Reversal Potential Research Articles

Conditional deletion of KCC2 impairs synaptic plasticity and both spatial and nonspatial memory.

The postsynaptic inhibition through GABAA receptors (GABAAR) relies on two mechanisms, a shunting effect due to an increase in the postsynaptic membrane conductance and, in mature neurons, a hyperpolarization effect due to an entry of chloride into postsynaptic neurons. The second effect requires the action of the K+-Cl- cotransporter KCC2 which extrudes Cl- from the cell and maintains its cytosolic concentration very low. Neuronal chloride equilibrium seems to be dysregulated in several neurological and psychiatric conditions such as epilepsy, anxiety, schizophrenia, Down syndrome, or Alzheimer's disease. In the present study, we used the KCC2 Cre-lox knockdown system to investigate the role of KCC2 in synaptic plasticity and memory formation in adult mice. Tamoxifen-induced conditional deletion of KCC2 in glutamatergic neurons of the forebrain was performed at 3 months of age and resulted in spatial and nonspatial learning impairment. On brain slices, the stimulation of Schaffer collaterals by a theta burst induced long-term potentiation (LTP). The lack of KCC2 did not affect potentiation of field excitatory postsynaptic potentials (fEPSP) measured in the stratum radiatum (dendrites) but increased population spike (PS) amplitudes measured in the CA1 somatic layer, suggesting a reinforcement of the EPSP-PS potentiation, i.e., an increased ability of EPSPs to generate action potentials. At the cellular level, KCC2 deletion induced a positive shift in the reversal potential of GABAAR-driven Cl- currents (EGABA), suggesting an intracellular accumulation of chloride subsequent to the downregulation of KCC2. After treatment with bumetanide, an antagonist of the Na+-K+-Cl- cotransporter NKCC1, spatial memory impairment, chloride accumulation, and EPSP-PS potentiation were rescued in mice lacking KCC2. The presented results emphasize the importance of chloride equilibrium and GABA-inhibiting ability in synaptic plasticity and memory formation.

Cardiac PDGFRα+ interstitial cells generate spontaneous inward currents that contribute to excitability in the heart.

The cell types and conductance that contribute to normal cardiac functions remain under investigation. We used mice that express an enhanced green fluorescent protein (eGFP)-histone 2B fusion protein driven off the cell-specific endogenous promoter for Pdgfra to investigate the distribution and functional role of PDGFRα+ cells in the heart. Cardiac PDGFRα+ cells were widely distributed within the endomysium of atria, ventricle, and sino-atrial node (SAN) tissues. PDGFRα+ cells formed a discrete network of cells, lying in close apposition to neighboring cardiac myocytes in mouse and Cynomolgus monkey (Macaca fascicularis) hearts. Expression of eGFP in nuclei allowed unequivocal identification of these cells following enzymatic dispersion of muscle tissues. FACS purification of PDGFRα+ cells from the SAN and analysis of gene transcripts by qPCR revealed that they were a distinct population of cells that expressed gap junction transcripts, Gja1 and Gjc1. Cardiac PDGFRα+ cells generated spontaneous transient inward currents (STICs) and spontaneous transient depolarizations (STDs) that reversed at 0 mV. Reversal potential was maintained when ECl = -40 mV. [Na+ ]o replacement and FTY720 abolished STICs, suggesting they were due to a non-selective cation conductance (NSCC) carried by TRPM7. PDGFRα+ cells also express β2 -adrenoceptor gene transcripts, Adrb2. Zinterol, a selective β2 -receptor agonist, increased the amplitude and frequency of STICs, suggesting these cells could contribute to adrenergic regulation of cardiac excitability. PDGFRα+ cells in cardiac muscles generate inward currents via an NSCC. STICs generated by these cells may contribute to the integrated membrane potentials of cardiac muscles, possibly affecting the frequency of pacemaker activity.

Responses in fast-spiking interneuron firing rates to parameter variations associated with degradation of perineuronal nets

The perineuronal nets (PNNs) are sugar coated protein structures that encapsulate certain neurons in the brain, such as parvalbumin positive (PV) inhibitory neurons. As PNNs are theorized to act as a barrier to ion transport, they may effectively increase the membrane charge-separation distance, thereby affecting the membrane capacitance. Tewari et al. (2018) found that degradation of PNNs induced a 25%-50% increase in membrane capacitance c_text {m} and a reduction in the firing rates of PV-cells. In the current work, we explore how changes in c_text {m} affects the firing rate in a selection of computational neuron models, ranging in complexity from a single compartment Hodgkin-Huxley model to morphologically detailed PV-neuron models. In all models, an increased c_text {m} lead to reduced firing, but the experimentally reported increase in c_text {m} was not alone sufficient to explain the experimentally reported reduction in firing rate. We therefore hypothesized that PNN degradation in the experiments affected not only c_text {m}, but also ionic reversal potentials and ion channel conductances. In simulations, we explored how various model parameters affected the firing rate of the model neurons, and identified which parameter variations in addition to c_text {m} that are most likely candidates for explaining the experimentally reported reduction in firing rate.

Abstract LB275: Transcriptomic reversal analysis yields cytokines and drugs mediating tumor microenvironmental reprogramming during cancer progression and therapy response

Abstract Recent computational work links therapy response to a drug based on the drug’s ability to reverse a tumor’s transcriptome towards a healthy state. This idea of reversal has been used to mine databases of cell-line transcriptional responses against drug libraries to prioritize anti-cancer drugs. However, there are two key shortcomings in these approaches: (1) though cytokines and their receptors are proposed as modulators of therapy response, there is no reversal-based method to prioritize cytokines as potential drugs or targets, and (2) responses of microenvironmental cell types to drugs, which dictate therapy response, have not been considered. We address these limitations by exploiting recent databases of cytokine transcriptional response and single-cell RNA-seq datasets of patient responses to cancer therapies. We first used a novel approach to derive drug response signatures from LINCS data that retained the coordinated nature of gene expression changes occurring during treatment. We then used these signatures to compute a transcriptional reversal score that ranks drugs by their ability to reverse TCGA RNA-seq profiles of a tumor towards its corresponding normal in GTEx. We found that FDA-approved drugs in prostate, lung, colorectal and breast adenocarcinomas have a significantly higher reversal potential than unapproved drugs, and that these drugs are more effective at in vitro cell killing amongst CTRP and GDSC cell viability measurements. Next, we extended our signature derivation and reversal computation approach to find cytokines in the CytoSig database that can reverse TCGA RNA-seq profiles. We found that the higher the reversal potential of a cytokine, the greater its association with better overall patient survival. In particular, our approach revealed the IL10 family and IL24 as potentially therapeutic cytokines based on their predicted ability to reverse cell states across tumor types and indeed found them to be associated with better overall and progression-free survival in the TCGA cohort. Finally, in two clinical cohorts where RNA-seq was carried out on breast cancer and multiple myeloma patients before and after therapy, cell types within the tumor microenvironment of responders showed a stronger reversal towards a healthy state compared to non-responders. Altogether, our work establishes the importance of transcriptional reversal in therapy response, particularly the role of microenvironmental transcriptional alterations. This allows us to refine the reversal principle based on pan-cell type transcriptional effects and to identify cytokines that mediate patient survival as a new class of drugs and drug targets. This research was supported by the Intramural Research Program of the NIH, Citation Format: Vishaka Gopalan, Sridhar Hannenhalli. Transcriptomic reversal analysis yields cytokines and drugs mediating tumor microenvironmental reprogramming during cancer progression and therapy response [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr LB275.

Acceleration of GABA-switch after early life stress changes mouse prefrontal glutamatergic transmission

Early life stress (ELS) alters the excitation-inhibition-balance (EI-balance) in various rodent brain areas and may be responsible for behavioral impairment later in life. The EI-balance is (amongst others) influenced by the switch of GABAergic transmission from excitatory to inhibitory, the so-called “GABA-switch”.Here, we investigated how ELS affects the GABA-switch in mouse infralimbic Prefrontal Cortex layer 2/3 neurons, using the limited-nesting-and-bedding model. In ELS mice, the GABA-switch occurred already between postnatal day (P) 6 and P9, as opposed to P15–P21 in controls. This was associated with increased expression of the inward chloride transporter NKCC1, compared to the outward chloride transporter KCC2, both of which are important for the intracellular chloride concentration and, hence, the GABA reversal potential (Erev). Chloride transporters are not only important for regulating chloride concentration postsynaptically, but also presynaptically. Depending on the Erev of GABA, presynaptic GABAA receptor stimulation causes a depolarization or hyperpolarization, and thereby enhanced or reduced fusion of glutamate vesicles respectively, in turn changing the frequency of miniature postsynaptic currents (mEPSCs). In accordance, bumetanide, a blocker of NKCC1, shifted the Erev GABA towards more hyperpolarized levels in P9 control mice and reduced the mEPSC frequency. Other modulators of chloride transporters, e.g. VU0463271 (a KCC2 antagonist) and aldosterone -which increases NKCC1 expression-did not affect postsynaptic Erev in ELS P9 mice, but did increase the mEPSC frequency. We conclude that the mouse GABA-switch is accelerated after ELS, affecting both the pre- and postsynaptic chloride homeostasis, the former altering glutamatergic transmission. This may considerably affect brain development.

High calcium transport by Polycystin-2 (TRPP2) induces channel clustering and oscillatory currents

The regulation by Ca2+ of Ca2+-permeable ion channels represents an important mechanism in the control of cell function. Polycystin-2 (PC2, TRPP2), a member of the TRP channel family (Transient Potential Receptor), is a Ca2+ permeable non-selective cation channel. Previous studies from our laboratory demonstrated that physiological concentrations of Ca2+ do not regulate in vitro translated PC2 (PC2iv) channel activity. However, the issue as to PC2′s Ca2+ permeability and regulation remain ill-defined, in particular because Ca2+ transport is usually observed in the presence of other ionic gradients. In this study, we assessed Ca2+ transport by PC2iv in a lipid bilayer reconstitution system in a high Ca2+ gradient (CaCl2 100 mM cis, CaCl2 10 mM trans) in the presence of either 3:7 or 7:3 1-palmitoyl-2-oleoyl-choline and ethanolamine lipid mixtures. Reconstituted PC2iv showed spontaneous Ca2+ currents in both lipid mixtures, with a maximum conductance of 63 ± 13 pS (n = 19) and 105 pS ± 9.8 (n = 9), respectively. In both cases, we best fitted the experimental data with the Goldman-Hodgkin-Katz equation, observing a reversal potential (Vrev ∼ −27 mV) consistent with strict Ca2+ selectivity. The R742X mutated PC2 (PC2R742X), lacking the carboxy terminal domain of the channel showed no differences with wild type PC2. Interestingly, we also observed the onset of spontaneous Ca2+ current oscillations whenever PC2-containing samples were reconstituted in the 3:7, but not 7:3 POPC:POPE lipid mixture. The amplitude and frequency of the ionic oscillations were highly dependent on the applied voltage, the imposed Ca2+ gradient, and the presence of high Ca2+, which induced PC2 channel clustering as observed by atomic force microscopy (AFM). We also used the QuB suite to kinetically model the PC2 channel Ca2+ oscillations based on the presence of subconductance states in the channel. The encompassed evidence supports a high Ca2+ permeability by PC2, and a novel oscillatory mechanism dependent on the presence of Ca2+ and phospholipids that provides the first evidence for the relation between stochasticity and deterministic processes mediated by ion channels.

A Mean-Field Firing-Rate Model for the Suprachiasmatic Nucleus

We present a mean-field formalism for modeling firing-rate statistics of brain regions whose neurons exhibit atypical firing patterns and heterogeneous electrophysiological properties. We apply the formalism to the suprachiasmatic nucleus (SCN)—the human circadian pacemaker—whose neurons can intrinsically exhibit depolarized low-amplitude membrane oscillations (DLAMOs), depolarization block (DB), and standard action potential firing at different times of day. Further, gamma-aminobutyric acid reversal potentials and molecular circadian phases of SCN neurons, among other properties, vary across the network and/or slowly over time. Our formalism consists of a system of integro-differential equations describing the time evolution of the mean and standard deviation of synaptic conductances across the network. Electrophysiological properties of SCN neurons are incorporated by computing responses to synaptic conductance inputs of a Hodgkin–Huxley-type SCN neuron model that exhibits DLAMOs and DB. Such responses are then averaged over distributions of relevant quantities and included in the differential equations. Results suggest mechanisms by which physiologically relevant changes to firing activities may arise, highlighting means by which the amplitude of firing rates may shrink, the standard deviation of firing rates may grow, and by which a mid-day dip in firing rates may appear. For instance, results show that a large spread in circadian phases across SCN neurons reduces the size of oscillations in SCN network firing activity across the 24-hour day, identifying a mechanism by which heterogeneities in neuron electrophysiology could influence circadian rhythms.

Ion exclusion, osmoregulation and management of oxidative stress improve salt tolerance in rice at seedling stage

Excess ion accumulation disturbs ionic homeostasis, creates an osmotic imbalance, and generates oxidative stress in plants under salinity stress. In the present experiment, the effect of salt stress at the seedling stage on the osmotic equilibrium and ROS scavenging potential was evaluated in ten differentially salt-sensitive rice genotypes. For this, the plants were grown hydroponically and salt stress equivalent to 12 dS m-1 was imposed at 3-4 leaf stages. The results showed that a few genotypes like FL478, AC41585, and AC39416A were able to maintain a lower Na+/K+ ratio in the leaf and thus proved more tolerant to salt stress than others. Additionally, these genotypes produced greater organic osmolytes (proline, glycine betaine, trehalose) and also had higher activities of key antioxidant enzymes (superoxide dismutase, catalase, peroxidase). On the contrary, Rashpanjor and CSR27 showed lesser ionic discrimination (higher leaf Na+/K+ ratio) but a moderate degree of salt tolerance, perhaps using Na+ effectively as an inorganic osmoticum to overcome stress. The susceptible genotypes like IR29 and Sabita were found extremely poor in restricting the upward movement of Na+, as well as the management of oxidative stress under saline conditions. From this study, we conclude that an efficient reactive oxygen species scavenging system along with greater osmotolerance helps to render salt tolerance at the seedling stage in rice.

Activation of mouse Otop3 proton channels by Zn2+

Otopetrins (Otop1-Otop3) belong to a newly identified family of proton (H+) channels activated by extracellular acidification. Here, we found that Zn2+ activates the mouse Otop3 (mOtop3) proton channels by using electrophysiological patch-clamp techniques. In mOtop3-expressing human embryonic kidney HEK293T cells, a biphasic inward mOtop3 H+ current comprising a fast transient current followed by a sustained current was observed upon extracellular acidification at pH 5.0. No significant activation of the mOtop3 channel was observed at pH 6.5 and 7.4, but interestingly, Zn2+ dose-dependently induced a sustained activation of mOtop3 under these pH conditions. Increasing the Zn2+ concentration had no effect on the reversal potential of the channel currents, suggesting that Zn2+ does not permeate through the mOtop3. The activation of the mOtop3 channel was specific to Zn2+ among divalent metal cations. Our findings reveal a novel modulatory mechanism of mOtop3 proton channels by Zn2+.

Bumetanide increases postsynaptic inhibition after chronic SCI and decreases presynaptic inhibition with step-training.

Current anti-spastic medication significantly compromises motor recovery after spinal cord injury (SCI), indicating a critical need for alternative interventions. Because a shift in chloride homeostasis decreases spinal inhibition and contributes to hyperreflexia after SCI, we investigated the effect of bumetanide, an FDA-approved sodium-potassium-chloride intruder (NKCC1) antagonist, on presynaptic and postsynaptic inhibition. We compared its effect with step-training as it is known to improve spinal inhibition by restoring chloride homeostasis. In SCI rats, a prolonged bumetanide treatment increased postynaptic inhibition but not presynaptic inhibition of the plantar H-reflex evoked by posterior biceps and semitendinosus (PBSt) group I afferents. By using in vivo intracellular recordings of motoneurons, we further show that a prolonged bumetanide increased postsynaptic inhibition by hyperpolarizing the reversal potential for inhibitory postsynaptic potentials (IPSPs) after SCI. However, in step-trained SCI rats an acute delivery of bumetanide decreased presynaptic inhibition of the H-reflex, but not postsynaptic inhibition. These results suggest that bumetanide might be a viable option to improve postsynaptic inhibition after SCI, but it also decreases the recovery of presynaptic inhibition with step-training. We discuss whether the effects of bumetanide are mediated by NKCC1 or by off-target effects. KEY POINTS: After spinal cord injury (SCI), chloride homeostasis is dysregulated over time in parallel with the decrease in presynaptic inhibition of Ia afferents and postsynaptic inhibition of motoneurons, and the development of spasticity. While step-training counteracts these effects, it cannot always be implemented in the clinic because of comorbidities. An alternative intervention is to use pharmacological strategies to decrease spasticity without hindering the recovery of motor function with step-training. Here we found that, after SCI, a prolonged bumetanide (an FDA-approved antagonist of the sodium-potassium-chloride intruder, NKCC1) treatment increases postsynaptic inhibition of the H-reflex, and it hyperpolarizes the reversal potential for inhibitory postsynaptic potentials in motoneurons. However, in step-trained SCI, an acute delivery of bumetanide decreases presynaptic inhibition of the H-reflex, but not postsynaptic inhibition. Our results suggest that bumetanide has the potential to decrease spastic symptoms related to a decrease in postsynaptic but not presynaptic inhibition after SCI.

Curcumin-stabilized silver nanoparticles encapsulated in biocompatible electrospun nanofibrous scaffold for sustained eradication of drug-resistant bacteria

Due to the misuse of antibiotics, the emerging drug-resistance of pathogenic microbes has aroused considerable concerns for the public health, which demands the continuous search for safe and efficient antimicrobial treatment. In this study, curcumin reduced and stabilized silver nanoparticles (C-Ag NPs) were successfully encapsulated into electrospun nanofiber membranes consisted of polyvinyl alcohol (PVA) cross-linked by citric acids (CA), which exhibited desirable biocompatibility and broad-spectrum antimicrobial activities. The homogeneously distributed and sustained release of C-Ag NPs in the constructed nanofibrous scaffolds yield prominent killing effect against Escherichia coli, Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus (MRSA), which involved the reactive oxygen species (ROS) generation. Outstanding elimination of bacterial biofilms and excellent antifungal activity against Candida albicans was also identified after treated with PVA/CA/C-Ag. Transcriptomic analysis on MRSA treated by PVA/CA/C-Ag revealed the antibacterial process is related to disrupting carbohydrate and energy metabolism, as well as destroying bacterial membranes. Significant down-regulation of the expression of multidrug-resistant efflux pump gene sdrM was observed pointing to the role of PVA/CA/C-Ag to overcome the bacterial resistance. Therefore, the constructed ecofriendly and biocompatible nanofibrous scaffolds provide a robust and versatile nanoplatform of reversal potential to eradicate drug-resistant pathogenic microbe in environmental as well as healthcare applications.

The mechanoelectrochemical effect on the electrochemical corrosion of austenitic stainless steel

In this paper, the mechanoelectrochemical effect on the electrochemical corrosion of 316 L austenitic stainless steel was investigated by theoretical, experimental and simulated methods. The modified piecewise formula φa,eq=φa,eq0−ΔPmVmzF−TRzFln(KN0εp+1) for calculating the strain on anode equilibrium potential of austenitic stainless steel was proposed. It is demonstrated that the corrosion current increases from 8.03×10−6A⋅cm−2 to 28.38×10−6A⋅cm−2 within 20% strain, especially within 10%, which is in better agreement with the experimental results. This modification not only made the physical meaning of parameters clearer, simplified the formula, but also made the calculation results more accurate. Moreover, the localized galvanic corrosion caused by non-uniform deformation had been verified by finite element model. The results revealed that the non-uniform strain at the initial stage of strain (εp<0.05) has a greater influence on the corrosion resistance.

Coexisting Behavior and Status Transition of the Hodgkin-Huxley Cardiac Purkinje Fiber Model Under External AC Injection

The membrane current I_m of the Hodgkin-Huxley (HH) cardiac Purkinje fiber (CPF) model is usually calculated as direct current (DC). In this paper, a conventional alternating current (AC) is used, namely I_{AC} = textrm{Asin}(2 pi ft), as the impressed injection. Dynamic characters of the model with different AC parameters and various initial conditions are observed through phase plane orbits, waveforms, bifurcation diagrams, and Lyapunov exponent spectra, which reveal multiple coexisting membrane action potential patterns, corresponding to the coexisting attractors of the same or different periods in the phase diagram. Meanwhile, the model undergoes period, quasi-period and local forward or inverse period-doubling bifurcations with changes in amplitude A or frequency f, which further proves the complex nonlinear property of the AC-injected model. In addition, by changing the external current I_m, the sodium-ion and potassium-ion equilibrium potentials, i.e. E_{Na} and E_{K}, respectively, the regularity of the CPF heartbeat frequency is observed. The state transformations of CPF are found between normal, abnormal and sudden cardiac arrest, and the method adjusting from the dangerous state to the normal heartbeat frequency range is investigated. This study may provide a reference for exploring the evolution of nonlinear dynamics in HH CPF model and protecting the health of life and heart.

The effect of oxygen partial pressure and humidification in proton exchange membrane fuel cells at intermediate temperature (80–120 °C)

The integration of proton exchange membrane fuel cells (PEMFCs) in heavy-duty vehicles would be facilitated if operating temperatures above 100 °C were possible. In this work, the effect of temperature in the intermediate range of 80–120 °C is investigated for a commercial membrane electrode assembly (MEA) through polarization curves and electrochemical impedance spectroscopy. The importance of oxygen partial pressure on voltage is systematically studied by decoupling it from humidity and temperature. The results show that adequate operation at intermediate temperature is achievable if the oxygen partial pressure is sufficient. Although the cathode kinetics is faster with rising temperatures, the voltage gain is counteracted by the decreasing equilibrium potential. At intermediate temperature, the water transport is enhanced, levelling out the relative humidity difference between anode and cathode. However, ionic conductivity in the polymer can become limiting at high currents, due to a smaller relative humidity increase at these temperatures. To conclude, a higher operating temperature does not inherently cause a decrease in obtained current density. Rather, the difficulty to simultaneously have sufficient oxygen partial pressure and high relative humidity causes limitations within the cathode that to some extent can be solved by pressurizing the cell.

Accurate descriptions of molecule-surface interactions in electrocatalytic CO2 reduction on the copper surfaces

Copper-based catalysts play a pivotal role in many industrial processes and hold a great promise for electrocatalytic CO2 reduction reaction into valuable chemicals and fuels. Towards the rational design of catalysts, the growing demand on theoretical study is seriously at odds with the low accuracy of the most widely used functionals of generalized gradient approximation. Here, we present results using a hybrid scheme that combines the doubly hybrid XYG3 functional and the periodic generalized gradient approximation, whose accuracy is validated against an experimental set on copper surfaces. A near chemical accuracy is established for this set, which, in turn, leads to a substantial improvement for the calculated equilibrium and onset potentials as against the experimental values for CO2 reduction to CO on Cu(111) and Cu(100) electrodes. We anticipate that the easy use of the hybrid scheme will boost the predictive power for accurate descriptions of molecule-surface interactions in heterogeneous catalysis.

Reversible Chlorite Anion/Chlorine Dioxide Redox Couple for Low-Cost Energy Storage

The ClO2–/ClO2 electrochemical reaction is shown to be highly reversible in acidic, near-neutral, and alkaline electrolytes while using low-cost carbon electrodes. Its equilibrium potential (0.954 V vs SHE) is pH-independent and enables high aqueous cell voltages of 1.38–2.15 V when used as a positive electrode with negative electrodes such as Zn, Fe, or S. This anion redox couple may enable low-cost aqueous rechargeable batteries free of resource-constrained metals, here demonstrated in prototype Zn–NaClO2 full cells. The rapid reaction kinetics and stability of the ClO2 phase at low temperatures also suggest that chlorite-based batteries may be favorable for applications in cold environments.

Activation of Kir4.1/Kir5.1 contributes to the cyclosporin A-induced stimulation of the renal NaCl cotransporter and hyperkalemic hypertension.

Cyclosporin A (CsA) is a widely used immunosuppressive drug that causes hypertension and hyperkalemia. Moreover, CsA-induced stimulation of the thiazide-sensitive NaCl cotransporter (NCC) in the kidney has been shown to be responsible for the development of hyperkalemic hypertension. In this study, we tested whether CsA induces the activation of NCC by stimulating the basolateral Kir4.1/Kir5.1 channel in the distal convoluted tubule (DCT). Electrophysiology, immunoblotting, metabolic cages, and radio-telemetry methods were used to examine the effects of CsA on Kir4.1/Kir5.1 activity in the DCT, NCC function, and blood pressure in wild-type (WT) and kidney-specific Kir4.1 knockout (KS-Kir4.1 KO) mice. The single-channel patch clamp experiment demonstrated that CsA stimulated the basolateral 40 pS K+ channel in the DCT. Whole-cell recording showed that short-term CsA administration (2h) not only increased DCT K+ currents but also shifted the K+ current (IK ) reversal potential to the negative range (hyperpolarization). Furthermore, CsA administration increased phosphorylated NCC (pNCC) levels and inhibited renal Na+ and K+ excretions in WT mice but not in KS-Kir4.1 KO mice, suggesting that Kir4.1 is required to mediate CsA effects on NCC function. Finally, long-term CsA infusion (14 days) increased blood pressure, plasma K+ concentration, and total NCC or pNCC abundance in WT mice, but these effects were blunted in KS-Kir4.1 KO mice. We conclude that CsA stimulates basolateral K+ channel activity in the DCT and that Kir4.1 is essential for CsA-induced NCC activation and hyperkalemic hypertension.

Analysis of heat generation due to open-circuit voltage hysteresis in lithium-ion cells

For the application of lithium-ion batteries, the thermal management is an important aspect, as the temperature is one of the main parameters for the behavior of Li-ion batteries. Several mechanisms influence the thermal behavior of Li-ion batteries, which can be split into reversible and irreversible losses. In literature, mostly Joule heat and reversible heat due to the entropy is discussed. With upcoming new active materials like silicon, which exhibits a significant charge-direction dependent voltage hysteresis, the losses due to the hysteresis become a more prominent factor. In this paper, we show the influence of the hysteresis losses for the most common active materials, which exhibit a voltage hysteresis. With a detailed analysis of calorimeter measurements, a true equilibrium potential can be calculated. The results show, that the equilibrium potential is not in the middle between both open-circuit voltage (OCV) curves as most of the heat due to hysteresis is generated during charging, which can have a significant influence on the thermal behavior of Li-ion batteries. For pure graphite cells 63 % and for silicon-graphite composite cells even 75 % of the hysteresis heat is generated while charging.

γ-Aminobutyric Acid-Ergic Development Contributes to the Enhancement of Electroencephalogram Slow-Delta Oscillations Under Volatile Anesthesia in Neonatal Rats.

General anesthetics (eg, propofol and volatile anesthetics) enhance the slow-delta oscillations of the cortical electroencephalogram (EEG), which partly results from the enhancement of (γ-aminobutyric acid [GABA]) γ-aminobutyric acid-ergic (GABAergic) transmission. There is a GABAergic excitatory-inhibitory shift during postnatal development. Whether general anesthetics can enhance slow-delta oscillations in the immature brain has not yet been unequivocally determined. Perforated patch-clamp recording was used to confirm the reversal potential of GABAergic currents throughout GABAergic development in acute brain slices of neonatal rats. The power density of the electrocorticogram and the minimum alveolar concentrations (MAC) of isoflurane and/or sevoflurane were measured in P4-P21 rats. Then, the effects of bumetanide, an inhibitor of the Na+-K+-2Cl- cotransporter (NKCC1) and K+-Cl- cotransporter (KCC2) knockdown on the potency of volatile anesthetics and the power density of the EEG were determined in vivo. Reversal potential of GABAergic currents were gradually hyperpolarized from P4 to P21 in cortical pyramidal neurons. Bumetanide enhanced the hypnotic effects of volatile anesthetics at P5 (for MACLORR, isoflurane: 0.63% ± 0.07% vs 0.81% ± 0.05%, 95% confidence interval [CI], -0.257 to -0.103, P < .001; sevoflurane: 1.46% ± 0.12% vs 1.66% ± 0.09%, 95% CI, -0.319 to -0.081, P < .001); while knockdown of KCC2 weakened their hypnotic effects at P21 in rats (for MACLORR, isoflurane: 0.58% ± 0.05% to 0.77% ± 0.20%, 95% CI, 0.013-0.357, P = .003; sevoflurane: 1.17% ± 0.04% to 1.33% ± 0.04%, 95% CI, 0.078-0.244, P < .001). For cortical EEG, slow-delta oscillations were the predominant components of the EEG spectrum in neonatal rats. Isoflurane and/or sevoflurane suppressed the power density of slow-delta oscillations rather than enhancement of it until GABAergic maturity. Enhancement of slow-delta oscillations under volatile anesthetics was simulated by preinjection of bumetanide at P5 (isoflurane: slow-delta changed ratio from -0.31 ± 0.22 to 1.57 ± 1.15, 95% CI, 0.67-3.08, P = .007; sevoflurane: slow-delta changed ratio from -0.46 ± 0.25 to 0.95 ± 0.97, 95% CI, 0.38-2.45, P = .014); and suppressed by KCC2-siRNA at P21 (isoflurane: slow-delta changed ratio from 16.13 ± 5.69 to 3.98 ± 2.35, 95% CI, -18.50 to -5.80, P = .002; sevoflurane: slow-delta changed ratio from 0.13 ± 2.82 to 3.23 ± 2.49, 95% CI, 3.02-10.79, P = .003). Enhancement of cortical EEG slow-delta oscillations by volatile anesthetics may require mature GABAergic inhibitory transmission during neonatal development.

Toward data‐ and mechanistic‐driven volcano plots in electrocatalysis

AbstractThe present application note summarizes an advanced methodology that allows for deriving potential‐dependent volcano curves for energy storage and conversion processes. The conventional approach relies on the combination of density functional theory calculations and scaling relations for a single mechanistic pathway as well as a discussion of electrocatalytic activity by means of the potential‐determining step, determined at the equilibrium potential of the reaction. Herein, it is illustrated how several reaction mechanisms can be factored into the volcano curve and how the rate‐determining step based on the descriptor Gmax(U) can be derived by a rigorous thermodynamic analysis of adsorption free energies fed by a data‐inspired methodology.