Este trabalho foi apoiado por números de atribuição da subvenção R01HL096593 e K23HL72825 do Instituto Nacional do Coração, Pulmão e Sangue. O conteúdo é de responsabilidade exclusiva de Thomas Jefferson University e não representam, necessariamente, a posição oficial do Instituto Nacional do Coração, Pulmão e Sangue ou do Instituto Nacional de Saúde.

Apresentação de Caso (exigida, se aplicável): NA, este ainda é um procedimento de pesquisa experimental, ainda não utilizados clinicamente. Avaliação, Diagnóstico e Plano (obrigatório, se aplicável): NA, este ainda é um procedimento de pesquisa experimental, ainda não utilizados clinicamente. Procedimento (obrigatório): Esta parte deve incluir uma descrição passo-a-passo dos procedimentos relevantes, atendendo as orientações abaixo. 1. Microscopia capilar (Figura 1) <ol><li> A nossa técnica capilaroscopia foi adaptado a partir serne e seus colegas 1. Um critério de exclusão para este procedimento é a doença vascular do colagénio, desde doenças do colágeno vascular produz conhecidas alterações capilares 2. </li><li> Após um jejum de 10 horas durante a noite e no mínimo 20 minutos de descanso sentada, medições microvasculares são realizados para uma meia hora entre 7e 11 horas, em uma sala com temperatura tranquila, controlada (mantida entre 21,5-22,5 ° C), com o assunto na posição sentada e da mão esquerda na altura do coração. </li><li> Periungueal capilares na pele dorsal do terceiro dedo são visualizadas em um estereomicroscópio (Olympus; Center Valley, PA), ligado a uma introspecção SPOT 4 megapixels monocromático câmera digital (Número do Modelo IN-1400: Instrumentos de Diagnóstico; Sterling Heights, MI), e um computador portátil (Dell Latitude D600: Dell; Austin, TX). Para limitar o movimento, a mão esquerda e antebraço são frouxamente tapado com um cobertor dobrado, e repousou no outro cobertor dobrado posicionado na base do microscópio. </li><li> Iluminação leito ungueal é conseguida com uma lâmpada de fibra de 250 W halogéneo óptica (KL 2500LCD: Schott-FOSTEC; Elmsford, NY); iluminação adicional a partir de uma fibra de 150W suplementar fonte de luz óptica halogéneo (B &amp; B microscópios, Ltd., Warrendale, PA) é usado em indivíduos pigmentadas. </li><li> Para visualizar os vasos capilares, o3.2x objectivo (Olympus 3.2/0.07) é usado com um sistema de amplificação total de 38.4x. </li><li> Usando o software de imagens SPOT fornecido com a câmera, luz / escuridão contraste nas fotografias capilar é reforçada com a função mesmo software padrão SPOT (alongamento dos níveis de claro e escuro) para maximizar a visibilidade dos capilares em todas as disciplinas. </li><li> Para quantificar a densidade capilar, fotomicrografias digitais são tiradas a cada 3-5 segundos, durante cada uma das três fases, na linha de base de descanso, durante a hiperemia reativa postocclusive, e durante a oclusão venosa. (A) No início do estudo descanso, fotomicrografias são tomadas durante um período de três minutos para detectar capilares perfundidos em repouso. (B) Durante a hiperemia reativa postocclusive, fotomicrografias são levados para quantificar capilares funcionalmente perfundidos (linha de base mais reserva de capilares), como segue. Em primeiro lugar, uma braçadeira de oclusão no braço esquerdo é inflada até 40 mm Hg acima da pressão arterial sistólica, durante 10 min. Fotomicrografias são então levados durante thprimeiro minuto e imediatamente após o lançamento de oclusão arterial, visualizando todos os capilares funcionalmente perfundido. Menor densidade capilar após hiperemia reativa indica prejudicada recrutamento funcional capilar, e, portanto, rarefação funcional. (C) Durante fotomicrografias oclusão venosa são levados para quantificar a densidade capilar máxima, que inclui tanto a perfusão (com glóbulo vermelho ativo (RBC) de movimento) e nonperfused (cheio de estagnado, sem movimento hemácias) capilares 3 da seguinte forma. Depois de dez minutos de descanso após o procedimento reativa postocclusive hiperemia, o manguito do braço é inflado a 60 mm Hg para 60 seg, passivamente forçando o sangue para todos os presentes capilares patente e fotomicrografias foram tomadas durante este tempo. Uma vez que a densidade capilar máxima inclui todos os capilares estruturalmente presente, uma redução máxima na densidade capilar indica rarefação estrutural. </li><li> A densidade capilar é definida como o número de capilares por quadrado millimeter de pele periungueal, e é calculado como a média de quatro medições obtidas a partir de quatro imagens mais claramente definidas, pelo menos distorcida pelo movimento. Nos nossos estudos, os valores típicos para as contagens capilar (capilares / mm 2) foram 55-80 para linha de base, 65-90 para o pós-isquémica, e 90-105 para a oclusão venosa. Os valores para o recrutamento por cento capilar são, tipicamente, entre 5% e 25% (média de ~ 10-15%), e para os capilares perfundidos por cento entre os 70% e 95% (média de ~ 80-90%), com valores sendo menor entre hipertensos do que normotensos. A reprodutibilidade do processo de contagem foi verificada com três observadores independentes que realizaram as avaliações manuais de fotografias de 10 indivíduos diferentes (Figura 2). Os observadores foram cegos para a pressão de identidade e do sangue destes assuntos. Após o treinamento, conta subseqüentes realizadas de forma independente mostrou um alto nível de concordância. Média de discrepâncias inter e intra-avaliadores eram da order de 2-3 capilares / mm 2, e os coeficientes de correlação intraclasse foram todos maiores do que 0,90. Curto prazo variação das medidas capilaroscópicos foram da mesma ordem de grandeza que as discrepâncias inter-e intra-observador (aproximadamente 2 capilares / mm 2), mas a longo prazo a variação observada ao longo de 2-3 anos era uma ordem de magnitude maior (cerca de 15 capilares / mm 2), indicando que as alterações longitudinais podem ser facilmente distinguidas das variações de avaliadores. Confiabilidade das duas medidas função capilar também foi alta (coeficiente de correlação intraclasse = 0,84 por cento para o recrutamento capilar e 0,82 por cento para os capilares perfundidos). </li><li> Os investigadores agora utilizar um método baseado em computador para quantificar a densidade capilar utilizando-Image Pro Plus software de imagem (Versão 6.2, Media Cybernetics, Inc., Bethesda, MD: Figura 3). Correlações de Pearson entre linha de base, pós-isquêmica, e venocontagens de nós de congestionamento feitas com o software e os correspondentes contagens manuais em 10 indivíduos foram de 0,78, 0,78, e 0,71, respectivamente (todos p &lt;0,05), indicando boa concordância entre os dois métodos. Fiabilidade das contagens baseados em computador é um pouco menor do que a contagem de manuais mas ainda elevada (coeficiente de correlação intraclasse = 0,91 para linha de base, 0,86 para o pós-isquémica, e de 0,84 para a oclusão venosa). Temos também dados inéditos que demonstram a associação de contagens automatizadas com múltiplos fatores de risco cardiovasculares, incluindo hipertensão, o que estamos preparando para publicação. </li><li> A Tabela 1 resume as medições da densidade capilar e cálculos. Recrutamento por cento capilar é avaliada através da divisão do aumento na densidade capilar induzido pela HR postocclusive (postocclusive densidade capilar menos hiperemia reativa de linha de base da densidade capilar), a densidade máxima de capilar (observado durante passivo venonos oclusão). Por cento capilares perfundidos representa a proporção de todas as que estão presentes capilares perfundidos (funcionalmente activa), e é calculada dividindo a densidade postocclusive HR capilar pela densidade máxima capilar. Tanto o recrutamento por cento por cento capilar e perfundidos capilares refletem o número de capilares funcionais. Os valores mais baixos para estas medidas indicam rarefação capilar funcional. </li></ol> 2. Avaliação da função endotelial <ol><li> A função endotelial é avaliada antes e após a hiperemia reativa postocclusive, utilizando medições não invasivas pletismografia de fluxo sanguíneo do antebraço, de acordo com o método de Sivertsson 4, que utiliza o estímulo endotélio-dependente de HR para induzir vasodilatação. </li><li> Com o tema na posição sentada após 10 minutos de descanso em decúbito dorsal, um de mercúrio em medidor de tensão de borracha esticado para 10% além do seu comprimento de repouso for repetido around o assunto do antebraço de 5 cm abaixo da fossa antecubital. </li><li> O calibre de tensão está ligado a um pletismógrafo (EC-4: DE Hokanson, Inc; Bellevue, WA), o qual por sua vez está ligado a um registador de Doppler (CW-1; DE Hokanson, Inc; Belleveue, WA). </li><li> Uma braçadeira de oclusão superior do braço é aplicado, e o braço é suspenso confortavelmente ao nível do coração utilizando uma atadura funda ligado a um pólo ajustável intravenosa. Sistólica e diastólica e freqüência cardíaca são obtidos com um 100 automático Dinamap ProCare manguito BP (GE Healthcare, Piscataway, NJ) colocado no braço oposto. </li><li> Um manguito pediátrico em torno do punho é inflado para 200 mm Hg para ocluir o fluxo para o lado. A braçadeira é insuflada superior a 50 mm Hg, esvaziado para 1,5 seg, e, em seguida, re-inflado rapidamente antes de cada medição do fluxo sanguíneo do antebraço, obtido por meio da expansão do medidor de tensão colocado ao redor do antebraço. </li><li> Fluxo de sangue do antebraço (FSA) é medido no início descansou (FBF base) e novamente umat vasodilatação induzida por hiperemia postocclusive máxima (FBF max). Para as medições da linha de base do fluxo sanguíneo, quatro curvas consecutivas FBF são obtidos dentro de 30 seg (FBF base). </li><li> A braçadeira de oclusão é então inflada até 40 mm Hg acima da pressão arterial sistólica, durante 10 min. Após a libertação da oclusão arterial (HR postocclusive), quatro curvas FBF consecutivos são obtidos dentro dos primeiros 30 segundos de fluxo (FBF max). </li><li> A proporção de base FBF max / FBF é calculado como uma estimativa da vasodilatação, dividindo-se a média dos quatro valores FBF max pela média dos quatro valores de base FBF. Antebraço 5 Resistência vascular na vasodilatação máxima (FVR max) é calculado como a pressão arterial média (PAM) dividido pela FBF max. FBF máximo durante hiperemia reativa está diretamente relacionada à FBF após a infusão máxima de intra-arterial de acetilcolina, um vasodilatador endotélio-dependente. 6 </sup&gt; Assim, FBF máximo ea relação FBF base de max / FBF são aceites medidas não invasivas da função endotelial. 6-8 Além disso, tanto a FBF max max e FVR refletir as mudanças de resistência das artérias estruturais (aumento do índice de parede / lúmen). 9 </li></ol>

<ol>
	<li>Serne, E. H., Gans, R. O., ter Maaten, J. C., ter Wee, P. M., Donker, A. J., Stehouwer, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Capillary+recruitment+is+impaired+in+essential+hypertension+and+relates+to+insulin's+metabolic+and+vascular+actions.">Capillary recruitment is impaired in essential hypertension and relates to insulin's metabolic and vascular actions.</a> Cardiovasc. Res. 49, 161-168 (2001).</li><li>Bollinger, A., Fagrell, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Clinical Capillaroscopy: A guide to its use in clinical research and practice. , (1990).</li><li>Antonios, T. F., Rattray, F. E., Singer, D. R., Markandu, N. D., Mortimer, P. S., MacGregor, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maximization+of+skin+capillaries+during+intravital+video-microscopy+in+essential+hypertension:+comparison+between+venous+congestion,+reactive+hyperaemia+and+core+heat+load+tests.">Maximization of skin capillaries during intravital video-microscopy in essential hypertension: comparison between venous congestion, reactive hyperaemia and core heat load tests.</a> Clin. Sci. (Lond). 97, 523-528 (1999).</li><li>Sivertsson, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+hemodynamic+importance+of+structural+vascular+changes+in+essential+hypertension.">The hemodynamic importance of structural vascular changes in essential hypertension.</a> Acta. Physiol. Scand. Suppl. 343, 1-56 (1970).</li><li>Raitakari, O. T., Celermajer, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Testing+for+endothelial+dysfunction.">Testing for endothelial dysfunction.</a> Ann. Med. 32, 293-304 (2000).</li><li>Tousoulis, D., Antoniades, C., Stefanadis, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluating+endothelial+function+in+humans:+a+guide+to+invasive+and+non-invasive+techniques.">Evaluating endothelial function in humans: a guide to invasive and non-invasive techniques.</a> Heart. 91, 553-558 (2005).</li><li>Higashi, Y., Sasaki, S., Nakagawa, K., Matsuura, H., Kajiyama, G., Oshima, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+noninvasive+measurement+of+reactive+hyperemia+that+can+be+used+to+assess+resistance+artery+endothelial+function+in+humans.">A noninvasive measurement of reactive hyperemia that can be used to assess resistance artery endothelial function in humans.</a> Am. J. Cardiol. 87, 121-125 (2001).</li><li>Higashi, Y., Yoshizumi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+methods+to+evaluate+endothelial+function:+method+for+assessingendothelial+function+in+humans+using+a+strain-gauge+plethysmography:+nitric+oxide-dependent+and+independent+vasodilation.">New methods to evaluate endothelial function: method for assessingendothelial function in humans using a strain-gauge plethysmography: nitric oxide-dependent and independent vasodilation.</a> J. Pharmacol. Sci. 93, 399-404 (2003).</li><li>Lind, L., Sarabi, M., Millgard, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methodological+aspects+of+the+evaluation+of+endothelium+dependent+vasodilatation+in+the+human+forearm.">Methodological aspects of the evaluation of endothelium dependent vasodilatation in the human forearm.</a> Clin. Physiol. 18, 81-87 (1998).</li><li>Cheng, C., Diamond, J. J., Falkner, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+capillary+rarefaction+in+mild+blood+pressure+elevation.">Functional capillary rarefaction in mild blood pressure elevation.</a> Clinical and Translational Science. 1, 75-79 (2008).</li><li>Cheng, C., Daskalakis, C., Falkner, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Capillary+rarefaction+in+treated+and+untreated+hypertensive+subjects.">Capillary rarefaction in treated and untreated hypertensive subjects.</a> Therapeutic Advances in Cardiovascular Disease. 2, 79-88 (2008).</li><li>Antonios, T. F., Singer, D. R., Markandu, N. D., Mortimer, P. S., MacGregor, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rarefaction+of+skin+capillaries+in+borderline+essential+hypertension+suggests+an+early+structural+abnormality.">Rarefaction of skin capillaries in borderline essential hypertension suggests an early structural abnormality.</a> Hypertension. 34, 655-658 (1999).</li><li>Noon, J. P., Walker, B. R., Webb, D. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+microvascular+dilatation+and+capillary+rarefaction+in+young+adults+with+a+predisposition+to+high+blood+pressure.">Impaired microvascular dilatation and capillary rarefaction in young adults with a predisposition to high blood pressure.</a> J. Clin. Invest. 99, 1873-1879 (1997).</li><li>Antonios, T. F., Rattray, F. M., Singer, D. R., Markandu, N. D., Mortimer, P. S., MacGregor, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rarefaction+of+skin+capillaries+in+normotensive+offspring+of+individuals+with+essential+hypertension.">Rarefaction of skin capillaries in normotensive offspring of individuals with essential hypertension.</a> Heart. 89, 175-178 (2003).</li><li>de Jongh, R. T., Ijzerman, R. G., Serne, E. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visceral+and+truncal+subcutaneous+adipose+tissue+are+associated+with+impaired+capillary+recruitment+in+healthy+individuals.">Visceral and truncal subcutaneous adipose tissue are associated with impaired capillary recruitment in healthy individuals.</a> J. Clin. Endocrinol. Metab. 91, 5100-5106 (2006).</li><li>de Jongh, R. T., Serne, E. H., Ijzerman, R. G., de Vries, G., Stehouwer, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Free+fatty+acid+levels+modulate+microvascular+function:+relevance+for+obesity-associated+insulin+resistance,+hypertension,+and+microangiopathy.">Free fatty acid levels modulate microvascular function: relevance for obesity-associated insulin resistance, hypertension, and microangiopathy.</a> Diabetes. 53, 2873-2882 (2004).</li><li>de Jongh, R. T., Serne, E. H., RG, I. J., Stehouwer, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microvascular+function:+a+potential+link+betweensalt+sensitivity,+insulin+resistance+and+hypertension.">Microvascular function: a potential link betweensalt sensitivity, insulin resistance and hypertension.</a> J. Hypertens. 25, 1887-1893 (2007).</li><li>Ijzerman, R. G., Voordouw, J. J., Van Weissenbruch, M. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TNF-alpha+levels+are+associated+withskin+capillary+recruitment+in+humans:+a+potential+explanation+for+the+relationship+between+TNF-alpha+and+insulin+resistance.">TNF-alpha levels are associated withskin capillary recruitment in humans: a potential explanation for the relationship between TNF-alpha and insulin resistance.</a> Clin. Sci. (Lond). 110, 361-368 (2006).</li><li>Yvonne-Tee, G. B., Rasool, A. H., Halim, A. S., Rahman, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Noninvasive+assessment+of+cutaneous+vascular+function+in+vivo+using+capillaroscopy,+plethysmography+and+laser-Doppler+instruments:+its+strengths+and+weaknesses.">Noninvasive assessment of cutaneous vascular function in vivo using capillaroscopy, plethysmography and laser-Doppler instruments: its strengths and weaknesses.</a> Clin. Hemorheol. Microcirc. 34, 457-473 (2006).</li><li>Wilkinson, I. B., Webb, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Venous+occlusion+plethysmography+in+cardiovascular+research:+methodology+and+clinical+applications.">Venous occlusion plethysmography in cardiovascular research: methodology and clinical applications.</a> Br. J. Clin. Pharmacol. 52, 631-646 (2001).</li><li>Thijssen, D. H., Bleeker, M. W., Smits, P., Hopman, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reproducibility+of+blood+flow+and+post-occlusive+reactive+hyperaemia+as+measured+by+venous+occlusion+plethysmography.">Reproducibility of blood flow and post-occlusive reactive hyperaemia as measured by venous occlusion plethysmography.</a> Clin. Sci. (Lond). 108, 151-157 (2005).</li><li>Leslie, S. J., Attina, T., Hultsch, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+two+plethysmography+systems+in+assessment+of+forearm+blood+flow.">Comparison of two plethysmography systems in assessment of forearm blood flow.</a> J. Appl. Physiol. 96, 1794-179 (2004).</li><li>James, M. A., Tullett, J., Hemsley, A. G., Shore, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+aging+and+hypertension+on+the+microcirculation.">Effects of aging and hypertension on the microcirculation.</a> Hypertension. 47, 968-9674 (2006).</li><li>Debbabi, H., Uzan, L., Mourad, J. J., Safar, M., Levy, B. I., Tibirica, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+skin+capillary+density+in+treated+essential+hypertensive+patients.">Increased skin capillary density in treated essential hypertensive patients.</a> Am. J. Hypertens. 19, 477-483 (2006).</li><li>Serne, E. H., Gans, R. O., ter Maaten, J. C., Tangelder, G. J., Donker, A. J., Stehouwer, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+skincapillary+recruitment+in+essential+hypertension+is+caused+by+both+functional+and+structural+capillary+rarefaction.">Impaired skincapillary recruitment in essential hypertension is caused by both functional and structural capillary rarefaction.</a> Hypertension. 38, 238-242 (2001).</li><li>Carpentier, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+techniques+for+clinical+assessment+of+the+peripheral+microcirculation.">New techniques for clinical assessment of the peripheral microcirculation.</a> Drugs. 58, 17-22 (2001).</li><li>Shore, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Capillaroscopy+and+the+measurement+of+capillary+pressure.">Capillaroscopy and the measurement of capillary pressure.</a> Br. J. Clin. Pharmacol. 50, 501-513 (2000).</li></ol>

Os autores não têm conflitos de interesses financeiros relacionados com a conduta ou publicação deste trabalho de divulgar.

Capilaroscopia (microscopia capilar) é uma medida não-padrão da estrutura capilar. No entanto, atualmente, não existem métodos padronizados para avaliação direta da structure.Furthermore capilar, microscopia capilar tem sido amplamente utilizado para a avaliação direta de capillarydensity em um corpo grande e crescente de trabalhos publicados 12, 13, 10, 11, 14-18 . Além disso, validamos a técnica de microscopia capilar, correlacionando as alterações capilaroscópicas com fluxo antebraço sangue,...

non-invasive assessment of microvascular and endothelial function

Os autores utilizaram capilaroscópicas e antebraço técnicas de fluxo de sangue para investigar o papel da disfunção microvascular na patogênese da doença cardiovascular. Capilaroscopia é uma metodologia não-invasiva, relativamente barato para a visualização direta da microcirculação. Recrutamento por cento capilar é avaliada através da divisão do aumento na densidade capilar induzida por postocclusive HR (HR postocclusive densidade capilar menos densidade basal capilar), a densidade máxima por capilar (observado durante a oclusão venosa passiva). Por cento capilares perfundidos representa a proporção de todas as que estão presentes capilares perfundidos (funcionalmente activa), e é calculada dividindo a densidade postocclusive HR capilar pela densidade máxima capilar. Tanto o recrutamento por cento por cento capilar e perfundidos capilares refletem o número de capilares funcionais. O fluxo de sangue do antebraço (FBF), técnica provides aceite não invasivos medidas da função endotelial: A relação de base FBF max / FBF é calculada como uma estimativa de vasodilatação, dividindo-se a média dos quatro valores FBF max por a média dos quatro valores de base FBF. Resistência vascular no antebraço vasodilatação máxima (FVR max) é calculado como a pressão arterial média (PAM), dividido pela FBF max. Ambos capilaroscopia e técnicas do antebraço são facilmente aceitável para os pacientes e pode ser aprendido rapidamente. A função endotelial microvascular e medidas obtidas utilizando as metodologias descritas neste documento podem ter utilidade futura em clínicas de pacientes de risco cardiovascular estratégias de redução. Como já publicados relatórios que demonstram que a disfunção endotelial microvascular e são encontrados em estágios iniciais de hipertensão, incluindo a pré-hipertensão, medidas de função endotelial microvascular e pode eventualmente ajudar na identificação precoce, Estratificação de risco e prevenção da fase final da patologia vascular, com suas consequências potencialmente fatais.

Diferenças na aparência da microvasculatura entre normotensos e hipertensos são prontamente evidentes ao comparar as Figuras 4 e 5. Figura 4 mostra a rede de capilares normal reto em filas bem organizadas em um indivíduo normotenso. Em contraste, a Figura 5 mostra um padrão mais desordenada de encolhido, enrolada capilares. Os autores têm interesse em curso no papel da disfunção microvascular na patogênese da doença cardiovascular....

Watch this Scientific Journal Video about Non-invasive Assessment of Microvascular and Endothelial Function at JoVE.com

Non-invasive Assessment of Microvascular and Endothelial Function

Capilaroscopia é uma metodologia não-invasiva, relativamente barato para a visualização direta da microcirculação. O antebraço técnica de fluxo de sangue proporciona aceitos não invasivos medidas da função endotelial.

Avaliação não invasiva da função microvascular e endotelial

The authors have utilized capillaroscopy and forearm blood flow techniques to investigate the role of microvascular dysfunction in pathogenesis of ...

non-invasive-assessment-of-microvascular-and-endothelial-function

Research

JoVE Journal

Medicine

16.0K visualizações.  Thomas Jefferson University. Capilaroscopia é uma metodologia não-invasiva, relativamente barato para a visualização direta da microcirculação. O antebraço técnica de fluxo de sangue proporciona aceitos não invasivos medidas da função endotelial.

Vídeo: Non-invasive Assessment of Microvascular and Endothelial Function

An in vivo Rodent Model of Contraction-induced Injury and Non-invasive Monitoring of Recovery

Muscle strains are one of the most common complaints treated by physicians. A muscle injury is typically diagnosed from the patient history and physical exam alone, however the clinical presentation can vary greatly depending on the extent of injury, the patient's pain tolerance, etc. In patients with muscle injury or muscle disease, assessment of muscle damage is typically limited to clinical signs, such as tenderness, strength, range of motion, and more recently, imaging studies. Biological markers, such as serum creatine kinase levels, are typically elevated with muscle injury, but their levels do not always correlate with the loss of force production. This is even true of histological findings from animals, which provide a "direct measure" of damage, but do not account for all the loss of function. Some have argued that the most comprehensive measure of the overall health of the muscle in contractile force. Because muscle injury is a random event that occurs under a variety of biomechanical conditions, it is difficult to study. Here, we describe an in vivo animal model to measure torque and to produce a reliable muscle injury. We also describe our model for measurement of force from an isolated muscle in situ. Furthermore, we describe our small animal MRI procedure.

An in vivo Rodent Model of Contraction-induced Injury and Non-invasive Monitoring of Recovery

An in vivo animal model of injury is described. The method takes advantage of the subcutaneous position of the fibular nerve. Velocity, timing of muscle activation, and arc of motion are all pre-determined and synchronized using commercial software. Post injury changes are monitored in vivo using MR imaging/spectroscopy.

Um In vivo Modelo de Contração de roedores induzida por lesões e não-invasivos acompanhamento da recuperação

Muscle strains are one of the most  common complaints treated by physicians. A muscle injury is typically diagnosed from the patient history and  physical ...

Medicina

Non-invasive Optical Measurement of Cerebral Metabolism and Hemodynamics in Infants

Perinatal brain injury remains a significant cause of infant mortality and morbidity, but there is not yet an effective bedside tool that can accurately screen for brain injury, monitor injury evolution, or assess response to therapy. The energy used by neurons is derived largely from tissue oxidative metabolism, and neural hyperactivity and cell death are reflected by corresponding changes in cerebral oxygen metabolism (CMRO2). Thus, measures of CMRO2 are reflective of neuronal viability and provide critical diagnostic information, making CMRO2 an ideal target for bedside measurement of brain health.
Brain-imaging techniques such as positron emission tomography (PET) and single-photon emission computed tomography (SPECT) yield measures of cerebral glucose and oxygen metabolism, but these techniques require the administration of radionucleotides, so they are used in only the most acute cases.
Continuous-wave near-infrared spectroscopy (CWNIRS) provides non-invasive and non-ionizing radiation measures of hemoglobin oxygen saturation (SO2) as a surrogate for cerebral oxygen consumption. However, SO2 is less than ideal as a surrogate for cerebral oxygen metabolism as it is influenced by both oxygen delivery and consumption. Furthermore, measurements of SO2 are not sensitive enough to detect brain injury hours after the insult 1,2, because oxygen consumption and delivery reach equilibrium after acute transients 3. We investigated the possibility of using more sophisticated NIRS optical methods to quantify cerebral oxygen metabolism at the bedside in healthy and brain-injured newborns. More specifically, we combined the frequency-domain NIRS (FDNIRS) measure of SO2 with the diffuse correlation spectroscopy (DCS) measure of blood flow index (CBFi) to yield an index of CMRO2 (CMRO2i) 4,5.
With the combined FDNIRS/DCS system we are able to quantify cerebral metabolism and hemodynamics. This represents an improvement over CWNIRS for detecting brain health, brain development, and response to therapy in neonates. Moreover, this method adheres to all neonatal intensive care unit (NICU) policies on infection control and institutional policies on laser safety. Future work will seek to integrate the two instruments to reduce acquisition time at the bedside and to implement real-time feedback on data quality to reduce the rate of data rejection.

We combined frequency-domain near-infrared spectroscopy measures of cerebral hemoglobin oxygenation with diffuse correlation spectroscopy measures of cerebral blood flow index to estimate an index of oxygen metabolism. We tested the utility of this measure as a bedside screening tool to evaluate the health and development of the newborn brain.

Não-invasivo de medição óptica do metabolismo cerebral e Hemodinâmica em Lactentes

Perinatal brain injury remains a significant cause of infant mortality  and morbidity, but there is not yet an effective bedside tool that can  accurately ...

Murine Spinotrapezius Model to Assess the Impact of Arteriolar Ligation on Microvascular Function and Remodeling

The murine spinotrapezius is a thin, superficial skeletal support muscle that extends from T3 to L4, and is easily accessible via dorsal skin incision. Its unique anatomy makes the spinotrapezius useful for investigation of ischemic injury and subsequent microvascular remodeling. Here, we demonstrate an arteriolar ligation model in the murine spinotrapezius muscle that was developed by our research team and previously published1-3. For certain vulnerable mouse strains, such as the Balb/c mouse, this ligation surgery reliably creates skeletal muscle ischemia and serves as a platform for investigating therapies that stimulate revascularization. Methods of assessment are also demonstrated, including the use of intravital and confocal microscopy. The spinotrapezius is well suited to such imaging studies due to its accessibility (superficial dorsal anatomy) and relative thinness (60-200 &mu;m). The spinotrapezius muscle can be mounted en face, facilitating imaging of whole-muscle microvascular networks without histological sectioning. We describe the use of intravital microscopy to acquire metrics following a functional vasodilation procedure; specifically, the increase in arterilar diameter as a result of muscle contraction. We also demonstrate the procedures for harvesting and fixing the tissues, a necessary precursor to immunostaining studies and the use of confocal microscopy.

We demonstrate a novel arterial ligation model in murine spinotrapezius muscle, including a step-by-step procedure and description of required instrumentation. We describe the surgery and relevant outcome measurements relating to vascular network remodeling and functional vasodilation using intravital and confocal microscopy.

Modelo Spinotrapezius murino para avaliar o impacto da ligadura arteriolar em função microvascular e Remodelação

The murine spinotrapezius is a thin, superficial skeletal support muscle that extends from T3 to L4, and is easily accessible via dorsal skin incision. ...

A Methodological Approach to Non-invasive Assessments of Vascular Function and Morphology

The endothelium is the innermost lining of the vasculature and is involved in the maintenance of vascular homeostasis. Damage to the endothelium may predispose the vessel to atherosclerosis and increase the risk for cardiovascular disease. Assessments of peripheral endothelial function are good indicators of early abnormalities in the vascular wall and correlate well with assessments of coronary endothelial function. The present manuscript details the important methodological steps necessary for the assessment of microvascular endothelial function using laser Doppler imaging with iontophoresis, large vessel endothelial function using flow-mediated dilatation, and carotid atherosclerosis using carotid artery ultrasound. A discussion on the methodological considerations for each of the techniques is also presented, and recommendations are made for future research.

The present article describes the methodological considerations for several non-invasive assessments of vascular function and morphology that are commonly used in medical research to assess different stages of atherosclerosis.

Uma abordagem metodológica para avaliações não-invasivas de função vascular e Morfologia

The endothelium is the innermost lining of the vasculature and is involved in the maintenance of vascular homeostasis.  Damage to the endothelium may ...

Non-invasive Assessment of the Efficacy of New Therapeutics for Intestinal Pathologies Using Serial Endoscopic Imaging of Live Mice

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic mechanisms that contribute to the onset and progression of intestinal diseases. The identification of new molecules that promote these pathologies has led to a flurry of activity focused on the development of potential new therapies to inhibit their function. As a result, various pre-clinical mouse models with an intact immune system and stromal microenvironment are now heavily used. Here we describe three experimental protocols to test the efficacy of new therapeutics in pre-clinical models of (1) acute mucosal damage, (2) chronic colitis and/or colitis-associated colon cancer, and (3) sporadic colorectal cancer. We also outline procedures for serial endoscopic examination that can be used to document the therapeutic response of an individual tumor and to monitor the health of individual mice. These protocols provide complementary experimental platforms to test the effectiveness of therapeutic compounds shown to be well tolerated by mice.

We describe methods for longitudinal monitoring of the efficacy of therapeutics for the treatment of colonic pathologies in mice using a rigid endoscope. This protocol can be readily used for the characterization of the therapeutic response of an individual tumor in live mice and also for monitoring potential disease relapse.

Avaliação não invasiva da eficácia de novos Therapeutics para Intestinal Patologias Usando imagem endoscópica série da Vivo Ratos

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic ...

Ultrasound Assessment of Endothelial Function: A Technical Guideline of the Flow-mediated Dilation Test

Cardiovascular disease is the primary cause of mortality and a major cause of disability worldwide. The dysfunction of the vascular endothelium is a pathological condition characterized mainly by a disruption in the balance between vasodilator and vasoconstrictor substances and is proposed to play an important role in the development of atherosclerotic cardiovascular disease. Therefore, a precise evaluation of endothelial function in humans represents an important tool that could help better understand the etiology of multiple cardio-centric pathologies.
Over the past twenty-five years, many methodological approaches have been developed to provide an assessment of endothelial function in humans. Introduced in 1989, the FMD test incorporates a forearm occlusion and subsequent reactive hyperemia that promotes nitric oxide production and vasodilation of the brachial artery. The FMD test is now the most widely utilized, non-invasive, ultrasonic assessment of endothelial function in humans and has been associated with future cardiovascular events.
Although the FMD test could have clinical utility, it is a physiological assessment that has inherited several confounding factors that need to be considered. This article describes a standardized protocol for determining FMD including the recommended methodology to help minimize the physiological and technical issues and improve the precision and reproducibility of the assessment.

The flow mediated dilation (FMD) test is the most commonly utilized, non-invasive, ultrasound assessment of endothelial function in humans. Although the FMD test has been related with the prediction of future cardiovascular disease and events, it is a physiological assessment with many inherent confounding factors that need to be considered.

Avaliação ultra-sons da função endotelial: A Orientação Técnica do Teste de dilatação fluxo-mediada

Cardiovascular disease is the primary cause of mortality and a major cause of disability worldwide. The dysfunction of the vascular endothelium is a ...

Assessment of Human Adipose Tissue Microvascular Function Using Videomicroscopy

While obesity is closely linked to the development of metabolic and cardiovascular disease, little is known about mechanisms that govern these processes. It is hypothesized that pro-atherogenic mediators released from fat tissues particularly in association with central/visceral adiposity may promote pathogenic vascular changes locally and systemically, and the notion that cardiovascular disease may be the consequence of adipose tissue dysfunction continues to evolve. Here, we describe a unique method of videomicroscopy that involves analysis of vasodilator and vasoconstrictor responses of intact small human arterioles removed from the adipose depot of living human subjects. Videomicroscopy is used to examine functional properties of isolated microvessels in response to pharmacological or physiological stimuli using a pressured system that mimics in vivo conditions. The technique is a useful approach to gain understanding of the pathophysiology and molecular mechanisms that contribute to vascular dysfunction locally within the adipose tissue milieu. Moreover, abnormalities in the adipose tissue microvasculature have also been linked with systemic diseases. We applied this technique to examine depot-specific vascular responses in obese subjects. We assessed endothelium-dependent vasodilation to both increased flow and acetylcholine in adipose arterioles (50 - 350 &#181;m internal diameter, 2 - 3 mm in length) isolated from two different adipose depots during bariatric surgery from the same individual. We demonstrated that arterioles from visceral fat exhibit impaired endothelium-dependent vasodilation compared to vessels isolated from the subcutaneous depot. The findings suggest that the visceral microenvironment is associated with vascular endothelial dysfunction which may be relevant to clinical observation linking increased visceral adiposity to systemic disease mechanisms. The videomicroscopy technique can be used to examine vascular phenotypes from different fat depots as well as compare findings across individuals with different degrees of obesity and metabolic dysfunction. The method can also be used to examine vascular responses longitudinally in response to clinical interventions.

Videomicroscopy systems are used to examine functional properties of isolated adipose tissue arterioles in response to physiological and pharmacological stimuli. This technique can be used to examine microvascular phenotypes in different adipose tissue domains in obese humans.

Avaliação da função Microvascular de tecido adiposo humano usando tele-microscopia

While obesity is closely linked to the development of metabolic and cardiovascular disease, little is known about mechanisms that govern these processes. ...

Skeletal Muscle Neurovascular Coupling, Oxidative Capacity, and Microvascular Function with 'One Stop Shop' Near-infrared Spectroscopy

Exercise represents a major hemodynamic stress that demands a highly coordinated neurovascular response in order to match oxygen delivery to metabolic demand. Reactive hyperemia (in response to a brief period of tissue ischemia) is an independent predictor of cardiovascular events and provides important insight into vascular health and vasodilatory capacity. Skeletal muscle oxidative capacity is equally important in health and disease, as it determines the energy supply for myocellular processes. Here, we describe a simple, non-invasive approach using near-infrared spectroscopy to assess each of these major clinical endpoints (reactive hyperemia, neurovascular coupling, and muscle oxidative capacity) during a single clinic or laboratory visit. Unlike Doppler ultrasound, magnetic resonance images/spectroscopy, or invasive catheter-based flow measurements or muscle biopsies, our approach is less operator-dependent, low-cost, and completely non-invasive. Representative data from our lab taken together with summary data from previously published literature illustrate the utility of each of these end-points. Once this technique is mastered, application to clinical populations will provide important mechanistic insight into exercise intolerance and cardiovascular dysfunction.

Skeletal Muscle Neurovascular Coupling, Oxidative Capacity, and Microvascular Function with &#039;One Stop Shop&#039; Near-infrared Spectroscopy

Here, we describe a simple, non-invasive approach using near-infrared spectroscopy to assess reactive hyperemia, neurovascular coupling and skeletal muscle oxidative capacity in a single clinic or laboratory visit.

Acoplamento de Neurovascular do músculo esquelético, capacidade oxidativa e função Microvascular com 'Balcão único' espectroscopia de infravermelho próximo

Exercise represents a major hemodynamic stress that demands a highly coordinated neurovascular response in order to match oxygen delivery to metabolic ...

Non-invasive Assessment of Dorsiflexor Muscle Function in Mice

Assessment of skeletal muscle contractile function is an important measurement for both clinical and research purposes. Numerous conditions can negatively affect skeletal muscle. This can result in a loss of muscle mass (atrophy) and/or loss of muscle quality (reduced force per unit of muscle mass), both of which are prevalent in chronic disease, muscle-specific disease, immobilization, and aging (sarcopenia). Skeletal muscle function in animals can be evaluated by a range of different tests. All tests have limitations related to the physiological testing environment, and the selection of a specific test often depends on the nature of the experiments. Here, we describe an in vivo, non-invasive technique involving a helpful and easy assessment of force frequency-curve (FFC) in mice that can be performed on the same animal over time. This permits monitoring of disease progression and/or efficacy&#160;of a potential therapeutic treatment.

Measurement of rodent skeletal muscle contractile function is a useful tool that can be used to track disease progression as well as efficacy of therapeutic intervention. We describe here the non-invasive, in vivo assessment of the dorsiflexor muscles that can be repeated over time in the same mouse.

Avaliação não-invasiva da função de músculos levantadores em ratos

Assessment of skeletal muscle contractile function is an important measurement for both clinical and research purposes. Numerous conditions can negatively ...

A Computerized Functional Skills Assessment and Training Program Targeting Technology Based Everyday Functional Skills

Today, many functional skills are technology-based, so development of a technology-based training program has broad importance. Here we present a computerized functional skills training program that was paired in half of the participants with a commercially available cognitive training (CCT) program.
Non-impaired older individuals (NC) aged 60+ (n=45) and similarly aged individuals with mild cognitive impairment (MCI; n=50) were randomized to receive 12 weeks of twice-weekly computerized functional skills training (CFST) or 12 weeks of twice-weekly sessions split between CCT and CFST. Skills trained were use of an ATM; internet banking; ticket kiosk; telephone and internet prescription refill; medication management; and internet shopping. As with previous functional capacity assessments, we focus on completion time for each simulation.
51 participants completed the training program, either by mastering all 6 tasks (34) or completing 12 weeks of training. 44 more participants completed 4 or more training sessions so they were also analyzed for improvement up to their last training session. Completion time for all 6 tests significantly improved from the baseline assessment to the final training session in both groups of participants (all p&#60;0.001 with an average improvement in task completion time of 45%). Further, there was no differential improvement in MCI and NC in the 6 tests from baseline to end of training (all t&#60;1.66, all p&#62;0.12). Finally, combined CCT plus CFST did not differ from CSFT alone on any of the percent-change score measures (all t&#60;1.64, all p&#62;0.11).
Both NC and MCI groups evidenced substantial improvements in performance. CCT supplementation led to similar functional gains with half as many training sessions. The NC participants proceeded through the training fairly rapidly even without CCT supplementation; MCI participants required more training but learned equivalently. These findings suggest that even in cases with memory impairments, functional skills can be efficiently learned with training.

This training protocol uses computerized training to teach technology-related everyday functional skills. These skills include financial skills, travel and transit, as well as medication management.