bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of caffeine on peripheral chemoreceptor activity in premature neonates: interaction with sleep stages
Authors: Chardon Karen , Bach Véronique , Telliez Frédéric , Cardot Virginie , Tourneux Pierre , Leke André , Libert Jean-Pierre
Published: February 2004

Caffeine is widely used for the treatment of apnea in premature neonates. However, the localization of caffeine's target site (central nervous system and/or peripheral chemoreceptors) is not well defined, especially for sleeping neonates whose sleep stages interact with respiratory control. The aim of this study was to assess the activity of the peripheral chemoreceptors in relation to sleep stages in premature neonates treated (or not) with caffeine for idiopathic apnea. Peripheral chemoreceptor activity was assessed in 22 neonates (postconceptional age of 36 ± 1 wk with birth weights ranging from 790 to 1,910 g) by performing a 30-s hyperoxic test during active and quiet sleep. Eleven neonates received caffeine treatment (4.0 ± 0.5 mg·kg-1·day-1) and 11 served as controls. For all neonates, the decrease in minute ventilation observed during hyperoxia was greater during active than during quiet sleep. Neonates receiving caffeine showed a significantly greater decrease in ventilation during hyperoxia in both sleep stages, compared with controls (caffeine; -29.7 ± 12.8% vs. control; -22.0 ± 7.4%; F1,15= 4.6, P = 0.04). We conclude that caffeine administration increases the effectiveness of chemoreceptor activity. Because sleep stage durations were not affected by the treatment, it is likely that the decrease in apneic episodes typically observed with caffeine therapy is only related to respiratory processes and is independent of the sleep stage organization.
Modeled microgravity-induced protein kinase C isoform expression in human lymphocytes
Authors: Sundaresan A. , Risin D. , Pellis N. R.
Published: February 2004

In long-term space travel, the crew is exposed to microgravity and radiation that invoke potential hazards to the immune system. T cell activation is a critical step in the immune response. Receptor-mediated signaling is inhibited in both microgravity and modeled microgravity (MMG) as reflected by diminished DNA synthesis in peripheral blood lymphocytes and their locomotion through gelled type I collagen. Direct activation of protein kinase C (PKC) bypassing cell surface events using the phorbol ester PMA rescues MMG-inhibited lymphocyte activation and locomotion, whereas the calcium ionophore ionomycin had no rescue effect. Thus calcium-independent PKC isoforms may be affected in MMG-induced locomotion inhibition and rescue. Both calcium-dependent isoforms and calcium-independent PKC isoforms were investigated to assess their expression in lymphocytes in 1 g and MMG culture. Human lymphocytes were cultured and harvested at 24, 48, 72, and 96 h, and serial samples were assessed for locomotion by using type I collagen and expression of PKC isoforms. Expression of PKC-α, -δ, and -ϵ was assessed by RT-PCR, flow cytometry, and immunoblotting. Results indicated that PKC isoforms δ and ϵ were downregulated by >50% at the transcriptional and translational levels in MMG-cultured lymphocytes compared with 1- g controls. Events upstream of PKC, such as phosphorylation of phospholipase Cγ in MMG, revealed accumulation of inactive enzyme. Depressed calcium-independent PKC isoforms may be a consequence of an upstream lesion in the signal transduction pathway. The differential response among calcium-dependent and calcium-independent isoforms may actually result from MMG intrusion events earlier than PKC, but after ligand-receptor interaction.
Men are more fatigable than strength-matched women when performing intermittent submaximal contractions
Authors: Hunter Sandra K. , Critchlow Ashley , Shin In-Sik , Enoka Roger M.
Published: February 2004

The purpose of this study was to compare the time to task failure for a series of intermittent submaximal contractions performed with the elbow flexor muscles by men and women who were matched for strength ( n = 20, 18–34 yr). The fatigue task comprised isometric contractions at 50% of maximal voluntary contraction (MVC) torque (6-s contraction, 4-s rest). The MVC torque was similar for the men and women [64.8 ± 9.2 (SD) vs. 62.2 ± 7.9 N·m; P > 0.05]. However, the time to task failure was longer for the women (1,408 ± 1,133 vs. 513 ± 194 s; P < 0.05), despite the similar torque levels. The mean arterial pressure, heart rate, and rating of perceived exertion started and ended at similar values for the men and women, but the rate of increase was less for the women. The rate of increase in the average of the rectified electromyogram (AEMG; % peak MVC) for the elbow flexor muscles was less for the women: the AEMG was greater for the men compared with the women at task failure (72 ± 28 vs. 50 ± 21%; P < 0.05), despite similar AEMG values at the start of the fatiguing contraction (32 ± 9 vs. 36 ± 13%). These results indicate that for intermittent contractions performed with the elbow flexor muscles 1) the sex difference in time to task failure was not explained by the absolute strength of the men and women, but involved another mechanism that is present during perfused conditions, and 2) men required a more rapid increase in descending drive to maintain a similar torque.
Opposing Electrophysiological Actions of 5-HT on Noncholinergic and Cholinergic Neurons in the Rat Ventral Pallidum In Vitro
Authors: Bengtson C. Peter , Lee David J. , Osborne Peregrine B.
Published: February 2004

The ventral pallidum in rat is a basal forebrain structure that contains neurons that project in the limbic striatopallidal circuitry and magnocellular cholinergic corticopetal neurons. Because 5-hydroxytryptamine (5-HT) terminals on dorsal raphe projections form close appositions with these neurons, we made patch-clamp recordings in immature rat brain slices to determine whether they are modulated by postsynaptic 5-HT receptors. Inward currents were predominantly induced by 5-HT in noncholinergic neurons, which were distinguished from cholinergic neurons by immunohistochemical and electrophysiological criteria. The inward current induced by 5-HT was mimicked and occluded when adenylyl cyclase was stimulated with forskolin, and was almost abolished when h-currents in noncholinergic neurons were blocked with cesium. Consistent with 5-HT7 receptor activation of h-curents by cAMP in other brain regions, we found inward currents were mimicked by the mixed 5-HT1/5-HT7 agonists 5-methoxytryptamine, and by 5-carboxamidotryptamine (5-CT), which was more potent than 5-HT. In contrast, 5-HT1 preferring 8-OH-DPAT was a weak partial agonist, and the 5-HT1–selective antagonist pindolol had no effect. However, despite this profile, antagonists that bind at the 5-HT7 receptor only partly reduced the agonist inward current (SB-269970 and clozapine), or had no effect (mianserin and pimozide). We found in cholinergic neurons that 5-HT predominantly induced hyperpolarizing currents, which were carried by potassium channels, and were smaller than currents induced by 8-OH-DPAT and 5-CT. We conclude from this study that ascending 5-HT projections from the dorsal raphe could have direct and opposite effects on the activities of neurons within the limbic striatopallidal and cholinergic corticopetal circuitry in the ventral pallidum.
Ocular Dominance Predicts Neither Strength Nor Class of Disparity Selectivity With Random-Dot Stimuli in Primate V1
Authors: Read Jenny C. A. , Cumming Bruce G.
Published: February 2004

We address two unresolved issues concerning the coding of binocular disparity in primary visual cortex. Experimental studies and theoretical models have suggested a relationship between a cell's ocular dominance, assessed with monocular stimuli, and its tuning to binocular disparity. First, the disparity energy model of disparity selectivity suggests that there should be a correlation between ocular dominance and the strength of disparity tuning. Second, several studies have reported a relationship between ocular dominance and the shape of the disparity tuning curve, with cells dominated by one eye more likely to have disparity tuning of the tuned-inhibitory type. We investigated both of these relationships in single neurons recorded from the primary visual cortex of awake fixating macaques, using dynamic random-dot patterns as a stimulus. To classify disparity tuning curves quantitatively, we develop a new measure of symmetry, which can be applied to any function. We find no evidence for any correlation between ocular dominance and the nature of disparity tuning. This places constraints on the circuitry underlying disparity tuning.
Integration of Retinal Disparity and Fixation-Distance Related Signals Toward an Egocentric Coding of Distance in the Posterior Parietal Cortex of Primates
Authors: Genovesio Aldo , Ferraina Stefano
Published: February 2004

For those movements that are directed toward objects located in extrapersonal space, it is necessary that visual inputs are first remapped from a retinal coordinate system to a body-centered one. The posterior parietal cortex (PPC) most likely integrates retinal and extraretinal information to determine the egocentric distance of an object located in three-dimensional (3-D) space. This determination requires both a retinal disparity signal and a parallel estimate of the fixation distance. We recorded from the lateral intraparietal area (LIP) to see if single neurons respond to both vergence angle and retinal disparity and if these two signals are integrated to encode egocentric distance. Monkeys were trained to make saccades to real targets in 3-D space. When both fixation distance and disparity of visual stimuli were varied, the disparity tuning of individual neurons display a fixation-distance modulation. We propose that the observed modulation contributes to a spatial coding domain intermediate between retinal and egocentric because the disparity tuning shifts in a systematic way with changes in fixation distance.
Maturation of Layer V Pyramidal Neurons in the Rat Prefrontal Cortex: Intrinsic Properties and Synaptic Function
Authors: Zhang Zhong-wei
Published: February 2004

Layer V pyramidal neurons in the rat medial prefrontal cortex (PFC) were examined with whole cell patch-clamp recording in acute slices from postnatal day 1 (P1) to P36. In the first few days after birth, layer V pyramidal neurons had low resting potentials, high-input resistance, and long membrane time constant. During the next 2 wk, the resting potential shifted by -14 mV, while the input resistance and time constant decreased by 15- and 4-fold, respectively. Between P3 and P21, the surface area of the cell body doubled, while the total lengths of apical and basal dendrites increased by 5- and 13-fold, respectively. Action potentials (APs) were observed at all aged tested. The peak amplitude of APs increased by 30 mV during the first 3 wk, while AP rise time and half-maximum duration shortened significantly. Compared with neurons at P21 or older, neurons in the first week required much smaller currents to reach their maximum firing frequencies, but the maximum frequencies were lower than those at older ages. Stimulation of layer II/III induced monosynaptic responses in neurons older than P5. Paired-pulse responses showed a short-term depression at P7, which shifted progressive to facilitation at older ages. These results demonstrate that, similar to other neurons in the brain, layer V pyramidal neurons in the PFC undergo a period of rapid development during the first 3 wk after birth. These findings suggest that the intrinsic properties of neurons and the properties of synaptic inputs develop concomitantly during early life.
Visual Cortex Neurons of Monkeys and Cats: Temporal Dynamics of the Spatial Frequency Response Function
Authors: Frazor Robert A. , Albrecht Duane G. , Geisler Wilson S. , Crane Alison M.
Published: February 2004

We measured the responses of striate cortex neurons as a function of spatial frequency on a fine time scale, over the course of an interval that is comparable to the duration of a single fixation (200 ms). Stationary gratings were flashed on for 200 ms and then off for 300 ms; the responses were analyzed at sequential 1-ms intervals. We found that 1) the preferred spatial frequency shifts through time from low frequencies to high frequencies, 2) the latency of the response increases as a function of spatial frequency, and 3) the poststimulus time histograms (PSTHs) are relatively shape-invariant across spatial frequency. The dynamic shifts in preferred spatial frequency appear to be a simple consequence of the latency shifts and the transient nature of the PSTH. The effects of these dynamic shifts on the coding of spatial frequency information are examined within the context of several different temporal integration strategies, and pattern-detection performance is determined as a function of the interval of integration, following response onset. The findings are considered within the context of related investigations as well as a number of functional issues: motion selectivity in depth, “coarse-to-fine” processing, direction selectivity, latency as a code for stimulus attributes, and behavioral response latency. Finally, we demonstrate that the results are qualitatively consistent with a simple feedforward model, similar to the one originally proposed in 1962 by Hubel and Wiesel, that incorporates measured differences in the response latencies and the receptive field sizes of different lateral geniculate nucleus inputs.
Retinal pH Reflects Retinal Energy Metabolism in the Day and Night
Authors: Dmitriev Andrey V. , Mangel Stuart C.
Published: February 2004

The extracellular pH of living tissue in the retina and elsewhere in the brain is lower than the pH of the surrounding milieu. We have shown that the pH gradient between the in vitro retina and the superfusion solution is regulated by a circadian (24-h) clock so that it is smaller in the subjective day than in the subjective night. We show here that the circadian changes in retinal pH result from a clock-mediated change in the generation of H+ that accompanies energy production. To demonstrate this, we suppressed energy metabolism and recorded the resultant reduction in the pH difference between the retina and superfusate. The magnitude of the reduction in the pH gradient correlated with the extent of energy metabolism suppression. We also examined whether the circadian-induced increase in acid production during the subjective night results from an increase in energy metabolism or from the selective activation of glycolysis compared with oxidative phosphorylation. We found that the selective suppression of either oxidative phosphorylation or glycolysis had almost identical effects on the dynamics and extent of H+ production during the subjective day and night. Thus the proportion of glycolysis and oxidative phosphorylation is maintained the same regardless of circadian time, and the pH difference between the tissue and superfusion solution can therefore be used to evaluate total energy production. We conclude that circadian clock regulation of retinal pH reflects circadian regulation of retinal energy metabolism.
Direct Physiological Evidence for Synaptic Connectivity Between Medium-Sized Spiny Neurons in Rat Nucleus Accumbens In Situ
Authors: Taverna Stefano , van Dongen Yvette C. , Groenewegen Henk J. , Pennartz Cyriel M.A.
Published: February 2004

Dual whole cell patch-clamp recordings in rat nucleus accumbens, the main component of the ventral striatum, were made to assess the presence of synaptic interconnections between medium-sized spiny neurons, a group of GABAergic and peptidergic neurons that constitute the principal cells of the striatum. Neurons were stained with biocytin for subsequent morphological analysis. Electrical activity of cells was recorded in current- and voltage-clamp mode; the characteristics of medium-sized spiny neurons were confirmed by electrophysiological and morphological properties. Thirteen of 38 medium-sized spiny neuron pairs (34%) showed a synaptic connection. In these pairs, suprathreshold stimulation with current injection evoked a train of action potentials in the presynaptic cell, which in turn elicited depolarizing postsynaptic potentials (dPSPs) in the postsynaptic cell. Twelve of these 13 pairs were connected unilaterally. The onset latency of the postsynaptic response was 1.7 ± 0.7 ms. dPSPs were blocked by 12.5 μM bicuculline, suggesting they were mediated by GABAA receptors. A linear fit of the current-voltage relationship of GABAergic currents crossed the voltage axis near the value of -20 mV, in agreement with the Cl- equilibrium potential predicted from the composition of the artificial cerebrospinal fluid and pipette medium. No evidence for electrotonic coupling was found. Paired-pulse facilitation and depression were induced when the amplitude of the first IPSC of a pair was relatively small and large, respectively. No clear dependence of paired-pulse facilitation or depression was found on the width of the spike interval, which ranged between 100 and 380 ms. Conversely, 1- to 2-s trains of dPSPs showed marked frequency facilitation at low presynaptic frequencies, but frequency depression at high firing rates. These data show that intra-accumbens synaptic communication between medium-sized spiny neurons exists, is mediated by GABAA receptors, and exhibits spike train-dependent short-term dynamics.
Effects of Muscimol Inactivation of the Cerebellar Nuclei on Precision Grip
Authors: Monzée Joël , Drew Trevor , Smith Allan M.
Published: February 2004

A single monkey was trained to perform a grasp, lift, and hold task in which a stationary hand- held object was sometimes subjected to brief, predictable force-pulse perturbations. The displacement, grip, and lifting forces were measured as well the three-dimensional forces and torques to quantify specific motor deficits after reversible inactivation of the cerebellar nuclei. A prior single-cell recording study in the same monkey provided the stereotaxic coordinates used to guide intranuclear injections of muscimol. In total, 34 penetrations were performed at 28 different loci throughout the cerebellar nuclei. On each penetration, two 1.0-μl injections of 5 μg/μl muscimol, were made 1.0 mm apart either within the nuclei or in the white matter just lateral or posterior to the dentate nucleus. Injections in the region corresponding to the anterior interpositus nucleus produced pronounced dynamic tremor and dysmetric movements of the ipsilateral arm when the animal performed unrestrained reaching and grasping movements. In contrast, no relatively short-latency (15-20 min.) deficits were observed after injection in the dentate nucleus, although some effects were observed after several hours. When tested in a primate chair with the forearm supported and restrained at the wrist and elbow, the monkey performed the lift and hold task without tremor or dysmetria. However, with the restraint removed, the forces and torques applied to the manipulandum were poorly controlled and erratic. The monkey's arm was ataxic and a 5-Hz intention tremor was clearly visible. In addition, the animal was generally unable to compensate for the predictable perturbations and the anticipatory grip force increases were absent. However, overall the results suggest that reversible cerebellar nuclear inactivation with muscimol has little effect on isolated distal movements of the wrist and fingers.
Predictive Smooth Ocular Pursuit During the Transient Disappearance of a Visual Target
Authors: Bennett Simon J. , Barnes Graham R.
Published: February 2004

When a moving target disappears and there is a complete absence of visual feedback signals, eye velocity decays rapidly but often recovers to previous levels if there is an expectation the target will reappear further along its trajectory Given that eye velocity cannot be maintained under such circumstances, the anticipatory recovery may function to minimize the developing velocity error. When there is a change in target velocity during a transient, any recovery should ideally be scaled and hence predictive of the expected target velocity at reappearance. This study confirmed that subjects did not maintain eye velocity close to target velocity for the duration of the inter-stimulus interval (ISI). The majority of subjects exhibited an initial reduction in eye velocity followed by a scaled recovery prior to target reappearance. Eye velocity during the ISI was, therefore, predictive of the expected change in target velocity. These behavioral data were simulated using a model in which gain applied to the visuomotor drive is reduced after the loss of visual feedback and then modulated depending on subject’s expectation regarding the target’s future trajectory.
Genomic profiling of the human heart before and after mechanical support with a ventricular assist device reveals alterations in vascular signaling networks
Authors: Hall Jennifer L. , Grindle Suzanne , Han Xinqiang , Fermin David , Park Soon , Chen Yingjie , Bache Robert J. , Mariash Ami , Guan Zhanjun , Ormaza Sofia , Thompson Jeanne , Graziano Judith , de Sam Lazaro Shireen E. , Pan Shuchong , Simari Robert D. , Miller Leslie W.
Published: February 2004

Mechanical unloading of the heart with a left ventricular assist device (LVAD) significantly decreases mortality in patients with heart failure. Moreover, it provides a human model to define the critical regulatory genes governing myocardial remodeling in response to significant reductions in wall stress. Statistical analysis of a gene expression library of 19 paired human heart samples harvested at the time of LVAD implant and again at explant revealed a set of 22 genes that were downregulated and 85 genes that were upregulated in response to mechanical unloading with a false discovery rate of less than 1%. The analysis revealed a high percentage of genes involved in the regulation of vascular networks including neuropilin-1 (a VEGF receptor), FGF9, Sprouty1, stromal-derived factor 1, and endomucin. Taken together these findings suggest that mechanical unloading alters the regulation of vascular organization and migration in the heart. In addition to vascular signaling networks, GATA-4 binding protein, a critical mediator of myocyte hypertrophy, was significantly downregulated following mechanical unloading. In summary, these findings may have important implications for defining the role of mechanical stretch and load on autocrine/paracrine signals directing vascular organization in the failing human heart and the role of GATA-4 in orchestrating reverse myocardial remodeling. This unbiased gene discovery approach in paired human heart samples has the potential to provide critical clues to the next generation of therapeutic treatments aimed at heart failure.
Genetic contributors to lipoprotein cholesterol levels in an intercross of 129S1/SvImJ and RIIIS/J inbred mice
Authors: Lyons Malcolm A. , Korstanje Ron , Li Renhua , Walsh Kenneth A. , Churchill Gary A. , Carey Martin C. , Paigen Beverly
Published: February 2004

To determine the genetic contribution to variation among lipoprotein cholesterol levels, we performed quantitative trait locus (QTL) analyses on an intercross between mouse strains RIIIS/J and 129S1/SvImJ. Male mice of the parental strains and the reciprocal F1and F2populations were fed a high-cholesterol, cholic acid-containing diet for 8–12 wk. At the end of the feeding period, plasma total, high-density lipoprotein (HDL), and non-HDL cholesterol were determined. For HDL cholesterol, we identified three significant QTLs on chromosomes (Chrs) 1 ( D1Mit507, 88 cM, 72–105 cM, 4.8 LOD), 9 ( D11Mit149, 14 cM, 10–25 cM, 9.4 LOD), and 12 ( D12Mit60, 20 cM, 0–50 cM, 5.0 LOD). These QTLs were considered identical to QTLs previously named Hdlq5, Hdlq17, and Hdlq18, respectively, in crosses sharing strain 129. For total cholesterol, we identified two significant QTLs on Chrs 1 and 9, which were named Chol10 ( D1Mit507, 88 cM, 10–105 cM, 3.9 LOD) and Chol11 ( D11Mit149, 14 cM, 0–30 cM, 4.4 LOD), respectively. In addition, for total cholesterol, we identified two suggestive QTLs on Chrs 12 (distal) and 17, which remain unnamed. For non-HDL cholesterol, we identified and named one new QTL on Chr 17, Nhdlq3 ( D17Mit221, 58 cM, 45–60 cM, 3.4 LOD). Nhdlq3 colocalized with orthologous human QTLs for lipoprotein phenotypes, and with Abcg5 and Abcg8. Overall, we detected eight QTLs for lipoprotein cholesterol concentrations on Chrs 1, 9, 12, and 17 (each two per chromosome), including a new QTL for non-HDL cholesterol, Nhdlq3, on Chr 17.
Identification of two novel shear stress responsive elements in rat angiotensin I converting enzyme promoter
Authors: Miyakawa Ayumi Aurea , de Lourdes Junqueira Maria , Krieger José Eduardo
Published: February 2004

Mechanical forces contribute to maintenance of cardiovascular homeostasis via the control of release and production of vasoactive substances. We demonstrated previously that shear stress decreases rat ACE activity and expression. Using a reporter gene approach and mutagenesis, we show now that the classic shear stress responsive element or SSRE (GAGACC) contained within 1,274 bp of this promoter is not functional in response to shear stress (15 dyn/cm2, 18 h) [for the wild-type ACE promoter (WLuc), static control (C) = 107 ± 6.5%, shear stress (SS) = 65.9 ± 9.4%, n = 8; for the promoter with the classic SSRE mutated (WSS-mut), C = 100 ± 8.2%, SS = 60.2 ± 5.2%, n = 10, respectively]. Analysis of progressive deletion mutants unraveled a 57-bp fragment, position −251 to −195, from the transcription start site, containing functional SSRE (for WLuc, C = 107 ± 6.5%, SS = 65.9 ± 9.4%, n = 8; for 378, C = 100 ± 6.4%, SS = 60.4 ± 4.3%, n = 11; for 251, C = 99.7 ± 2.6%, SS = 63.2 ± 5.5%, n = 7; for 194, C = 104.6 ± 8.1%, SS = 92.4 ± 6.9%, n = 9). This fragment responded to shear stress even in the context of a heterologous promoter. Finally, functional analysis of mutated candidate regulatory elements identified by gel shift, DNase I footprint, and conservation of aligned sequences revealed that only the double mutant (Barbie/GAGA-mut) but not isolated disruption of the Barbie (WBarbie-mut) or the GAGA (WGAGA-mut) prevented the shear-stress-induced response (for Barbie/GAGA-mut, C = 97.9 ± 5%, SS = 99.4 ± 7.2%, n = 6; for WBarbie-mut, C = 106.1 ± 8.6%, SS = 65.9 ± 9.4%, n = 6; for WGAGA-mut, C = 100.1 ± 2.9%, SS = 66.7 ± 1.6, n = 6;). Taken together, these data provide direct evidence for the new role of Barbie and GAGA boxes in mediating the shear-stress-induced downregulation of rat ACE expression and demonstrate that the classic SSRE (GAGACC) is not functional under the experimental conditions tested.
Clinical and Biochemical Results of the Metalloproteinase Inhibition with Subantimicrobial Doses of Doxycycline to Prevent Acute Coronary Syndromes (MIDAS) Pilot Trial
Authors: Brown David L. , Desai Kavita K. , Vakili Babak A. , Nouneh Chadi , Lee Hsi-Ming , Golub Lorne M.
Published: February 2004

Background—Vulnerable plaque demonstrates intense inflammation in which macrophages secrete matrix metalloproteinases (MMPs) that degrade the fibrous cap, ultimately leading to rupture, in situ thrombosis, and an associated clinical event. Thus, inhibition of MMP activity or more general suppression of vascular inflammation are attractive targets for interventions intended to reduce plaque rupture. We hypothesized that subantimicrobial doses of doxycycline (SDD) (20 mg twice daily) would benefit patients with coronary artery disease by reducing inflammation and MMP activity and thus possibly prevent coronary plaque rupture events.

Methods and Results—We conducted a prospective, randomized, double-blind, placebo-controlled pilot study of 6 months of SDD or placebo treatment to reduce inflammation and prevent plaque rupture events. A total of 50 patients were enrolled, of whom 24 were randomized to placebo and 26 to SDD. At 6 months, there was no difference in the composite endpoint of sudden death, fatal myocardial infarction (MI), non-fatal MI, or troponin-positive unstable angina in SDD compared with placebo-treated patients (8.4% versus 0%,P=0.491). Biochemical markers of inflammation were assessed in plasma at study entry and after 6 months of therapy in 30 patients. In SDD-treated patients, high-sensitivity C-reactive protein (CRP) was reduced by 46% from 4.8±0.6 μg/mL to 2.6±0.4 μg/mL (P=0.007), whereas CRP was not significantly reduced in placebo patients. Interleukin (IL)-6 decreased from 22.1±3.7 pg/mL at baseline to 14.7±1.8 pg/mL at 6 months in SDD-treated patients (P=0.025) but did not decrease significantly in placebo-treated patients. On zymography, pro-MMP-9 activity was reduced 50% by SDD therapy (P=0.011), whereas it was unchanged by placebo treatment.

Conclusion—SDD appears to exert potentially beneficial effects on inflammation that could promote plaque stability. These findings should be investigated in a larger study.
Cholesterol 7α-Hydroxylase Deficiency in Mice on an APOE*3-Leiden Background Impairs Very-Low-Density Lipoprotein Production
Authors: Post Sabine M. , Groenendijk Martine , Solaas Karianne , Rensen Patrick C. N. , Princen Hans M. G.
Published: February 2004

Objective—Cholesterol 7α-hydroxylase (cyp7a1) catalyzes the rate-limiting step in conversion of cholesterol to bile acids. To study the relationship between bile acid biosynthesis and triglyceride metabolism, we cross-bred mice lackingcyp7a1on a hyperlipidemic APOE*3-Leiden background.

Methods and Results—Female mice received a chow or lipogenic diet. On both diets, fecal bile acid excretion was 70% decreased concomitantly with a 2-fold increased neutral sterol output. The differences in bile acid biosynthesis did not change plasma cholesterol levels. However, plasma triglyceride levels decreased by 41% and 38% in thecyp7a1−/−. APOE*3-Leiden mice as compared with APOE*3-Leiden mice on chow and lipogenic diet, respectively. Mechanistic studies showed that very-low-density lipoprotein (VLDL)–apolipoprotein B and VLDL–triglyceride production rates were reduced incyp7a1−/−. APOE*3-Leiden mice as compared with APOE*3-Leiden mice (−34% and −35%, respectively).Cyp7a1deficiency also increased the hepatic cholesteryl ester and triglyceride content (2.8-fold and 2.5-fold, respectively). In addition, hepatic anti-oxidative vitamin content, which can influence VLDL-production, was lower. Hepatic mRNA analysis showed decreased expression of genes involved in lipogenesis includingsrebf1.

Conclusions—Cyp7a1deficiency in APOE*3-Leiden mice decreases the VLDL particle production rate, as a consequence of a strongly reduced bile acid biosynthesis, leading to a decrease in plasma triglycerides. These data underscore the close relationship between bile acid biosynthesis and triglyceride levels.
In-Depth Haplotype Analysis of ABCA1 Gene Polymorphisms in Relation to Plasma ApoA1 Levels and Myocardial Infarction
Authors: Tregouet David-Alexandre , Ricard Sylvain , Nicaud Viviane , Arnould Isabelle , Soubigou Stéphane , Rosier Marie , Duverger Nicolas , Poirier Odette , Macé Sandrine , Kee Frank , Morrison Caroline , Denèfle Patrice , Tiret Laurence , Evans Alun , Deleuze Jean-Francois , Cambien Francois
Published: February 2004

Objective—

By regulating the cellular cholesterol efflux from peripheral cells to high-density lipoprotein, the ABCA1 protein is suspected to play a key role in lipid homeostasis and atherosclerosis. Twenty-six polymorphisms of the ABCA1 gene were genotyped and tested for association with plasma levels of ApoA1 and myocardial infarction (MI) in the ECTIM study.

Methods and Results—

In addition to single-locus analysis, a systematic exploration of all possible haplotype effects was performed, with this exploration being performed on a minimal set of “tag” polymorphisms that define the haplotype structure of the gene. Two polymorphisms were associated with plasma levels of ApoA1, 1 in the promoter (C-564T) and 1 in the coding (R1587K) regions, whereas only 1 polymorphism (R219K) was associated with the risk of MI. However, no haplotype effect was detected on ApoA1 variability or on the risk of MI.

Conclusion—

ABCA1 gene polymorphisms but not haplotypes are involved in the variability of plasma ApoA1 and the susceptibility to coronary artery disease.
Angiotensin II Type 1a Receptor Is Involved in Cell Infiltration, Cytokine Production, and Neovascularization in Infarcted Myocardium
Authors: Toko Haruhiro , Zou Yunzeng , Minamino Tohru , Sakamoto Masaya , Sano Masanori , Harada Mutsuo , Nagai Toshio , Sugaya Takeshi , Terasaki Fumio , Kitaura Yasushi , Komuro Issei
Published: February 2004

Objective—Angiotensin II is critically involved in left ventricular remodeling after myocardial infarction. Neovascularization has been thought to prevent the development of left ventricular remodeling and deterioration to heart failure. To elucidate the role of angiotensin II in neovascularization during cardiac remodeling, we induced myocardial infarction in angiotensin II type1a receptor (AT1) knockout (KO) mice.

Methods and Results—There were more vessels in the border zone of infarcted hearts of wild-type (WT) mice and AT1KO mice at 14 days after operation, compared with in the left ventricle of sham-operated mice, and the number was larger in WT mice than in AT1KO mice. Consistent with these observations, the infarcted heart of AT1KO mice expressed lower levels of matrix metalloproteinase and endothelial nitric oxide synthase activity. More inflammatory cells such as granulocytes and macrophages were infiltrated in the infarcted hearts of WT mice than AT1KO mice at 4 days. A variety of cytokines and chemokines were increased in infarcted hearts of WT and AT1KO mice, and many of them were more remarkable in WT mice than in AT1KO mice at 14 days.

Conclusions—AT1 plays a critical role in inflammatory cell infiltration, cytokine production, and neovascularization in infarcted hearts.
Patients With Type 3 Severe von Willebrand Disease Are Not Protected Against Atherosclerosis
Authors: Šrámek A. , Bucciarelli P. , Federici A.B. , Mannucci P.M. , De Rosa V. , Castaman G. , Morfini M. , Mazzucconi M.G. , Rocino A. , Schiavoni M. , Scaraggi F.A. , Reiber J.H.C. , Rosendaal F.R.
Published: February 2004

Background—

The results of a number of studies in pigs and mice suggest that absence of von Willebrand factor (vWF) protects against the development of atherosclerosis. We studied whether patients with a complete deficiency of vWF (type 3 von Willebrand disease [vWD]) develop fewer atherosclerotic vessel wall changes than healthy controls.

Methods and Results—

This study included 47 individuals with type 3 vWD and 84 healthy controls. Early atherosclerotic changes were assessed by measuring the thickness of the intima-media in the carotid and femoral arteries by B-mode ultrasonography. Advanced atherosclerotic changes were quantified by summing the maximal thickness of atherosclerotic plaques in the carotid and femoral arteries and were expressed as a plaque score. Established risk factors were determined to adjust for possible differences between the groups. We found no substantial difference in intima-media thickness between vWD patients and controls (adjusted difference for carotid artery 0.007 mm, 95% CI −0.022 to 0.036 mm; femoral artery 0.069 mm, 95% CI −0.056 to 0.19 mm). Similar proportions of patients and controls had atherosclerotic plaques (19% and 17%, respectively). No difference was found in the plaque score between groups (adjusted difference −0.22 mm, 95% CI −0.69 to 0.26). Among vWD patients, we found no effect of treatment with vWF concentrates on intima-media thickness or plaque score.

Conclusions—

The results of this study indicate that vWF does not play a substantial role in human atherogenesis.
Therapy of Ischemic Cardiomyopathy With the Immunomodulating Agent Pentoxifylline
Authors: Sliwa Karen , Woodiwiss Angela , Kone Veuvette N. , Candy Geoffrey , Badenhorst Danelle , Norton Gavin , Zambakides Chris , Peters Ferande , Essop Rafique
Published: February 2004

Background—

Inflammatory immune activation commonly occurs in heart failure and may perpetuate this syndrome. We sought to determine whether the immunomodulating agent pentoxifylline enhances left ventricular function in patients with ischemic cardiomyopathy. We also investigated the effect of therapy on levels of brain natriuretic peptide (NT-pro BNP), C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), and the marker of apoptosis, Fas/Apo-1.

Methods and Results—

In a single-center, prospective, randomized, double-blind, placebo-controlled study, 38 patients with ischemic cardiomyopathy received pentoxifylline 400 mg TID or placebo in addition to standard therapy. Clinical assessment, radionuclide ventriculography, echocardiography, and blood analyses were performed at baseline and after 6 months. There were no differences in baseline characteristics between the groups. Five patients died (4 in the placebo group). Pentoxifylline treatment resulted in an improvement in functional class (
P
<0.005) and an increase in systolic blood pressure (
P
<0.05) and left ventricular radionuclide ejection fraction (
P
<0.05) compared with the placebo-treated group. There were reductions in plasma concentrations of CRP, NT-pro BNP, TNF-α, and Fas/Apo-1 in the pentoxifylline compared with the placebo-treated group.

Conclusions—

In patients with heart failure due to ischemic left ventricular dysfunction, the addition of pentoxifylline to standard therapy results in improvements in clinical status and radionuclide ejection fraction, which are accompanied by reductions in plasma markers of inflammation, prognosis, and apoptosis.
Single-Wire Pressure and Flow Velocity Measurement to Quantify Coronary Stenosis Hemodynamics and Effects of Percutaneous Interventions
Authors: Siebes Maria , Verhoeff Bart-Jan , Meuwissen Martijn , de Winter Robbert J. , Spaan Jos A.E. , Piek Jan J.
Published: February 2004

Background—

Lack of high-fidelity simultaneous measurements of pressure and flow velocity distal to a coronary artery stenosis has hampered the study of stenosis pressure drop–velocity (ΔP-v) relationships in patients.

Methods and Results—

A novel 0.014-inch dual-sensor (pressure and Doppler velocity) guidewire was used in 15 coronary lesions to obtain per-beat averages of pressure drop and velocity after an intracoronary bolus of adenosine. ΔP-v relations from resting to maximal hyperemic velocity were constructed before and after stepwise executed percutaneous coronary intervention (PCI). Before PCI, half of the ΔP-v relations revealed the presence of a compliant stenosis, which was stabilized by angioplasty. Fractional flow reserve (FFR), coronary flow reserve (CFVR), and velocity-based indices of stenosis resistance (h-SRv) and microvascular resistance (h-MRv) at maximal hyperemia were compared. Stepwise PCI significantly lowered h-SRv, with an initial marked reduction in hyperemic pressure drop followed by further gains in velocity. A concomitant significant reduction of h-MRv accounted for half of the gain in velocity after PCI. The average magnitude of absolute incremental hemodynamic changes was highest for h-SRv (56.8±39.2%) compared with CFVR (35.3±34.5%,
P
<0.005) or FFR (19.5±25.2%,
P
<0.0001).

Conclusions—

ΔP-v relations comprehensively visualize improvements in coronary hemodynamics after PCI. h-SRv is a powerful and sensitive descriptor of the functional gain achieved by PCI, combining information about both pressure gradient and velocity, which are oppositely affected by PCI. Simultaneous assessment of stenosis and microvascular resistance may provide a valuable tool for guidance of PCI.
Reduced Myocardial Ischemia-Reperfusion Injury in Toll-Like Receptor 4-Deficient Mice
Authors: Oyama Jun-ichi , Blais Charles , Liu Xiaoli , Pu Minying , Kobzik Lester , Kelly Ralph A. , Bourcier Todd
Published: February 2004

Background—

Myocardial ischemia and reperfusion-induced tissue injury involve a robust inflammatory response, but the proximal events in reperfusion injury remain incompletely defined. Toll-like receptor 4 (TLR4) is a proximal signaling receptor in innate immune responses to lipopolysaccharide of Gram-negative pathogens. TLR4 is also expressed in the heart and vasculature, but a role for TLR4 in the myocardial response to injury separate from microbial pathogens has not been examined. This study assessed the role of TLR4 in myocardial infarction and inflammation in a murine model of ischemia-reperfusion injury.

Methods and Results—

Myocardial ischemia-reperfusion (MIR) was performed on 2 strains of TLR4-deficient mice (C57/BL10 ScCr and C3H/HeJ) and controls (C57/BL10 ScSn and C3H/OuJ). Mice were subjected to 1 hour of coronary ligation, followed by 24 hours of reperfusion. TLR4-deficient mice sustained significantly smaller infarctions compared with control mice given similar areas at risk. Fewer neutrophils infiltrated the myocardium of TLR4-deficient Cr mice after MIR, indicated by less myeloperoxidase activity and fewer CD45/GR1-positive cells. The myocardium of TLR4-deficient Cr mice contained fewer lipid peroxides and less complement deposition compared with control mice after MIR. Serum levels of interleukin-12, interferon-γ, and endotoxin were not increased after ischemia-reperfusion. Neutrophil trafficking in the peritoneum was similar in all strains after injection of thioglycollate.

Conclusions—

TLR4-deficient mice sustain smaller infarctions and exhibit less inflammation after myocardial ischemia-reperfusion injury. The data suggest that in addition to its role in innate immune responses, TLR4 serves a proinflammatory role in murine myocardial ischemia-reperfusion injury.
Pitavastatin Downregulates Expression of the Macrophage Type B Scavenger Receptor, CD36
Authors: Han Jihong , Zhou Xiaoye , Yokoyama Toru , Hajjar David P. , Gotto Antonio M. , Nicholson Andrew C.
Published: February 2004

Background—

Pitavastatin (NK-104) is a novel inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, the rate-limiting enzyme for cholesterol biosynthesis. In clinical trials, pitavastatin has been shown to significantly decrease serum LDL cholesterol and triglyceride levels and increase HDL cholesterol. Scavenger receptor-mediated accumulation of oxidized LDL (OxLDL)-derived cholesteryl ester is considered to be a critical step in the development of atherosclerotic foam cell formation. We studied the effect of pitavastatin on CD36 (a class B scavenger receptor) expression by murine macrophages.

Methods and Results—

Treatment of J774 cells and murine peritoneal macrophages with pitavastatin decreased CD36 mRNA expression in a dose-dependent manner. Decreased CD36 mRNA was associated with decreased CD36 cell surface protein expression in human THP-1 cells and human monocyte-derived macrophages. Pitavastatin also reduced the increase in CD36 mRNA, cell surface protein, and binding/uptake of OxLDL induced by peroxisome proliferator-activated receptor-γ (PPARγ) ligands and/or OxLDL. Pitavastatin did not alter the half-life of CD36 mRNA, which suggests pitavastatin downregulates CD36 expression by reducing CD36 transcription. In addition, pitavastatin significantly decreased PPARγ mRNA and protein expression. Finally, pitavastatin increased p44/42 mitogen-activated protein kinase activity and PPARγ phosphorylation and increased the ratio of phosphorylated PPARγ to nonphosphorylated PPARγ.

Conclusions—

The present data demonstrate that pitavastatin prevents OxLDL uptake by macrophages through PPARγ-dependent inhibition of CD36 expression and suggest that pitavastatin could modulate CD36-mediated atherosclerotic foam cell formation.
Intramuscular Gene Transfer of Soluble Tumor Necrosis Factor-α Receptor 1 Activates Vascular Endothelial Growth Factor Receptor and Accelerates Angiogenesis in a Rat Model of Hindlimb Ischemia
Authors: Sugano Masahiro , Tsuchida Keiko , Makino Naoki
Published: February 2004

Background—

In a pathological setting, tumor necrosis factor (TNF)-α inhibits the proliferative response of endothelial cells through inactivation of receptors for vascular endothelial growth factor (VEGF). Soluble TNF-α receptor 1 (sTNFR1) is an extracellular domain of TNFR1 and an antagonist to TNF-α. In the present study, we examined the effect of sTNFR1 expression plasmid on receptor for VEGF (KDR/flk-1) and angiogenesis in a rat model of hindlimb ischemia.

Methods and Results—

The left femoral artery was exposed and excised to induce limb ischemia. A total of 400 μg of sTNFR1 or LacZ plasmid was injected into 3 different sites of the adductor muscle immediately after the induction of ischemia. TNF-α bioactivity in ischemic adductors increased in rats receiving LacZ plasmid compared with sham-operated rats. However, sTNFR1 plasmid significantly suppressed the increase in TNF-α bioactivity. KDR/flk-1 mRNA and tyrosine phosphorylation of KDR/flk-1 were significantly increased in the muscles injected with sTNFR1 plasmid compared with those injected with LacZ plasmid. VEGF increased both in muscles injected with sTNFR1 plasmid and in muscles injected with LacZ plasmid but did not differ significantly between them. At 21 days after the induction of ischemia, the sTNFR1 plasmid-transfected muscles showed significantly increased capillary density compared with LacZ plasmid-transfected muscles.

Conclusions—

In a rat model of hindlimb ischemia, VEGF increased but activation of KDR/flk-1 was suppressed, possibly by TNF-α, which might impair angiogenesis. Suppression of TNF-α with sTNFR1 plasmid upregulated KDR/flk-1 and accelerated angiogenesis. Local transfection of the sTNFR1 gene can be a new strategy for therapeutic angiogenesis in peripheral ischemic diseases.
Relationship of High and Low Ankle Brachial Index to All-Cause and Cardiovascular Disease Mortality
Authors: Resnick Helaine E. , Lindsay Robert S. , McDermott Mary McGrae , Devereux Richard B. , Jones Kristina L. , Fabsitz Richard R. , Howard Barbara V.
Published: February 2004

Background—

The associations of low (<0.90) and high (>1.40) ankle brachial index (ABI) with risk of all-cause and cardiovascular disease (CVD) mortality have not been examined in a population-based setting.

Methods and Results—

We examined all-cause and CVD mortality in relation to low and high ABI in 4393 American Indians in the Strong Heart Study. Participants had bilateral ABI measurements at baseline and were followed up for 8.3±2.2 years (36 589 person-years). Cox regression was used to quantify mortality rates among participants with high and low ABI relative to those with normal ABI (0.90 ≤ABI ≤1.40). Death from all causes occurred in 1022 participants (23.3%; 27.9 deaths per 1000 person-years), and of these, 272 (26.6%; 7.4 deaths per 1000 person-years) were attributable to CVD. Low ABI was present in 216 participants (4.9%), and high ABI occurred in 404 (9.2%). Diabetes, albuminuria, and hypertension occurred with greater frequency among persons with low (60.2%, 44.4%, and 50.1%) and high (67.8%, 49.9%, and 45.1%) ABI compared with those with normal ABI (44.4%, 26.9%, and 36.5%), respectively (
P
<0.0001). Adjusted risk estimates for all-cause mortality were 1.69 (1.34 to 2.14) for low and 1.77 (1.48 to 2.13) for high ABI, and estimates for CVD mortality were 2.52 (1.74 to 3.64) for low and 2.09 (1.49 to 2.94) for high ABI.

Conclusions—

The association between high ABI and mortality was similar to that of low ABI and mortality, highlighting a U-shaped association between this noninvasive measure of peripheral arterial disease and mortality risk. Our data suggest that the upper limit of normal ABI should not exceed 1.40.