Ca(2+)-activated Cl(-) current in sheep lymphatic smooth muscle.

Freshly dispersed sheep mesenteric lymphatic smooth muscle cells were studied at 37 degrees C using the perforated patch-clamp technique with Cs(+)- and K(+)-filled pipettes. Depolarizing steps evoked currents that consisted of L-type Ca(2+) [I(Ca(L))] current and a slowly developing current. The slow current reversed at 1 +/- 1.5 mV with symmetrical Cl(-) concentrations compared with 23.2 +/- 1.2 mV (n = 5) and -34.3 +/- 3.5 mV (n = 4) when external Cl(-) was substituted with either glutamate (86 mM) or I(-) (125 mM). Nifedipine (1 microM) blocked and BAY K 8644 enhanced I(Ca(L)), the slow-developing sustained current, and the tail current. The Cl(-) channel blocker anthracene-9-carboxylic acid (9-AC) reduced only the slowly developing inward and tail currents. Application of caffeine (10 mM) to voltage-clamped cells evoked currents that reversed close to the Cl(-) equilibrium potential and were sensitive to 9-AC. Small spontaneous transient depolarizations and larger action potentials were observed in current clamp, and these were blocked by 9-AC. Evoked action potentials were triphasic and had a prominent plateau phase that was selectively blocked by 9-AC. Similarly, fluid output was reduced by 9-AC in doubly cannulated segments of spontaneously pumping sheep lymphatics, suggesting that the Ca(2+)-activated Cl(-) current plays an important role in the electrical activity underlying spontaneous activity in this tissue. PMID: 11029279 [PubMed - indexed for MEDLINE]

Tetrodotoxin-sensitive sodium current in sheep lymphatic smooth muscle.

1. Fast inward currents were elicited in freshly isolated sheep lymphatic smooth muscle cells by depolarization from a holding potential of -80 mV using the whole-cell patch-clamp technique. The currents activated at voltages positive to -40 mV and peaked at 0 mV. 2. When sodium chloride in the bathing solution was replaced isosmotically with choline chloride inward currents were abolished at all potentials. 3. These currents were very sensitive to tetrodotoxin (TTX). Peak current was almost abolished at 1 microM with half-maximal inhibition at 17 nM. 4. Examination of the voltage dependence of steady state inactivation showed that more than 90% of the current was available at the normal resting potential of these cells (-60 mV). 5. The time course of recovery from inactivation was studied using a double-pulse protocol and showed that recovery was complete within 100 ms with a time constant of recovery of 20 ms. 6. Under current clamp, action potentials were elicited by depolarizing current pulses. These had a rapid upstroke and a short duration and could be blocked with 1 microM TTX. 7. Spontaneous contractions of isolated rings of sheep mesenteric lymphatic vessels were abolished or significantly depressed by 1 microM TTX.

The calcium channel blocker used with cyclosporin has an effect on gingival overgrowth

Background/aims, To investigate whether the choice of calcium channel blocker, used in conjunction with cyclosporin A, affected the prevalence of gingival overgrowth.

Survival of Swiss-Webster mouse cerebellar granule neurons is promoted by a combination of potassium channel blockers

<p>Cultured cerebellar granule neurons (CGN) are commonly used to assess neurotoxicity, but are routinely maintained in supraphysiological (25 mM) extracellular K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup> concentrations [K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup>]<sub style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">o</sub>. We investigated the effect of potassium channel blockade on survival of CGN derived from Swiss-Webster mice in supraphysiological (25 mM) and physiological (5.6 mM) [K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup>]<sub style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">o</sub>. CGN were cultured for 5 days in 25 mM K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup>, then in 5.6 mM K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup> or 25 mM K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup> (control). Viability, assayed 24 h later by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) reduction and by lactate dehydrogenase (LDH) release, was ∼50% in 5.6 mM K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup> versus 25 mM K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup> (<em>p</em> &lt; .001). Potassium channel blockers, 2 mM 4-aminopyridine (4-AP), 2 mM tetraethylammonium (TEA) or 1 mM Ba<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">2+</sup>, individually afforded limited protection in 5.6 mM K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup>. However, survival in 5.6 mM K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup> with a combination of 4-AP, TEA and Ba<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">2+</sup> was similar to survival in 25 mM K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup> without blockers (<em>p</em> &lt; .001 versus 5.6 mM K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup> alone). CGN survival in 25 mM K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup> was attenuated 25% by 2 μM nifedipine (<em>p</em> &gt; .001), but nifedipine did not attenuate neuroprotection by K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup> channel blockers. Together, these results suggest that the survival of CGN depends on the K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup> permeability of the membrane rather than the activity of a particular type of K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup> channel, and that the mechanism of neuroprotection by K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup> channel blockers is different from that of elevated [K<sup style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">+</sup>]<sub style="border: 0px; font-size: 0.75em; margin: 0px; padding: 0px; line-height: 0; color: rgb(46, 46, 46); font-family: 'Arial Unicode MS', 'Arial Unicode', Arial, 'URW Gothic L', Helvetica, Tahoma, sans-serif; text-align: justify; word-spacing: -1.03587377071381px; background-color: rgb(255, 255, 255);">o</sub>.</p>

GLP-1 and related peptides cause concentration-dependent relaxation of rat aorta through a pathway involving K_ATP and cAMP

increasing evidence from both clinical and experimental studies indicates that the insulin-releasing hormone, glucagon-like peptide-1 (GLP-1) may exert additional protective/reparative effects on the cardiovascular system. The aim of this study was to examine vasorelaxant effects of GLP-1(7-36)amide, three structurally-related peptides and a non-peptide GLP-1 agonist in rat aorta. Interestingly, all GLP-1 compounds, including the established GLP-1 receptor antagonist, exendin (9-39) caused concentration-dependent relaxation. Mechanistic studies employing hyperpolarising concentrations of potassium or glybenclamide revealed that these relaxant effects are mediated via specific activation of ATP-sensitive potassium channels. Further experiments using a specific membrane-permeable cyclic AMP (cAMP) antagonist, and demonstration of increased cAMP production in response to GLP-1 illustrated the critical importance of this pathway. These data significantly extend previous observations suggesting that GLP-1 may modulate vascular function, and indicate that this effect may be mediated by the GLP-1 receptor. However, further studies are required in order to establish whether GLP-1 related agents may confer additional cardiovascular benefits to diabetic patients. (c) 2008 Elsevier Inc. All rights reserved.

Inward rectifier potassium channels in the HL-1 cardiomyocyte-derived cell line.

HL-1 is a line of immortalized cells of cardiomyocyte origin that are a useful complement to native cardiomyocytes in studies of cardiac gene regulation. Several types of ion channel have been identified in these cells, but not the physiologically important inward rectifier K(+) channels. Our aim was to identify and characterize inward rectifier K(+) channels in HL-1 cells. External Ba(2+) (100?µM) inhibited 44?±?0.05% (mean?±?s.e.m., n?=?11) of inward current in whole-cell patch-clamp recordings. The reversal potential of the Ba(2+)-sensitive current shifted with external [K(+)] as expected for K(+)-selective channels. The slope conductance of the inward Ba(2+)-sensitive current increased with external [K(+)]. The apparent Kd for Ba(2+) was voltage dependent, ranging from 15?µM at -150 ?mV to 148?µM at -75 ?mV in 120 ?mM external K(+). This current was insensitive to 10?µM glybenclamide. A component of whole-cell current was sensitive to 150?µM 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), although it did not correspond to the Ba(2+)-sensitive component. The effect of external 1 mM Cs(+) was similar to that of Ba(2+). Polymerase chain reaction using HL-1 cDNA as template and primers specific for the cardiac inward rectifier K(ir)2.1 produced a fragment of the expected size that was confirmed to be K(ir)2.1 by DNA sequencing. In conclusion, HL-1 cells express a current that is characteristic of cardiac inward rectifier K(+) channels, and express K(ir)2.1 mRNA. This cell line may have use as a system for studying inward rectifier gene regulation in a cardiomyocyte phenotype.

Low SNR Capacity for MIMO Rician and Rayleigh-Product Fading Channels with Single Co-channel Interferer and Noise

This paper studies the ergodic capacity of multiple-input multiple-output (MIMO) systems with a single co-channel interferer in the low signal-to-noise-ratio (SNR) regime. Two MIMO models namely Rician and Rayleigh-product channels are investigated. Exact analytical expressions for the minimum energy per information bit, Eb/N0min, and wideband slope, S0, are derived for both channels. Our results show that the minimum energy per information bit is the same for both channels while their wideband slopes differ significantly. Further, the impact of the numbers of transmit and receive antennas, the Rician K factor, the channel mean matrix and the interference-to-noise-ratio (INR) on the capacity, is addressed. Results indicate that interference degrades the capacity by increasing the required minimum energy per information bit and reducing the wideband slope. Simulation results validate our analytical results.

How does the short-wavelength-sensitive contrast sensitivity function for detection and resolution change with age in the periphery?

To determine the age-related change in the peripheral short-wavelength-sensitive (SWS) grating contrast sensitivity function (CSF), cut-off spatial frequency (acuity) and contrast sensitivity for both a detection and resolution task were measured at 8 degrees eccentricity under conditions of SWS-cone isolation for 51 subjects (19-72 years). The acuity for both the detection and resolution task declined with age, the detection acuity being significantly higher than the resolution acuity at all ages (p

Lack of horizontal gene transfer of methicillin-resitance genetic determinants from PBP2a-positive, coagulase-negative Staphylococci to methicillin-sensitive Staphylococcus aureus using transcutaneous electric nerve stimulation (TENS),

Previous research shows that approximately half of the coagulase-negative staphylococci (CNS) isolated from patients in the intensive care unit (ICU) at Belfast City Hospital were resistant to methicillin. The presence of this relatively high proportion of methicillin-resistance genetic material gives rise to speculation that these organisms may act as potential reservoirs of methicillinresistance genetic material to methicillin-sensitive Staphylococcus aureus (MSSA). Mechanisms of horizontal gene transfer from PBP2a-positive CNS to MSSA, potentially transforming MSSA to MRSA, aided by electroporation-type activities such as transcutaneous electrical nerve stimulation (TENS), should be considered. Methicillin-resistant CNS (MR-CNS) isolates are collected over a two-month period from a variety of clinical specimen types, particularly wound swabs. The species of all isolates are confirmed, as well as their resistance to oxacillin by standard disc diffusion assays. In addition, MSSA isolates are collected over the same period and confirmed as PBP2a-negative. Electroporation experiments are designed to mimic the time/voltage combinations used commonly in the clinical application of TENS. No transformed MRSA were isolated and all viable S. aureus cells remained susceptible to oxacillin and PBP2a-negative. Experiments using MSSA pre-exposed to sublethal concentrations of oxacillin (0.25 µg/mL) showed no evidence of methicillin gene transfer and the generation of an MRSA. The study showed no evidence of horizontal transfer of methicillin resistance genetic material from MR-CNS to MSSA. These data support the belief that TENS and the associated time/voltage combinations used do not increase conjugational transposons or facilitate horizontal gene transfer from MR-CNS to MSSA.

Human odontoblasts express functional thermo-sensitive TRP channels: implications for dentin sensitivity

Odontoblasts form the outermost cellular layer of the dental pulp where they have been proposed to act as sensory receptor cells. Despite this suggestion, evidence supporting their direct role in mediating thermo-sensation and nociception is lacking. Transient receptor potential (TRP) ion channels directly mediate nociceptive functions, but their functional expression in human odontoblasts has yet to be elucidated. In the present study, we have examined the molecular and functional expression of thermo-sensitive TRP channels in cultured odontoblast-like cells and in native human odontoblasts obtained from healthy wisdom teeth. PCR and western blotting confirmed gene and protein expression of TRPV1, TRPA1 and TRPM8 channels. Immunohistochemistry revealed that these channels were localised to odontoblast-like cells as determined by double staining with dentin sialoprotein (DSP) antibody. In functional assays, agonists of TRPV1, TRPA1 and TRPM8 channels elicited [Ca2+]i transients that could be blocked by relevant antagonists. Application of hot and cold stimuli to the cells also evoked rises in [Ca2+]i which could be blocked by TRP-channel antagonists. Using a gene silencing approached we further confirmed a role for TRPA1 in mediating noxious cold responses in odontoblasts. We conclude that human odontoblasts express functional TRP channels that may play a crucial role in mediating thermal sensation in teeth. Cultured and native human odontoblasts express functional TRP channels that may play a crucial role in mediating thermal sensation in teeth.

Ion channel gates: comparative analysis of energy barriers.

The energetic profile of an ion translated along the axis of an ion channel should reveal whether the structure corresponds to a functionally open or closed state of the channel. In this study, we explore the combined use of Poisson–Boltzmann electrostatic calculations and evaluation of van der Waals interactions between ion and pore to provide an initial appraisal of the gating state of a channel. This approach is exemplified by its application to the bacterial inward rectifier potassium channel KirBac3.1, where it reveals the closed gate to be formed by a ring of leucine (L124) side chains. We have extended this analysis to a comparative survey of gating profiles, including model hydrophobic nanopores, the nicotinic acetylcholine receptor, and a number of potassium channel structures and models. This enables us to identify three gating regimes, and to show the limitation of this computationally inexpensive method. For a (closed) gate radius of 0.4 nm