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PubMed2004

Clinically normal atopic skin vs. non-atopic skin as seen through electrical impedance.

BACKGROUND/AIMS: In an earlier study, we have shown that the electrical impedance (IMP) is dependent on the lipid content of the stratum corneum as studied by lipid extraction. Therefore, we now employ the IMP technique to compare the properties of clinically normal atopic skin with that of non-atopic skin. We looked at the same time at concomitant alterations in transepidermal water loss (TEWL) and skin moisture results. METHODS: Twenty-two healthy participants and 26 patients with atopic dermatitis were recruited. One test site on each volar forearm was used in the test procedure, of cyclohexane swabbing, skin stripping, and lipid extraction using a mixture of hexane:isopropanol. Responses were evaluated by measuring TEWL, skin moisture using electrical capacitance, and finally IMP spectroscopy in the frequency range 1 kHz to 1 MHz before the start of the study, after each step of the test procedure, and also during a 2 weeks recovery period. RESULTS: Only the IMP results showed significant changes between baseline values of clinically normal atopic skin and healthy skin. Furthermore, IMP and TEWL showed significantly larger reactivity for the atopic group after the skin stripping procedure, and IMP after lipid extraction as well. CONCLUSION: Our results show that IMP is a useful method for detection of changes in the lipid content of the stratum corneum by discriminating healthy looking atopic skin from healthy control skin, and to detect changes in reactivity between the two groups.

Adult↗
PubMed1994

Human newborn skin: the effect of isopropanol on skin surface hydrophobicity.

The development of a hydrophobic skin surface in newborn mammals such as the rat plays an important role in promoting adaptation to the abrupt change in the environment that occurs at birth. To determine whether the skin surface plays a similar role in the human neonate, we performed tests of water sorption and desorption on the chest wall of 13 term newborns. These tests were performed within the first 24 h of life on unperturbed skin (controls) and after perturbation of a contralateral site with isopropanol. The degree of surface hydration was determined by measurement of skin surface electrical capacitance, and desorption rates were calculated by 1st-order kinetic analysis. The unperturbed surface of the newborn skin exhibited a peak sorption value (change from baseline after water loading) of 435 +/- 83 pF (mean +/- SEM) and a desorption rate of 0.048 +/- 0.009 s-1. After exposure to isopropanol, the peak sorption value increased to 594 +/- 79 pF (p < 0.05) and the desorption rate decreased to 0.024 +/- 0.004 s-1 (p < 0.01). Paired sorption values were positively correlated (r2 = 0.8, p < 0.001). These results support the hypothesis that the skin surface of the human newborn, by limiting the sorption of water (or amniotic fluid) on the skin, may play a role in the adaptation to the change in environment at birth.

1-Propanol↗
PubMed2000

Rhythmic opening and closing of vesicles during constitutive exo- and endocytosis in chromaffin cells.

Constitutive exo- and endocytic events are expected to increase and diminish the cell surface area in small spontaneous steps. Indeed, cell-attached patch-clamp measurements in resting chromaffin cells revealed spontaneous upward and downward steps in the electrical capacitance of the plasma membrane. The most frequent step size indicated cell surface changes of <0.04 microm(2), corresponding to vesicles of <110 nm diameter. Often downward steps followed upward steps within seconds, and vice versa, as if vesicles transiently opened and closed their lumen to the external space. Transient openings and closings sometimes alternated rhythmically for tens of seconds. The kinase inhibitor staurosporine dramatically increased the occurrence of such rhythmic episodes by making vesicle closure incomplete and by inhibiting fission. Staurosporine also promoted transient closures of large endocytic vesicles possibly representing remnants of secretory granules. We suggest that staurosporine blocks a late step in the endocytosis of both small and large vesicles, and that endocytosis involves a reaction cascade that can act as a chemical oscillator.

Animals↗
PubMed1987

Effect of insulin on lipid bilayer viscoelasticity.

Changes in the Young elasticity modulus in perpendicular direction to the membrane surface E perpendicular, in the coefficient of dynamic viscosity eta, in the electric capacitance C, in the surface charge U1, in the conductivity g and in the coefficient of non-linearity beta of current-voltage characteristic caused by insulin were studied in bilayer lipid membranes (BLM) prepared from a mixture of egg lecithin and cholesterol (4:1, w/w) in n-heptane. Even relatively small concentrations of insulin in electrolyte (ci approximately 4.8 x 10(-11) mol/l) caused a diminution in parameters E perpendicular and eta. Negative surface charge emerged on the membrane due to the insulin absorption, and U1 gradually increased depending on the concentration of the hormone in the electrolyte. Addition of insulin was also followed by an increase in membrane conductivity and affected the value of the coefficient of non-linearity beta of current-voltage characteristic. The effect of insulin on the BLM structure was discussed on the basis of the results obtained.

Elasticity↗
PubMed1997

Acute irritant contact dermatitis: recovery time in man.

Our understanding of the details of the recovery time of acute irritant contact dermatitis (ICD) is limited. We examined skin reactivity to a model surfactant, sodium lauryl sulfate (SLS), on previous acute ICD and normal sites over time with visual grading and noninvasive instruments. Acute ICD was induced on the upper arms of 18 volunteers (aged 30 to 51 years) by occluded application of 1% SLS for 24 h. Previous ICD and normal sites were provoked by occluded application of 2% or 7.5% SLS 30 min daily 4 consecutive days. Skin reactivity was assessed daily by visual erythema scoring (VES), transepidermal water loss (TEWL), skin color reflectance (SCR) and electrical capacitance (EC). Skin function of previous ICD sites assessed by VES, TEWL, SCR, and EC did not normalize until 2 weeks later; all parameters of previous ICD returned to normal after 3 weeks. While skin reactivity to 2% and 7.5% SLS showed no differences between previous ICD and normal sites at 4 weeks, differences of irritant reactivity especially 7.5% SLS between previous ICD and normal sites were significant at 3 weeks post-provocation. Our results demonstrate that irritation evaluated with irritant provocation was long-lasting, even though skin functional parameters assessed by various bioengineering instruments returned to normal. Complete recovery of skin function including irritability after acute ICD induced by 1% SLS was achieved approximately 4 weeks later. The date were generated with a model surfactant; it remains to be determined whether similar responses will be noted with chemicals of different physiochemical properties.

Acute Disease↗
PubMedDate not supplied

The pattern of the electrodermal activity as indicator of stress related reaction.

The aim of the present study was to characterize the patterns of signals associated with noninvasively detected skin sudomotor transient reflexes in a group of 221 apparently healthy people, aged between 16 and 72 years. By means of two pairs of surface electrodes applied respectively to the palm of the hand and to the sole of the foot skin impedance changes were recorded as two electrodermal parameters: skin electrical resistance and skin electrical capacitance. The recordings were obtained under standard conditions initially in the basal resting state of the subject and then, after the application of some standardized stimuli (evoked electrodermal responses). In terms of the basal (spontaneous) electrodermal activity and the evoked responses the subjects were divided into three categories: I--those with a high threshold of sympathetic activation (quiet basal trace and ample evoked responses); II--those with a medium threshold for sympathetic activation (basal trace with some spontaneous electrodermal activity, but obviously of a lower amplitude than the evoked responses); III--those with a low threshold for sympathetic activation (a tracing showing frequent, larger spontaneous sympathetic discharges from which the evoked responses cannot be distinguished). Of the 221 subjects, 22.2% in group I, 69.2% were included in group II, and 8.6% in group III. Our data show that each individual presents a characteristic electrodermal pattern which expresses the degree of stability/instability of the sympathetic nervous system, the most sensitive component of the physiological stress reaction system.

Adolescent↗
PubMed1993

Surfactant-induced stratum corneum hydration in vivo: prediction of the irritation potential of anionic surfactants.

After repeated contact, many surfactants will cause skin irritation and, especially, dryness and scaliness. Earlier in vitro investigations suggested that the irritation potential of anionic surfactants was related to the induction of hydration of isolated stratum corneum membranes. We have now investigated early surfactant-stratum corneum interaction in vivo. Sodium salts of n-alkyl sulfates with variable carbon chain length (n = 8-14) were tested for the promotion of stratum corneum hydration by measurements of skin surface water loss and electrical capacitance measurements in healthy adult human volunteers. The surfactant-induced increase in water uptake was confirmed in vitro by means of isolated stratum corneum samples and surfactant solutions labeled with tritiated water. In a parallel experiment the irritation potential of these compounds was investigated by 24-h patch testing in human volunteers. The irritant responses were quantified non-invasively by erythema (skin color reflectance measurements) and transepidermal water loss measurements. Hydration of stratum corneum exposed for 5 min to surfactant solutions significantly exceeded that of controls (phosphate-buffered saline). It increased with application time and was concentration dependent, saturable with increasing concentration, and rapidly reversible. Baseline hydration was re-established only 10-15 min after treatment termination. Induction of hydration was closely correlated with the irritation potential of the investigated compounds. It initially increased with increasing carbon chain length. The maximum response was obtained for the C12 analogue (sodium lauryl sulfate). With further increases in molecular size induction of stratum corneum hydration subsequently decreased. We have demonstrated that anionic surfactants increase stratum corneum hydration in vivo. The present results suggest that the mechanisms responsible for the hydration are related to the irritation properties of these compounds.

Adolescent↗
PubMed2004

Membrane cholesterol modulates dihydropyridine receptor function in mice fetal skeletal muscle cells.

Caveolae and transverse (T-) tubules are membrane structures enriched in cholesterol and glycosphingolipids. They play an important role in receptor signalling and myogenesis. The T-system is also highly enriched in dihydropyridine receptors (DHPRs), which control excitation-contraction (E-C) coupling. Recent results have shown that a depletion of membrane cholesterol alters caveolae and T-tubules, yet detailed functional studies of DHPR expression are lacking. Here we studied electrophysiological and morphological effects of methyl-beta-cyclodextrin (MbetaCD), a cholesterol-sequestering drug, on freshly isolated fetal skeletal muscle cells. Exposure of fetal myofibres to 1-3 mM MbetaCD for 1 h at 37 degrees C led to a significant reduction in caveolae and T-tubule areas and to a decrease in cell membrane electrical capacitance. In whole-cell voltage-clamp experiments, the L-type Ca(2+) current amplitude was significantly reduced, and its voltage dependence was shifted approximately 15 mV towards more positive potentials. Activation and inactivation kinetics were slower in treated cells than in control cells and stimulation by a saturating concentration of Bay K 8644 was enhanced. In addition, intramembrane charge movement and Ca(2+) transients evoked by a depolarization were reduced without a shift of the midpoint, indicating a weakening of E-C coupling. In contrast, T-type Ca(2+) current was not affected by MbetaCD treatment. Most of the L-type Ca(2+) conductance reduction and E-C coupling weakening could be explained by a decrease of the number of DHPRs due to the disruption of caveolae and T-tubules. However, the effects on L-type channel gating kinetics suggest that membrane cholesterol content modulates DHPR function. Moreover, the significant shift of the voltage dependence of L-type current without any change in the voltage dependence of charge movement and Ca(2+) transients suggests that cholesterol differentially regulates the two functions of the DHPR.

Action Potentials↗
PubMed1969

Interaction of a bacterial adenosine triphosphatase with phospholipid bilayers.

A solubilized nonparticulate adenosine triphosphatase from Streptococcus fecalis spheroplast membranes (Abrams, A., and C. Baron, Biochemistry, 7,501 (1968)) interacts at pH 7.5 with lipid bilayer membranes to produce a 10(2)- to 10(4)-fold increase in the electric conductance of the bilayer. In addition, a small decrease in the electrical capacitance of the interactant system is also observed. Interaction is obtained with bilayers formed from a solution in n-decane of either purified egg phosphatidyl-choline, synthetic diphytanoyl phosphatidyl-choline, or a total extract of the membrane lipid of S. fecalis. The magnitude of the increase in conductance is dependent on the presence of Mg(++) and upon the concentrations of both Na(+) and K(+) in the range of 10(-2) to 10(-1)M. An additional tenfold increase in conductance is obtained when ATP is added to the ambient aqueous phase surrounding the bilayer ATPase interactant system. No conductance change is obtained with ATPase which has been treated with pronase. ATPase activity is dependent upon Mg(++) and upon the concentrations of Na(+) and K(+) in the region of 10(-1)M. The function of the ATPase in the intact bacterial membrane is apparently associated with the active transport of cations. The increased conductance in the bilayer which results from its interaction with the ATPase, together with the similarity between the dependence of this interaction and the dependence of ATPase activity on Mg(++), ATP, and the Na(+), K(+) concentrations suggests that the bilayer-ATPase interactant complex may be similar in structure and properties to the membrane-ATPase complex in the intact organism.

Adenosine Triphosphatases↗
PubMed1998

Apical Cl- channels in A6 cells.

Short-circuit current (Isc), transepithelial conductance (Gt), electrical capacitance (CT) and the fluctuation in Isc were analyzed in polarized epithelial cells from the distal nephron of Xenopus laevis (A6 cell line). Tissues were incubated with Na+- and Cl--free solutions on the apical surface. Basolateral perfusate was NaCl-Ringer. Agents that increase cellular cAMP evoked increases in Gt, CT, Isc and generated a Lorentzian Isc-noise. The responses could be related to active, electrogenic secretion of Cl-. Arginine-vasotocin and oxytocin caused a typical peak-plateau response pattern. Stimulation with a membrane-permeant nonhydrolyzable cAMP analogue or forskolin showed stable increases in Gt with only moderate peaking of Isc. Phosphodiesterase inhibitors also stimulated Cl- secretion with peaking responses in Gt and Isc. All stimulants elicited a spontaneous Lorentzian noise, originating from the activated apical Cl- channel, with almost identical corner frequency (40-50 Hz). Repetitive challenge with the hormones led to a refractory behavior of all parameters. Activation of the cAMP route could overcome this refractoriness. All agents caused CT, a measure of apical membrane area, to increase in a manner roughly synchronous with Gt. These results suggest that activation of the cAMP-messenger route may, at least partly, involve exocytosis of a vesicular Cl- channel pool. Apical flufenamate depressed Cl- current and conductance and apparently generated blocker-noise. However, blocking kinetics extracted from noise experiments could not be reconciled with those obtained from current inhibition, suggesting the drug does not act as simple open-channel inhibitor.

Animals↗
PubMed2001

Structure suggests function: the case for synaptic ribbons as exocytotic nanomachines.

Synaptic ribbons, the organelles identified in electron micrographs of the sensory synapses involved in vision, hearing, and balance, have long been hypothesized to play an important role in regulating presynaptic function because they associate with synaptic vesicles at the active zone. Their physiology and molecular composition have, however, remained largely unknown. Recently, a series of elegant studies spurred by technical innovation have finally begun to shed light on the ultrastructure and function of ribbon synapses. Electrical capacitance measurements have provided sub-millisecond resolution of exocytosis, evanescent-wave microscopy has filmed the fusion of single 30 nm synaptic vesicles, electron tomography has revealed the 3D architecture of the synapse, and molecular cloning has begun to identify the proteins that make up ribbons. These results are consistent with the ribbon serving as a vesicle "conveyor belt" to resupply the active zone, and with the suggestion that ribbon and conventional chemical synapses have much in common.

Animals↗
PubMed2003

Towards on-line monitoring of cell growth in microporous scaffolds: Utilization and interpretation of complex permittivity measurements.

Here we demonstrate the ability to characterize microporous scaffolds and evaluate cell concentration variation via the utilization and interpretation of complex permittivity measurements (CP), a direct and nondestructive method. Polymer-based microporous scaffolds are of importance to tissue engineering, particularly in the promotion of cell adhesion, proliferation, and differentiation in predefined shapes. Chitosan gel scaffolds were seeded with increasing concentrations of macrophages to simulate cell growth. Complex permittivity measurements were performed using a dielectric probe and a vector network analyzer over a frequency ranging from 200 MHz to 2 GHz. An effective medium theory was applied to interpret the data obtained; respectively, Looyenga and Maxwell-Wagner-Hanai functions were used to retrieve the porosity and the variation of the cell concentration from the CP measurements. Calculated porosities were in agreement with experimental evaluation-porosity ranged from 81-96%. Changes in cell concentration inside the scaffolds upon injection of differing cell concentrations into the scaffold were detected distinguishably. Variations resulting from the cumulative injection of 400-1800 microL of 10(6) cells/mL solution into the scaffold were monitored. Results suggest that CP measurements in combination with an appropriate effective medium approximation can enable on-line monitoring of cell growth within scaffolds.

Algorithms↗
PubMed1992

A double-tuned probe for metabolic NMR studies.

A double-tuned single coil probe for high field metabolic NMR studies has been designed and tested. The resonant coil is a single-turn of copper foil wound on a quartz cylinder. It is fed by a short length of parallel strip line made of the same material; the feeder is also used as a tuning capacitance. Electrical parameters (quality factors, efficiencies at proton and carbon frequencies) and magnetic resonance data (90 degrees pulse lengths, minimum decoupler power) for lossy samples have been measured. The probe allows for the optimal use of the limited high homogeneity region of a vertical magnet. It shows good sensitivity and efficient decoupling.

Copper↗
PubMed2004

Expression of functional kainate and AMPA receptors in developing lateral superior olive neurons of the rat.

A functional analysis of AMPA and kainate receptors (AMPARs and KARs) in the lateral superior olive (LSO), a major nucleus in the auditory brainstem, has not been performed so far, to our knowledge. Here we investigated the presence and characteristics of such receptors in the rat LSO by means of whole-cell patch-clamp recordings in combination with pharmacology. Current responses evoked by 200 microM AMPA were completely blocked by the specific AMPAR antagonist GYKI 52466 (100 microM). Properties of the AMPAR-mediated currents (latency, activation time constant, and peak amplitude) remained constant between postnatal day 3 (P3) and P10. Current responses evoked by 100 microM KA were not completely blocked by 100 microM GYKI 52466, indicating that the residual component was mediated by KARs. Throughout development, two groups of KAR-mediated currents (fast I(KA) and slow I(KA)) were distinguished because they had significantly different mean activation time constants. Moreover, the mean peak amplitude of fast I(KA) was significantly higher than that of slow I(KA). The differentiation into fast I(KA) and slow I(KA) can be explained by the existence of two groups of LSO neurons displaying different KAR densities, distributions, and/or diverse types with differences in conductance. Application of the specific KAR subunit agonists SYM 2081 (10 microM), ATPA (10 microM), or iodowillardiine (1 microM) evoked currents in almost all cells tested, showing that GluR5 subunits are a component of functional KARs in LSO neurons. Electrical stimulation of ipsilateral input fibers in the presence of KAR antagonists (NS-102 and GAMS), modulators (WGA), or GYKI 52466 revealed the presence of synaptic KARs in LSO neurons.

Age Factors↗
PubMed2003

Heuristic charge assignment for deconvolution of electrospray ionization mass spectra.

We propose a new algorithm for deconvolution of electrospray ionization mass spectra based on direct assignment of charge to the measured signal at each mass-to-charge ratio (m/z). We investigate two heuristics for charge assignment: the entropy-based heuristic is adapted from a deconvolution algorithm by Reinhold and Reinhold;10 the multiplicative-correlation heuristic is adapted from the multiplicative-correlation deconvolution algorithm of Hagen and Monnig.6 The entropy-based heuristic is insensitive to overestimates of z(max), the maximum ion charge. We test the deconvolution algorithm on two single-component samples: the measured spectrum of human beta-endorphin has two prominent and one very weak line whereas myoglobin has a well-developed quasi-gaussian envelope of 17 peaks. In both cases, the deconvolution algorithm gives a clean deconvoluted spectrum with one dominant peak and very few artefacts. The relative heights of the peaks due to the parent molecules in the deconvoluted spectrum of a mixture of two peptides, which are expected to ionize with equal efficiency, give an accurate measure of their relative concentration in the sample.

Algorithms↗
PubMed1997

Reduced engraftment and wound closure of cryopreserved cultured skin substitutes grafted to athymic mice.

Cryopreservation of cultured skin substitutes is a requirement for establishment of banks of alternative materials for treatment of acute and chronic skin wounds. To determine whether cryopreservation of skin substitutes that contain cultured cells reduces their efficacy for wound closure, cell-biopolymer grafts were frozen, recovered into culture, and grafted to wounds on athymic mice. Grafts consisted of cultured human keratinocytes and fibroblasts attached to collagen-glycosaminoglycan substrates that were frozen in cell culture medium with 20% serum and 10% DMSO at a controlled rate and stored overnight in liquid nitrogen. After recovery into culture for 24 h, frozen or unfrozen (control) skin substitutes were grafted to full-thickness wounds on athymic mice. Wound area and surface electrical capacitance were measured at 2, 3, and 4 weeks after grafting at which time animals were sacrificed. Wounds were scored for presence of human cells by direct immunofluorescence staining with a monoclonal antibody to HLA-ABC. The data demonstrate that cell-biopolymer grafts are less efficacious after controlled-rate cryopreservation using 10% DMSO as a cryoprotectant. Frozen grafts at 4 weeks after surgery have significantly smaller wound areas, higher capacitance (wetter surface), and fewer healed wounds that contain human cells. The results suggest that these conditions for cryopreservation of cultured grafts reduce graft viability. Improved conditions for cryopreservation are required to maintain viability and efficacy of cultured skin substitutes after frozen storage.

Animals↗
PubMed2000

Calmodulin and immunophilin are required as functional partners of a ryanodine receptor in ascidian oocytes at fertilization.

Fertilization of oocytes incites numerous changes relying on Ca(2+) signaling. In inseminated ascidian eggs, an increase in the egg surface membrane, monitored by a change in electrical capacitance, is recorded at the onset of meiosis resumption. This membrane addition to the cell surface is controlled by calcium release through a ryanodine receptor (RyR), sensitive to cyclic ADP-ribose. Using confocal microscopy analysis of ascidian oocytes immunostained with anti-RyR antibody, we show here that this calcium channel is asymmetrically located in the vegetal cortical zone. Interestingly, the increase in cell capacitance occurring at fertilization is correlated with a fluorescent signal, imaged by the marker of vesicle trafficking FM 1-43, located close to the RyR region. Two putative partners of RyR, namely an FKBP-like protein and a calmodulin, are identified in these oocyte extracts by detection of enzyme activity and PCR amplification. Both are necessary to sustain ryanodine receptor activity in these oocytes since the membrane insertion triggered by fertilization is inhibited by the FKBP ligand rapamycin and by a calmodulin antagonist peptide. These findings suggest that exocytosis in ascidian eggs is triggered at fertilization by a functional Ca(2+) release unit operating as a complex of several proteins, including a calmodulin and an immunophilin, around the intracellular calcium channel itself.

Amino Acid Sequence↗
PubMed1985

Optical and electrical properties of thin monoolein lipid bilayers.

Monoolein lipid bilayers were formed using a monolayer transfer technique and from dispersions of monoolein in squalene, triolein, 1-chlorodecane and 1-bromodecane. Measurements of optical reflectance and electrical capacitance were used to determine the thickness and dielectric constant of the bilayers. The thickness of the hydrocarbon region of the five bilayer systems ranged from 2.5 to 3.0 nm. Two of the bilayer systems (made from 1-chlorodecane and 1-bromodecane solvents) had a high dielectric constant (2.8 to 2.9) whereas the other bilayer systems had dielectric constants close to that of pure hydrocarbons (2.2). The charge-pulse technique was used to study the transport kinetics of three lipophilic ions and two ion carrier complexes in the bilayers. For the low dielectric constant bilayers, the transport of the lipophilic ions tetraphenylborate, tetraphenylarsonium and dipicrylamine was governed mainly by the thickness of the hydrocarbon region of the bilayer whereas the transport of the ion-carrier complexes proline valinomycin-K+ and valinomycin-Rb+ was nearly independent of thickness. This is consistent with previous studies on thicker monoolein bilayers. The transport of lipophilic anions across bilayers with a high dielectric constant was 20 to 50 times greater than expected on the basis of thickness alone. This agrees qualitatively with predictions based on Born charging energy calculations. High dielectric constant bilayers were three times more permeable to the proline valinomycin-K+ complex than were low dielectric constant bilayers but were just as permeable as low dielectric constant bilayers to the valinomycin-Rb+ complex.

Glycerides↗