Pascal Rousseau

Microvascular endothelial function in obstructive sleep apnea: Impact of continuous positive airway pressure and mandibular advancement.

OBJECTIVESnEndothelial dysfunction has been proposed as a potential mechanism implicated in the pathogenesis of cardiovascular complications of obstructive sleep apnea syndrome (OSAS). This study aimed to evaluate the microvascular endothelial function (MVEV) in OSAS and the impact on MVEF of 2 months of treatment with continuous positive airway pressure (CPAP) and mandibular advancement device (MAD).nnnMETHODSnMicrovascular reactivity was assessed using laser Doppler flowmetry combined with acetylcholine (Ach) and sodium nitroprusside (SNP) iontophoresis in 24 OSAS patients and 9 control patients. In 12 of the 24 OSAS patients, microvascular reactivity was reassessed after 2 months of CPAP and MAD using a randomized cross-over design.nnnRESULTSnAch-induced vasodilation was significantly lower in OSAS patients than in matched controls and correlated negatively with apnea hypopnea index (r=-0.49, p<0.025) and nocturnal oxygen desaturations (r=-0.63, p<0.002). Ach-induced vasodilation increased significantly with both CPAP and MAD. The increase in Ach-induced vasodilation under OSAS treatment correlated with the decrease in nocturnal oxygen desaturations (r=0.48, p=0.016).nnnCONCLUSIONnOur study shows an impairment of MVEF in OSAS related to OSAS severity. Both CPAP and MAD treatments were associated with an improvement in MVEF that could contribute to improve cardiovascular outcome in OSAS patients.

Increasing the "region of interest" and "time of interest", both reduce the variability of blood flow measurements using laser speckle contrast imaging.

OBJECTIVEnBoth spatial variability and temporal variability of skin blood flow are high. Laser speckle contrast imagers (LSCI) allow non-contact, real-time recording of cutaneous blood flow on large skin surfaces. Thereafter, the observer can define different sizes for the region of interest (ROI) in the images to decrease spatial variability and different durations over which the blood flow values are averaged (time of interest, TOI) to decrease temporal variability. We aimed to evaluate the impact of the choices of ROI and TOI on the analysis of rest blood flow and post occlusive reactive hyperemia (PORH).nnnMETHODSnCutaneous blood flow (CBF) was assessed at rest and during PORH. Three different sizes of ROI (1mm(2), 10mm(2) and 100mm(2)), and three different TOI (CBF averaged over 1s, 15s, and 30s for rest, and over 1s, 5s and 10s for PORH peak) were evaluated. Inter-subjects and intra-subjects coefficient of variations (inter-CV and intra-CV) were studied.nnnRESULTSnThe inter-subject variability of CBF is about 25% at rest and is moderately improved when the size of the ROI increases (inter-CV=31%, for 1s and 1mm(2) versus inter-CV=23%, for 15s and 100mm(2)). However, increasing the TOI does not improve the results. The variability of the PORH peak is lower with an inter-CV varying between 11.4% (10s and 100mm(2)) and 21.6% (5s and 1mm(2)). The lowest intra-CV for the CBF at rest was 7.3% (TOI of 15s on a ROI of 100mm(2)) and was 3.1% for the PORH peak (TOI of 10s on a ROI of 100mm(2)).nnnCONCLUSIONnWe suggest that a size of ROI larger than 10mm(2) and a TOI longer than 1s are required to reduce the variability of CBF measurements both at rest and during PORH peak evaluations at the forearm level. Many technical aspects such as comparison of laser speckle contrast imaging and laser Doppler imaging or the effect of skin to head distance on recorded values with LCSI are required to improve future studies using this fascinating clinical tool.

Laser speckle contrast imaging accurately measures blood flow over moving skin surfaces

Cutaneous blood flow (CBF) can be assessed non-invasively with lasers. Unfortunately, movement artefacts in the laser skin signal (LS(sk)) might sometimes compromise the interpretation of the data. To date, no method is available to remove movement artefacts point-by-point. Using a laser speckle contrast imager, we simultaneously recorded LS(sk) and the signal backscattered from an adjacent opaque surface (LS(os)). The completion of a first protocol allowed a definition of a simple equation to calculate the CBF from movement artefact-affected traces of LS(sk) and LS(os). We then recorded LS(sk) and LS(os) before, during and for 5 min after the tourniquet ischemia, both when subjects (n=8) were immobile or submitted to external passive movements of random intensity throughout the test. The typical post-occlusive reactive hyperemia trace was not identifiable within the LS(sk) recordings, with LS(sk) being 2 to 3 times higher during movements than in the immobile situation. After the calculation of CBF, traces in the immobile versus movement conditions were comparable, with the r cross-correlation coefficient being 0.930+/-0.010. Our method might facilitate future investigations in microvascular physiology and pathophysiology, specifically in subjects who have frequent or continuous involuntary movements.

Aging-Associated Sensory Neuropathy Alters Pressure-Induced Vasodilation in Humans

Healthy skin is protected from pressure-induced ischemic damage because of the presence of pressure-induced vasodilation (PIV). PIV relies on small sensory nerve fibers and endothelial function. Since aging alters both nervous and vascular functions, we hypothesized that PIV is altered with aging. We compared PIV in non-neuropathic and neuropathic older subjects (60-75 years) with that of young subjects (20-35 years). Laser Doppler flowmetry was used to evaluate the cutaneous responses to local pressure application, acetylcholine, and local heating. Quantitative sensory tests were used to evaluate sensory-nerve-fiber function. The non-neuropathic older subjects had an impaired PIV (12+/-7% increase in blood flow with pressure) compared with young subjects (62+/-4%, P<0.001). In the presence of peripheral neuropathy, the older subjects were totally deprived of PIV, leading to early pressure-induced cutaneous ischemia (-31+/-10%, P<0.001). This inability of the skin to adapt to localized pressure in older subjects is related to the severity of the sensory-fiber dysfunction rather than to endothelial dysfunction, which was comparable between the non-neuropathic (141+/-19% increased blood flow with acetylcholine, P<0.05) and neuropathic older subjects (145+/-28% increase, P<0.05) compared with young subjects (234+/-25% increase).

Distance between laser head and skin does not influence skin blood flow values recorded by laser speckle imaging.

BACKGROUNDnLaser Speckle contrast imaging (LSCI) allows non-contact, real-time recording of cutaneous blood flow (CBF). Different distances from laser-head to skin (distances(L-S)) can be chosen by the operator to perform these recordings. We aimed to evaluate the impact of different Distances(L-S) on the analysis of rest blood flow and post-occlusive reactive hyperemia (PORH).nnnMETHODSnFour distances(L-S) (10, 15, 20, and 30 cm) were evaluated in a random order in 11 healthy subjects. We analyzed the concordance between each recording at each distance(L-S). We compared CBF results (absolute values and cutaneous vascular conductance (CBF divided by mean arterial pressure)) obtained for each distance(L-S). The intra-subject coefficients of variation due to distances(L-S) (intra-CV, in%) were also studied.nnnRESULTSnThe mean r (standard deviation) cross-correlation coefficient was 0.99 (0.00) between each CBF trace issued from different distance(L-S). Both kinds of CBF results, at rest and for PORH peak, show non-significant differences when the distance(L-S) is modified. The intra-CV varies from 5.9% to 8.6% at rest and from 5.6% to 9.1% for the PORH peak.nnnCONCLUSIONnDistance(L-S) neither influences SBFR at rest, nor at peak post-occlusive hyperemia in the 10-30 cm interval using LSCI.

Multifractality, sample entropy, and wavelet analyses for age‐related changes in the peripheral cardiovascular system: Preliminary results

Using signal processing measures we evaluate the effect of aging on the peripheral cardiovascular system. Laser Doppler flowmetry (LDF) signals, reflecting the microvascular perfusion, are recorded on the forearm of 27 healthy subjects between 20-30, 40-50, or 60-70 years old. Wavelet-based representations, Hölder exponents, and sample entropy values are computed for each time series. The results indicate a possible modification of the peripheral cardiovascular system with aging. Thus, the endothelial-related metabolic activity decreases, but not significantly, with aging. Furthermore, LDF signals are more monofractal for elderly subjects than for young people for whom LDF signals are weakly multifractal: the average range of Holder exponents computed with a parametric generalized quadratic variation based estimation method is 0.13 for subjects between 20 and 30 years old and 0.06 for subjects between 60 and 70 years old. Moreover, the average mean sample entropy value of LDF signals slightly decreases with age: it is 1.34 for subjects between 20 and 30 years old and 1.19 for subjects between 60 and 70 years old. Our results could assist in gaining knowledge on the relationship between microvascular system status and age and could also lead to a more accurate age-related nonlinear modeling.

Reproducibility of non-invasive assessment of skin endothelial function using laser Doppler flowmetry and laser speckle contrast imaging.

Background Endothelial dysfunction precedes atherosclerosis. Vasodilation induced by acetylcholine (ACh) is a specific test of endothelial function. Reproducibility of laser techniques such as laser-Doppler-flowmetry (LDF) and Laser-speckle-contrast-imaging (LSCI) to detect ACh vasodilation is debated and results expressions lack standardization. We aimed to study at a 7-day interval (i) the inter-subject reproducibility, (ii) the intra-subjects reproducibility, and (iii) the effect of the results expressions over variability. Methods and Results Using LDF and LSCI simultaneously, we performed two different ACh-iontophoresis protocols. The maximal ACh vasodilation (peak-ACh) was expressed as absolute or normalized flow or conductance values. Inter-subject reproducibility was expressed as coefficient of variation (inter-CV,%). Intra-subject reproducibility was expressed as within subject coefficients of variation (intra-CV,%), and intra-class correlation coefficients (ICC). Fifteen healthy subjects were included. The inter-subject reproducibility of peak-ACh depended upon the expression of the results and ranged from 55% to 162% for LDF and from 17% to 83% for LSCI. The intra-subject reproducibility (intra-CV/ICC) of peak-ACh was reduced when assessed with LSCI compared to LDF no matter how the results were expressed and whatever the protocol used. The highest intra-subject reproducibility was found using LSCI. It was 18.7%/0.87 for a single current stimulation (expressed as cutaneous vascular conductance) and 11.4%/0.61 for multiple current stimulations (expressed as absolute value). Conclusion ACh-iontophoresis coupled with LSCI is a promising test to assess endothelial function because it is reproducible, safe, and non-invasive. N°: NCT01664572.

Air movements interfere with laser speckle contrast imaging recordings.

Laser speckle contrast imaging (LSCI) allows the noncontact monitoring of changes in cutaneous blood flow (CBF) [1, 2]. Recent evidence shows that the LSCI technique dramatically reduces the variability in clinical measurements as compared to laser Doppler, making the technique a fascinating tool to facilitate microvascular studies in clinical routine [2, 3]. LSCI records CBF using a camera placed at a distance ranging from 10 cm to 30 cm from the area of interest of the skin [4, 5]. As such, we hypothesized that movements of air particulate matter between the skin and the laser head could interfere with the LSCI backscattered signal during microvascular tests. This is of particular interest because microvascular studies are generally performed in air-conditioned rooms and the fan of the air-conditioner might induce rapid air movements. Further, an open window, or displacement of objects (door opening) or personnel in the room are a few of the events that could induce air movements. We tested the influence of air movements induced by a fan with different CBF levels: at rest, during ischaemia, and during postocclusive reactive hyperaemia (PORH).

Central Role of P2Y6 UDP Receptor in Arteriolar Myogenic Tone

Objective—Myogenic tone (MT) of resistance arteries ensures autoregulation of blood flow in organs and relies on the intrinsic property of smooth muscle to contract in response to stretch. Nucleotides released by mechanical strain on cells are responsible for pleiotropic vascular effects, including vasoconstriction. Here, we evaluated the contribution of extracellular nucleotides to MT. Approach and Results—We measured MT and the associated pathway in mouse mesenteric resistance arteries using arteriography for small arteries and molecular biology. Of the P2 receptors in mouse mesenteric resistance arteries, mRNA expression of P2X1 and P2Y6 was dominant. P2Y6 fully sustained UDP/UTP-induced contraction (abrogated in P2ry6−/− arteries). Preventing nucleotide hydrolysis with the ectonucleotidase inhibitor ARL67156 enhanced pressure-induced MT by 20%, whereas P2Y6 receptor blockade blunted MT in mouse mesenteric resistance arteries and human subcutaneous arteries. Despite normal hemodynamic parameters, P2ry6−/− mice were protected against MT elevation in myocardial infarction–induced heart failure. Although both P2Y6 and P2Y2 receptors contributed to calcium mobilization, P2Y6 activation was mandatory for RhoA–GTP binding, myosin light chain, P42–P44, and c-Jun N-terminal kinase phosphorylation in arterial smooth muscle cells. In accordance with the opening of a nucleotide conduit in pressurized arteries, MT was altered by hemichannel pharmacological inhibitors and impaired in Cx43+/− and P2rx7−/− mesenteric resistance arteries. Conclusions—Signaling through P2 nucleotide receptors contributes to MT. This mechanism encompasses the release of nucleotides coupled to specific autocrine/paracrine activation of the uracil nucleotide P2Y6 receptor and may contribute to impaired tissue perfusion in cardiovascular diseases.

Platelet inhibition by low-dose aspirin but not by clopidogrel reduces the axon-reflex current-induced vasodilation in humans.

We previously showed a prolonged inhibition of current-induced vasodilation (CIV) after a single oral high dose of aspirin. In this study, we tested the hypothesis of platelet involvement in CIV. Nine healthy volunteers took 75 mg aspirin/day, 98 mg of clopidogrel bisulfate/day, or placebo for 4 days. CIV was induced by two consecutive 1-min anodal current applications (0.08 mA/cm(2)) through deionized water with a 10-min interval. CIV was measured with laser Doppler flowmetry and expressed as a percentage of baseline cutaneous vascular conductance: %C(b). In a second experiment in 10 volunteers, aspirin and placebo were given as in experiment 1, but a 26-h delay from the last aspirin intake elapsed before ACh iontophoresis and postocclusive hyperemia were studied in parallel to CIV. In experiment 1, the means +/- SE amplitude of CIV was 822 +/- 314, 313 +/- 144, and 746 +/- 397%C(b) with placebo, aspirin (P < 0.05 from placebo and clopidogrel), and clopidogrel (NS from placebo), respectively. In experiment 2, CIV impairment with aspirin was confirmed: CIV amplitudes were 300 +/- 99, and 916 +/- 528%C(b) under aspirin and placebo, respectively (P < 0.05), whereas vasodilation to ACh iontophoresis (322 +/- 74 and 365 +/- 104%C(b)) and peak postocclusive hyperemia (491 +/- 137 and 661 +/- 248%C(b)) were not different between aspirin and placebo, respectively. Low-dose aspirin, even 26 h after oral administration, impairs CIV, while ACh-mediated vasodilation and postocclusive hyperemia are preserved. If platelets are involved in the neurovascular mechanism triggered by galvanic current application in humans, it is likely to occur through the cyclooxygenase but not the ADP pathway.