Modulation of BK Channels by Small Endogenous Molecules and Pharmaceutical Channel Openers.

Voltage- and Ca(2+)-activated K(+) channels of big conductance (BK channels) are abundantly found in various organs and their relevance for smooth muscle tone and neuronal signaling is well documented. Dysfunction of BK channels is implicated in an array of human diseases involving many organs including the nervous, pulmonary, cardiovascular, renal, and urinary systems. In humans a single gene (KCNMA1) encodes the pore-forming α subunit (Slo1) of BK channels, but the channel properties are variable because of alternative splicing, tissue- and subcellular-specific auxiliary subunits (ß, γ), posttranslational modifications, and a multitude of endogenous signaling molecules directly affecting the channel function. Initiatives to develop drugs capable of activating BK channels (channel openers) therefore need to consider the tissue-specific variability of BK channel structure and the potential interference with endogenously produced regulatory factors. The atomic structural basis of BK channel function is only beginning to be revealed. However, building on detailed knowledge of BK channel function, including its single-channel characteristics, voltage- and Ca(2+) dependence of channel gating, and modulation by diffusible messengers, a multi-tier allosteric model of BK channel gating (Horrigan and Aldrich (HA) model) has become a valuable tool in studying modulation of the channel. Using the conceptual framework of the HA model, we here review the functional impact of endogenous modulatory factors and select small synthetic compounds that regulate BK channel activity. Furthermore, we devise experimental approaches for studying BK channel-drug interactions with the aim to classify BK-modulating substances according to their molecular mode of action.

Population dynamics of Armigeres subalbatus (Diptera: Culicidae) across a temperate altitudinal gradient.

Understanding the impacts of weather fluctuations, and environmental gradients, on the abundance of vectors is fundamental to grasp the dynamic nature of the entomological risk for disease transmission. The mosquito Armigeres subalbatus (Coquillet) is a common vector of filariasis. Nevertheless, its population dynamics have been relatively poorly studied. Here, we present results from a season long study where we studied spatio-temporal abundance patterns of Ar. subalbatus across the altitudinal gradient of Mt. Konpira in Nagasaki, Japan. Spatially, we found that abundance of adult Ar. subalbatus decreased with altitude and increased in areas where the ground was rich in leaf litter. Similarly, adult activity was observed only when relative humidity was over 65%. Temporally, we found that peaks in abundance followed large rainfall events. Nevertheless, this mosquito was under significant density dependence regulation. Our results suggest that Ar. subalbatus population peaks following large rainfall events could reflect the recruitment of individuals that were dormant as dry eggs. We did not find a clear signal of temperature on abundance changes of this mosquito, but only on its phenology. Since ground cover seemed more critical than temperature to its spatial distribution, we propose that this mosquito might have some degree of autonomy to changes in temperature.

UltraPulse--simulating a human arterial pulse with focussed airborne ultrasound.

Medical simulators provide a risk-free environment for trainee doctors to practice and improve their skills. UltraPulse is a new tactile system designed to utilise focussed airborne ultrasound to mimic a pulsation effect such as that of a human arterial pulse. In this paper, we focus on the construction of the haptics component, which can later be integrated into a variety of medical procedure training simulators.

Transduction of voltage and Ca2+ signals by Slo1 BK channels.

Large-conductance Ca2+ -and voltage-gated K+ channels are activated by an increase in intracellular Ca2+ concentration and/or depolarization. The channel activation mechanism is well described by an allosteric model encompassing the gate, voltage sensors, and Ca2+ sensors, and the model is an excellent framework to understand the influences of auxiliary ß and γ subunits and regulatory factors such as Mg2+. Recent advances permit elucidation of structural correlates of the biophysical mechanism.

An order-N electronic structure theory with generalized eigenvalue equations and its application to a ten-million-atom system.

A linear algebraic theory called the 'multiple Arnoldi method' is presented and realizes large-scale (order-N) electronic structure calculations with generalized eigenvalue equations. A set of linear equations, in the form of (zS - H)x = b, are solved simultaneously with multiple Krylov subspaces. The method is implemented in a simulation package ELSES (www.elses.jp) with tight-binding-form Hamiltonians. A finite-temperature molecular dynamics simulation is carried out for metallic and insulating materials. A calculation with 10(7) atoms was realized by a workstation. The parallel efficiency is shown up to 1024 CPU cores.

Serum inflammatory proteins and frontal lobe dysfunction in patients with cardiovascular risk factors.

BACKGROUND: Recent studies have shown that the levels of circulating inflammatory markers are associated with cognitive decline and cerebral small-vessel disease. Frontal lobe dysfunction is believed to be a relatively characteristic neuropsychological symptom in vascular cognitive impairment caused by cerebral small-vessel disease. The purpose of this study was to investigate whether the levels of serum inflammatory markers are associated with frontal lobe dysfunction, particularly executive dysfunction. METHODS: Between January 2003 and September 2007, 388 patients who had one or more atherosclerotic risk factors and subsequently underwent brain MRI and neuropsychological testing including mini-mental state examination (MMSE), frontal assessment battery (FAB), and modified Stroop test were enrolled in this study. We evaluated the effect of serum levels of inflammatory markers and white matter lesions on frontal lobe function. RESULTS: The FAB score was negatively correlated with serum inflammatory marker levels (hsCRP; r = -0.170, IL-6; r = -0.143, IL-18; r = -0.175) and white matter lesions. In the modified Stroop test, interference measure was positively correlated with the levels of hsCRP (r = -0.198), and IL-18 (r = -0.152), and white matter lesions. However, the MMSE score was not correlated with either inflammatory marker levels. The association between hsCRP and FAB score or interference measure remained significant when controlling for other confounding factors and MRI findings. CONCLUSIONS: The circulating level of hsCRP is associated with frontal lobe dysfunction in patients with cardiovascular risk factors independent of white matter lesions in brain MRI.

Noncontact Tactile Display Based on Radiation Pressure of Airborne Ultrasound.

This paper describes a tactile display which provides unrestricted tactile feedback in air without any mechanical contact. It controls ultrasound and produces a stress field in a 3D space. The principle is based on a nonlinear phenomenon of ultrasound: Acoustic radiation pressure. The fabricated prototype consists of 324 airborne ultrasound transducers, and the phase and intensity of each transducer are controlled individually to generate a focal point. The DC output force at the focal point is 16 mN and the diameter of the focal point is 20 mm. The prototype produces vibrations up to 1 kHz. An interaction system including the prototype is also introduced, which enables users to see and touch virtual objects.

Novel algorithm of large-scale simultaneous linear equations.

We review our recently developed methods of solving large-scale simultaneous linear equations and applications to electronic structure calculations both in one-electron theory and many-electron theory. This is the shifted COCG (conjugate orthogonal conjugate gradient) method based on the Krylov subspace, and the most important issue for applications is the shift equation and the seed switching method, which greatly reduce the computational cost. The applications to nano-scale Si crystals and the double orbital extended Hubbard model are presented.

Domain boundary formation in helical multishell gold nanowires.

Helical multishell gold nanowires are studied theoretically for the formation mechanism of the helical domain boundary. Nanowires with a wire length of more than 10 nm are relaxed by quantum mechanical molecular dynamics simulation with a tight-binding form Hamiltonian. In the results, non-helical nanowires are transformed into helical ones with the formation of atom pair defects at the domain boundary, where the defective atom pair is moved from an inner shell. Analysis of local electronic structure shows a competitive feature of the energy gain of reconstruction on the wire surface and the energy loss of the defect formation. A simple energy scaling theory gives a general explanation of domain boundary formation.

Development of the simulation package 'ELSES' for extra-large-scale electronic structure calculation.

An early-stage version of the simulation package 'ELSES' (extra-large-scale electronic structure calculation) is developed for simulating the electronic structure and dynamics of large systems, particularly nanometer-scale and ten-nanometer-scale systems (see www.elses.jp). Input and output files are written in the extensible markup language (XML) style for general users. Related pre-/post-simulation tools are also available. A practical workflow and an example are described. A test calculation for the GaAs bulk system is shown, to demonstrate that the present code can handle systems with more than one atom species. Several future aspects are also discussed.

Accuracy control in ultra-large-scale electronic structure calculations.

Numerical aspects are investigated in ultra-large-scale electronic structure calculations. Accuracy control methods in process (molecular-dynamics) calculations are focused upon. Flexible control methods are proposed so as to control variational freedoms, automatically at each time step, within the framework of generalized Wannier state theory. The method is demonstrated in a silicon cleavage simulation with 10(2)-10(5) atoms. The idea is of general importance among process calculations and is also used in Krylov subspace theory, which is another large-scale calculation theory.

Oxygen sensing in the body.

This review is divided into three parts: (a) The primary site of oxygen sensing is the carotid body which instantaneously respond to hypoxia without involving new protein synthesis, and is historically known as the first oxygen sensor and is therefore placed in the first section (Lahiri, Roy, Baby and Hoshi). The carotid body senses oxygen in acute hypoxia, and produces appropriate responses such as increases in breathing, replenishing oxygen from air. How this oxygen is sensed at a relatively high level (arterial PO2 approximately 50 Torr) which would not be perceptible by other cells in the body, is a mystery. This response is seen in afferent nerves which are connected synaptically to type I or glomus cells of the carotid body. The major effect of oxygen sensing is the increase in cytosolic calcium, ultimately by influx from extracellular calcium whose concentration is 2 x 10(4) times greater. There are several contesting hypotheses for this response: one, the mitochondrial hypothesis which states that the electron transport from the substrate to oxygen through the respiratory chain is retarded as the oxygen pressure falls, and the mitochondrial membrane is depolarized leading to the calcium release from the complex of mitochondria-endoplasmic reticulum. This is followed by influx of calcium. Also, the inhibitors of the respiratory chain result in mitochondrial depolarization and calcium release. The other hypothesis (membrane model) states that K(+) channels are suppressed by hypoxia which depolarizes the membrane leading to calcium influx and cytosolic calcium increase. Evidence supports both the hypotheses. Hypoxia also inhibits prolyl hydroxylases which are present in all the cells. This inhibition results in membrane K(+) current suppression which is followed by cell depolarization. The theme of this section covers first what and where the oxygen sensors are; second, what are the effectors; third, what couples oxygen sensors and the effectors. (b) All oxygen consuming cells have a built-in mechanism, the transcription factor HIF-1, the discovery of which has led to the delineation of oxygen-regulated gene expression. This response to chronic hypoxia needs new protein synthesis, and the proteins of these genes mediate the adaptive physiological responses. HIF-1alpha, which is a part of HIF-1, has come to be known as master regulator for oxygen homeostasis, and is precisely regulated by the cellular oxygen concentration. Thus, the HIF-1 encompasses the chronic responses (gene expression in all cells of the body). The molecular biology of oxygen sensing is reviewed in this section (Semenza). (c) Once oxygen is sensed and Ca(2+) is released, the neurotransmittesr will be elaborated from the glomus cells of the carotid body. Currently it is believed that hypoxia facilitates release of one or more excitatory transmitters from glomus cells, which by depolarizing the nearby afferent terminals, leads to increases in the sensory discharge. The transmitters expressed in the carotid body can be classified into two major categories: conventional and unconventional. The conventional neurotransmitters include those stored in synaptic vesicles and mediate their action via activation of specific membrane bound receptors often coupled to G-proteins. Unconventional neurotransmitters are those that are not stored in synaptic vesicles, but spontaneously generated by enzymatic reactions and exert their biological responses either by interacting with cytosolic enzymes or by direct modifications of proteins. The gas molecules such as NO and CO belong to this latter category of neurotransmitters and have unique functions. Co-localization and co-release of neurotransmitters have also been described. Often interactions between excitatory and inhibitory messenger molecules also occur. Carotid body contains all kinds of transmitters, and an interplay between them must occur. But very little has come to be known as yet. Glimpses of these interactions are evident in the discussion in the last section (Prabhakar).

BKCa channels activating at resting potential without calcium in LNCaP prostate cancer cells.

Large-conductance Ca2+-dependent K+ (BK(Ca)) channels are activated by intracellular Ca2+ and membrane depolarization in an allosteric manner. We investigated the pharmacological and biophysical characteristics of a BK(Ca)-type K+ channel in androgen-dependent LNCaP (lymph node carcinoma of the prostate) cells with novel functional properties, here termed BK(L). K+ selectivity, high conductance, activation by Mg2+ or NS1619, and inhibition by paxilline and penitrem A largely resembled the properties of recombinant BK(Ca) channels. However, unlike conventional BK(Ca) channels, BK(L) channels activated in the absence of free cytosolic Ca2+ at physiological membrane potentials; the half-maximal activation voltage was shifted by about -100 mV compared with BK(Ca) channels. Half-maximal Ca2+-dependent activation was observed at 0.4 microM: for BK(L) (at -20 mV) and at 4.1 microM: for BK(Ca) channels (at +50 mV). Heterologous expression of hSlo1 in LNCaP cells increased the BK(L) conductance. Expression of hSlo-beta1 in LNCaP cells shifted voltage-dependent activation to values between that of BK(L) and BK(Ca) channels and reduced the slope of the P (open) (open probability)-voltage curve. We propose that LNCaP cells harbor a so far unknown type of BK(Ca) subunit, which is responsible for the BK(L) phenotype in a dominant manner. BK(L)-like channels are also expressed in the human breast cancer cell line T47D. In addition, functional expression of BK(L) in LNCaP cells is regulated by serum-derived factors, however not by androgens.

Comparative effects of xenon and nitrous oxide on diaphragmatic contractility in dogs.

BACKGROUND: Xenon at two different concentrations (30%, 60%) has no effect on diaphragmatic contractility. This study was undertaken to compare the effects of xenon and nitrous oxide (N2O), a commonly used and well-established gas anesthetic, on diaphragmatic contractility in dogs. METHODS: Twenty-one pentobarbitone-anesthetized dogs were randomly divided into three groups of seven each: group 1 received xenon 30% (0.25 MAC) in oxygen; group 2 received N2O 47% (0.25 MAC) in oxygen; and group 3 received N2O 60% (0.32 MAC) in oxygen. Diaphragmatic contractility was assessed by transdiaphragmatic pressure (Pdi) at low- (20-Hz) and high-frequency (100-Hz) stimulation, after maintaining 60 min of stable condition. The integrated electrical activity of diaphragm (Edi) to each stimulus was measured. RESULTS: With an inhalation of xenon 30%, N2O 47%, or N2O 60%, Pdi and Edi at both stimuli did not change. No difference in Pdi or Edi was observed among the groups. CONCLUSION: When used at clinical concentration, xenon or N2O does not affect contractility and electrical activity of the diaphragm in dogs.

Disordered cellular migration and angiogenesis in cd39-null mice.

BACKGROUND: Nucleoside triphosphate diphosphohydrolase-1 (NTPDase1)/CD39 is the major ectonucleotidase of endothelial cells and monocytes and catalyzes phosphohydrolysis of extracellular nucleoside diphosphates (NDP) and triphosphates (NTP, eg, ATP and UTP). Deletion of cd39 causes perturbations in the hydrolysis of NTP and NDP in the vasculature. Activation of P2 receptors appears to influence endothelial cell chemotactic and mitogenic responses in vitro. Therefore, aberrant regulation of nucleotide P2 receptors may influence angiogenesis in cd39-null mice. Methods and Results- In control mice, implanted Matrigel plugs containing growth factors were rapidly populated by monocyte/macrophages, endothelial cells, and pericytes, with the development of new vessels over days. In cd39-null mice, migrating cells were completely confined to the tissue-Matrigel interface in a clearly stratified manner. Absolute failure of new vessel ingrowth was consistently observed in the mutant mice. Linked to these findings, chemotaxis of cd39-null monocyte/macrophages to nucleotides was impaired in vitro. This abnormality was associated with desensitization of nucleotide receptor P2Y-mediated signaling pathways. CONCLUSIONS: Our findings demonstrate a role for NTPDase1 and phosphohydrolysis of extracellular nucleotides in the regulation of the cellular infiltration and new vessel growth in a model of angiogenesis.

Activation of ATP-sensitive K(+) channels by epoxyeicosatrienoic acids in rat cardiac ventricular myocytes.

1. We examined the effects of epoxyeicosatrienoic acids (EETs), which are cytochrome P450 metabolites of arachidonic acid (AA), on the activities of the ATP-sensitive K(+) (K(ATP)) channels of rat cardiac myocytes, using the inside-out patch-clamp technique. 2. In the presence of 100 microM cytoplasmic ATP, the K(ATP) channel open probability (P(o)) was increased by 240 +/- 60 % with 0.1 microM 11,12-EET and by 400 +/- 54 % with 5 microM 11,12-EET (n = 5-10, P < 0.05 vs. control), whereas neither 5 microM AA nor 5 microM 11,12-dihydroxyeicosatrienoic acid (DHET), which is the epoxide hydrolysis product of 11,12-EET, had any effect on P(o). 3. The half-maximal activating concentration (EC(50)) was 18.9 +/- 2.6 nM for 11,12-EET (n = 5) and 19.1 +/- 4.8 nM for 8,9-EET (n = 5, P = n.s. vs. 11,12-EET). Furthermore, 11,12-EET failed to alter the inhibition of K(ATP) channels by glyburide. 4. Application of 11,12-EET markedly decreased the channel sensitivity to cytoplasmic ATP. The half-maximal inhibitory concentration of ATP (IC(50)) was increased from 21.2 +/- 2.0 microM at baseline to 240 +/- 60 microM with 0.1 microM 11,12-EET (n = 5, P < 0.05 vs. control) and to 780 +/- 30 microM with 5 microM 11,12-EET (n = 11, P < 0.05 vs. control). 5. Increasing the ATP concentration increased the number of kinetically distinguishable closed states, promoting prolonged closure durations. 11,12-EET antagonized the effects of ATP on the kinetics of the K(ATP) channels in a dose- and voltage-dependent manner. 11,12-EET (1 microM) reduced the apparent association rate constant of ATP to the channel by 135-fold. 6. Application of 5 microM 11,12-EET resulted in hyperpolarization of the resting membrane potential in isolated cardiac myocytes, which could be blocked by glyburide. 7. These results suggest that EETs are potent activators of the cardiac K(ATP) channels, modulating channel behaviour by reducing the channel sensitivity to ATP. Thus, EETs could be important endogenous regulators of cardiac electrical excitability.

[Three-dimensional computed tomography angiography of coronary artery bypass graft with electron beam tomography].

OBJECTIVES: Assessment of coronary artery bypass graft patency by three-dimensional reconstructed computed tomography angiography (3D-CTA) derived from electrocardiography-gated contrast-enhanced electron beam tomography (EBT) was evaluated. METHODS: Thirty-nine patients with 99 grafts (45 arterial grafts and 54 venous grafts) underwent 3D-CTA and selective coronary angiography within a 3-week interval. 3D-CTA images of the coronary bypass grafts were compared with the coronary angiography images used as the control. RESULTS: 3D-CTA defined 42 of 44 arterial grafts as patent (sensitivity: 95%), all 47 venous grafts as patent (sensitivity: 100%) and all 7 venous grafts as occlusive (specificity: 100%). The overall sensitivity and specificity were 98% and 88%, respectively. CONCLUSIONS: 3D-CTA is an useful noninvasive technique with adequate sensitivity and specificity to assess coronary artery bypass graft patency.

"Schistotaxis" in cerebral blood flow on 99mTc-HMPAO single-photon emission computed tomography during a seizure following a stroke.

We describe a patient with seizures following a stroke in whom on ictal 99mTc-HMPAO single-photon emission computed tomography demonstrated cerebral blood flow "schistotaxis", i.e., focal hyperaemia corresponding to an epileptogenic focus together with an extensive hypoperfused area in the same hemisphere. This phenomenon may have been caused by haemodynamic alternation and a remote transneural effect during the seizures.