Values Of Membrane Potential Research Articles

Assessing the Relationship between proAKAP4 Level and Longevity of Sexed Sperm Quality after Thawing.

ProAKAP4 is a sperm structural protein that regulates motility through the PKA-dependent cAMP signaling pathway, which is synthesized as an X chromosome-linked member of the gene family. This study aims to determine the optimal level of proAKAP4 for evaluating sexed semen through investigating its relationship with the longevity of sperm quality in sexed Holstein bull sperm. A total of 30 sexed sperm samples (bearing X chromosomes) from 30 distinct Holstein bulls (n = 30) were analyzed. The frozen bull sperm samples were assessed for their proAKAP4 levels, mitochondrial membrane potential, plasma membrane and acrosome integrity (PMAI), and spermatozoa movement parameters at hours 0 and 3 after thawing. The proAKAP4 levels in the sexed sperm samples ranged from 16.35 to 72.10 ng/10 M spz, with an average of 37.18 ± 15.1 ng/10 M spz. A strong positive correlation was observed between proAKAP4 levels and total motility, progressive motility, PMAI, high mitochondrial membrane potential, VAP, and VCL values after 3 h of incubation, when compared to post-thaw analyses. The results also reveal that spermatozoa with proAKAP4 levels of ≥40 ng/10 M spz exhibit higher quality. In conclusion, the level of proAKAP4 in sexed sperm aligns with previous studies and shows potential as a biomarker for assessing the longevity of sexed sperm quality.

Thiacalix[4]arene chalcone amides effect on myometrium contraction

Calixarenes are macrocyclic compounds, the biochemical effects of which are actively studied. In this study we synthesized thiacalix[4]arene chalcone amides С-1191 and С-1192, which have a sulfur atom in their structure and different spatial arrangement of chalcone amide groups, and studied their effect on myometrium functioning. Experiments were conducted with the use of rat uterine smooth muscles preparations, isolated myometrial mitochondria and permeabilized myometrial cells. The relative value of mitochondria membrane potential (Δψ) was assayed with a voltage-sensitive fluorescent probe TMRM. The spontaneous contractive activity was studied by tenzometric method followed by mechanokinetic analysis. It was shown that C-1191 and C-1192 induced mitochondria hyperpolarization and increased the basal tension of myometrium smooth muscle preparation. Thiacalix[4]arene С-1191 did not change the uterine cycle, but increased the force, velocity and impulse parameters of muscle contractive activity. On the contrary, С-1192 modified the uterine cycle considerably, increased the total efficiency of the myometrium spontaneous contractive activity and decreased the force, time and impulse parameters. It is concluded that changes in the mechanokinetic parameters of myometrial contractile activity induced by С-1191 and С-1192 are determined by the functional activity of mitochondria. Keywords: mechanokinetic analysis, mitochondria, mitochondria membrane potential, myometrial contractile activity, thiacalix[4]arene chalcone amides

Changes in bioenergetic characteristics of the murine lymphoma cells under the action of a thiazole derivative in complex with polymeric nanoparticles

Background. Mitochondria can influence cancer cells both indirectly via reactive oxygen species mediation and directly through mitochondrial biogenesis. Energy production in mitochondria is crucial as it facilitates the synthesis of essential molecules needed for cellular biosynthesis, growth, and proliferation. The development of new anticancer drugs that target the energy metabolism of tumor cells shows promise in cancer treatment. Our study aimed to investigate how the thiazole derivative N-(5-benzyl-1,3-thiazol-2-yl)-3,5-dimethyl-1-benzofuran-2-carboxamide (BF1), the polymeric nanoparticles based on the polyethylene glycol (PEG-PN, Th5), and their complex with BF1 (Th6) affect respiration and mitochondrial membrane potential in murine NK/Ly tumor cells. Materials and Methods. The study was performed on white wild-type male mice with grafted NK/Ly lymphoma. The test substances were added to the cell suspension at a final concentration of 10 μM and incubated for 15 min at 37 °C. Oxygen uptake rates in NK/Ly cells were measured using a polarographic method with Clark electrode. Changes in mitochondrial membrane potential were assessed using the tetramethylrhodamine methyl ester fluorescence dye. The fluorescence intensity was evaluated using the ImageJ computer program. Results. After incubating NK/Ly cells with BF1 (10 µM), Th5, or the BF1 + PEG-PN complex (Th6) for 15 min, no changes were observed in glucose-fueled basal respiration. However, the Th6 complex significantly activated FCCP-stimulated respiratory processes in NK/Ly lymphoma cells. Fluorescent microscopy data indicated that BF1 or Th5 alone did not affect mitochondrial membrane potential values. However, the Th6 complex significantly decreased mitochondrial membrane potential, suggesting a reduction in NK/Ly cell viability. Conclusions The investigated complex of thiazole derivative BF1 with PEG-based polymeric nanoparticles may realize its cytotoxic effect by depolarization of mitochondrial membrane in NK/Ly lymphoma cells.

Analysis of the dynamical behavior of discrete memristor-coupled scale-free neural networks

The synchronization of neural networks is crucial for neural information processing and represents a key feature of various functional brain diseases. Memristors are ideal electronic components for mimicking biological synapses, among which discrete memristors have the advantage of fast computing speed and are often used in memristor-based neural networks. For these reasons, this paper proposes a novel discrete memristor-coupled Scale-Free neural network (DMSNN). Phase diagrams and time series of membrane potential are employed to analyze the firing pattern coexistence of individual neurons in the network. Furthermore, Spatiotemporal patterns, heat maps of the Spearman correlation coefficient matrix and the values of neuron membrane potential at a particular time point are adopted to declare the spatio-temporal dynamics of the complex neural network, encompassing asynchronization, chimeric state, synchronization and synchronization transition. The study also identifies the phenomenon of topology-induced coexistence and elucidates the underlying reasons for the emergence of chimeric states in the DMSNN as the coupling strength increases. Finally, a hardware implementation platform is constructed using a highly integrated SSD202 processor to validate the accuracy of the DMSNN. The results are consistent with the numerical simulations.

Label-free, non-contact determination of resting membrane potential using dielectrophoresis

Measurement of cellular resting membrane potential (RMP) is important in understanding ion channels and their role in regulation of cell function across a wide range of cell types. However, methods available for the measurement of RMP (including patch clamp, microelectrodes, and potential-sensitive fluorophores) are expensive, slow, open to operator bias, and often result in cell destruction. We present non-contact, label-free membrane potential estimation which uses dielectrophoresis to determine the cytoplasm conductivity slope as a function of medium conductivity. By comparing this to patch clamp data available in the literature, we have demonstratet the accuracy of this approach using seven different cell types, including primary suspension cells (red blood cells, platelets), cultured suspension cells (THP-1), primary adherent cells (chondrocytes, human umbilical mesenchymal stem cells), and adherent (HeLa) and suspension (Jurkat) cancer cell lines. Analysis of the effect of ion channel inhibitors suggests the effects of pharmaceutical agents (TEA on HeLa; DMSO and neuraminidase on red blood cells) can also be measured. Comparison with published values of membrane potential suggest that the differences between our estimates and values recorded by patch clamp are accurate to within published margins of error. The method is low-cost, non-destructive, operator-independent and label-free, and has previously been shown to allow cells to be recovered after measurement.

A mandibular advancement device attenuates the abnormal morphology and function of mitochondria from the genioglossus in obstructive sleep apnea-hypopnea syndrome rabbits.

Obstructive sleep apnea hypopnea syndrome (OSAHS) is a serious and potentially life-threatening disease. Mandibular advancement device (MAD) has the characteristics of non-invasive, comfortable, portable and low-cost, making it the preferred treatment for mild-to-moderate OSAHS. Our previous studies found that abnormal contractility and fibre type distribution of the genioglossus could be caused by OSAHS. However, whether the mitochondria participate in these tissue changes is unclear. The effect of MAD treatment on the mitochondria of the genioglossus in OSAHS is also uncertain. To examine the morphology and function of mitochondria from the genioglossus in a rabbit model of obstructive sleep apnea-hypopnea syndrome (OSAHS), as well as these factors after insertion of a mandibular advancement device (MAD). Thirty male New Zealand white rabbits were randomised into three groups: control, OSAHS and MAD, with 10 rabbits in each group. Animals in Group OSAHS and Group MAD were induced to develop OSAHS by injection of gel into the submucosal muscular layer of the soft palate. The rabbits in Group MAD were fitted with a MAD. The animals in the control group were not treated. Further, polysomnography (PSG) and cone-beam computed tomography (CBCT) scan were used to measure MAD effectiveness. CBCT of the upper airway and PSG suggested that MAD was effective. Rabbits in the three groups were induced to sleep for 4-6 h per day for eight consecutive weeks. The genioglossus was harvested and detected by optical microscopy and transmission electron microscopy. The mitochondrial membrane potential was determined by laser confocal microscopy and flow cytometry. Mitochondrial complex I and IV activities were detected by mitochondrial complex assay kits. OSAHS-like symptoms were induced successfully in Group OSAHS and rescued by MAD treatment. The relative values of the mitochondrial membrane potential, mitochondrial complex I activity and complex IV activity were significantly lower in Group OSAHS than in the control group; however, there was no significant difference between Group MAD and the control group. The OSAHS-induced injury and the dysfunctional mitochondria of the genioglossus muscle were reduced by MAD treatment. Damaged mitochondrial structure and function were induced by OSAHS and could be attenuated by MAD treatment.

Mechanosensor channels of the lens and ciliary body: patch clamp studies

Epithelia from the lens and the ciliary body possess mechanosensors, including Piezo1 and transient receptor potential (TRP) channels, that may partake in the regulation of lens and aqueous humor volume. Although these mechanosensor channels commonly allow calcium entry, the outcomes of elevated intracellular calcium vary with the type of channel activated. For instance, separate activation of TRPV4 and TRPV1 increases Na-K-ATPase and Na/K/2Cl cotransporter activities, respectively. To further understand the role of mechanosensitivity in lens and ciliary body, we are currently using patch clamp techniques to characterize the response of primary cultured cells to chemical agonists and antagonists, as well as direct mechanical stimulation. Previously, we found that in mouse lens epithelial cells (mLEC), TRPV4 activation by agonist GSK1016790A (GSK) is followed by disappearance of a fast, transient K+-like outward current (Ipeak), decrease of a steady state outward current (Iss), fast depolarization of resting membrane potential (RMP) values from near -50 mV to near 0 mV, increase of Cx43 hemichannel-like events, and increase of overall membrane conductance ( gm) at holding potential (HP) = -50mV. More recently, we found that in mLEC, shear stress (streaming of external solution from a pipette tip over the cell membrane) reduced Ipeak but did not cause large RMP changes. Porcine non-pigmented epithelial cells (pNPE) in normal conditions display no Ipeak, small Iss, small RMP near -20 mV and spontaneous Cx43 hemichannel-like event activity. Nevertheless, GSK caused further depolarization of pNPE cells, and while no gm changes were seen when GSK was applied while holding cells at -50 mV, gm did increase when GSK exposure occurred during repeated ±80 mV pulses. Preliminary experiments show that activation of TRPM3 by its agonist pregnenolone causes a minimal hyperpolarization in mLEC. In comparison, in a line of human LEC (HLE B3) expressing TRPM3 channels, and which appear generally depolarized, pregnenolone caused a proportionally larger shift toward more negative RMP values; more intriguing, GSK has no measurable effect on the gm of HLE B3 cells. Taken together, and apart from considerations of cell- and species-specific differences, the data suggest that mechanosensor channels have different and/or opposite effects on the regulation of membrane ion channel activity, and thus on cell function. R01EY029171 and R01EY009532. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

A red-emitting carborhodamine for monitoring and measuring membrane potential

Biological membrane potentials, or voltages, are a central facet of cellular life. Optical methods to visualize cellular membrane voltages with fluorescent indicators are an attractive complement to traditional electrode-based approaches, since imaging methods can be high throughput, less invasive, and provide more spatial resolution than electrodes. Recently developed fluorescent indicators for voltage largely report changes in membrane voltage by monitoring voltage-dependent fluctuations in fluorescence intensity. However, it would be useful to be able to not only monitor changes but also measure values of membrane potentials. This study discloses a fluorescent indicator which can address both. We describe the synthesis of a sulfonated tetramethyl carborhodamine fluorophore. When this carborhodamine is conjugated with an electron-rich, methoxy (-OMe) containing phenylenevinylene molecular wire, the resulting molecule, CRhOMe, is a voltage-sensitive fluorophore with red/far-red fluorescence. Using CRhOMe, changes in cellular membrane potential can be read out using fluorescence intensity or lifetime. In fluorescence intensity mode, CRhOMe tracks fast-spiking neuronal action potentials (APs) with greater signal-to-noise than state-of-the-art BeRST 1 (another voltage-sensitive fluorophore). CRhOMe can also measure values of membrane potential. The fluorescence lifetime of CRhOMe follows a single exponential decay, substantially improving the quantification of membrane potential values using fluorescence lifetime imaging microscopy (FLIM). The combination of red-shifted excitation and emission, mono-exponential decay, and high voltage sensitivity enable fast FLIM recording of APs in cardiomyocytes. The ability to both monitor and measure membrane potentials with red light using CRhOMe makes it an important approach for studying biological voltages.

Dynamical analysis of the FitzHugh–Nagumo model with memristive synapse

Understanding the transition mechanism between different activity patterns in realistic neuron models brings forward fundamental challenges for the dynamical systems theory. The knowledge about the mechanisms can present precious predictions and intuitions for determining fundamental principles of neuron functioning. This paper aims to present a dynamic analysis of the FitzHugh–Nagumo neuron model with a memristive synapse. In this article, the effects of memristive synapse's conductance gain, applied current, and the speed of electrical signal propagation along axons and dendrites on the system's behavior and the neuron's membrane potential are investigated. Analyzes are performed by investigating bifurcation and Lyapunov exponent diagrams versus various model parameters. The physiological role of some parameters and the effect of parameter changes on the model behaviors are discussed. The results demonstrate that when no current is applied to the system, the maximum value of membrane potential is larger, and also, a lower memristive synapse conductance gain leads to a larger maximum value of membrane potential. So, it can be concluded that the neuron in an amplitude death state may be triggered and start firing by decreasing the conductance gain or not applying current.

Harmful effects of true-to-life nanoplastics derived from PET water bottles in human alveolar macrophages.

The increasing presence of secondary micro/nanoplastics (MNPLs) in the environment requires knowing if they represent a real health concern. To such end, an important point is to test representative MNPLs such as the denominated true-to-life MNPLs, resulting from the degradation of plastic goods in lab conditions. In this study, we have used polyethylene terephthalate (PET) NPLs resulting from the degradation of PET water bottles. Since inhalation is an important exposure route to environmental MNPLS, we have used mouse alveolar macrophages (MH-S) as a target cell, and the study focused only on the cells that have internalized them. This type of approach is novel as it may capture the realistic adverse effects of PETNPLs only in the internalized cells, thereby mitigating any biases while assessing the risk of these MNPLs. Furthermore, the study utilized a set of biomarkers including intracellular reactive oxygen species (ROS) levels, variations on the mitochondrial membrane potential values, and the macrophage polarization to M1 (pro-inflammatory response) and M2 (anti-proinflammatory response) as possible cellular effects due to PETNPLs in only the cells that internalized PETNPLs. After exposures lasting for 3 and 24 h to a range of concentrations (0, 25, 50, and 100 μg/mL) the results indicate that no toxicity was induced despite the 100% internalization observed at the highest concentration. Significant intracellular levels of ROS were observed, mainly at exposures lasting for 24 h, in an indirect concentration-effect relationship. Interestingly, a reduction in the mitochondrial membrane potential was observed, but only at exposures lasting for 24 h, but without a clear concentration-effect relationship. Finally, PETNPL exposure shows a significant polarization from M0 to M1 and M2 subtypes. Polarization to M1 (pro-inflammatory stage) was more marked and occurred at both exposure times. Polarization to M2 (anti-inflammatory stage) was only observed after exposures lasting for 24 h. Due to the relevance of the described biomarkers, our results underscore the need for further research, to better understand the health implications associated with MNPL exposure.

Optical Estimation of Membrane Potential Values Using Fluorescence Lifetime Imaging Microscopy and Hybrid Chemical-Genetic Voltage Indicators.

Membrane potential (Vm), the voltage across a cell membrane, is an important biophysical phenomenon, central to the physiology of cells, tissues, and organisms. Voltage-sensitive fluorescent indicators are a powerful method for interrogating membrane potential in living systems, but most indicators are best suited for detecting changes in membrane potential rather than measuring values of the membrane potential. One promising approach is to use fluorescence lifetime imaging microscopy (FLIM) in combination of chemically synthesized dyes to estimate a value of membrane potential. However, a drawback is that chemically synthesized dyes show poor specificity of staining. To address this problem, we applied a chemical-genetic voltage imaging approach to FLIM to enable optical estimation of membrane potential values from genetically defined cells. In this report, we detail the characterization and evaluation of two of these systems in mammalian cells. We further validate the use of a FLIM-based chemical genetic voltage indicator in mammalian neurons. Finally, we discuss opportunities for future improvements to chemical-genetic FLIM-based voltage indicators.

Mass Spectrometric Identification of Metabolites after Magnetic-Pulse Treatment of Infected Pyrus communis L. Microplants

The major goal of this study is to create a venue for further work on the effect of pulsed magnetic fields on plant metabolism. It deals with metabolite synthesis in the aforementioned conditions in microplants of Pyrus communis L. So far, there have been glimpses into the governing factors of plant biochemistry in vivo, and low-frequency pulsed magnestatic fields have been shown to induce additional electric currents in plant tissues, thus perturbing the value of cell membrane potential and causing the biosynthesis of new metabolites. In this study, sixty-seven metabolites synthesized in microplants within 3–72 h after treatment were identified and annotated. In total, thirty-one metabolites were produced. Magnetic-pulse treatment caused an 8.75-fold increase in the concentration of chlorogenic acid (RT = 8.33 ± 0.0197 min) in tissues and the perturbation of phenolic composition. Aucubin, which has antiviral and antistress biological activity, was identified as well. This study sheds light on the effect of magnetic fields on the biochemistry of low-molecular-weight metabolites of pear plants in vitro, thus providing in-depth metabolite analysis under optimized synthetic conditions. This study utilized high-resolution gas chromatography-mass spectrometry, metabolomics methods, stochastic dynamics mass spectrometry, quantum chemistry, and chemometrics, respectively. Stochastic dynamics uses the relationships between measurands and molecular structures of silylated carbohydrates, showing virtually identical mass spectra and comparable chemometrics parameters.

Minimal model for the dependence of stresses in the wall of a cerebral vessel on the parameters of a smooth muscle cell

A minimal mathematical model of the wall of a small arterial vessel is described. This model is created based on the published results of experiments performed on rat cerebral vessels. It is assumed that the active stress has only a circumferential component and depends on the circumferential stretch, calcium concentration in the cytoplasm, and the membrane potential of smooth muscle cells. The presented model for a small artery can qualitatively reproduce the results of more sophisticated models for other vessels under normal physiological conditions. Unlike a similar model, that accounts for only one cellular parameter, the addition of membrane potential as one of the main parameters was crucial to reveal a qualitative change in the dependency of circumferential stress on stretch and the radial coordinate with an alteration in vascular tone. At fixed values of membrane potential and calcium concentration in the phase of a development of vascular tone, stress decreases as it approaches the outer wall of the vessel and increases as stretch increases, and after it is formed, the direction of changes in the circumferential stress reverses.

Tannic Acid Interfacial Modification of Prochloraz Ethyl Cellulose Nanoparticles for Enhancing the Antimicrobial Effect and Biosafety of Fungicides.

With the poorly soluble and intrinsically unstable feature, prochloraz (Pro) was confronted with lower bioavailability in the crop defense against fungal erosion. Therefore, it was a challenging project to explore the innovative antifungal compound delivery system for improving bioavailability. The superior adhesive fungicide formulation was supposed to be an efficient pathway to enhance transmembrane permeability and biological activity. According to abundant phenolic hydroxyl groups, tannic acid (TA) was an ideal modified adhesive biomaterial to improve interfacial interactions. The fundamental purpose of this research was focused on the synergistic mechanism of TA-interfacial-modified Pro-ethyl cellulose (EC) nanoparticles for improving bioavailability and biosafety. In the stability test, TA-modified Pro-EC nanoparticles had the capacity to reduce Pro initial release burst, extending a persistent validity and improving anti-photodegradation property. The toxicity index of Pro-EC and Pro-EC-TA was approximately 2.93-fold and 4.96-fold that of Pro technical against Fusarium graminearum (F. graminearum), respectively. Compared with nonmodified EC nanoparticles, TA-modified EC nanoparticles obtained eminent transmembrane permeability and superior adherence ability to F. graminearum, for hydroxyl and carboxyl groups of TA to enhance interaction with target cell membranes. The contents of cellular reactive oxygen species induced by Pro-EC and Pro-EC-TA nanoparticles were about 2.31 times and 3.00 times that of the control check (CK), respectively. Compared to the CK group, the membrane potential and ergosterol values of F. graminearum treated with Pro-EC-TA nanoparticles were drastically reduced by 74.91 and 56.20%, respectively. In the biosafety assay, the maximum half-lethal concentration value of the TA-modified Pro-EC nanoparticles indicated that the acute toxicity of the Pro-EC-TA nanoparticles to adult zebrafish was approximately 8.34-fold reduced compared to that of the Pro technical. These findings demonstrated that the successful interfacial modification of Pro-EC nanoparticles with TA was a highly efficient, environmentally safe, and promising alternative for sustainable agricultural application, thus making the fungicide formulation process more simplified, easier fabrication, and lower cost.

Electrochemical Characterization of Charged Membranes from Different Materials and Structures via Membrane Potential Analysis.

Electrochemical characterization of positively and negatively charged membranes is performed by analyzing membrane potential values on the basis of the Teorell-Meyer-Sievers (TMS) model. This analysis allows the separate estimation of Donnan (interfacial effects) and diffusion (differences in ions transport through the membrane) contributions, and it permits the evaluation of the membrane's effective fixed charge concentration and the transport number of the ions in the membrane. Typical ion-exchange commercial membranes (AMX, Ionics or Nafion) are analyzed, though other experimental and commercial membranes, which are derived from different materials and have diverse structures (dense, swollen or nanoporous structures), are also considered. Moreover, for some membranes, changes associated with different modifications and other effects (concentration gradient or level, solution stirring, etc.) are also analyzed.

On the use of Kolmogorov equations for determining ion channels characteristics

The paper proposes a method for determination of the dynamic properties of conduction within voltage-gated ion channels with several sequentially located energy barriers named gate particles using a theory of Markov random processes in continuous time and with a discrete state space. The number of states was taken equal to the number of energy barriers in the channel plus one. If the hypothesis that random flows switching gate particles between an open and a closed are Poisson process, is true, then mathematical description of the system’s state of similar channels can be a system of Kolmogorov linear differential equations for state probabilities. Using this model and based on Volt-Clamp test results, published in open access journals, it is possible to find out how fixed values of membrane potential depend on the intensities of Poisson process for different types of potassium channels. Function parameters, that describe the intensity of ion transition across the channel, in the Kolmogorov equation can be restored using the generalized least squares method. This paper contains the examples of determining the intensities of transitions for two types of voltage-dependent potassium channels known as «delayed rectifiier potassium channels» (IKdr, two identical activation gate particles) and channels with fast activation and inactivation processes (IKa, three identical activation and one inactivation gate particles). It is shown that channel activation and deactivation is described by solving the general Kolmogorov equation.

Neuroprotective and Antiherpetic Properties of Polyphenolic Compounds from Maackia amurensis Heartwood.

In this study, we isolated a new isoflavanostilbene maackiapicevestitol (1) as a mixture of two stable conformers 1a and 1b as well as five previously known dimeric and monomeric stilbens: piceatannol (2), maackin (3), scirpusin A (4), maackiasine (5), and maackolin (6) from M. amurensis heartwood, using column chromatography on polyamide, silicagel, and C-18. The structures of these compounds were elucidated by NMR, HR-MS, and CD techniques. Maksar® obtained from M. amurensis heartwood and polyphenolics 1-6 possessed moderate anti-HSV-1 activity in cytopathic effect (CPE) inhibition and RT-PCR assays. A model of PQ-induced neurotoxicity was used to study the neuroprotective potential of polyphenolic compounds from M. amurensis. Maksar® showed the highest neuroprotective activity and increased cell viability by 18% at a concentration of 10 μg/mL. Maackolin (6) also effectively increased the viability of PQ-treated Neuro-2a cells and the value of mitochondrial membrane potential at concentrations up to 10 μΜ. Maksar® and compounds 1-6 possessed higher FRAP and DPPH-scavenging effects than quercetin. However, only compounds 1 and 4 at concentrations of 10 μM as well as Maksar® (10 μg/mL) statistically significantly reduced the level of intracellular ROS in PQ-treated Neuro-2a cells.

Bioenergetic characteristics of the murine Nemeth-Kellner lymphoma cells exposed to thiazole derivative in complex with polymeric nanoparticles

The development of a new anticancer drugs targeted at energy metabolism of tumor cells is a promising­ approach for cancer treatment. The aim of our study was to investigate the action of thiazole derivative N-(5-benzyl-1,3-thiazol-2-yl)-3,5-dimethyl-1-benzofuran-2-carboxamide (BF1) and its complex with PEG based polymeric nanoparticle (PEG-PN) on respiration and mitochondrial membrane potential in murine NK/Ly tumor cells. The rate of oxygen uptake in NK/Ly cells was recorded by a polarographic method using a Clark electrode. The mitochondrial potential relative values were registered using fluorescence TMRM dye. No changes in glucose-fuelled basal respiration or maximal FCCP-stimulated respiration was detected after 15-min incubation of cells with BF1 (10 µM), PEG-PN or BF1 + PEG-PN complex Fluorescent microscopy data showed that BF1 or PEG-PN separately had no effect on the value of mitochondrial membrane potential, while BF1 + PEG-PN complex caused a significant decrease in mitochondrial membrane potential, indicating­ on the decrease of NK/Ly cells viability. Keywords: cell respiration, mitochondrial membrane potential, NK/Ly tumor cells, PEG, polymeric nanoparticles, thiazole derivative

Fluorescence Imaging of Cell Membrane Potential: From Relative Changes to Absolute Values.

Membrane potential is a fundamental property of biological cells. Changes in membrane potential characterize a vast number of vital biological processes, such as the activity of neurons and cardiomyocytes, tumorogenesis, cell-cycle progression, etc. A common strategy to record membrane potential changes that occur in the process of interest is to utilize organic dyes or genetically-encoded voltage indicators with voltage-dependent fluorescence. Sensors are introduced into target cells, and alterations of fluorescence intensity are recorded with optical methods. Techniques that allow recording relative changes of membrane potential and do not take into account fluorescence alterations due to factors other than membrane voltage are already widely used in modern biological and biomedical studies. Such techniques have been reviewed previously in many works. However, in order to investigate a number of processes, especially long-term processes, the measured signal must be corrected to exclude the contribution from voltage-independent factors or even absolute values of cell membrane potential have to be evaluated. Techniques that enable such measurements are the subject of this review.

ПУТИ РЕАЛИЗАЦИИ АПОПТОЗА ЛИМФОЦИТОВ ЧЕЛОВЕКА, ИНДУЦИРОВАННОГО ПЕРОКСИДОМ ВОДОРОДА, И ИХ РЕГУЛИРОВАНИЕ

The changes in marker indicators of apoptotic cell death were studied: the level of functional activity of initiating caspases-8, -9, -12, membrane death receptors Fas, intracellular reactive oxygen species, cytosolic calcium, mitochondrial potential of peripheral blood lymphocytes of donors induced by exposure to hydrogen peroxide at final concentrations of 10-6 and 10-5 mol/l. Activation of initiating caspase-8 associated with membrane death receptors Fas was detected after exposure to hydrogen peroxide lymphocytes at a concentration of 10-6 mol/l. There was a significant increase in the functional activity of initiating caspase-9, the level of Fas receptors and intracellular reactive oxygen species, as well as a decrease in the concentration of cytosolic calcium and the value of the mitochondrial membrane potential 1 and 2 hours after modification of cells with hydrogen peroxide at a concentration of 10-5 mol/l compared with those for intact immunocytes. It was found that apoptosis of lymphocytes under the influence of exogenous hydrogen peroxide is realized with the participation of receptor-mediated caspase, mitochondrial (with activation of caspase-9) and p53-dependent mechanisms. The protective effect of resveratrol (10-6 mol/l) on lymphocytes after exposure to hydrogen peroxide (10-5 mol / l), which is due to its ability to reduce the level of reactive oxygen species in intact and modified hydrogen peroxide immunocytes, as well as deactivate hydrogen peroxide.