Biomedical subjects

Ignacio Moreno

Publications and source records attributed to Ignacio Moreno.

15 recordsLinked to original sources

PubMed2004

Diltiazem inhibits hKv1.5 and Kv4.3 currents at therapeutic concentrations.

OBJECTIVE: In the present study we examined the effects of diltiazem, an L-type Ca(2+) channel blocker widely used for the control of the ventricular rate in patients with supraventricular arrhythmias, on hKv1.5 and Kv4.3 channels that generate the cardiac ultrarapid delayed rectifier (I(Kur)) and the 4-aminopyridine sensitive transient outward (I(to)) K(+) currents, respectively. METHODS: hKv1.5 and Kv4.3 channels were stably and transiently expressed in mouse fibroblast and Chinese hamster ovary cells, respectively. Currents were recorded using the whole-cell patch clamp. RESULTS: Diltiazem (0.01 nM-500 muM) blocked hKv1.5 channels, in a frequency-dependent manner exhibiting a biphasic dose-response curve (IC(50)=4.8+/-1.5 nM and 42.3+/-3.6 muM). Diltiazem delayed the initial phase of the tail current decline and shifted the midpoint of the activation (Vh=-16.5+/-2.1 mV vs -20.4+/-2.6 mV, P<0.001) and inactivation (Vh=-22.4+/-0.7 mV vs. -28.2+/-1.9 mV, P<0.001) curves to more negative potentials. The analysis of the development of the diltiazem-induced block yielded apparent association (k) and dissociation (P) rate constants of (1.6+/-0.2) x 10(6) M(-1)s(-1) and 46.8+/-4.8 s(-1), respectively. Diltiazem (0.1 nM-100 muM) also blocked Kv4.3 channels in a frequency-dependent manner exhibiting a biphasic dose-response curve (IC(50)=62.6+/-11.1 nM and 109.9+/-12.8 muM). Diltiazem decreased the peak current and, at concentrations > or =0.1 microM, accelerated the inactivation time course. The apparent association and dissociation rate constants resulted (1.7+/-0.2) x 10(6) M(-1)s(-1) and 258.6+/-38.1 s(-1), respectively. Diltiazem, 10 nM, shifted to more negative potentials the voltage-dependence of Kv4.3 channel inactivation (Vh=-33.1+/-2.3 mV vs -38.2+/-3.5 mV, n=6, Plt;0.05) the blockade increasing at potentials at which the amount of inactivated channels increased. CONCLUSION: The results demonstrated for the first time that diltiazem, at therapeutic concentrations, decreased hKv1.5 and Kv4.3 currents by binding to the open and the inactivated state of the channels.

Action Potentials↗

PubMed2004

Programmable birefringent lenses with a liquid-crystal display.

We study the properties of a novel birefringent lens constructed by combination of a regular glass lens and a programmable diffractive lens addressed to a liquid-crystal display (LCD). The LCD affects only the vertical polarization state. Consequently the birefringent lens produces two images of an input object with different locations and magnifications for the two orthogonal polarization states. Using a properly oriented analyzer polarizer produces interference fringes. We then show how the imaging system acts as a common-path polarization interferometer for wave-front analysis of objects in the input plane. Finally, we subtract the two images to produce an edge-enhanced version of the input image. All these effects can be controlled because we can program lenses with different focal lengths onto the LCD.

Journal Article↗

PubMed2004

Modulation light efficiency of diffractive lenses displayed in a restricted phase-mostly modulation display.

We present an analysis of the diffraction efficiency of diffractive lenses displayed on spatial light modulators that depends on the modulation response of the display. An ideal display would produce continuous phase-only modulation, reaching a maximum phase-modulation depth of 2pi. We introduce the concept of modulation diffraction efficiency that accounts for the effect of nonlinearities only in the phase modulation of the display. We review a diffractive model with which to evaluate this modulation efficiency, including modulation defects such as nonlinear phase modulation, coupled amplitude modulation, phase quantization, and a limited modulation depth. We apply this diffractive model to Fresnel lenses and show that these modulation defects produce a lens multiplex effect. Finally we demonstrate that the application of a minimum Euclidean projection principle leads to high modulation diffraction efficiency even if the phase-modulation depth is much less than 2pi. We demonstrate that the modulation efficiency can exceed 90% for a modulation depth of 1.4pi and can exceed 40% (the equivalent for a binary phase element) for a modulation depth of only 0.7pi. Experimental results from use of a twisted nematic liquid-crystal display are presented to confirm these conclusions.

Journal Article↗

PubMed2004

Interaction of angiotensin II with the angiotensin type 2 receptor inhibits the cardiac transient outward potassium current.

OBJECTIVE: The Ca2+ -independent transient outward K+ current (Ito) plays a crucial role in shaping the cardiac action potential. In the present study, we examined whether angiotensin II (AngII) regulated the Ito as well as the putative intracellular cascade responsible for the effects. METHODS: Ito was recorded in rat ventricular myocytes using the nystatin-perforated patch-clamp configuration. RESULTS: AngII (0.1 microM) inhibited Ito (21.9+/-4.8% at +40 mV), but not the IK1, in a voltage- and time-independent manner. The inhibition decreased at concentrations higher than 1 microM resulting in a bell-shaped dose-response curve (IC50 = 3.1+/-1.5 microM). The blocking effects were abolished in the presence of the type 2 AngII receptor (AT2R) antagonist PD123319, but not in the presence of the selective type 1 AngII receptor (AT1R) antagonist candesartan. Moreover, the selective AT2R agonist CGP42112A completely reproduced the effects of AngII (20.5+/-2.4% of block at +40 mV), indicating that AngII-induced Ito block was mediated via stimulation of AT2R. Furthermore, selective stimulation of AT2R by CGP42112A significantly prolonged the rat atrial action potentials recorded using conventional microelectrode techniques. The AngII-induced inhibition of I(to) was not modified by either Npi-nitro-L-arginine-methyl ester (L-NAME) or eicosatetrayonic acid (ETYA), indicating that neither the nitric oxide (NO)-guanosine 3',5'-cyclic monophosphate (cGMP) system nor the arachidonic acid cascade was implicated in the effects of AngII on Ito. However, the AngII-induced Ito inhibition was completely abolished by the serine/threonine phosphatase type 2A (PP2A) inhibitors, okadaic acid and cantharidin, but not by the inactive analog of okadaic acid, 1-norokadaone. Intracellular application of PP2A decreased Kv4.2 currents recorded in transiently transfected Chinese hamster ovary cells (CHO). CONCLUSION: These results indicate that AngII activates PP2A through the stimulation of the AT2R, resulting in a decrease of the Ito amplitude.

Action Potentials↗

PubMed2004

Optical processing with vortex-producing lenses.

We discuss two types of optical processing using vortex-producing angular phase plates. In the most common spatial-filtering operation, an input object is Fourier transformed (either by Fraunhofer diffraction or with a lens system). The Fourier transform is then multiplied by an angular phase pattern, and the product is again Fourier transformed. The output is a space-invariant, edge-enhanced version of the input object. Alternatively we can directly image the object using a lens multiplied by the angular phase. The space-variant image is severely distorted along the optical axis of the system. We encode the phase plates onto a liquid-crystal display and present experimental results on both systems.

Journal Article↗

PubMed2003

Fractional Fourier transform optical system with programmable diffractive lenses.

We present experimental results for a fractional Fourier transform (FRFT) system implemented with programmable lenses written onto a liquid-crystal spatial light modulator (LCSLM). Because the focal length can be changed, different orders of the FRFT can be obtained without changing the optical setup. The LCSLM can very easily implement more complicated operations, including the realization of simultaneous orders of the FRFT and anamorphic transforms.

Journal Article↗

PubMed2003

Phase-only filtering on the three-dimensional Fourier spectrum of color images.

We propose what is to our knowledge a novel technique for filtering a two-dimensional multichannel color image based on its interpretation as a three-dimensional (3D) signal and the application of a 3D Fourier transform. The color image is considered to have the two usual spatial coordinates with the color distribution as the third dimension of the signal. This approach permits the 3D generalization of the usual phase-only filter and phase correlation, and color pattern recognition can be performed by means of a 3D correlation. We analyze two cases: whitening only along the color Fourier spectrum and whitening the whole 3D Fourier spectrum. We study the transformation induced in the color content of the image by the whitening of the color Fourier spectrum and how it affects the correlation distribution. In particular it is demonstrated that the whitening of the 3D Fourier spectrum improves the recognition performance of the correlation.

Journal Article↗

PubMed2003

Spironolactone and its main metabolite, canrenoic acid, block human ether-a-go-go-related gene channels.

BACKGROUND: It has been demonstrated that spironolactone (SP) decreases the QT dispersion in chronic heart failure. In this study, the effects of SP and its metabolite, canrenoic acid (CA), on human ether-a-go-go-related gene (HERG) currents were analyzed. METHODS AND RESULTS: HERG currents elicited in stably transfected Chinese hamster ovary cells were measured with the whole-cell patch-clamp technique. SP decreased HERG currents in a concentration-dependent manner (IC50=23.0+/-1.5 micromol/L) and shifted the midpoint of the activation curve to more negative potentials (Vh=-13.1+/-3.4 versus -18.9+/-3.6 mV, P<0.05) without modifying the activation and deactivation kinetics. SP-induced block (1 micromol/L) appeared at the range of membrane potentials coinciding with that of channel activation, and thereafter, it remained constant, reaching 24.7+/-3.8% at +60 mV (n=6, P<0.05). CA (0.01 nmol/L to 500 micromol/L) blocked HERG channels in a voltage- and frequency-independent manner. CA at 1 nmol/L shifted the midpoint of the activation curve to -19.9+/-1.8 mV and accelerated the time course of channel activation (tau=1064+/-125 versus 820+/-93 ms, n=11, P<0.01). The envelope of the tail test demonstrated that at the very beginning of the pulses to +40 mV (25 ms), a certain amount of block was apparent (31.3+/-9.9%). CA did not modify the voltage-dependence of HERG channel inactivation (Vh=-60.8+/-5.6 versus -62.9+/-3.1 mV, n=6, P>0.05) or the kinetics of the reactivation process at any potential tested. CA and aldosterone also blocked the native I(Kr) in guinea-pig ventricular myocytes. CONCLUSIONS: At concentrations reached after administration of therapeutic doses of SP, CA blocked the HERG channels by binding to both the closed and open states of the channel.

Aldosterone↗

PubMed2003

Effects of propafenone and its main metabolite, 5-hydroxypropafenone, on HERG channels.

OBJECTIVES: Propafenone is a class Ic antiarrhythmic drug used to maintain sinus rhythm in patients with atrial fibrillation. During chronic therapy, it undergoes extensive first-pass hepatic metabolism to 5-hydroxypropafenone. In the present study we have analysed the effects of propafenone and 5-hydroxypropafenone on HERG current. METHODS: The whole-cell configuration of the patch-clamp technique was used in CHO cells stably transfected with the gene encoding HERG channels. RESULTS: Propafenone and 5-hydroxypropafenone (2 microM) inhibited HERG current by 78.7+/-2.3% (n=7) and 71.1+/-4.1% (n=7, P>0.05) when measured at the end of 5-s depolarizing pulses to -10 mV. Block measured at the maximum peak of tail currents recorded at -60 mV was similar for propafenone (78.3+/-2.0%, n=7, P>0.05) and higher for 5-hydroxypropafenone (79.3+/-1.5%, n=7, P<0.05). Propafenone and 5-hydroxypropafenone shifted the midpoint of the activation curve by -10.2+/-0.9 mV (n=7, P<0.01) and -7.4+/-1.1 mV (n=10, P<0.01), respectively. Both drugs accelerated the deactivation and the inactivation process of HERG current. Propafenone, but not 5-hydroxypropafenone, inhibited to a higher extent HERG current at the end of 5-s depolarizing pulses to 0 mV than after promoting the transition of HERG channels from the inactivated to the opened state. CONCLUSIONS: These results indicate that propafenone and its main active metabolite, 5-hydroxypropafenone, block HERG channels to a similar extent by binding predominantly to the open state of the channel.

Animals↗

PubMed2003

Effects of irbesartan on cloned potassium channels involved in human cardiac repolarization.

We studied the effects of irbesartan, a selective angiotensin II type 1 receptor antagonist, on human ether-a-go-go-related gene (HERG), KvLQT1+minK, hKv1.5, and Kv4.3 channels using the patch-clamp technique. Irbesartan exhibited a low affinity for HERG and KvLQT1+minK channels (IC(50) = 193.0 +/- 49.8 and 314.6 +/- 85.4 microM, respectively). In hKv1.5 channels, irbesartan produced two types of block, depending on the concentration tested. At 0.1 microM, irbesartan inhibited the current in a time-dependent manner (22 +/- 3.9% at +60 mV). The blockade increased steeply with channel activation increasing at more positive potentials. However, at 10 microM, irbesartan induced a time-independent blockade that occurred in the range of potentials of channel opening, reaching its maximum at approximately 0 mV, and remaining unchanged at more positive potentials (24.0 +/- 1.0% at +60 mV). In Kv4.3 currents, irbesartan produced a concentration-dependent block, which resulted in two IC(50) values (1.0 +/- 0.1 nM and 7.2 +/- 0.6 microM). At 1 microM, it inhibited the peak current and accelerated the time course of inactivation, decreasing the total charge crossing the membrane (36.6 +/- 7.8% at +50 mV). Irbesartan shifted the inactivation curve of Kv4.3 channels, the blockade increasing as the amount of inactivated channels increased. Molecular modeling was used to define energy-minimized dockings of irbesartan to hKv1.5 and HERG channels. In conclusion, irbesartan blocks Kv4.3 and hKv1.5 channels at therapeutic concentrations, whereas the blockade of HERG and KvLQT1+minK channels occurred only at supratherapeutic levels. In hKv1.5, a receptor site is apparent on each alpha-subunit of the channel, whereas in HERG channels a common binding site is present at the pore.

Animals↗

PubMed2002

Convergent optical correlator aligment based on frequency filtering.

The convergent correlator is widely used but it presents the drawback of the alignment requirements: fine focusing of the input scene Fourier transform on the filter plane, filter centering, scaling the scene Fourier transform to match the filter size, and azimuth matching of the filter with the input scene. We propose a set of tests to obtain a precise alignment of the convergent correlator. These methods are based on frequency filtering properties and they are applicable either for amplitude input or for phase-encoded input. The tests we present allow us to fulfill all the alignment requirements. The theory on which these tests are based is explained. The experimental results obtained during the alignment procedure are presented. We show some additional verifications of the correct alignment of the convergent correlator.

Journal Article↗

PubMed2002

Putative binding sites for benzocaine on a human cardiac cloned channel (Kv1.5).

OBJECTIVES: It has been demonstrated that at nanomolar concentrations benzocaine increased, whereas at micromolar concentrations, it blocked hKv1.5 channels in a voltage-dependent manner and modified the voltage-dependence of channel activation. The present study was undertaken to localize the putative binding sites involved in the 'agonists' and blocking effects of benzocaine. METHODS: Experiments were carried out on wild-type and site directed mutated hKv1.5 channels stably expressed on Ltk(-) cells using the whole-cell patch-clamp. RESULTS: At 35 mM [K+](i) the voltage-dependent unblock produced by 500 microM benzocaine was preserved at both 4 and 140 mM [K+](o). Mutations located in the inner mouth of the pore (T477S, T505A, L508M and V512M) abolished the agonist but increased the blocking effects of benzocaine. Intracellular application of tetraethylammonium (3 mM) abolished the 'agonist' effects whereas the blocking effects of benzocaine remained unaltered. Block induced by benzocaine and intracellular tetraethylammonium was additive. In contrast, the combination of benzocaine and bupivacaine (>25 microM) produced less blockade than bupivacaine alone. However, mutation of the extracellular residue R485Y did not modify the effects of benzocaine. Extracellular application of tetraethylammonium (100 mM) did not modify the agonist effects of benzocaine, but abolished the voltage- and time-dependence of benzocaine-induced block. CONCLUSIONS: The results suggested that benzocaine binds with high affinity to an intracellular binding site to produce 'agonist' effects and to a low affinity subsite, which is also located in the inner mouth, to produce the blocking effects. Furthermore, benzocaine and extracellular K(+) interact to modify the voltage-dependence of channel opening.

Animals↗

PubMed2002

Effects of levobupivacaine, ropivacaine and bupivacaine on HERG channels: stereoselective bupivacaine block.

1 Levobupivacaine and ropivacaine are the pure S(-) enantiomers of N-butyl- and N-propyl-2',6'-pipecoloxylidide, developed as less cardiotoxic alternatives to bupivacaine. In the present study, we have analysed the effects of levobupivacaine, ropivacaine and bupivacaine on HERG channels stably expressed in CHO cells. 2 The three drugs blocked HERG channels in a concentration-, time- and state-dependent manner. Block measured at the end of 5 s pulses to -10 mV induced by 20 microM bupivacaine (52.7+/-2.0%, n=15) and ropivacaine (55.5+/-2.7%, n=13) was similar (P>0.05) and both lower than that induced by levobupivacaine (67.5+/-4.2%, n=11) (P<0.05). 3 Dextrobupivacaine (20 microM) was less potent (47.2+/-5.2%, n=10) than levobupivacaine (P<0.05), indicating stereoselective HERG channel block. 4. Block induced by the three local anaesthetics exhibited a steep voltage dependence in the range of channel activation. In all cases, block measured at the maximum peak current at a test potential of 0 mV after promoting recovery from inactivation (I-->O) was lower than that observed at the end of 5-s pulses (I+O). 5. Levobupivacaine, ropivacaine and bupivacaine accelerated HERG inactivation kinetics, slowed the recovery from inactivation and shifted the inactivation curve towards more negative membrane potentials. The three local anaesthetics induced a rapid time-dependent decline after using a protocol that quickly activates HERG channels. 6. All these results suggest that: (1) these drugs bind to the open and the inactivated states of HERG channels, (2) they stabilize HERG channels in the inactivated state, and (3) block induced by bupivacaine enantiomers is stereoselective.

Amides↗

PubMed2002

The rate and character of spontaneous mutation in an RNA virus.

Estimates of spontaneous mutation rates for RNA viruses are few and uncertain, most notably due to their dependence on tiny mutation reporter sequences that may not well represent the whole genome. We report here an estimate of the spontaneous mutation rate of tobacco mosaic virus using an 804-base cognate mutational target, the viral MP gene that encodes the movement protein (MP). Selection against newly arising mutants was countered by providing MP function from a transgene. The estimated genomic mutation rate was on the lower side of the range previously estimated for lytic animal riboviruses. We also present the first unbiased riboviral mutational spectrum. The proportion of base substitutions is the same as that in a retrovirus but is lower than that in most DNA-based organisms. Although the MP mutant frequency was 0.02-0.05, 35% of the sequenced mutants contained two or more mutations. Therefore, the mutation process in populations of TMV and perhaps of riboviruses generally differs profoundly from that in populations of DNA-based microbes and may be strongly influenced by a subpopulation of mutator polymerases.

Base Sequence↗

PubMed2002

Assembly with the Kvbeta1.3 subunit modulates drug block of hKv1.5 channels.

The assembly of voltage-gated potassium (Kv) channels with beta subunits modifies the electrophysiological characteristics of the alpha subunits. Kvbeta1.3 subunits shift the midpoint of the activation curve toward more negative voltages and slow the deactivation process. In addition, the Kvbeta1.3 subunit converts hKv1.5 from a delayed rectifier with a modest degree of slow inactivation to a channel with both fast and slow components of inactivation. In the present study, we have analyzed the effects of bupivacaine and a permanently charged analog [R(+)-N-methyl-bupivacaine (RB(+)1C)] on Kvalpha1.5 and Kvalpha1.5+Kvbeta1.3 channels expressed in human embryonic kidney 293 cells using the whole-cell configuration of the patch-clamp technique. Block induced by RB(+)1C binding to its external receptor site was not modified by the presence of this beta subunit. However, hKvalpha1.5+Kvbeta1.3 channels were ~4-fold less sensitive to bupivacaine than hKv1.5 channels in the absence of beta subunits (IC(50) = 47.5 +/- 5.1 versus 13.1 +/- 0.8 microM, respectively, p < 0.01). Quinidine was also less potent to block Kvalpha1.5+Kvbeta1.3 channels than Kvalpha1.5 channels (IC(50) = 49.6 microM versus 6.2 microM, respectively). These results suggest that the Kvbeta1.3 subunit does not modify the affinity of the charged bupivacaine for its external receptor site but markedly reduces the affinity of bupivacaine and quinidine for their internal receptor site in hKv1.5 channels.

Anesthetics, Local↗