after LPS and Yoda1 stimulation, and immunofluorescence and alizarin red staining showed similar results. Inhibition of TLR4 or Piezo1 attenuated the osteogenic differentiation of AVICs. Cellular immunofluorescence results showed that TLR4 could be transported from the Golgi to the cell membrane under Yoda stimulation, and Golgi-cell membrane transport of TLR4 was attenuated after Rab3b knockdown by siRNA. After first pre-stimulation with Yoda1 and then stimulation with LPS, Western blot and immunofluorescence results showed that the protein levels of ALP and Runx2 as well as NF-κB phosphorylation were to be significantly increased. Conclusion: Stimulation of Piezo1 and TLR4 can enhance the osteogenic response of AVICs. Stimulation of Piezo1 enhances the transport of TLR4 from the Golgi to the cell membrane via Rab3b, and Piezo1 synergistically enhances the osteogenic response of AVICs with TLR4, thus promoting the development of CAVD.

Activation of Hedgehog pathway by circEEF2/miR-625-5p/TRPM2 axis promotes prostate cancer cell proliferation through mitochondrial stress.

The purpose of this study was to identify the role played by circEEF2 (has-circ-0048559) in prostate cancer (PCa) development and to determine the potential mechanism involved. circEEF2, miR-625-5p, and the transient receptor potential M2 channel protein (TRPM2) were determined using RT-qPCR in PCa. Cell proliferation was determined by CCK-8 assay and colony formation assay, whereas migration and invasion were assessed by Transwell assay, and apoptosis was evaluated by flow cytometry after annexin V-FITC and propidium iodide staining. The interactions between circEEF2 and miRNAs were investigated through the Circular RNA Interactome database, and the downstream targets of miR-625-5p were forecasted using TargetScan. The interaction was confirmed using both the dual luciferase reporter gene assay and RNA pull-down assay. TRPM2, Hedgehog signaling pathway proteins (GLI1 and GLI2), ubiquinone oxidase subunit B8, and cytochrome C oxidase subunit IV (COX4) were analyzed by protein blotting. JC-1 fluorescence detection was applied for mitochondrial membrane potential changes, fluorescent probe assay for intracellular ROS levels, and immunofluorescence staining for γ-H2AX expression. The role of circEEF2 in PCa tumor growth was tested by xenograft experiments. CircEEF2 expression was upregulated in PCa (p<0.05). Cells of PCa were inhibited in proliferation, migration, invasion, and enhanced in apoptosis by depleting circEEF2 (p<0.05). circEEF2 directly targeted adsorbed miR-625-5p. TRPM2 bound to miR-625-5p. Upregulating TRPM2 likewise reversed the therapeutic effect of depleting circEEF2 on cancer development in PCa cells. circEEF2 activated the Hedgehog pathway through the miR-625-5p/TRPM2 axis, promotes mitochondrial stress, and promotes PCa development in vivo. circEEF2 upregulates mitochondrial stress to promote PCa by activating the Hedgehog pathway through the miR-625-5p/TRPM2 axis.

Multicellular adaptation to electrophysiological perturbations analyzed by deterministic and stochastic bioelectrical models

Cells can compensate a disruptive change in one ion channel by compensatory changes in other channels. We have simulated the adaptation of a multicellular aggregate of non-excitable cells to the electrophysiological perturbation produced by the external blocking of a cation channel. In the biophysical model employed, we consider that this blocking provokes a cell depolarization that opens a voltage-gated calcium channel, thus allowing toxic Ca2+ levels. The cell adaptation to this externally-induced perturbation is ascribed to the multiplicity of channels available to keep the cell membrane potential within a physiological window. We propose that the cell depolarization provokes the upregulated expression of a compensatory channel protein that resets the cell potential to the correct polarized value, which prevents the calcium entry. To this end, we use two different simulation algorithms based on deterministic and stochastic methods. The simulations suggest that because of the local correlations coupling the cell potential to transcription, short-term bioelectrical perturbations can trigger long-term biochemical adaptations to novel stressors in multicellular aggregates. Previous experimental data on planarian flatworms’ adaptation to a barium-containing environment is also discussed.

Numerical simulation study on discharge characteristics of the Dual hollow cathode plasma source using magnetic mirror field

The Dual hollow cathode plasma source uses hollow cathode cylindrical sputtering and ionizing metal elements, which has a high material utilization rate and has potential application in the field of material modification. Its performance index requires the cooperation of electrode discharge parameters and the rational utilization of the inner surface of the hollow cathode. In this paper, a two-dimensional fluid model of a Dual hollow cathode structure is developed to simulate the basic distribution of the Dual hollow cathode discharge, which fills the central region of the Dual hollow cathode structure and the electrons oscillate on the axis. The effects of electrode parameters and hollow cathode radius on the discharge were investigated. The results show that the radial electron density of the hollow cathode increases with increasing heated cathode voltage, but as the hot cathode voltage continues to expand beyond 1200 V this trend becomes less obvious. The enhancement of the wall ion current by the heated cathode voltage is very pronounced, and the hollow cathode discharge is the result of the mutual enhancement of the heated cathode voltage and the hollow cathode voltage. Hollow cathode sputtering needs to reach a voltage threshold of 300 V, and the existence of an optimal radius (r = 4 mm−6 mm) to achieve a high plasma density and at the same time favor hollow cathode sputtering.

Rhythmic Contractions of Lymph Vessels and Lymph Flow Are Disrupted in Hypertensive Rats.

Hypertension increases the risk of lymphedema in patients with comorbidities, but whether hypertension directly compromises lymph vessel (LV) function and lymph flow is unclear. We compared the contractions of mesenteric LVs ex vivo and lymph flow in vivo between normotensive and Ang II (angiotensin II)-induced hypertensive rats and explored the ionic basis of contractile patterns. Key studies were recapitulated in spontaneously hypertensive rats and control Wistar-Kyoto rats. Video microscopy continuously recorded the diameters of cannulated rat mesenteric LVs, and high-speed optical imaging estimated mesenteric lymph flow in vivo. Jess capillary Western electrophoresis evaluated expression levels of ion channel proteins. Isolated LVs from Ang II-induced hypertensive rats exhibited dysrhythmic contractions, whereas LVs from both Ang II-induced hypertensive rats and spontaneously hypertensive rats exhibited reduced diastolic diameters and cross-sectional flow. Mesenteric lymph flow in vivo was 2.9-fold lower in Ang II-induced hypertensive rats compared with normotensive rats. Surprisingly, the LVs from Ang II-induced hypertensive rats expressed fewer intact L-type Ca2+ channel pore proteins and more modulatory cleaved C-terminal fragments. However, pharmacological block of voltage-gated K+ channels but not other K+ channel types in control LVs established the pattern of contractile dysfunction observed in hypertension. Jess capillary Western electrophoresis analysis confirmed a loss of Shaker-type KV1.2 channels in LVs from hypertensive rats. We provide initial evidence of lymphatic contractile dysfunction and compromised lymph flow in hypertensive rats, which may be caused by a loss of KV1.2 channels in the lymphatic muscle cells.

Identification of hypoxene metabolites in urine samples using gas chromatography–tandem mass spectrometry for anti-doping control

Objectives. Hypoxen is a drug which possesses antioxidant and antihypoxic effects. It achieves this by increasing the utilization of oxygen by mitochondria, intensifying oxidative phosphorylation, and as a result, improving tissue respiration. Athletes take it during prolonged exercise, in order to increase efficiency and reduce physical overwork. Since 2023, the World Anti-Doping Agency has included the drug in the monitoring program, in the belief that it can be used to gain a competitive advantage. It is thus a candidate for inclusion in the Prohibited List as a potential regulator of the human metabolism. Currently, there are no studies or scientific publications focusing on the identification of hypoxene in biofluids for the purpose of anti-doping control. The aim of this study is to determine the possible metabolites of the drug and their chromato-mass spectrometric characteristics in urine samples using gas chromatography–tandem mass spectrometry (GC–MS/MS) for doping control screening purposes.Methods. Sample preparation of urine samples was carried out using enzymatic hydrolysis, liquid–liquid extraction and derivatization. The GC–MS/MS method was used for analysis. Screening of hypoxene metabolites was carried out in the mode of total ion current after fragmentation of selected parent ions.Results. Three specific metabolites of hypoxene (m/z 342, 300, and 346, including trimethylsilyl derivatives) were identified in urine samples of volunteers (n = 3). They can act as markers for taking the target antihypoxant, and their possible structural formulas are given. The excretion curves of two metabolites with an m/z of 300 and 346 respectively in urine were studied. The maximum concentration is reached after 8–14 and 1.5–6 h, respectively. It was established, that these metabolites are reliably identified in urine 90 h or more after a single dose of the drug.Conclusions. Possible structures of hypoxene metabolites in urine samples from volunteers were determined for the first time and their chromato-mass spectrometric characteristics were established. The approach developed in this study can be used for screening analysis of hypoxene for the purpose of anti-doping control.

Molecular dynamics simulations reveal concentration-dependent blockage of graphene quantum dots to water channel protein openings

Graphene quantum dots (GQDs) have attracted significant attention across various scientific research areas due to their exceptional properties. However, studies on the potential toxicity of GQDs have yielded conflicting results. Therefore, a comprehensive evaluation of the toxicity profile of GQDs is essential for a thorough understanding of their biosafety. In this work, employing a molecular dynamics (MD) simulation approach, we investigate the interactions between GQDs and graphene oxide quantum dots (GOQDs) with the AQP1 water channel protein, aiming to explore the potential biological influence of GQDs/GOQDs. Our MD simulation results reveal that GQDs can adsorb to the loop region around the openings of AQP1 water channels, resulting in the blockage of these channels and potential toxicity. Interestingly, this blockage is concentration-dependent, with higher GQD concentrations leading to a greater likelihood of blockage. Additionally, GOQDs show a lower probability of blocking the openings of AQP1 water channels compared to GQDs, due to the hydrophobicity of the loop regions around the openings, which ultimately leads to lower interaction energy. Therefore, these findings provide new insights into the potential adverse impact of GQDs on AQP1 water channels through the blockage of their openings, offering valuable molecular insights into the toxicity profile of GQD nanomaterials.