Kinetic studies of Ca2+ release from sarcoplasmic reticulum of normal and malignant hyperthermia susceptible pig muscles.

The time-course of Ca2+ release from sarcoplasmic reticulum isolated from muscles of normal pigs and those of pigs susceptible to malignant hyperthermia were investigated using stopped-flow spectrophotometry and arsenazo III as a Ca2+ indicator. Several methods were used to trigger Ca2+ release: (a) addition of halothane (e.g., 0.2 mM); (b) an increase of extravesicular Ca2+ concentration ([Ca2+0]); (c) a combination of (a) and (b), and (d) replacement of ions (potassium gluconate with choline chloride) to produce membrane depolarization. The initial rates of Ca2+ release induced by either halothane or Ca2+ alone, or both, are at least 70% higher in malignant hyperthermic sarcoplasmic reticulum than in normal. The amount of Ca2+ released by halothane at low [Ca2+0] in malignant hyperthermic sarcoplasmic reticulum is about twice as large as in normal sarcoplasmic reticulum. Membrane depolarization led to biphasic Ca2+ release in both malignant hyperthermic and normal sarcoplasmic reticulum, the rate constant of the rapid phase of Ca2+ release induced by membrane depolarization being significantly higher in malignant hyperthermic sarcoplasmic reticulum (k = 83 s-1) than in normal (k = 37 s-1). Thus, all types of Ca2+ release investigated (a, b, c and d) have higher rates in malignant hyperthermic sarcoplasmic reticulum than normal sarcoplasmic reticulum. These results suggest that the putative Ca2+ release channels located in the sarcoplasmic reticulum are altered in malignant hyperthermic sarcoplasmic reticulum.

Dantrolene prevents the malignant hyperthermic syndrome by reducing free intracellular calcium concentration in skeletal muscle of susceptible swine.

Malignant hyperthermia (MH) is a pharmacogenetic disorder of skeletal muscle triggered when susceptible subjects are exposed to volatile anesthetic agents and/or depolarizing muscle relaxants. We have used Ca2+ selective microelectrodes to measure in vivo the intracellular free [Ca2+] in skeletal muscle of MH susceptible swine before and after the administration of dantrolene. We have investigated the effectiveness of this muscle relaxant in preventing clinical MH and the relationship between the resting intracellular free [Ca2+] and the probability of inducing the MH syndrome. The resting intracellular free [Ca2+] was 0.41 +/- 0.01 microM (M +/- SEM), which agrees with our previous measurements in susceptible swine. The administration of 0.5, 1, 2, 2.5 and 3 mg/Kg Dantrolene, reduced the intracellular free [Ca2+] to 0.31, 0.21, 0.09, 0.08, 0.08 microM respectively. The 0.5 mg/Kg dose induced a moderate decrease of [Ca2+]i and failed to prevent the MH syndrome after exposure to halothane (2%). The 1 mg/Kg dose produced a further reduction in [Ca2+]i and was sufficient to prevent the clinical syndrome in 2 out of 3 animals. The 2.5 mg/Kg dose was uniformly protective in all animals. These results suggest that the mechanism by which dantrolene protects susceptible animals exposed to triggering agents is by reducing the intracellular free [Ca2+] in skeletal muscle.

Malignant eccrine poroma: report of three cases.

The characteristic clinical and histological features of three cases of malignant eccrine poroma are discussed, in addition to the metastatic disease that had occurred in two cases. These cases were compared with previously reported cases of malignant eccrine poroma that had metastasised, and it is suggested that a strict classification of malignant eccrine sweat gland tumours should be made.

Immunohistochemical analysis of mononuclear cell subsets in inflammatory and non-inflammatory myopathies.

Immunohistochemical procedures were used to analyse the subpopulations of mononuclear cells in muscle biopsies from 24 patients with polymyositis. The character of the cellular infiltrate was similar at the perivascular, perimysial, and endomysial sites, with cytotoxic-suppressor T lymphocytes (T8+) and macrophages being the dominant elements. Helper T lymphocytes (T4+) and B lymphocytes were present in smaller numbers. A control series of 17 muscle biopsies from normal subjects and patients with non-inflammatory myopathies and neurogenic conditions was also studied: the numbers of mononuclear cells present were much smaller than in polymyositis, but the ratio of T4:T8 lymphocytes was similar to that found in biopsies affected by polymyositis. We conclude that both cytotoxic-suppressor T lymphocytes and macrophages are important in the pathogenesis of inflammatory myopathy.

Spatio-temporal contrast sensitivity, coherent motion, and visible persistence in developmental dyslexia.

Three experiments measured spatio-temporal contrast sensitivity, coherent motion, and visible persistence in a single group of children with developmental dyslexia and a matched control group. The findings were consistent with a transient channel disorder in the dyslexic group which showed a reduction in contrast sensitivity at low spatial frequencies, a significant reduction in sensitivity for coherent motion, and a significantly longer duration of visible persistence. The results were also examined by classifying the dyslexic group into dyseidetic, dysphonetic, and mixed (dysphoneidetic) subgroups. There were no differences between the control and dyseidetic groups in contrast sensitivity, in coherent motion and in visible persistence. In comparison to the control group, the mixed (dysphoneidetic) dyslexic subgroup was found to have a significant reduction in contrast sensitivity at low spatial frequencies, a significant reduction in sensitivity for coherent motion, and a significantly longer duration of visible persistence. In comparison to the control group, the dysphonetic group only showed a reduction in contrast sensitivity at low spatial frequencies. Comparisons between the dyseidetic, dysphonetic and mixed dyslexic subgroups showed that there were no substantive differences in contrast sensitivity, coherent motion, and visible persistence. The results support the proposal and findings by Borsting et al. (Borsting E, Ridder WH, Dudeck K, Kelley C, Matsui L, Motoyama J. Vis Res 1996;36:1047-1053) that a transient channel disorder may only be present in a dysphoneidetic dyslexic subgroup. Psychometric assessment revealed that all the children with dyslexia appear to have a concurrent disorder in phonological coding, temporal order processing, and short-term memory.

Safety of amide local anesthetics in patients susceptible to malignant hyperthermia.

Earlier reports on malignant hyperthermia warned against the use of local anesthetics in the amide class in persons susceptible to the syndrome. The preponderance of data supports the safety of amide local anesthetics in such patients, and these agents should not be withheld from persons at risk for developing the syndrome.

Sudden unexplained death in a patient with a family history of malignant hyperthermia.

A 23-year-old healthy white male in excellent physical condition developed cardiac arrest and rigidity during moderate exercise. He had a strong family history of malignant hyperthermia (MH). Two hours postmortem, his temperature was noted to be markedly elevated [41 degrees C (106 degrees F)]. A review of the possible differential diagnoses point to MH as a reasonable etiology.

Doping of carbon nanotubes with nitrogen improves protein coverage whilst retaining correct conformation.

Relevant parameters for non-covalent protein functionalization of carbon nanotubes are explored. Multiwalled carbon nanotubes are carboxylated and functionalized with metalloproteins. Using atomic force microscopy (AFM) we quantitatively determine that coverage with nitrogen-doped multiwalled carbon nanotubes is superior compared to coverage with un-doped multiwalled carbon nanotubes, due to enhanced carboxylation. Conformational analysis using a combination of AFM, antibody binding assays, circular dichroism and UV-visible spectroscopy demonstrates that the metalloproteins retain their native structure when adsorbed to nitrogen-doped multiwalled carbon nanotubes irrespective of their size, charge or folding motif.

Electrostatic and steric interactions determine bacteriorhodopsin single-molecule biomechanics.

Bacteriorhodopsin (bR) is a haloarchaeal membrane protein that converts the energy of single photons into large structural changes to directionally pump protons across purple membrane. This is achieved by a complex combination of local dynamic interactions controlling bR biomechanics at the submolecular level, producing efficient amplification of the retinal photoisomerization. Using single molecule force spectroscopy at different salt concentrations, we show that tryptophan (Trp) residues use steric specific interactions to create a rigid scaffold in bR extracellular region and are responsible for the main unfolding barriers. This scaffold, which encloses the retinal, controls bR local mechanical properties and anchors the protein into the membrane. Furthermore, the stable Trp-based network allows ion binding to two specific sites on the extracellular loops (BC and FG), which are involved in proton release and lateral transport. In contrast, the cytoplasmic side of bR is mainly governed by relatively weak nonspecific electrostatic interactions that provide the flexibility necessary for large cytoplasmic structural rearrangements during the photocycle. The presence of an extracellular Trp-based network tightly enclosing the retinal seems common to most haloarchaeal rhodopsins, and could be relevant to their exceptional efficiency.

Differential stiffness and lipid mobility in the leaflets of purple membranes.

Purple membranes (PM) are two-dimensional crystals formed by bacteriorhodopsin and a variety of lipids. The lipid composition and density in the cytoplasmic (CP) leaflet differ from those of the extracellular (EC) leaflet. A new way of differentiating the two sides of such asymmetric membranes using the phase signal in alternate contact atomic force microscopy is presented. This method does not require molecular resolution and is applied to study the stiffness and intertrimer lipid mobility in both leaflets of the PM independently over a broad range of pH and salt concentrations. PM stiffens with increasing salt concentration according to two different regimes. At low salt concentration, the membrane Young's normal modulus grows quickly but differentially for the EC and CP leaflets. At higher salt concentration, both leaflets behave similarly and their stiffness converges toward the native environment value. Changes in pH do not affect PM stiffness; however, the crystal assembly is less pronounced at pH > or = 10. Lipid mobility is high in the CP leaflet, especially at low salt concentration, but negligible in the EC leaflet regardless of pH or salt concentration. An independent lipid mobility study by solid-state NMR confirms and quantifies the atomic force microscopy qualitative observations.

Nonequilibrium quasiparticle relaxation in the vortex state of La2-xSrxCuO4.

We have measured the charge dynamics in the vortex state of La(2-x)Sr(x)CuO(4) by femtosecond time-resolved reflectance, which we demonstrate to be a direct probe of low-energy quasiparticle states. Application of a c-axis magnetic field induces regions surrounding vortex cores that display pseudogap charge dynamics. We determine the characteristic width approximately 130 A in optimally doped material and we show that it increases with decreasing doping. These results confirm a new experimental method of probing the microscopic properties of vortices in the cuprates.

Longitudinal photocurrent spectroscopy of a single GaAs/AlGaAs v-groove quantum wire.

Modulation-doped GaAs v-groove quantum wires (QWRs) have been fabricated with novel electrical contacts made to two-dimensional electron-gas (2DEG) reservoirs. Here, we present longitudinal photocurrent (photoconductivity/PC) spectroscopy measurements of a single QWR. We clearly observe conductance in the ground-state one-dimensional subbands; in addition, a highly temperature-dependent response is seen from other structures within the v-groove. The latter phenomenon is attributed to the effects of structural topography and localization on carrier relaxation. The results of power-dependent PC measurements suggest that the QWR behaves as a series of weakly interacting localized states, at low temperatures.