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Biomedical subjects

D Choquet

Publications and source records attributed to D Choquet.

At least 19 recordsLinked to original sources
PubMed2005

Noninvasive assessment of the tension-time index of inspiratory muscles at rest in obese male subjects.

OBJECTIVE: The aim of this study was to investigate the effect of excessive mechanical load caused by obesity on the inspiratory muscle performance in obese men at rest. METHODS: We therefore measure at rest spirometric flows and the noninvasive tension time index of inspiratory muscle (TTmus = PI/PImax x TI/TTOT) in eight obese male subjects (body mass index (BMI) > 30) and 10 controls. RESULTS: Spirometric flow (FEV1% pred, FVC% pred) and maximal inspiratory pressure (PImax) were significantly lower in obese subjects compared to controls (P < 0.001). The mean TTmus was significantly higher in obese subjects than in controls (0.136 +/- 0.003 vs 0.045 +/- 0.01). The increase in TTmus was primarily due to an increase in the ratio of mean inspiratory pressure to maximal inspiratory pressure (PI/PImax) and the duty cycle (TI/TTOT). We found a significant negative relationship between PImax and BMI (r = -0.74, P < 0.001), a positive correlation between TTmus and BMI (r = 0.80, P < 0.001) and a negative correlation between TTmus and forced expiratory volume in 1 s (r = -0.85, P < 0.001). CONCLUSION: Excessive mechanical load caused by obesity imposes a great burden on the inspiratory muscle, which may predispose such subjects to respiratory muscle weakness at rest.

Adult↗
PubMed2003

Single metallic nanoparticle imaging for protein detection in cells.

We performed a visualization of membrane proteins labeled with 10-nm gold nanoparticles in cells, using an all-optical method based on photothermal interference contrast. The high sensitivity of the method and the stability of the signals allows 3D imaging of individual nanoparticles without the drawbacks of photobleaching and blinking inherent to fluorescent markers. A simple analytical model is derived to account for the measurements of the signal amplitude and the spatial resolution. The photothermal interference contrast method provides an efficient, reproducible, and promising way to visualize low amounts of proteins in cells by optical means.

Animals↗
PubMed2003

Aerobic capacity and peripheral skeletal muscle function in coronary artery disease male patients.

The aim of this study was to determine if the diminished aerobic capacity of coronary artery disease male (CAD) patients is accompanied by an impaired skeletal muscle function compared to healthy control subjects. Thirteen CAD patients and 9 healthy subjects performed both a maximal laboratory exercise testing and an assessment of the peripheral skeletal muscle function on an isokinetic apparatus. The cardiorespiratory and mechanical parameters were measured at ventilatory threshold and at maximal effort during a maximal exercise testing. The peripheral skeletal muscle function of the quadriceps was assessed from the maximal voluntary isometric force (MVIF) and from the static endurance time (SET) at an intensity of 50 % of the MVIF. The CAD patients showed a diminished aerobic capacity compared to healthy control subjects at maximal effort (maximal VO(2) uptake: p < 0.0001, maximal ventilation: p < 0.01; maximal heart rate: p < 0.0001, maximal power: p < 0.001) but also at VT (VO(2) uptake VT: p < 0.0001, Power VT: p < 0.001). No difference was found on the MVIF (p < 0.90) between the CAD patients and the control subjects whereas the SET was lower in the CAD patients (p < 0.01). The CAD patients had a lower aerobic capacity and an impaired skeletal muscle endurance compared to healthy subjects.

Anthropometry↗
PubMed2001

On the elastic properties of tetramethylrhodamine F-actin.

(Iodoacetamido)tetramethylrhodamine disrupts F-actin. At the 1:1 fluorophore to actin (as monomer) ratio approximately 80% of the protein becomes non-sedimentable. The fluorescent, non-sedimentable actin copolymerizes with G-actin to yield fluorescent filaments. The tensile strength of these filaments changes with the ratio of the fluorescent non-sedimentable actin to the G-actin, being 1.6 pN, 2.9 pN and 3.6 pN at the 1/4, 2/3 and 1/1 ratios, respectively. These tensile strengths are approximately two orders of magnitude lower than those obtained by decoration of F-actin with phalloidin.

Actins↗
PubMed2001

Fast and reversible trapping of surface glycine receptors by gephyrin.

Variations in receptor number at a given synapse are known to contribute to synaptic plasticity, but methods used to establish this idea usually do not allow for the determination of the dynamics of these phenomena. We used single-particle tracking to follow in real time, on the cell surface, movements of the glycine receptor (GlyR) with or without the GlyR stabilizing protein gephyrin. GlyR alternated within seconds between diffusive and confined states. In the absence of gephyrin, GlyR were mostly freely diffusing. Gephyrin induced long confinement periods spatially associated with submembranous clusters of gephyrin. However, even when most receptors were stabilized, they still frequently made transitions through the diffusive state. These data show that receptor number in a cluster results from a dynamic equilibrium between the pools of stabilized and freely mobile receptors. Modification of this equilibrium could be involved in regulation of the number of receptors at synapses.

Animals↗
PubMed2000

Effects of aging on cardiorespiratory responses to brief and intense intermittent exercise in endurance-trained athletes.

The aim of this study was to investigate the effects of aging on athletes' cardiorespiratory responses to a brief intense intermittent effort, using the force-velocity test as an exercise model. Twelve young athletes (24.8 +/- 1.3 years) and twelve master athletes (65.1 +/- 1.2 years) with similar heights, body masses, and endurance training schedules participated in this study. They performed both a maximal graded exercise and the force-velocity tests. The force-velocity test consisted of the repetition of 6-second sprints against increasing braking forces with 5-minute recovery periods. None of the subjects presented abnormal electrocardiogram responses to the tests. During the force-velocity test, the heart rate magnitudes of response in all subjects were correlated to the corresponding sprint power output (p < .001), with higher values for the young athletes (p < .001). Both groups had similar systolic blood pressure peaks of response during the force-velocity test. Both groups had similar preexercise and end-of-recovery oxygen consumption (VO2), but the young athletes had higher peaks of response (p < .001). The VO2 magnitudes of response increased during the test (p < .01) in all subjects, with higher values for the young athletes (p < .001). There was a positive correlation between the VO2 magnitude of response and (1) the corresponding sprint power output (R = .58,p < .001) and (2) the corresponding number of sprint repetitions (R = .29, p < .02). The young athletes had higher end-of-recovery and peak carbon dioxide production (VCO2) responses than the master athletes (p < .001). Pulmonary ventilation (V(E)) peaks of response to the sprints were higher in the young athletes (p < .001). There was a positive relation between the V(E) and VCO2 peaks of response (R = 84,p < .001). In both groups the peak heart rate, VO2, VCO2, and V(E) values attained during the force-velocity test represented similar percentages of the maximal values reached at exhaustion of maximal graded exercise. These results showed that aging does not alter the percentage of the cardiorespiratory response to a brief intense intermittent exercise such as the force-velocity test. Moreover, the arterial blood pressure response is not significantly altered, whereas the vasodilatatory response is.

Adolescent↗
PubMed1999

Rhodamine phalloidin F-actin: critical concentration versus tensile strength.

The mechanic and elastic properties of rhodamine phalloidin F-actin were investigated as a function of the ionic strength and in the absence of Mg2+. By increasing ionic strength from 3 to 19 mM, critical concentration decreased from 146 to 36 nM and the yield strength increased from 5.6 pN to 28.6 pN. At the ionic strength of 12-13 mM, the elastic modulus by stretching increased by 330-430 kP. nm-1 up to the break point, where it was 38-44.2 MP. The work required to break the filament, 403-439 kJ.M-1 provides an estimate of the free energy of annealing of rhodamine phalloidin F-actin, the annealing constant being 2.8 x 1074 M-1.

Actins↗
PubMed1999

Cell migration as a five-step cycle.

The migration of cells over substrata is a fundamental and critical function that requires the co-ordination of several cellular processes which operate in a cycle. At the level of the light microscope, the cycle can be divided into five steps: (1) extension of the leading edge; (2) adhesion to matrix contacts; (3) contraction of the cytoplasm; (4) release from contact sites; and (5) recycling of membrane receptors from the rear to the front of the cell. Each step is dependent upon one or more cyclical biochemical processes. The development of many in vitro and subcellular assays for the fundamental biochemical processes involved has increased our understanding of each cycle dramatically in the last several years to include a definition of many of the protein and enzymic components, the role of the position of extracellular-matrix receptors on the cell, and the contribution of physical force. The next generation of questions are directed at resolving the roles of the many individual proteins in each step of the cell migration process. In this chapter we will examine each of the migration steps and discuss the biochemical mechanisms that may underlie them.

Cell Adhesion↗
PubMed1998

[Transesophageal echocardiography before electric cardioversion for supraventricular arrhythmia].

OBJECTIVES: The aim of this prospective study was to assess the risks of electrical shock cardio-version in the treatment of supraventricular rhythm disorders when administered under effective-dose but short duration anticoagulation in patients with no intracavitary thrombus detectable by transesophageal echocardiography. PATIENTS AND METHODS: One hundred nineteen patients, mean age 66 years, with permanent arrhythmia due to atrial fibrillation (n = 102), atrial flutter (n = 16) or atrial tachycardia (n = 1) and taking no long-term anticoagulant therapy were treated by electrical shock cardioversion. The patients were given heparin at an effective dose 72 hours prior to cardioversion. A transthoracic and a transesophageal echocardiography were performed less than 24 hours prior to cardioversion. RESULTS: Twenty-one thrombi were evidenced in 16 patients (14.6%) including 18 in the left auricle, 1 in the left atrium and 2 in the right atrium. A spontaneous contrast was visualized in 38 patients (32%). Cardioversion was performed in 103 patients without thrombus and later in 9 of the 16 patients with thrombus after absorption under anticoagulant therapy as evidenced on the control transesophageal echocardiography. A sinus rhythm was obtained in 82% of the cases. All patients were given anti-vitamin K anticoagulants for one month. There were no clinical manifestation of ischemic vascular events during cardioversion nor during the one-month follow-up. CONCLUSION: Early use of electrical shock cardioversion in patients with supraventricular rhythm disorders can be proposed without long-term anticoagulation therapy if the absence of thrombi is demonstrated by transesophageal echocardiography and short-term heparin is given followed by oral anticoagulants for at least 4 weeks. A large-scale randomized prospective study comparing the conventional strategy with the protocol used in this study would be required to definitively validate this approach and determine its possible advantages.

Adult↗
PubMed1998

Effects of interval training at the ventilatory threshold on clinical and cardiorespiratory responses in elderly humans.

This study assessed clinical and cardiorespiratory responses after an interval training programme in sedentary elderly adults using the ventilatory threshold (Vth) as the index of exercise training intensity. A selection of 22 subjects were randomized into two groups: 11 subjects served as the training group (TG) and the others as controls (CG). Maximal exercise tests were performed on a treadmill before (T0), each month (T1, T2) and after the 3-month interval training programme period (T3). The TG subjects were individually trained at the heart rate corresponding to Vth measured at T0, T1 and T2 as the breakpoint in the oxygen uptake-carbon dioxide production relationship. Their training programme consisted of walking/jogging sessions on a running track twice a week. The sessions consisted of varying durations of exercise alternating with active recovery in such a way that the subjects slowly increased their total exercise time from an initial duration of 30 min to a final duration of 1 h. During training the heart rate was continuously monitored by a cardiofrequency meter. Compared with the daily activities of the controls, no training programme-related injuries were observed in TG. Moreover, programme adherence (73%) and attendance (97.3%) were high. The maximal oxygen uptake and Vth were increased in TG, by 20% (P<0.05) and 26% (P<0.01), respectively. Interval training at Vth also significantly increased maximal O2 pulse (P<0.05) and maximal ventilation (P<0.01). A significant decrease in submaximal ventilation (P<0.05) and heart rate (P<0.01) was also noted. These results would suggest that for untrained elderly adults, an interval training programme at the intensity of Vth may be well-tolerated clinically and may significantly improve both maximal aerobic power and submaximal exercise tolerance.

Adaptation, Physiological↗
PubMed1997

Extracellular matrix rigidity causes strengthening of integrin-cytoskeleton linkages.

To move forward, migrating cells must generate traction forces through surface receptors bound to extracellular matrix molecules coupled to a rigid structure. We investigated whether cells sample and respond to the rigidity of the anchoring matrix. Movement of beads coated with fibronectin or an anti-integrin antibody was restrained with an optical trap on fibroblasts to mimic extracellular attachment sites of different resistance. Cells precisely sense the restraining force on fibronectin beads and respond by a localized, proportional strengthening of the cytoskeleton linkages, allowing stronger force to be exerted on the integrins. This strengthening was absent or transient with antibody beads, but restored with soluble fibronectin. Hence, ligand binding site occupancy was required. Finally, phenylarsine oxide inhibited strengthening of cytoskeletal linkages, indicating a role for dephosphorylation. Thus, the strength of integrin-cytoskeleton linkages is dependent on matrix rigidity and on its biochemical composition. Matrix rigidity may, therefore, serve as a guidance cue in a process of mechanotaxis.

3T3 Cells↗
PubMed1996

Ligand binding regulates the directed movement of beta1 integrins on fibroblasts.

To enable cells to crawl, adhesion receptors such as integrins must bind to extracellular molecules and simultaneously interact with force-generating components of the cytoskeleton. We show here that the binding of extracellular ligand in living cells induces the attachment of beta1 integrins to the retrograde-moving cytoskeleton. Unliganded integrins are not associated with the rearward-moving cytoskeleton: gold particles attached to beta1 integrin by a monoclonal antibody diffuse in the membrane. However, addition of soluble RGD peptide (single-letter amino-acid code) or the use of fibronectin-coated gold particles causes the attachment of integrins to the rearward-moving cytoskeleton. Deletion of the beta1 cytoplasmic tail blocks cytoskeletal attachment. The directed movement of integrins in response to ligand indicates that ligand binding is the critical step in regulating organized receptor movement on the cell surface and the migration of adherent cells.

3T3 Cells↗
PubMed1996

Regulation of ITAM signaling by specific sequences in Ig-beta B cell antigen receptor subunit.

B cell antigen receptors (BCR) are composed of an antigen binding subunit, the membrane Ig, and Ig-alpha/Ig-beta heterodimers, that contain a transducing motif named ITAM for "immuno-receptor tyrosine-based activation motif." Ig-alpha and Ig-beta ITAMs only differ by four amino acids located before the second conserved tyrosine (DCSM in Ig-alpha and QTAT in Ig-beta), which determine the in vitro association of Ig-alpha with the src kinase fyn. We have previously shown that Ig-alpha and Ig-beta BCR subunits activate different signaling pathways by expressing, in B cells, FcgammaRII chimeras containing the cytoplasmic tails of Ig-alpha or Ig-beta. We report here that the signaling capacity of Ig-beta ITAM is regulated by peptide sequences located inside (QTAT region) or outside the ITAM (flanking sequences). Furthermore, when isolated, Ig-alpha and Ig-beta ITAM have similar abilities as the entire Ig-alpha tail and the whole BCR in triggering tyrosine kinase activation, an increase of intracellular calcium concentration as well as late events of cell activation as assessed by cytokine secretion. These data show that alterations that modify the ability of Ig-alpha and Ig-beta to interact in vitro with the src kinase fyn (switch between QTAT and DCSM) also determine signal transduction capabilities of these molecules expressed in B cells.

Amino Acid Sequence↗
PubMed1995

Chemical synthesis, structural and functional characterisation of noxiustoxin, a powerful blocker of lymphocyte voltage-dependent K+ channels.

Two forms of the Centrudoides noxius scorpion noxiustoxin, containing an amidated and an acid C-terminus, were synthesized on a solid support by using Fmoc-chemistry and 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU) coupling. Comparison of the two synthetic forms with the native toxin by tryptic mapping and CD spectroscopy shows that noxiustoxin possesses an amidated C-terminus and the same fold as all short scorpion toxins. Patch-clamp assays on B lymphocytes demonstrate that noxiustoxin inhibits the voltage-dependent K+ channels with 2 nM affinity, but does not affect the Ca(2+)-activated K+ channels. This toxin, because of its high affinity and specificity for voltage-gated K+ channel, may provide a powerful tool in the investigation of the role(s) of these channels in the T and B lymphocyte activation and proliferation.

Amino Acid Sequence↗
PubMed1995

A primary T-cell immunodeficiency associated with defective transmembrane calcium influx.

We investigated a T-cell activation deficiency in a 3-month-old boy with protracted diarrhea, serious cytomegalovirus pneumonia, and a family history (in a brother) of cytomegalovirus infection and toxoplasmosis. In spite of detection of normal number of peripheral lymphocytes, T cells did not proliferate after activation by anti-CD3 and anti-CD2 antibodies, although proliferation induced by antigens was detectable. We sought to determine the origin of this defect as it potentially represented a valuable tool to analyze T-cell physiology. T-cell activation by anti-CD3 antibody or phytohemagglutinin (PHA) led to reduced interleukin-2 (IL-2) production and abnormal nuclear factor-activated T cell (NF-AT; a complex regulating the IL-2 gene transcription) binding activity to a specific oligonucleotide. T-cell proliferation was restored by IL-2. Early events of T-cell activation, such as anti-CD3 antibody-induced cellular protein tyrosine phosphorylation, p59fyn and p56lck kinase activities, and phosphoinositide turnover, were found to be normal. In contrast, anti-CD3 antibody-induced Ca2+ flux was grossly abnormal. Release from endoplasmic reticulum stores was detectable as tested in the presence of anti-CD3 antibody or thapsigargin after cell membrane depolarization in a K+ rich medium, whereas extracellular entry of Ca2+ was defective. The latter abnormality was not secondary to defective K+ channel function, which was found to be normal. A similar defect was found in other hematopoietic cell lineages and in fibroblasts as evaluated by both cytometry and digital video imaging experiments at a single-cell level. This primary T-cell immunodeficiency appears, thus, to be due to defective Ca2+ entry through the plasma membrane. The same abnormality did not alter B-cell proliferation, platelet function, and polymorphonuclear neutrophil (PMN) function. Elucidation of the mechanism underlying this defect would help to understand the physiology of Ca2+ mobilization in T cells.

Antigens, CD↗
PubMed1994

The calcium current activated by T cell receptor and store depletion in human lymphocytes is absent in a primary immunodeficiency.

Stimulation of antigen receptors of lymphocytes triggers a transitory release of Ca2+ from internal stores and the opening of a transmembrane Ca2+ conductive pathway. The latter underlies the sustained increase of intracellular free calcium concentration, and it seems to be a key event in the Ca(2+)-dependent biochemical cascade leading to T cell proliferation. Alternatively, pharmacological depletion of internal stores by itself activates Ca2+ influx. This has led to the hypothesis that antigen-triggered Ca2+ influx is secondary to Ca2+ release from internal stores. However, the precise relationship between antigen and Ca2+ release-activated Ca2+ currents remains unclear, particularly since neither of them has been electrophysiologically recorded in normal lymphocytes. Using the whole-cell and the perforated configurations of the patch clamp technique on peripheral blood lymphocytes, we found that a low amplitude Ca(2+)-selective current was triggered when intracellular stores were depleted by stimuli such as the intracellular perfusion of inositol triphosphate or thapsigargin and the extracellular perfusion of ionomycin. A similar current was elicited by the cross-linking of the T cell receptor-CD3 complex. This current displayed an inward rectification below 0 mV and was completely blocked by the divalent cation Cd2+. It was very selective for Ca2+ over Na+ and insensitive to changes in chloride concentration. The physiological relevance of this conductance was investigated with the analysis of abnormal Ca2+ signaling in lymphocytes from a patient suffering from a primary immunodeficiency associated with a defective T cell proliferation. Using fura-2 video imaging, an absence of Ca2+ influx was established in the patient's lymphocytes, whereas the Ca2+ release from internal stores was normal. This was the case whether cells were stimulated physiologically through their antigen receptors or with store depleting pharmacological agents. Most importantly, no Ca(2+)-selective current was elicited in these cells. Our data strongly suggest that the Ca2+ release-activated current underlies the sustained Ca2+ influx during antigenic stimulation and that it plays a key role in the immune function.

Biological Transport↗
PubMed1994

Different patterns of calcium signaling triggered through two components of the B lymphocyte antigen receptor.

The engagement of the B cell antigen receptor is the first step of antigenic stimulation of B lymphocytes. This step is followed by a series of biochemical events, including the activation of protein-tyrosine kinases, phosphoinositide turnover, and multiple patterns of calcium mobilization, which lead to the regulation of gene transcription and cellular responses. The B cell antigen receptor complex is composed of membrane immunoglobulins (as antigen recognition subunits) and associated chains (Ig-alpha and Ig-beta) that couple the receptor to cytoplasmic protein kinases. To investigate independently the relative signaling capacity of Ig-alpha and Ig-beta, chimeric proteins containing their cytoplasmic domains were expressed in a B cell line. We found that Ig-alpha and Ig-beta activate two distinct intracellular signaling pathways. The engagement of Ig-alpha chimeras induces a complete release of calcium from intracellular stores, followed by transmembrane calcium influx and late cell activation signals, detected by lymphokine secretion. In contrast, Ig-beta chimeras do not induce lymphokine secretion or calcium influx, but induce short oscillatory release of calcium, dependent on the activity of the Ca-ATPase pump of the endoplasmic reticulum. These results provide a structural basis for the diversity of B cell responses.

B-Lymphocytes↗