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PubMed2002

Electret-thermal analysis of blood.

The thermally stimulated discharge (TSD) method, intended for the analysis of charged dielectrics, was used for human blood research. Above-room-temperature TSD spectra of blood consist of three peaks. There are indications that the low-temperature peak (40-50 degrees C) corresponds to the thermally stimulated destruction of hydrate shells surrounding blood components while the mid-temperature peak (70-90 degrees C) is related to thermal denaturation of blood proteins. The intensive high-temperature peak (105-120 degrees C) is observed when a phase transition of blood, accompanied by the formation of a firm dry film of blood, occurs. The position of the high-temperature peak depends on the blood group. Data is discussed which suggests that the spontaneous "quasi-electret effect" of blood relates to the character of the biochemical processes taking place in human organisms. The TSD method might be used as simple and informative means of diagnostics in cooperation with medical and physical investigations.

Adult↗
PubMed2003

Interaction of amoebapores and NK-lysin with symmetric phospholipid and asymmetric lipopolysaccharide/phospholipid bilayers.

Amoebapores from protozoan parasite Entamoeba histolytica and NK-lysin of porcine cytotoxic lymphocytes belong to the same family of saposin-like proteins. In addition to the structural similarity, amoebapores and NK-lysin are both highly effective against prokaryotic and eukaryotic target cells in that they permeabilize the target cell membranes. Here, we have investigated in detail the protein/lipid interaction for the three isoforms of amoebapore and NK-lysin. Results obtained from electrical measurements on planar bilayer membranes, including reconstitution models of the lipid matrix of the outer membrane of Escherichia coli and phospholipid membranes, fluorescence energy transfer spectroscopy with liposomes, and monolayer measurements on a Langmuir trough, provided information on lipid preferences, pH dependences, and membrane interaction mechanisms. The three amoebapores led to the formation of transient pores with similar characteristics in conductance, sublevels, and lifetime for the different isoforms. The conductance of the pores was dependent on the polarity of the applied clamp voltage, and the distribution of the sublevels was affected by the value of the clamp voltage. The size of the pores and distribution of conductance sublevels differed between symmetric phospholipid and asymmetric lipopolysaccharide/phospholipid bilayers. Notably, NK-lysin caused the formation of well-defined pores, which were lipid- and voltage-dependent, and their characteristics differed from those induced by amoebapores; e.g., the protein concentration necessary to induce pore formation was 20 times higher. The biophysical data give important information on the mode of action of these small effector proteins, which may further lead to a better understanding of peptide-membrane interactions in general.

Animals↗
PubMed2005

Negative dipole potentials of uncharged langmuir monolayers due to fluorination of the hydrophilic heads.

The dipole potential, affecting the structure, functions, and interactions of biomembranes, lipid bilayers, and Langmuir monolayers, is positive toward the hydrocarbon moieties. We show that uncharged Langmuir monolayers of docosyl trifluoroethyl ether (DFEE) exhibit large negative dipole potentials, while the nonfluorinated docosyl ethyl ether (DEE) forms films with positive dipole potentials. Comparison of the Delta V values for these ethers with those of the previously studied(37-39) monolayers of trifluoroethyl ester (TFEB) and ethyl ester of behenic acid (EB) shows that the reversal of the sign of Delta V causes the same change Delta(Delta V) = -706 +/- 16 mV due to fluorination of heads. The Delta V values of both TFEB and EB films differ by -122 +/- 16 mV from those of DFEE and DEE monolayers, respectively, with the same density. Such quantitative coincidence points to a common mechanism of reversal of the sign of the dipole potential for the ether and ester films despite the different structure of their heads. The mechanical properties and phase behaviors of these monolayers show that both fluorinated heads are less hydrated, suggesting that the change of the sign of Delta V could, at least partially, be related to different hydration water structure. The same negative contribution of the carbonyl bond in both TFEB and EB films contrasts with the generally accepted positive contribution of the C(delta+)=O(delta-) bond in condensed Langmuir monolayers of fatty acids, their alcohol esters, glycerides, and phospholipids but concurs with the theoretical analysis of Delta V of stearic acid monolayers. Both results question the literature values of the molecular dipole moments of these substances calculated via summation of bonds and atomic group contributions. Mixed monolayers of DFEE and DEE show smooth monotonic variation of Delta V from +450 to -235 mV, indicating a way for adjustment of the sign and magnitude of the dipole potential at the membrane-water boundary and regulation of such membrane behaviors as binding and translocation rate of hydrophobic ions and ion-carriers, adsorption and penetration of amphiphilic peptides, polarization of hydration water, and short-range repulsion. The interaction of the hydrophobic ions tetraphenylboron TPhB- and tetraphenylphosphonium TPhP+ with DFEE and DEE monolayers qualitatively follows the theory of binding of such ions to lipid bilayers, but the shifts Delta(Delta V) from the values obtained on water are much smaller than those for DPPC monolayers. This difference seems to be due to the solid (polycrystalline) character of the DFEE and DEE films that hampers the penetration of TPhB- and TPhP+ in the monolayers and reduces the attractive interaction with the hydrophobic moiety. This conclusion orients the future synthesis of amphiphiles with fluorinated heads to those which could form liquid-expanded Langmuir monolayers.

Chemical Phenomena↗
PubMed2005

Electrooxidation of carbon monoxide on gold nanoparticle ensemble electrodes: effects of particle coverage.

Gold nanoparticles were attached to amine-functionalized indium tin oxide substrate to form particle ensemble electrodes with controlled particle coverage. Electrooxidation of carbon monoxide (CO) on these particle ensemble electrodes was studied in CO-saturated alkaline solutions by means of cyclic voltammetry, with an emphasis on the effects of particle coverage. CO oxidation half-wave potential was found to decrease with increasing particle density. However, the current density was significantly larger at lower particle coverage electrodes. On the basis of a model for electron transfer on a partially covered electrode, the observations were explained in terms of the change in reactant mass transport pattern with varying particle coverages: CO diffusion is predominantly mixed spherical and linear at low particle coverages and changes to mostly linear at high particle coverages. The possibility of contributions from particle agglomeration is also briefly discussed.

Carbon Monoxide↗
PubMedDate not supplied

An estimator for the electrotonic size of neurons independent of charge equalization time constants.

Electrotonic properties are important aspects of neuronal function but have been difficult to estimate without accurate morphological reconstruction. The complexity of the branching dendritic cables often gives charging curves composed of a very large number of exponential functions, making it difficult to distinguish the time constants that are needed for electrotonic estimates. We describe an estimator P for the electrotonic size of neurons based on simple measures from voltage and current clamp recordings that does not rely on the higher rank exponential components of the response. Our estimator gives a bounded scale for the electrotonic size of the cell and can be used for categorization and comparison when morphology is not available.

Action Potentials↗
PubMedDate not supplied

A novel mechanism for irregular firing of a neuron in response to periodic stimulation: irregularity in the absence of noise.

Irregular firing of action potentials (AP's) is a characteristic feature of neurons in the brain. The variability has been attributed to noise from various sources. This study illustrates an alternative mechanism, namely, deterministic irregularity within a model of ionic conductances. Specifically, a model based on modern measurements of the Na+ and K+ current components from the squid giant axon fires irregularly in response to a continuous train of near-threshold current pulses. The interspike interval histogram from these simulations is multi-modal, a result which in other systems has been attributed to stochastic resonance. Moreover, the simulations exhibited short burst of spikes followed by relatively long quiescent periods, a result suggestive of patterned input to the model even though the input consisted of a train of regularly spaced current pulses. The variability of firing is attributable to variations in AP parameters, in particular AP amplitude. The action potential for squid giant axons is not all-or-none. Rather, it is fundamentally a continuous function of stimulus amplitude. That is, the membrane lacks a threshold. Variation in AP amplitude, and to a lesser extent, AP duration, can produce variations in the time to a subsequent AP, which represents a paradigm shift for understanding irregular neuronal firing. The emphasis is not as much on events prior to an AP as it is on the AP's themselves.

Acoustic Stimulation↗
PubMed2001

Changes of biophysical properties of the skin measured by non-invasive techniques after Q-switched Nd-YAG laser therapy in patients with nevus of Ota.

BACKGROUND/AIMS: The aim of the study was to objectify the effect of a Q-switched Nd-YAG laser on Ota's nevus in view of barrier function, surface contour changes, dermal blood flow, surface color changes and sebum production rate. METHODS: Fifteen Korean patients with nevus of Ota (between 14 and 54 years of age) were enrolled. All the patients were treated by Q-switched Nd-YAG laser and followed up for 12 weeks. A reflectance spectrophotometer, a colorimeter, laser Doppler flowmetry, a Tewameter, a Corneometer and a Sebumeter were used to make measurements. RESULTS: Pvalues of less than 0.05 were regarded as statistically significant. In skin reflectance measurements, L* values declined for 4 weeks and increased at 12 weeks, indicating that the brightness of the lesions improved. a* values showed a significant increase at 2 and 4 weeks. On the contrary, b* values decreased at 2 weeks. Transepidermal water loss and dermal blood flux showed identical patterns, showing increases at 2 weeks. The water holding capacity decreased at 2 and 4 weeks, and thereafter showed a delayed recovery. Casual sebum production increased at 4, 8 and 12 weeks. The loss of skin roughness was observed at 4 weeks, recovering at 12 weeks, although statistically insignificant. Reflex spectrophotometry did not reflect the changing properties of the skin. CONCLUSIONS: Our data suggest a useful model for evaluating physiologic skin changes after laser surgery in patients with nevus of Ota.

Adolescent↗
PubMed2005

A tctex1-Ca2+ channel complex for selective surface expression of Ca2+ channels in neurons.

Voltage-gated Ca(2+) channels (VGCCs) are important in regulating a variety of cellular functions in neurons. It remains poorly understood how VGCCs with different functions are sorted within neurons. Here we show that the t-complex testis-expressed 1 (tctex1) protein, a light-chain subunit of the dynein motor complex, interacts directly and selectively with N- and P/Q-type Ca(2+) channels, but not L-type Ca(2+) channels. The interaction is insensitive to Ca(2+). Overexpression in hippocampal neurons of a channel fragment containing the binding domain for tctex1 significantly decreases the surface expression of endogenous N- and P/Q-type Ca(2+) channels but not L-type Ca(2+) channels, as determined by immunostaining. Furthermore, disruption of the tctex1-Ca(2+) channel interaction significantly reduces the Ca(2+) current density in hippocampal neurons. These results underscore the importance of the specific tctex1-channel interaction in determining sorting and trafficking of neuronal Ca(2+) channels with different functionalities.

Animals↗
PubMed2005

An experimental study of melting of CCl4 in carbon nanotubes.

We report dielectric relaxation spectroscopy measurements of the melting point of carbon tetrachloride confined within open-tip multi-walled carbon nanotubes with two different pore diameters, 4.0 and 2.8 nm. In both cases, a single transition temperature well above the bulk melting point was obtained for confined CCl4. These results contrast with what was obtained in our previous measurements using carbon nanotubes with a pore diameter of 5.0 nm, where multiple transition temperatures both above and below the bulk melting point of CCl4 were observed. Our experimental measurements are consistent with our recent molecular simulation results (F. R. Hung, B. Coasne, E. E. Santiso, K. E. Gubbins, F. R. Siperstein and M. Sliwinska-Bartkowiak, J. Chem. Phys., 2005, 122, 144706). Although the simulations overestimate the temperatures in which melting upon confinement occurs, both simulations and experiments suggest that all regions of adsorbate freeze at the same temperature, and that freezing occurs at higher temperatures upon reduction of the pore diameter.

Carbon Tetrachloride↗
PubMed2003

Electrophysiological properties of the utricular primary transducer are modified during development under hypergravity.

The electrophysiological development of hair cells between birth and the eight postnatal day (P8) was studied in the utricular macula of rats gestated in nest boxes mounted upon a centrifuge, subjecting the animals to a gravitational force of 2G. Whole-cell voltage-clamp recordings were made on cells in the acutely isolated epithelium. Cells were accessed through a tear in the epithelium, no enzymatic dissociation procedures were employed. Under artificially enhanced gravity, the whole cell conductance was dramatically altered in the two types of hair cells. Significant increases occurred from P3-4 in the type I cells while in the type II cells, the effect was delayed until P7-8. Fourfold and threefold increases of the mean slope conductance were observed at P7-8 in the type I and type II hair cells, respectively. These results indicate that the electrophysiological properties of a primary transducer such as utricle may be modified by variation of the primary stimulus during development.

Aging↗
PubMed2001

Rise of defibrillation energy requirement under carvedilol therapy.

Carvedilol, a nonselective beta 1-, beta 2-adrenoreceptor blocking agent with alpha 1-adrenoreceptor blocking activity, is often prescribed as an adjunctive pharmacological therapy in patients who received an ICD. Despite the new ICD technology, concomitant antiarrhythmic therapy still represents the most important concern in patients with an ICD. As illustrated by this case, carvedilol may also increase the energy requirement for internal defibrillation.

Aged↗
PubMed2002

AMPA glutamate receptor-mediated calcium signaling is transiently enhanced during development of oligodendrocytes.

Cells of the oligodendroglial lineage express Ca2+-permeable alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate-preferring glutamate receptors (AMPA-GluR) during development. Prolonged activation of their AMPA-GluR causes Ca2+ overload, resulting in excitotoxic death. Prior studies have shown that oligodendroglial progenitors and immature oligodendrocytes are susceptible to excitotoxicity, whereas mature oligodendrocytes are resistant. An unresolved issue has been why Ca2+-permeability of AMPA-GluR varies so markedly with oligodendroglial development, although the level of expression of edited GluR2, an AMPA-GluR subunit which blocks Ca2+ entry, is relatively constant. To address this question, we performed Ca2+ imaging, molecular and electrophysiological analyses using purified cultures of the rat oligodendroglial lineage. We demonstrate that transient up-regulation of expression of GluR3 and GluR4 subunits in oligodendroglial progenitors and immature oligodendrocytes results in the assembly by these cells, but not by oligodendroglial pre-progenitors or mature oligodendrocytes, of a population of AMPA-GluR which lack GluR2. This stage-specific up-regulation of edited GluR2-free, and hence Ca2+-permeable, AMPA-GluR explains the selective susceptibility to excitotoxicity of cells at these stages of oligodendroglial differentiation, and is likely to be important to these cells in the trans-synaptic Ca2+-signaling from glutamatergic neurons, which occurs in hippocampus

Animals↗
PubMed2002

Wave analysis of implant screw loosening using an air cylindrical cyclic loading device.

STATEMENT OF PROBLEM: The mechanics of implant screw loosening or fracture are well understood in the field of engineering. They have not been as widely explored in dentistry. PURPOSE: This study investigated the effects of simulated mastication on implant components and used wave analysis to document the basic mechanisms of screw loosening in a simulated oral environment. MATERIAL AND METHODS: A pneumatic cylindrical cyclic loading device was fabricated to simulate masticatory movement. Thirteen standard abutments were connected on external hexagonal implants with titanium abutment screws tightened to 20 Newton centimeters (Ncm), and single crowns were retained with gold screws tightened to 10 Ncm on each abutment, respectively. Ten single-implant crowns were loaded with the use of a cyclic loading device with 100 N of force at 30 degrees angles to the long-axis for 0.2 seconds of contact time with a frequency of 1 Hz. Three crowns were loaded vertically under the same conditions to serve as the control group. The effects of up to 1 million cyclic loads and various tightening torque forces (2, 4, 6, 8, 10, and 12 Ncm) on screw loosening were evaluated by wave analysis. A software program was written to record every wave mode and to stop the machine automatically if the amount of horizontal displacement of the crown was more than 0.5 mm, which was designated to represent perceptible loosened implant crown mobility clinically. The general wave patterns and characteristics of loosened and stable screws and the effect of various tightening torques were analyzed by comparing the differences in wave patterns. RESULTS: The wave mode was divided into 4 stages for loosened gold screws: initial displacement, initial vibration, elastic deformation, and recovery stage. However, the initial displacement and initial vibration stages were not discernible for stable gold screws. Of the 10 gold occlusal screws, 4 loosened before the 1 million cyclic loads in the 10 single crowns tested. There was no screw loosening in the control group. There was no effect of screw loosening on the elastic deformation stage. CONCLUSION: Within the limitations of this study, tightening torque had a significant effect on screw loosening. It would appear that more than 10 Ncm of tightening torque should be recommended for the gold screws in this external hexagon implant system.

Crowns↗
PubMed2005

Neuronal calcium sensor-1 potentiates glucose-dependent exocytosis in pancreatic beta cells through activation of phosphatidylinositol 4-kinase beta.

Cytosolic free Ca2+ plays an important role in the molecular mechanisms leading to regulated insulin secretion by the pancreatic beta cell. A number of Ca2+-binding proteins have been implicated in this process. Here, we define the role of the Ca2+-binding protein neuronal Ca2+ sensor-1 (NCS-1) in insulin secretion. In pancreatic beta cells, NCS-1 increases exocytosis by promoting the priming of secretory granules for release and increasing the number of granules residing in the readily releasable pool. The effect of NCS-1 on exocytosis is mediated through an increase in phosphatidylinositol (PI) 4-kinase beta activity and the generation of phosphoinositides, specifically PI 4-phosphate and PI 4,5-bisphosphate. In turn, PI 4,5-bisphosphate controls exocytosis through the Ca2+-dependent activator protein for secretion present in beta cells. Our results provide evidence for an essential role of phosphoinositide synthesis in the regulation of glucose-induced insulin secretion by the pancreatic beta cell. We also demonstrate that NCS-1 and its downstream target, PI 4-kinase beta, are critical players in this process by virtue of their capacity to regulate the release competence of the secretory granules.

1-Phosphatidylinositol 4-Kinase↗
PubMed2002

Inositol hexakisphosphate promotes dynamin I- mediated endocytosis.

Membrane homeostasis is maintained by exocytosis and endocytosis. The molecular mechanisms regulating the interplay between these two processes are not clear. Inositol hexakisphosphate (InsP(6)) is under metabolic control and serves as a signal in the pancreatic beta cell stimulus-secretion coupling by increasing Ca(2+)-channel activity and insulin exocytosis. We now show that InsP(6) also promotes dynamin I-mediated endocytosis in the pancreatic beta cell. This effect of InsP(6) depends on calcineurin-induced dephosphorylation and is accounted for by both activation of protein kinase C and inhibition of the phosphoinositide phosphatase synaptojanin and thereby formation of phosphatidylinositol 4,5-bisphosphate. In regulating both exocytosis and endocytosis, InsP(6) thus may have an essential integral role in membrane trafficking.

Animals↗
PubMed2003

Rapid association of protein kinase C-epsilon with insulin granules is essential for insulin exocytosis.

Glucose-dependent exocytosis of insulin requires activation of protein kinase C (PKC). However, because of the great variety of isoforms and their ubiquitous distribution within the beta-cell, it is difficult to predict the importance of a particular isoform and its mode of action. Previous data revealed that two PKC isoforms (alpha and epsilon) translocate to membranes in response to glucose (Zaitzev, S. V., Efendic, S., Arkhammar, P., Bertorello, A. M., and Berggren, P. O. (1995) Proc. Natl. Acad. Sci. U. S. A. 92, 9712-9716). Using confocal microscopy, we have now established that in response to glucose, PKC-epsilon but not PKC-alpha associates with insulin granules and that green fluorescent protein-tagged PKC-epsilon changes its distribution within the cell periphery upon stimulation of beta-cells with glucose. Definite evidence of PKC-epsilon requirement during insulin granule exocytosis was obtained by using a dominant negative mutant of this isoform. The presence of this mutant abolished glucose-induced insulin secretion, whereas transient expression of the wild-type PKC-epsilon led to a significant increase in insulin exocytosis. These results suggest that association of PKC-epsilon with insulin granule membranes represents an important component of the secretory network because it is essential for insulin exocytosis in response to glucose.

Animals↗
PubMed2002

Tracking voltage-dependent conformational changes in skeletal muscle sodium channel during activation.

The primary voltage sensor of the sodium channel is comprised of four positively charged S4 segments that mainly differ in the number of charged residues and are expected to contribute differentially to the gating process. To understand their kinetic and steady-state behavior, the fluorescence signals from the sites proximal to each of the four S4 segments of a rat skeletal muscle sodium channel were monitored simultaneously with either gating or ionic currents. At least one of the kinetic components of fluorescence from every S4 segment correlates with movement of gating charge. The fast kinetic component of fluorescence from sites S216C (S4 domain I), S660C (S4 domain II), and L1115C (S4 domain III) is comparable to the fast component of gating currents. In contrast, the fast component of fluorescence from the site S1436C (S4 domain IV) correlates with the slow component of gating. In all the cases, the slow component of fluorescence does not have any apparent correlation with charge movement. The fluorescence signals from sites reflecting the movement of S4s in the first three domains initiate simultaneously, whereas the fluorescence signals from the site S1436C exhibit a lag phase. These results suggest that the voltage-dependent movement of S4 domain IV is a later step in the activation sequence. Analysis of equilibrium and kinetic properties of fluorescence over activation voltage range indicate that S4 domain III is likely to move at most hyperpolarized potentials, whereas the S4s in domain I and domain II move at more depolarized potentials. The kinetics of fluorescence changes from sites near S4-DIV are slower than the activation time constants, suggesting that the voltage-dependent movement of S4-DIV may not be a prerequisite for channel opening. These experiments allow us to map structural features onto the kinetic landscape of a sodium channel during activation.

Algorithms↗
PubMed2006

Phenotypic changes in NG2+ cells after spinal cord injury.

Following contusive spinal cord injury (SCI), 50% of oligodendrocytes in the residual white matter are lost within 24 h. NG2-expressing cell proliferation is maximal 3 days after SCI, and may be the source of mature oligodendrocytes and astrocytes that chronically replace those that were lost. We studied NG2(+) cells dissociated from the 3-day injured spinal cord for comparison with those from uninjured adult and early postnatal cords. After 24 h in serum-containing medium, we performed patch clamp analysis and immunocytochemistry for NG2 in combination with nestin (progenitors), and A2B5, O4, and O1 (oligodendrocyte lineage markers). We observed an NG2(+)/A2B5-/O4-/O1- population in both adult preparations. More than double the normal number of NG2(+) cells was isolated from the injured cord, but OX42(+) microglia/macrophages were the predominant cell type after injury. Most cells isolated at P7 were NG2-/A2B5(+), whereas those from the normal adult were NG2(+)/A2B5-. NG2(+) cells after SCI displayed altered voltage-gated potassium current profiles compared to normal adult and P7 animals. Additionally, less than 25% of adult cells (normal and injured) responded to GABA and glutamate, compared to 100% of P7 cells. Our results indicate that the adult NG2(+) cell pool is antigenically and physiologically different than the early postnatal pool, and that contusive injury induces changes in adult NG2(+) cells.

Animals↗