A dual voltage clamp technique to study gap junction hemichannels in astrocytes cultured from neonatal rodent spinal cords

Astrocytes express surface channels involved in purinergic signaling, and among these channels, pannexin-1 (Px1) and connexin-43 (Cx43) hemichannels (HCs) mediate ATP release that acts directly, or through its derivatives, on neurons and glia via purinergic receptors. Although HCs are functional, i.e., open and close, under physiological and pathological conditions, single channel conductance of Px1 HCs is not well defined. Here, we developed a dual voltage clamp technique in HeLa cells overexpressing human Px1-YFP, and then applied this system to rodent spinal astrocytes. Single channels were recorded in cell attached patches and evoked with ramp cycles of 2 s duration and -/+ 80-100 mV amplitude or rectangular pulses through another pipette in whole cell clamp. Conductance of Px1 HC openings recorded during ramp stimuli ranged 25-110 pS. Based on their single channel conductances, Px1 HCs could be distinguished from Cx43 HCs and P2X7 receptors (P2X7Rs) in spinal astrocytes during dual voltage clamp experiments. Furthermore, we found that single channel activity of Cx43 HCs and P2X7Rs was increased, and that of Px1 HCs was decreased, in spinal astrocytes treated for 7h with FGF-1, a growth factor implicated in neurodevelopment, repair and inflammation.

Comb filters for the removal of transcranial current stimulation artifacts from single channel EEG recordings

The combination of transcranial alternating current stimulation (tACS) and electroencephalogram (EEG) for mobile and home-based interventions offers the potential for control and adaptation of stimulation parameters. Yet, during stimulation, the EEG is heavily affected by stimulation artifacts. Spatial filters are often unsuited because too few channels are recorded and hardware capabilities are limited. Due to their speed and as they can be used for single channels, we explore the performance of single-channel weighted comb filters on artifact removal. At any given time point t, the recording r(t) is a superposition of a neurophysiological signal n(t), the stimulation artifact a(t) and noise e(t). Now, we can estimate the artifact a(t) based on the recording from an earlier (or later) time-point shifted by the artifacts period. A weighted estimate based on multiple time points has the potential to improve the signal recovery. Therefore, we explored several approaches and evaluated their performance on simulated and real data. The comb kernel filters were implemented in Matlab (https://github.com/agricolab/ARtACS) and Python (https://github.com/agricolab/pyARtACS), and the code is open access under an X11-license. We found that independent of the weighting function, all comb filters exhibit similarity in their suppression of the DC component, the artifacts frequency, and its harmonics. Yet, different weighting functions exhibit different pass-band performance, evident as ringing and amplification, and their induction of time-domain echoes. Interestingly, we note that a causal uniform filter is comparable to more complex approaches, while offering the option for real-time filtering. Comb filters are able to remove tACS artifacts even if only a single channel is available. As comb filters require no assumptions about the shape of the artifact, they might also be useful for filtering of non-sinusoidal, e.g. pulsed or saw-tooth, transcranial current stimulation.

Experimental Investigation of Trapped Oil Mobilization with Ferrofluid

Summary Nanoparticles have great potential to mobilize trapped oil in reservoirs because of their chemical, thermal, and interfacial properties. However, the direct application of magnetic forces on superparamagnetic nanoparticles in reservoir engineering applications has not been extensively investigated. We demonstrate the enhanced oil recovery (EOR) potential of hydrophilic superparamagnetic nanoparticles in oil production by direct observation using microfluidics. We studied the mobilization of oil blobs by a ferrofluid (a suspension of hydrophilic superparamagnetic nanoparticles in water) both in a converging/diverging micromodel channel and in a foot-long pore network micromodel, both with varying depth (so-called 2.5D micromodels). The water-based ferrofluid in all cases was the wetting fluid. Initial ferrofluid flooding experiments in single channels were performed without and then with a static magnetic field. This magnetic field caused oil droplet deformation, dynamic breakup into smaller droplets, and subsequent residual oil saturation reduction. During the flooding, after the magnetic field was applied, significant oil displacement was observed within 2 hours [6 pore volumes injected (PVI)], and 86.2% of the oil that was not mobilized without a magnetic field was mobilized within 64 hours (192 PVI). Then, in experiments in the micromodel and in a Hele-Shaw cell without flooding, we observed self-assembly of oil droplets, indicating the formation of the hydrophilic magnetic nanoparticle microstructures (chains under the magnetic field) and their interaction with the oil blobs. Further ferrofluid flooding experiments were performed in a foot-long micromodel under a rotating magnetic field. The oil saturation was reduced from 44.6 to 33.3% after 17 hours (8.5 PVI) of ferrofluid flooding after the rotating magnetic field was applied. Finally, a discussion of field application of ferrofluid flooding is presented.

Failure Modes Of Cold Formed Steel Angle Sections

Tension members consisting of single and double angles, single channels and similar sections are frequently used for lateral bracing and as truss elements. Such members normally have eccentric connections which results in bending of tension member. It is often permitted, by current design specifications, to neglect this eccentricity in the design of the member. The present study is focus on mode of failure on cold formed steel angle. This analysis carries single angles and double angles sections of 2mm under plain (without Lipped) and with Lipped conditions subjected to tension. Analyses were carried out for thirty-six numbers of angle sections under condition such as double angle were connected same side to gusset plate and connected to opposite side. Figure shows connection failure and mode of failure. Local buckling, global buckling, tearing failure, net section failure, and block shear failure.

Two-pore domain potassium channels (K2P) in GtoPdb v.2021.3

The 4TM family of K channels mediate many of the background potassium currents observed in native cells. They are open across the physiological voltage-range and are regulated by a wide array of neurotransmitters and biochemical mediators. The pore-forming α-subunit contains two pore loop (P) domains and two subunits assemble to form one ion conduction pathway lined by four P domains. It is important to note that single channels do not have two pores but that each subunit has two P domains in its primary sequence; hence the name two-pore domain, or K2P channels (and not two-pore channels). Some of the K2P subunits can form heterodimers across subfamilies (e.g. K2P3.1 with K2P9.1). The nomenclature of 4TM K channels in the literature is still a mixture of IUPHAR and common names. The suggested division into subfamilies, described in the More detailed introduction, is based on similarities in both structural and functional properties within subfamilies and this explains the "common abbreviation" nomenclature in the tables below.

Motor Unit Discharges from Multi-Kernel Deconvolution of Single Channel Surface Electromyogram

Surface electromyogram (EMG) finds many applications in the non-invasive characterization of muscles. Extracting information on the control of motor units (MU) is difficult when using single channels, e.g., due to the low selectivity and large phase cancellations of MU action potentials (MUAPs). In this paper, we propose a new method to face this problem in the case of a single differential channel. The signal is approximated as a sum of convolutions of different kernels (adapted to the signal) and firing patterns, whose sum is the estimation of the cumulative MU firings. Three simulators were used for testing: muscles of parallel fibres with either two innervation zones (IZs, thus, with MUAPs of different phases) or one IZ and a model with fibres inclined with respect to the skin. Simulations were prepared for different fat thicknesses, distributions of conduction velocity, maximal firing rates, synchronizations of MU discharges, and variability of the inter-spike interval. The performances were measured in terms of cross-correlations of the estimated and simulated cumulative MU firings in the range of 0–50 Hz and compared with those of a state-of-the-art single-kernel algorithm. The median cross-correlations for multi-kernel/single-kernel approaches were 92.2%/82.4%, 98.1%/97.6%, and 95.0%/91.0% for the models with two IZs, one IZ (parallel fibres), and inclined fibres, respectively (all statistically significant differences, which were larger when the MUAP shapes were of greater difference).

Membrane channel hypothesis of lysosomal permeabilization by beta-amyloid

Alzheimer's disease (AD) is the most common cause of dementia affecting millions of people. Neuronal death in AD is initiated by oligomeric amyloid-β (Aβ) peptides. Recently, we proposed the amyloid degradation toxicity hypothesis, which explains multiple major observations associated with AD - such as autophagy failure and a decreased metabolism. According to the hypothesis, the key event in the cellular toxicity of amyloid is the formation of non-selective membrane channels in lysosomal membranes by amyloid fragments that are produced by the digestion of Aβ previously absorbed by endocytosis. Electrophysiological data suggest that amyloid-formed channels have different sizes, which can be explained by the fact that barrel-shaped amyloid aggregates which create channels can consist of different number of monomers. To estimate the ability of channels to leak molecules of various molecular weights, we modeled the channels as saline-filled cylinders in non-conductive membranes that pass spheres with a density of average globular proteins. As a basis, we used the conductance distribution taken from the previously published experimental dataset, in which single channels with a conductance reaching one nanosiemens were registered. Our calculations show that channels with a giant conductance can allow for passing macromolecules such as lysosomal cathepsins implicated in the activation of apoptosis. The formation of giant channels is disproportionally promoted in an acidic environment. Also, amyloid fragments leaking from permeabilized lysosomes can reach the internal leaflet of the plasma membrane and permeabilize it. We conclude that while dissipation of the proton gradient by any - even the smallest amyloid channel - readily explains lysosomal failure, the relatively rare events of lysosomal permeabilization to large macromolecules can be an alternative mechanism of cellular death induced by exposure to Aβ.

Two P domain potassium channels in GtoPdb v.2021.2

The 4TM family of K channels mediate many of the background potassium currents observed in native cells. They are open across the physiological voltage-range and are regulated by a wide array of neurotransmitters and biochemical mediators. The pore-forming α-subunit contains two pore loop (P) domains and two subunits assemble to form one ion conduction pathway lined by four P domains. It is important to note that single channels do not have two pores but that each subunit has two P domains in its primary sequence; hence the name two P domain, or K2P channels (and not two-pore channels). Some of the K2P subunits can form heterodimers across subfamilies (e.g. K2P3.1 with K2P9.1). The nomenclature of 4TM K channels in the literature is still a mixture of IUPHAR and common names. The suggested division into subfamilies, described in the More detailed introduction, is based on similarities in both structural and functional properties within subfamilies and this explains the "common abbreviation" nomenclature in the tables below.

Performance analysis of beam divergence propagation through rainwater and snow pack in free space optical communication

In the present work the future communication requirements need to fulfill with high data rate, FSO (free space optic) with it is tremendous potential is the solution. This research observed the effectiveness analysis of FSO systems by modifying one of the most important FSO parameters beam divergence, under the most affected weather attenuating condition Rainwater and snow pack. The simulation is obtained and analyzed under single channels CSRZ-FSO (carrier-suppressed return-to-zero/free space optical) systems having capacity of 40 Gbps between two transceivers with variable distance. The connection is presently under 5 meteorological turbulences (light rain, medium rain, wet snow, heavy rain and dry snow). The results show the heavy rain and dry snow have a very high attenuation carried out in terms of Q-factor. this result led us to conclude that small divergence offers significant performance improvement for FSO link and this performance decrease every time the beam divergence increase, Therefore, to build inexpensive and reliable transmission media, we go with new method that still in the experiment area called hybrid RF/FSO (radio frequency/free space optical) that compatible with atmospherically status.

Heat Transfer Measurements Using Multiple Thermochromic Liquid Crystals in Symmetric Cooling Channels

The transient Thermochromic Liquid Crystal (TLC) method is applied to determine the distribution of the local heat transfer coefficients using a configuration with parallel cooling channels at an engine relevant Reynolds number. The rectangular channels with a moderate aspect ratio and a high length-to-diameter ratio are equipped with one-sided oblique ribs with high blockage, which is a promising configuration for turbine near wall cooling applications. In this arrangement, the three inner channels should experience same flow and thermal conditions. Numerical simulations are performed to substantiate this assumption. The symmetric single channels are sprayed with narrowband TLC with various indication temperatures. Multiple experiments were conducted. All start at ambient conditions before the fluid is heated up to several temperatures between 46°C and 73°C. The results show that the determined local heat transfer coefficients and therefore the Nusselt numbers vary significantly for the different experimental conditions especially at locations of high heat transfer coefficient behind the ribs. A simplified procedure with respect to measurement uncertainties is applied to enable an easy and fast valuation on the data quality. This might be used within the data reduction analysis for such experiments directly. The approach is illustrated using the obtained experimental data.