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      Reproducibility of Cutaneous Microvascular Function Assessment Using Laser Doppler Flowmetry and Acetylcholine Iontophoresis

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          Abstract

          Aim: To investigate the reproducibility of direct and axon-reflex vasodilator responses to increasing doses of acetylcholine administered transcutaneously by iontophoresis. Methods: Increasing doses of acetylcholine ions were introduced into the skin of the volar forearm with weak electric currents. The elicited vasodilatation was measured with laser Doppler flowmetry at the direct site of acetylcholine administration and approximately 5 mm from this site (within the distribution of axon-reflex vasodilatation). To investigate the reproducibility of these measures, iontophoresis was carried out twice at one site after a 50 min interval and once at another site on the forearm of 16 healthy females. Raw scores and data expressed as percentages of baseline, maximum, and response to heating were analysed to determine the most reliable form. Results: The dose-response curve was significantly greater at the upper site on the forearm than at the lower site for direct vasodilator responses, but did not differ between the upper and lower forearm for axon-reflex responses. The various forms of data expression yielded weak to moderate correlations for vasodilatation between sites, and moderate to strong correlations when the iontophoresis was repeated at the same site. Direct responses were most reproducible when expressed in proportion to levels recorded after 5–6 min of local 42°C heating (percentage of shared variance was 54.6% within sites and 61.0% between sites). Conclusion: Cutaneous vasodilatation to acetylcholine iontophoresis is reproducible over time at a single site but varies across sites. This will allow future research to assess microvascular function before and after a laboratory manipulation in a single session.

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          Most cited references25

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          Skin blood flow in adult human thermoregulation: how it works, when it does not, and why.

          The thermoregulatory control of human skin blood flow is vital to the maintenance of normal body temperatures during challenges to thermal homeostasis. Sympathetic neural control of skin blood flow includes the noradrenergic vasoconstrictor system and a sympathetic active vasodilator system, the latter of which is responsible for 80% to 90% of the substantial cutaneous vasodilation that occurs with whole body heat stress. With body heating, the magnitude of skin vasodilation is striking: skin blood flow can reach 6 to 8 L/min during hyperthermia. Cutaneous sympathetic vasoconstrictor and vasodilator systems also participate in baroreflex control of blood pressure; this is particularly important during heat stress, when such a large percentage of cardiac output is directed to the skin. Local thermal control of cutaneous blood vessels also contributes importantly--local warming of the skin can cause maximal vasodilation in healthy humans and includes roles for both local sensory nerves and nitric oxide. Local cooling of the skin can decrease skin blood flow to minimal levels. During menopause, changes in reproductive hormone levels substantially alter thermoregulatory control of skin blood flow. This altered control might contribute to the occurrence of hot flashes. In type 2 diabetes mellitus, the ability of skin blood vessels to dilate is impaired. This impaired vasodilation likely contributes to the increased risk of heat illness in this patient population during exposure to elevated ambient temperatures. Raynaud phenomenon and erythromelalgia represent cutaneous microvascular disorders whose pathophysiology appears to relate to disorders of local and/or reflex thermoregulatory control of the skin circulation.
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            Methodological issues in the assessment of skin microvascular endothelial function in humans.

            The study of microvascular function can be performed in humans using laser Doppler flowmetry of the skin. This technology lends itself to a wide range of applications for studying the endothelial function of skin blood vessels. We review the advantages and limitations of postocclusive hyperemia, local thermal hyperemia, acetylcholine iontophoresis, flowmotion and association with microdialysis as tools with which to investigate skin microvascular endothelial function in humans. Postocclusive hyperemia, thermal hyperemia and acetylcholine iontophoresis provide integrated indexes of microvascular function rather than specific endothelial markers. However, they are valuable tools and can be used as surrogate endpoints in clinical trials in which the assessment of microvascular function in humans is required.
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              Early morning attenuation of endothelial function in healthy humans.

              Cardiovascular events such as myocardial infarction, sudden death, and stroke have a peak incidence in the early hours after waking. The mechanisms involved in this circadian variation are not clear. Endothelial dysfunction is associated with increased risk for cardiovascular events. We tested the hypothesis that endothelial function is reduced in the early morning, around the time of waking, compared with measurements obtained both before sleep and later in the day in healthy humans. We studied 30 subjects (19 men, 11 women; mean age, 41.6 years). All participants underwent polysomnography to exclude obstructive sleep apnea or other sleep disorders. Brachial artery flow-mediated endothelium-dependent vasodilation (FMD) and endothelium-independent dilation (non-FMD) were measured on 3 different occasions: before subjects went to sleep (9 PM), the next morning immediately after waking (6 AM), and during the late morning 5 hours after waking (11 AM). All subjects had normal sleep with good sleep efficiency of 84+/-2%. Compared with before sleep, FMD decreased markedly in the early morning after waking and recovered by late morning (9 pm, 7.5+/-1%; 6 am, 4.4+/-0.7%; 11 am, 7.7+/-1%; P=0.02). Non-FMD was similar for the 3 periods of observation (9 pm, 17.3+/-1.6%; 6 am, 17.2+/-1.3%; 11 am, 18.5+/-1.7%). FMD is blunted in the early morning in healthy subjects. Decreased endothelial function in the early morning may have implications for our understanding of the morning peak in cardiac and vascular events.
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                Author and article information

                Journal
                SPP
                Skin Pharmacol Physiol
                10.1159/issn.1660-5527
                Skin Pharmacology and Physiology
                S. Karger AG
                1660-5527
                1660-5535
                2009
                November 2009
                25 September 2009
                : 22
                : 6
                : 313-321
                Affiliations
                School of Psychology, Murdoch University, Perth, W.A., Australia
                Article
                241301 Skin Pharmacol Physiol 2009;22:313–321
                10.1159/000241301
                19786825
                695d40d3-19f8-462d-a834-0c27f7889136
                © 2009 S. Karger AG, Basel

                Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher. Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug. Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

                History
                : 16 February 2009
                : 10 July 2009
                Page count
                Figures: 3, Tables: 4, References: 33, Pages: 9
                Categories
                Original Paper

                Oncology & Radiotherapy,Pathology,Surgery,Dermatology,Pharmacology & Pharmaceutical medicine
                Dose-response curve,Laser Doppler flowmetry,Acetylcholine,Iontophoresis,Endothelium,Axon-reflex

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