Caracterização bidimensional de um canal rádio Wimax

Com o crescimento previsível e exponencial das redes de comunicações móveis motivado pela mobilidade, flexibilidade e também comodidade do utilizador levam a que este se torne na fatia mais importante do mundo das telecomunicações dos dias que correm. Assim é importante estudar e caracterizar canais rádio para as mais diversas gamas de frequências utilizadas nas mais variadas tecnologias. O objectivo principal desta dissertação de Mestrado é caracterizar um canal rádio para a tecnologia sem fios Worldwide Inter-operability for Microwave Access (Wimax para as frequências de 3,5 GHz e 5 GHz) actualmente vista pela comunidade científica como a tecnologia sem fios com maiores perspectivas de sucesso. Para tal, determinaram-se o Perfil de Atraso de Potência (PAP) e também a Potência em Função da Distância (PFD) recorrendo ao método computacional de simulação Finite-Difference Time-Domain (FDTD). De forma a estudar e caracterizar o canal rádio, em termos de desvanecimento relativo ao espalhamento de atraso, usaram-se dois métodos alternativos que têm como entrada o PAP. Para caracterizar o canal quanto ao desvanecimento baseado em espalhamento de Doppler, recorreu-se também a duas técnicas alternativas tendo como entrada o PFD. Em ambas as situações os dois métodos alternativos convergiram para os mesmos resultados. A caracterização é feita em dois cenários diferentes: um em que consideramos que a maioria dos obstáculos são condutores eléctricos perfeitos (CEP) e que passaremos a designar Cenário PEC, e um segundo cenário em que os obstáculos têm propriedades electromagnéticas diferentes, e que passará a ser designado por Cenário MIX. Em ambos os cenários de análise concluiu-se que o canal é plano, lento e sem ISI.

Nuevas Técnicas de Transmisión de Muy Alta Velocidad para Redes de Computadores: Una aproximación mediante ondículas.

Indoor multpropagation channel is modeled by the Kaiser electromagnetic wavelet. A method for channel characterization is proposed by modeling all the reflections of indoor propagation in a kernel function instead of its impulse response. This led us to consider a fractal modulation scheme in which Kaiser wavelets substitute the traditional sinusoidal carrier.

Contribución a la caracterización espacial de canales con sistemas MIMO-OFDM en la banda de 2,45 Ghz

La tecnología de múltiples antenas ha evolucionado para dar soporte a los actuales y futuros sistemas de comunicaciones inalámbricas en su afán por proporcionar la calidad de señal y las altas tasas de transmisión que demandan los nuevos servicios de voz, datos y multimedia. Sin embargo, es fundamental comprender las características espaciales del canal radio, ya que son las características del propio canal lo que limita en gran medida las prestaciones de los sistemas de comunicación actuales. Por ello surge la necesidad de estudiar la estructura espacial del canal de propagación para poder diseñar, evaluar e implementar de forma más eficiente tecnologías multiantena en los actuales y futuros sistemas de comunicación inalámbrica. Las tecnologías multiantena denominadas antenas inteligentes y MIMO han generado un gran interés en el área de comunicaciones inalámbricas, por ejemplo los sistemas de telefonía celular o más recientemente en las redes WLAN (Wireless Local Area Network), principalmente por la mejora que proporcionan en la calidad de las señales y en la tasa de transmisión de datos, respectivamente. Las ventajas de estas tecnologías se fundamentan en el uso de la dimensión espacial para obtener ganancia por diversidad espacial, como ya sucediera con las tecnologías FDMA (Frequency Division Multiplexing Access), TDMA (Time Division Multiplexing Access) y CDMA (Code Division Multiplexing Access) para obtener diversidad en las dimensiones de frecuencia, tiempo y código, respectivamente. Esta Tesis se centra en estudiar las características espaciales del canal con sistemas de múltiples antenas mediante la estimación de los perfiles de ángulos de llegada (DoA, Direction-of- Arrival) considerando esquemas de diversidad en espacio, polarización y frecuencia. Como primer paso se realiza una revisión de los sistemas con antenas inteligentes y los sistemas MIMO, describiendo con detalle la base matemática que sustenta las prestaciones ofrecidas por estos sistemas. Posteriormente se aportan distintos estudios sobre la estimación de los perfiles de DoA de canales radio con sistemas multiantena evaluando distintos aspectos de antenas, algoritmos de estimación, esquemas de polarización, campo lejano y campo cercano de las fuentes. Así mismo, se presenta un prototipo de medida MIMO-OFDM-SPAA3D en la banda ISM (Industrial, Scientific and Medical) de 2,45 Ghz, el cual está preparado para caracterizar experimentalmente el rendimiento de los sistemas MIMO, y para caracterizar espacialmente canales de propagación, considerando los esquemas de diversidad espacial, por polarización y frecuencia. Los estudios aportados se describen a continuación. Los sistemas de antenas inteligentes dependen en gran medida de la posición de los usuarios. Estos sistemas están equipados con arrays de antenas, los cuales aportan la diversidad espacial necesaria para obtener una representación espacial fidedigna del canal radio a través de los perfiles de DoA (DoA, Direction-of-Arrival) y por tanto, la posición de las fuentes de señal. Sin embargo, los errores de fabricación de arrays así como ciertos parámetros de señal conlleva un efecto negativo en las prestaciones de estos sistemas. Por ello se plantea un modelo de señal parametrizado que permite estudiar la influencia que tienen estos factores sobre los errores de estimación de DoA, tanto en acimut como en elevación, utilizando los algoritmos de estimación de DOA más conocidos en la literatura. A partir de las curvas de error, se pueden obtener parámetros de diseño para sistemas de localización basados en arrays. En un segundo estudio se evalúan esquemas de diversidad por polarización con los sistemas multiantena para mejorar la estimación de los perfiles de DoA en canales que presentan pérdidas por despolarización. Para ello se desarrolla un modelo de señal en array con sensibilidad de polarización que toma en cuenta el campo electromagnético de ondas planas. Se realizan simulaciones MC del modelo para estudiar el efecto de la orientación de la polarización como el número de polarizaciones usadas en el transmisor como en el receptor sobre la precisión en la estimación de los perfiles de DoA observados en el receptor. Además, se presentan los perfiles DoA obtenidos en escenarios quasiestáticos de interior con un prototipo de medida MIMO 4x4 de banda estrecha en la banda de 2,45 GHz, los cuales muestran gran fidelidad con el escenario real. Para la obtención de los perfiles DoA se propone un método basado en arrays virtuales, validado con los datos de simulación y los datos experimentales. Con relación a la localización 3D de fuentes en campo cercano (zona de Fresnel), se presenta un tercer estudio para obtener con gran exactitud la estructura espacial del canal de propagación en entornos de interior controlados (en cámara anecóica) utilizando arrays virtuales. El estudio analiza la influencia del tamaño del array y el diagrama de radiación en la estimación de los parámetros de localización proponiendo, para ello, un modelo de señal basado en un vector de enfoque de onda esférico (SWSV). Al aumentar el número de antenas del array se consigue reducir el error RMS de estimación y mejorar sustancialmente la representación espacial del canal. La estimación de los parámetros de localización se lleva a cabo con un nuevo método de búsqueda multinivel adaptativo, propuesto con el fin de reducir drásticamente el tiempo de procesado que demandan otros algoritmos multivariable basados en subespacios, como el MUSIC, a costa de incrementar los requisitos de memoria. Las simulaciones del modelo arrojan resultados que son validados con resultados experimentales y comparados con el límite de Cramer Rao en términos del error cuadrático medio. La compensación del diagrama de radiación acerca sustancialmente la exactitud de estimación de la distancia al límite de Cramer Rao. Finalmente, es igual de importante la evaluación teórica como experimental de las prestaciones de los sistemas MIMO-OFDM. Por ello, se presenta el diseño e implementación de un prototipo de medida MIMO-OFDM-SPAA3D autocalibrado con sistema de posicionamiento de antena automático en la banda de 2,45 Ghz con capacidad para evaluar la capacidad de los sistemas MIMO. Además, tiene la capacidad de caracterizar espacialmente canales MIMO, incorporando para ello una etapa de autocalibración para medir la respuesta en frecuencia de los transmisores y receptores de RF, y así poder caracterizar la respuesta de fase del canal con mayor precisión. Este sistema incorpora un posicionador de antena automático 3D (SPAA3D) basado en un scanner con 3 brazos mecánicos sobre los que se desplaza un posicionador de antena de forma independiente, controlado desde un PC. Este posicionador permite obtener una gran cantidad de mediciones del canal en regiones locales, lo cual favorece la caracterización estadística de los parámetros del sistema MIMO. Con este prototipo se realizan varias campañas de medida para evaluar el canal MIMO en términos de capacidad comparando 2 esquemas de polarización y tomando en cuenta la diversidad en frecuencia aportada por la modulación OFDM en distintos escenarios. ABSTRACT Multiple-antennas technologies have been evolved to be the support of the actual and future wireless communication systems in its way to provide the high quality and high data rates required by new data, voice and data services. However, it is important to understand the behavior of the spatial characteristics of the radio channel, since the channel by itself limits the performance of the actual wireless communications systems. This drawback raises the need to understand the spatial structure of the propagation channel in order to design, assess, and develop more efficient multiantenna technologies for the actual and future wireless communications systems. Multiantenna technologies such as ‘Smart Antennas’ and MIMO systems have generated great interest in the field of wireless communications, i.e. cellular communications systems and more recently WLAN (Wireless Local Area Networks), mainly because the higher quality and the high data rate they are able to provide. Their technological benefits are based on the exploitation of the spatial diversity provided by the use of multiple antennas as happened in the past with some multiaccess technologies such as FDMA (Frequency Division Multiplexing Access), TDMA (Time Division Multiplexing Access), and CDMA (Code Division Multiplexing Access), which give diversity in the domains of frequency, time and code, respectively. This Thesis is mainly focus to study the spatial channel characteristics using schemes of multiple antennas considering several diversity schemes such as space, polarization, and frequency. The spatial characteristics will be study in terms of the direction-of-arrival profiles viewed at the receiver side of the radio link. The first step is to do a review of the smart antennas and MIMO systems technologies highlighting their advantages and drawbacks from a mathematical point of view. In the second step, a set of studies concerning the spatial characterization of the radio channel through the DoA profiles are addressed. The performance of several DoA estimation methods is assessed considering several aspects regarding antenna array structure, polarization diversity, and far-field and near-field conditions. Most of the results of these studies come from simulations of data models and measurements with real multiantena prototypes. In the same way, having understand the importance of validate the theoretical data models with experimental results, a 2,4 GHz MIMO-OFDM-SPAA2D prototype is presented. This prototype is intended for evaluating MIMO-OFDM capacity in indoor and outdoor scenarios, characterize the spatial structure of radio channels, assess several diversity schemes such as polarization, space, and frequency diversity, among others aspects. The studies reported are briefly described below. As is stated in Chapter two, the determination of user position is a fundamental task to be resolved for the smart antenna systems. As these systems are equipped with antenna arrays, they can provide the enough spatial diversity to accurately draw the spatial characterization of the radio channel through the DoA profiles, and therefore the source location. However, certain real implementation factors related to antenna errors, signals, and receivers will certainly reduce the performance of such direction finding systems. In that sense, a parameterized narrowband signal model is proposed to evaluate the influence of these factors in the location parameter estimation through extensive MC simulations. The results obtained from several DoA algorithms may be useful to extract some parameter design for directing finding systems based on arrays. The second study goes through the importance that polarization schemes can have for estimating far-field DoA profiles in radio channels, particularly for scenarios that may introduce polarization losses. For this purpose, a narrowband signal model with polarization sensibility is developed to conduct an analysis of several polarization schemes at transmitter (TX) and receiver (RX) through extensive MC simulations. In addition, spatial characterization of quasistatic indoor scenarios is also carried out using a 2.45 GHz MIMO prototype equipped with single and dual-polarized antennas. A good agreement between the measured DoA profiles with the propagation scenario is achieved. The theoretical and experimental evaluation of polarization schemes is performed using virtual arrays. In that case, a DoA estimation method is proposed based on adding an phase reference to properly track the DoA, which shows good results. In the third study, the special case of near-field source localization with virtual arrays is addressed. Most of DoA estimation algorithms are focused in far-field source localization where the radiated wavefronts are assume to be planar waves at the receive array. However, when source are located close to the array, the assumption of plane waves is no longer valid as the wavefronts exhibit a spherical behavior along the array. Thus, a faster and effective method of azimuth, elevation angles-of-arrival, and range estimation for near-field sources is proposed. The efficacy of the proposed method is evaluated with simulation and validated with measurements collected from a measurement campaign carried out in a controlled propagation environment, i.e. anechoic chamber. Moreover, the performance of the method is assessed in terms of the RMSE for several array sizes, several source positions, and taking into account the effect of radiation pattern. In general, better results are obtained with larger array and larger source distances. The effect of the antennas is included in the data model leading to more accurate results, particularly for range rather than for angle estimation. Moreover, a new multivariable searching method based on the MUSIC algorithm, called MUSA (multilevel MUSIC-based algorithm), is presented. This method is proposed to estimate the 3D location parameters in a faster way than other multivariable algorithms, such as MUSIC algorithm, at the cost of increasing the memory size. Finally, in the last chapter, a MIMO-OFDM-SPAA3D prototype is presented to experimentally evaluate different MIMO schemes regarding antennas, polarization, and frequency in different indoor and outdoor scenarios. The prototype has been developed on a Software-Defined Radio (SDR) platform. It allows taking measurements where future wireless systems will be developed. The novelty of this prototype is concerning the following 2 subsystems. The first one is the tridimensional (3D) antenna positioning system (SPAA3D) based on three linear scanners which is developed for making automatic testing possible reducing errors of the antenna array positioning. A set of software has been developed for research works such as MIMO channel characterization, MIMO capacity, OFDM synchronization, and so on. The second subsystem is the RF autocalibration module at the TX and RX. This subsystem allows to properly tracking the spatial structure of indoor and outdoor channels in terms of DoA profiles. Some results are draw regarding performance of MIMO-OFDM systems with different polarization schemes and different propagation environments.

Modulation of insect Cav channels by peptidic spider toxins

Insects have a much smaller repertoire of voltage-gated calcium (Ca-v) channels than vertebrates. Drosophila melanogaster harbors only a single ortholog of each of the vertebrate Ca(v)1, Ca(v)2, and Ca(v)3 subtypes, although its basal inventory is expanded by alternative splicing and editing of Ca-v channel transcripts. Nevertheless, there appears to be little functional plasticity within this limited panel of insect Ca-v channels, since severe loss-of-function mutations in genes encoding the pore-forming a, subunits in Drosophila are embryonic lethal. Since the primary role of spider venom is to paralyze or kill insect prey, it is not surprising that most, if not all, spider venoms contain peptides that potently modify the activity of these functionally critical insect Ca-v channels. Unfortunately, it has proven difficult to determine the precise ion channel subtypes recognized by these peptide toxins since insect Ca-v channels have significantly different pharmacology to their vertebrate counterparts, and cloned insect Ca-v channels are not available for electrophysiological studies. However, biochemical and genetic studies indicate that some of these spider toxins might ultimately become the defining pharmacology for certain subtypes of insect Ca-v channels. This review focuses on peptidic spider toxins that specifically target insect Ca-v channels. In addition to providing novel molecular tools for ion channel characterization, some of these toxins are being used as leads to develop new methods for controlling insect pests. (c) 2006 Elsevier Ltd. All rights reserved.

Bi-Dimensional Characterization of a Wimax Radio Channel at 3.5GHz

This paper presents the characterization of an indoor Wimax radio channel using the Finite-Difference Time-Domain (FDTD) [1] method complemented with the Convolutional Perfect Matched Layer (CPML) technique [2]. An indoor 2D scenario is simulated in the 3.5GHz band (IEEE 802.16d-2004 and IEEE 802.16e-2005 [3]). In this study, we used two complementary techniques in both analysis, technique A and B for fading based on delay spread and technique C and D for fading based on Doppler spread. Both techniques converge to the same result. Simulated results define the channel as flat, slow and without inter-symbolic interference (ISI), making the application of the spatial diversity the most appropriate scheme.

Functional Characterization of a Novel Frameshift Mutation in the C-terminus of the Nav1.5 Channel Underlying a Brugada Syndrome with Variable Expression in a Spanish Family.

INTRODUCTION We functionally analyzed a frameshift mutation in the SCN5A gene encoding cardiac Na(+) channels (Nav1.5) found in a proband with repeated episodes of ventricular fibrillation who presented bradycardia and paroxysmal atrial fibrillation. Seven relatives also carry the mutation and showed a Brugada syndrome with an incomplete and variable expression. The mutation (p.D1816VfsX7) resulted in a severe truncation (201 residues) of the Nav1.5 C-terminus. METHODS AND RESULTS Wild-type (WT) and mutated Nav1.5 channels together with hNavβ1 were expressed in CHO cells and currents were recorded at room temperature using the whole-cell patch-clamp. Expression of p.D1816VfsX7 alone resulted in a marked reduction (≈90%) in peak Na(+) current density compared with WT channels. Peak current density generated by p.D1816VfsX7+WT was ≈50% of that generated by WT channels. p.D1816VfsX7 positively shifted activation and inactivation curves, leading to a significant reduction of the window current. The mutation accelerated current activation and reactivation kinetics and increased the fraction of channels developing slow inactivation with prolonged depolarizations. However, late INa was not modified by the mutation. p.D1816VfsX7 produced a marked reduction of channel trafficking toward the membrane that was not restored by decreasing incubation temperature during cell culture or by incubation with 300 μM mexiletine and 5 mM 4-phenylbutirate. CONCLUSION Despite a severe truncation of the C-terminus, the resulting mutated channels generate currents, albeit with reduced amplitude and altered biophysical properties, confirming the key role of the C-terminal domain in the expression and function of the cardiac Na(+) channel.

Deposits of the sandy braided South Saskatchewan River: Implications for the use of modern analogs in reconstructing channel dimensions in reservoir characterization

Estimation of the dimensions of fluvial geobodies from core data is a notoriously difficult problem in reservoir modeling. To try and improve such estimates and, hence, reduce uncertainty in geomodels, data on dunes, unit bars, cross-bar channels, and compound bars and their associated deposits are presented herein from the sand-bed braided South Saskatchewan River, Canada. These data are used to test models that relate the scale of the formative bed forms to the dimensions of the preserved deposits and, therefore, provide an insight as to how such deposits may be preserved over geologic time. The preservation of bed-form geometry is quantified by comparing the Alluvial architecture above and below the maximum erosion depth of the modem channel deposits. This comparison shows that there is no significant difference in the mean set thickness of dune cross-strata above and below the basal erosion surface of the contemporary channel, thus suggesting that dimensional relationships between dune deposits and the formative bed-form dimensions are likely to be valid from both recent and older deposits. The data show that estimates of mean bankfull flow depth derived from dune, unit bar, and cross-bar channel deposits are all very similar. Thus, the use of all these metrics together can provide a useful check that all components and scales of the alluvial architecture have been identified correctly when building reservoir models. The data also highlight several practical issues with identifying and applying data relating to cross-strata. For example, the deposits of unit bars were found to be severely truncated in length and width, with only approximately 10% of the mean bar-form length remaining, and thus making identification in section difficult. For similar reasons, the deposits of compound bars were found to be especially difficult to recognize, and hence, estimates of channel depth based on this method may be problematic. Where only core data are available (i.e., no outcrop data exist), formative flow depths are suggested to be best reconstructed using cross-strata formed by dunes. However, theoretical relationships between the distribution of set thicknesses and formative dune height are found to result in slight overestimates of the latter and, hence, mean bankfull flow depths derived from these measurements. This article illustrates that the preservation of fluvial cross-strata and, thus, the paleohydraulic inferences that can be drawn from them, are a function of the ratio of the size and migration rate of bed forms and the time scale of aggradation and channel migration. These factors must thus be considered when deciding on appropriate length:thickness ratios for the purposes of object-based modeling in reservoir characterization.

Cloning and characterization of a zebrafish homologue of human AQP1: a bifunctional water and gas channel

Chen LM, Zhao J, Musa-Aziz R, Pelletier MF, Drummond IA, Boron WF. Cloning and characterization of a zebrafish homologue of human AQP1: a bifunctional water and gas channel. Am J Physiol Regul Integr Comp Physiol 299: R1163-R1174, 2010. First published August 25, 2010; doi:10.1152/ajpregu.00319.2010.-The mammalian aquaporins AQP1, AQP4, and AQP5 have been shown to function not only as water channels but also as gas channels. Zebrafish have two genes encoding an AQP1 homologue, aqp1a and aqp1b. In the present study, we cloned the cDNA that encodes the zebrafish protein Aqp1a from the 72-h postfertilization (hpf) embryo of Danio rerio, as well as from the swim bladder of the adult. The deduced amino-acid sequence of aqp1a consists of 260 amino acids and is 59% identical to human AQP1. By analyzing the genomic DNA sequence, we identified four exons in the aqp1a gene. By in situ hybridization, aqp1a is expressed transiently in the developing vasculature and in erythrocytes from 16 to 48 h of development. Later, at 72 hpf, aqp1a is expressed in dermal ionocytes and in the swim bladder. Western blot analysis of adult tissues reveals that Aqp1a is most highly expressed in the eye and swim bladder. Xenopus oocytes expressing aqp1a have a channel-dependent (*) osmotic water permeability (P(f)*) that is indistinguishable from that of human AQP1. On the basis of the magnitude of the transient change in surface pH (Delta pHS) that were recorded as the oocytes were exposed to either CO(2) or NH(3), we conclude that zebrafish Aqp1a is permeable to both CO(2) and NH(3). The ratio (Delta pHS*)CO2/P(f)* is about half that of human AQP1, and the ratio (Delta pHS*)NH3/P(f)* is about one-quarter that of human AQP1. Thus, compared with human AQP1, zebrafish Aqp1a has about twice the selectivity for CO(2) over NH(3).

Characterization of selectivity and pharmacophores of type 1 sea anemone toxins by screening seven Na-v sodium channel isoforms

During their evolution, animals have developed a set of cysteine-rich peptides capable of binding various extracellular sites of voltage-gated sodium channels (VGSC). Sea anemone toxins that target VGSCs delay their inactivation process, but little is known about their selectivities. Here we report the investigation of three native type 1 toxins (CGTX-II, delta-AITX-Bcg1a and delta-AITX-Bcg1b) purified from the venom of Bunodosoma cangicum. Both delta-AITX-Bcg1a and delta-AITX-Bcg1b toxins were fully sequenced. The three peptides were evaluated by patch-clamp technique among Nav1.1-1.7 isoforms expressed in mammalian cell lines, and their preferential targets are Na(v)1.5 > 1.6 > 1.1. We also evaluated the role of some supposedly critical residues in the toxins which would interact with the channels, and observed that some substitutions are not critical as expected. In addition, CGTX-II and delta-AITX-Bcg1a evoke different shifts in activation/inactivation Boltzmann curves in Nav1.1 and 1.6. Moreover, our results suggest that the interaction region between toxins and VGSCs is not restricted to the supposed site 3 (S3-54 linker of domain IV), and this may be a consequence of distinct surface of contact of each peptide vs. targeted channel. Our data suggest that the contact surfaces of each peptide may be related to their surface charges, as CGTX-II is more positive than delta-AITX-Bcg1a and delta-AITX-Bcg1b. (C) 2011 Elsevier Inc. All rights reserved.

Looking over Toxin-K+ Channel Interactions. Clues from the Structural and Functional Characterization of alpha-KTx Toxin Tc32, a Kv1.3 Channel Blocker

alpha-KTx toxin Tc32, from the Amazonian scorpion Tityus cambridgei, lacks the dyad motif; including Lys27, characteristic of the family and generally associated with channel blockage. The toxin has been cloned and expressed for the first time. Electrophysiological experiments, by showing that the recombinant form blocks Kv1.3 channels of olfactory bulb periglomerular cells like the natural Tc32 toxin, when tested on the Kv1.3 channel of human T lymphocytes, confirmed it is in an active fold. The nuclear magnetic resonance-derived structure revealed it exhibits an alpha/beta scaffold typical of the members of the alpha-KTx family. TdK2 and TdK3, all belonging to the same alpha-KTx 18 subfamily, share significant sequence identity with Tc32 but diverse selectivity and affinity for Kv1.3 and Kv1.1 channels. To gain insight into the structural features that may justify those differences, we used the recombinant Tc32 nuclear magnetic resonance-derived structure to model the other two toxins, for which no experimental structure is available. Their interaction with Kv1.3 and Kv1.1 has been investigated by means of docking simulations. The results suggest that differences in the electrostatic features of the toxins and channels, in their contact surfaces, and in their total dipole moment orientations govern the affinity and selectivity of toxins. In addition, we found that, regardless of whether the dyad motif is present, it is always a Lys side chain that physically blocks the channels, irrespective of its position in the toxin sequence.

Characterization of L-type Calcium Channel Binding-Site of a new class of Calcium modulators by a Multidisciplinary approach

Many potential diltiazem related L-VDCC blockers were developed using a multidisciplinary approach. This current study was to investigate and compare diltiazem with to the newly developed compounds by mouse Langendorff-perfused heart, Ca2+-transient and on recombinant L-VDCC. Twenty particular compounds were selected by the ligand-based virtual screening procedure (LBVS). From these compounds, five of them (5b, M2, M7, M8 and P1) showed a potent and selective inotropic activity on guinea-pig left atria driven 1 Hz. Further assays displayed an interesting negative inotropic effect of M2, M8, P1 and M7 on guinea pig isolated left papillary muscle driven at 1 Hz, a relevant vasorelaxant activity of 5b, M2, M7, M8 and P1 on K+-depolarized guinea-pig ileum longitudinal smooth muscle and a significant inhibition of contraction of 5b, M2, M8 and P1 on carbachol stimulated ileum longitudinal smooth muscle. Wild-type human heart and rabbit lung α1 subunits were expressed (combined with the regulatory α2δ and β3 subunits) in Xenopus Leavis oocytes using a two-electrode voltage clamp technique. Diltiazem is a benzothiazepine Ca2+ channel blocker used clinically for its antihypertensive and antiarrhythmic effects. Previous radioligand binding assays revealed a complex interaction with the benzothiazepine binding site for M2, M7 and M8. (Carosati E. et al. J. Med Chem. 2006, 49; 5206). In agreement with this findings, the relative order of increased rates of contraction and relaxation at lower concentrations s(≤10-6M) in unpaced hearts was M7>M2>M8>P1. Similar increases in Ca2+ transient were observed in cardiomyocytes. Diltiazem showed negative inotropic effects whereas 5b had no significant effect. Diltiazem blocks Ca2+current in a use-dependent manner and facilitates the channel by accelerating the inactivation and decelerating the recovery from inactivation. In contrast to diltiazem, the new analogs had no pronounced use-dependence. Application of 100 μM M8, M2 showed ~ 10% tonic block; in addition, M8, M2 and P1 shifted the steady state inactivation in hyperpolarized direction and the current inactivation time was significantly decreased compared with control (219.6 ± 11.5 ms, 226 ± 14.5 vs. 269 ± 12.9 vs. 199.28 ± 8.19 ms). Contrary to diltiazem, the recovery from the block by M8 and M2 was comparable to control. Only P1 showed a significantly decrease of the time for the recovery from inactivation. All of the compounds displayed the same sensitivity on the Ca2+ channel rabbit lung α1 except P1. Taken together, these findings suggest that M8, M2 and P1 might directly decrease the binding affinity or allow rapid dissociation from the benzothiazepine binding site.

Design, synthesis and pharmacological characterization of analogs of 2-aminoethyl diphenylborinate (2-APB), a known store-operated calcium channel blocker, for inhibition of TRPV6-mediated calcium transport

2-Aminoethyl diphenylborinate (2-APB) is a known modulator of the IP3 receptor, the calcium ATPase SERCA, the calcium release-activated calcium channel Orai and TRP channels. More recently, it was shown that 2-APB is an efficient inhibitor of the epithelial calcium channel TRPV6 which is overexpressed in prostate cancer. We have conducted a structure-activity relationship study of 2-APB congeners to understand their inhibitory mode of action on TRPV6. Whereas modifying the aminoethyl moiety did not significantly change TRPV6 inhibition, substitution of the phenyl rings of 2-APB did. Our data show that the diaryl borinate moiety is required for biological activity and that the substitution pattern of the aryl rings can influence TRPV6 versus SOCE inhibition. We have also discovered that 2-APB is hydrolyzed and transesterified within minutes in solution.

The characterization of an epithelial chloride channel and purification of a channel component

A membrane fraction (M$\sb{\rm PS}$), enriched in Cl$\sp-$ channels, has been isolated from bovine tracheal epithelia and renal cortex homogenates by hydrophobic chromatography. The tracheal fraction shows a 37 fold enrichment of Cl$\sp-$ channels over crude tracheal homogenates by net Cl$\sp-$ measurements in membrane vesicles. Alkaline phosphatase and (Na$\sp+$ + K$\sp+$)-ATPase are not found in these membranes, suggesting that they are not apical or basolateral plasma membranes. The M$\sb{\rm PS}$ fraction exhibits a protein profile unlike that of other membrane fractions with major proteins of 200 kDa and 42 kDa, proteins of 30 to 35 kDa, and lesser amounts of other proteins. Reconstitution of M$\sb{\rm PS}$ fractions from both trachea and kidney into planar lipid bilayers demonstrates the presence of a single type of anion channel. The current-voltage relationship of this channel is linear with a slope conductance of 84 pS in symmetrical 400 mM KCl, and is identical to that of the predominant anion channel observed in tracheal apical membranes under similar conditions (Valdivia, Dubinsky, and Coronado. Science, 1988). In addition, the voltage dependence, selectivity sequence of Cl$\sp- >$ Br$\sp- \ge$ I$\sp-$, and inhibition by low concentrations of the Cl$\sp-$ channel blocker, DIDS, correspond to those of the predominant apical membrane channel. Thus, although the M$\sb{\rm PS}$ fraction appears to be of subcellular origin, it may be functionally related to an apical membrane Cl$\sp-$ permeability. When renal M$\sb{\rm PS}$ membranes were treated with the detergent octyl-glucoside (OG, 2%) and centrifuged, the supernatant, sM$\sb{\rm PS}$, showed a 2 to 7-fold enrichment in specific Cl$\sp-$ flux activity compared with the detergent treated M$\sb{\rm PS}$. These solubilized proteins were then size fractionated on a Superose 12 HPLC gel filtration column, followed by fractionation on a Mono Q HPLC anion exchange column. Fractions that eluted in high salt consistently exhibited significant Cl$\sp-$ flux activity. These fractions had protein profiles consisting of a major band at 34 kDa, a band at 66 kDa, and variable faint bands. Fractions eluting in lower salt had protein profiles consisting of a single band at 34 kDa, and often had little or no Cl$\sp-$ flux activity. However, co-reconstitution of the low salt, solely 34 kDa protein-containing Mono Q fractions with sM$\sb{\rm PS}$ resulted in an enhancement of flux activity compared to that of sM$\sb{\rm PS}$ reconstituted alone. Flux assays of active Mono Q fractions showed that the channel retained its DIDS sensitivity. Applying sM$\sb{\rm PS}$ to a DIDS-affinity column and eluting with salt resulted in fractions with protein profiles again consisting of at least one major band at 34 kDa, a band at 66 kDa, and variable faint bands. Co-reconstitution with sM$\sb{\rm PS}$ again resulted in an enhancement of activity. Thus, the 34 kDa protein appears to be a component of the M$\sb{\rm PS}$ Cl$\sp-$ channel. ^

Time-frequency characterization of a sound propagation channel as an educational tool

This paper discusses the use of sound waves to illustrate multipath radio propagation concepts. Specifically, a procedure is presented to measure the time-varying frequency response of the channel. This helps demonstrate how a propagation channel can be characterized in time and frequency, and provides visualizations of the concepts of coherence time and coherence bandwidth. The measurements are very simple to carry out, and the required equipment is easily available. The proposed method can be useful for wireless or mobile communication courses.