Esta pesquisa foi apoiada pelo Programa intramuros Pesquisa do NIH, Instituto Nacional sobre Envelhecimento e do Estudo Longitudinal Baltimore sobre Envelhecimento. Uma parte do que o apoio foi através de um contrato de R &amp; D com o Instituto de Pesquisa MedStar. Protocolo padrão e perícia técnica foi fornecida por AtCor.

1. Procedimento <ol><li> Abra o software através do atalho na área de trabalho do Windows, clicando duas vezes sobre o ícone. </li><li> Clique Paciente Para ativar a tela do Paciente. </li><li> Para introduzir um novo paciente no banco de dados, clique no botão Novo na área de pesquisa paciente e, em seguida, clique no botão Sim para confirmar que você gostaria de introduzir um novo paciente. </li><li> Digite os detalhes do paciente, como sobrenome, nome, data de nascimento e sexo. Clique no botão Salvar para avançar para a próxima etapa. </li><li> Clique no botão PWV PWA ou para realizar a medição. Se apenas um destes modos está disponível, não há necessidade de clicar no botão, basta avançar para a tela do estudo. </li></ol> 2. Medida da Pressão Arterial Central Usando pulso Análise Wave (PWA) <ol><li> Clique no botão Estudo para entrar parâmetros de estudo. Introduza a pressão braquial tirado de um esfigmomanômetro calibrado e quaisquer outros detalhes que você deseja incluir. </li><li> Click no botão Dados Captura de proceder com a medição. Fixe os três eletrodos de ECG e leva para o paciente em uma configuração de chumbo II modificado ou usando membros do paciente, como indicado nos cabos. </li><li> Palpe artéria radial do paciente. Este está localizado na lateral do punho, imediatamente acima da articulação do pulso, perto da base do polegar. Pressione suavemente a artéria radial contra o osso subjacente com o meio e dedos indicadores. </li><li> Coloque o tonômetro sobre o ponto de pulso forte. Pressione suavemente o tonômetro para baixo até ver uma onda de pressão consistente exibido na tela de captura de dados. </li><li> Se Captura Automática está habilitada (ver nota abaixo), o sistema irá salvar automaticamente a medição. Para salvar manualmente a medição para a análise, pressione a barra de espaço no teclado (ou pisar no pedal, se utilizado). </li><li> Se uma medição repetição é necessária com os mesmos parâmetros do estudo, clique no botão Repetir para voltar à capturaTela de dados e repetir a medição como descrito nas etapas 2,4-2,5 acima. </li><li> Para realizar uma medição em um novo paciente, clique no botão do paciente e volte ao passo 1.3 acima. </li></ol> 3. Rigidez Arterial Medição Usando Onda de Pulso Velocity (PWV) <ol><li> Clique no botão Estudo para entrar parâmetros de estudo. </li><li> Digite pressão braquial tirado de um esfigmomanômetro calibrado Insira a distância medida para o distal (notch supra-esternal de pulso femoral) e proximal (notch supra-esternal de pulso carotídeo) sites. Campos de medicação, anotações e operador de rede podem ser inseridos, se desejado </li><li> Clique no botão Capturar para proceder com a medição. </li><li> Verifique se o sinal de ECG é de qualidade aceitável. </li><li> Palpar o pulso da artéria carótida. Este situa-se entre a laringe e o esternocleidomastoideu no pescoço. Ela pode ser sentida por empurrando o músculo para os lados e pressiona a artéria contra a laringe. </li><li&gt; Coloque o tonômetro na artéria carótida e pressione suavemente o tonômetro para baixo até que você receber uma onda de pressão consistente exibido na tela de captura de dados. </li><li> Quando você tem um mínimo de 13 segundos de ondas consistentes, clique no botão OK ou (imprensa sobre o pedal, se utilizado). </li><li> Palpar o pulso da artéria femoral. Ele está localizado ao longo da coxa ventral a meio caminho entre a sínfise púbica e espinha ilíaca ântero-superior. </li><li> Coloque o tonômetro na artéria femoral e pressione suavemente o tonômetro para baixo até que você receber uma onda de pressão consistente exibido na tela de captura. Quando você tem um mínimo de 13 segundos de ondas consistentes, clique no botão OK (ou pressione o no pedal, se utilizado). </li><li> O relatório (s) podem agora ser revistas. Estas medições são tipicamente realizados em triplicado. Se o usuário deseja medir a VOP entre artérias carótidas e radial, seria repetido estes passos apenas mostrados, exceto que em vez de medir a fartéria emoral, o usuário coloque o tonômetro na artéria radial. Para realizar uma medição em um novo paciente, clique no botão de Estudos e repita este procedimento ou usar o recurso de pesquisa para procurar um paciente existente. </li></ol> 4. Desligar <ol><li> O software salva automaticamente todos os relatórios. Para fechar o software, clique em Sistema no menu principal e clique em Sair. Coloque o tonômetro na bandeja do módulo de armazenamento. </li></ol>

<ol>
	<li>Najjar, S. S., Scuteri, A., 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=Pulse+wave+velocity+is+an+independent+predictor+of+the+longitudinal+increase+in+systolic+blood+pressure+and+of+incident+hypertension+in+the+Baltimore+Longitudinal+Study+of+Aging.">Pulse wave velocity is an independent predictor of the longitudinal increase in systolic blood pressure and of incident hypertension in the Baltimore Longitudinal Study of Aging.</a> J. Am. Coll. Cardiol. 51 (14), 1377-1383 (2008).</li><li>Doonan, R. J., Scheffler, P., 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=Altered+arterial+stiffness+and+subendocardial+viability+ratio+in+young+healthy+light+smokers+after+acute+exercise.">Altered arterial stiffness and subendocardial viability ratio in young healthy light smokers after acute exercise.</a> PloS one. 6 (10), (2011).</li><li>Laurent, S., Cockcroft, 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=Expert+consensus+document+on+arterial+stiffness:+methodological+issues+and+clinical+applications.">Expert consensus document on arterial stiffness: methodological issues and clinical applications.</a> Eur. Heart J. 27 (21), 2588-2605 (2006).</li><li>Mitchell, G. F., Hwang, S. -. 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=Arterial+stiffness+and+cardiovascular+events:+the+Framingham+Heart+Study.">Arterial stiffness and cardiovascular events: the Framingham Heart Study.</a> Circulation. 121 (4), 505-511 (2010).</li><li>Abbatecola, A. M., Chiodini, P., 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=Pulse+wave+velocity+is+associated+with+muscle+mass+decline:+Health+ABC+study.">Pulse wave velocity is associated with muscle mass decline: Health ABC study.</a> Age. , (2011).</li><li>Watson, N. L., Sutton-Tyrrell, K., 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=Arterial+stiffness+and+gait+speed+in+older+adults+with+and+without+peripheral+arterial+disease.">Arterial stiffness and gait speed in older adults with and without peripheral arterial disease.</a> Am. J. Hypertens. 24 (1), 90-95 (2011).</li><li>Mackenzie, I. S., Wilkinson, I. B., Cockcroft, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+arterial+stiffness+in+clinical+practice.">Assessment of arterial stiffness in clinical practice.</a> QJM. 95 (2), 67-74 (2002).</li><li>O'Rourke, M. F., Pauca, A., Jiang, X. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pulse+wave+analysis.">Pulse wave analysis.</a> Br. J. Clin. Pharmacol. 51 (6), 507-522 (2001).</li><li>Stoner, L., Young, J. M., Fryer, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessments+of+arterial+stiffness+and+endothelial+function+using+pulse+wave+analysis.">Assessments of arterial stiffness and endothelial function using pulse wave analysis.</a> Int. J. Vasc. Med. 2012, 903107 (2012).</li><li>Farro, I., Bia, D., 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=Pulse+wave+velocity+as+marker+of+preclinical+arterial+disease:+reference+levels+in+a+uruguayan+population+considering+wave+detection+algorithms,+path+lengths,+aging,+and+blood+pressure.">Pulse wave velocity as marker of preclinical arterial disease: reference levels in a uruguayan population considering wave detection algorithms, path lengths, aging, and blood pressure.</a> Int. J. Hypertens. 2012, (2012).</li><li>Lakatta, E. G., Wang, M., Najjar, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arterial+aging+and+subclinical+arterial+disease+are+fundamentally+intertwined+at+macroscopic+and+molecular+levels.">Arterial aging and subclinical arterial disease are fundamentally intertwined at macroscopic and molecular levels.</a> Med. Clin. N. Am. 93 (3), 583-604 (2009).</li><li>Ambrose, M. <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 Manual (CVMS) Rev. 2, 1-51 (2011).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reference+Values+for+Arterial+Stiffness'+Collaboration+Determinants+of+pulse+wave+velocity+in+healthy+people+and+in+the+presence+of+cardiovascular+risk+factors:+'establishing+normal+and+reference+values.">Reference Values for Arterial Stiffness' Collaboration Determinants of pulse wave velocity in healthy people and in the presence of cardiovascular risk factors: 'establishing normal and reference values.</a> Eur. Heart J. 31 (19), 2338-2350 (2010).</li><li>van Bortel, L. M., Laurent, S., 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=Expert+consensus+document+on+the+measurement+of+aortic+stiffness+in+daily+practice+using+carotid-femoral+pulse+wave+velocity.">Expert consensus document on the measurement of aortic stiffness in daily practice using carotid-femoral pulse wave velocity.</a> J. Hypertens. 30 (3), 445-448 (2012).</li><li>Scuteri, A., Brancati, A., Gianni, W., Assisi, A., Volpe, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arterial+stiffness+is+an+independent+risk+factor+for+cognitive+impairment+in+the+elderly:+a+pilot+study.">Arterial stiffness is an independent risk factor for cognitive impairment in the elderly: a pilot study.</a> J. Hypertens. 23 (6), 1211 (2005).</li><li>Khoshdel, A. R., Carney, S. L., Nair, B. R., Gillies, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Better+management+of+cardiovascular+diseases+by+pulse+wave+velocity:+combining+clinical+practice+with+clinical+research+using+evidence-based+medicine.">Better management of cardiovascular diseases by pulse wave velocity: combining clinical practice with clinical research using evidence-based medicine.</a> Clin. Med. Res. 5 (1), 45-52 (2007).</li></ol>

Os autores declaram que não têm interesses financeiros concorrentes. Os autores receberam assistência com descrição e estudo de desempenho de Sphygmocor que fabrica este produto descrito protocolo.

A medição da velocidade de onda de pulso (PWV) é um método não invasivo para analisar a rigidez arterial central de uma maneira relativamente simples e reprodutível 3. Para avaliar a VOP, a carótida (como um substituto para a aorta descendente) e formas de onda de pulso femoral são adquiridos através de um tonômetro de alta fidelidade, o tempo de atraso na chegada de onda de pulso na referenciação entre os dois locais é estimada uma ondas de ECG simultaneamente gravadas pelo design personalizado ...

A correction was made to <a href="http://www.jove.com/video/50817">Pulse Wave Velocity Testing in the Baltimore Longitudinal Study of Aging</a>. There was an error with the author, Majd AlGhatrif's, name. The author's name has been corrected to:
Majd AlGhatrif
instead of:
Majd al Ghatrif

A correction to the author list has been made for article <a href="http://www.jove.com/video/50817">Pulse Wave Velocity Testing in the Baltimore Longitudinal Study of Aging</a>.

Erratum: Pulse Wave Velocity Testing in the Baltimore Longitudinal Study of Aging

Onda de Pulso Velocity (VOP) é considerada o padrão-ouro na avaliação da rigidez arterial 2. Apesar de não ser utilizado clinicamente nos Estados Unidos, tem sido validado e amplamente utilizados na prática clínica em toda a Europa 3. Mudanças no PWV têm sido correlacionados com fatores de risco cardiovascular, como hipertensão, obesidade, intolerância à glicose e dislipidemia. Rigidez aórtica Superior tem sido associado com aumento do risco de um primeiro evento cardiovascular 4, o declínio da massa muscular 5, e menor velocidade de marcha em indivíduos com doença arterial periférica 6. Avaliação da PWV é realizado através da análise da velocidade com a qual um pulso de sangue viaja a partir da artéria carótida para a artéria femoral 7. É um método relativamente simples, com resultados altamente reprodutíveis. Os avanços na tecnologia de continuar a simplificar o procedimento, reduzindo a dependência do operador e diminuindo a quantidade de tempo necessária para oexame. O sistema utilizado em nosso laboratório é produzido por SphygmoCorand utiliza um eletrocardiograma e um tonômetro de alta fidelidade para adquirir formas de onda da carótida e pulsos femorais. O software analisa, então essas variáveis ​​para determinar a velocidade de 1) estimar o tempo que a onda de pulso leva para se deslocar entre a carótida e sítios femorais referenciando o momento da chegada da onda de pressão em cada local arterial em relação às formas de onda de eletrocardiograma 2) dividindo a distância medida entre os dois locais por o tempo estimado. O aproximar os locais de amostragem, menos precisa do PWV, limitando o uso deste método para vasos maiores, como as artérias femorais e carótidas. Além de determinar a VOP, o sistema pode calcular a pressão na aorta ascendente através da aplicação de uma função de transferência com a forma de onda produzida por pulso radial 8, que é a &quot;pressão central&quot; e a pressão que a maior parte tem um impacto directo no coração em vez de ejecção. Esta informação pode ajudar na avaliação do efeito do sistema arterial do ventrículo esquerdo de maneira fiável e não invasivo 9. Técnicas alternativas utilizadas para o exame da rigidez vascular, como distensibilidade carotídea através de rastreamento do eco, requerem uma grande quantidade de conhecimentos técnicos e são mais demoradas do que a VOP. Rastreamento Eco também utiliza geralmente a análise de vídeo-imagem e, portanto, a precisão dessas medições são limitados 3. Outras técnicas de análise das alterações vasculares incluem avaliação da carótida íntima espessura da camada média (IMT), que fornece resultados mais estreitamente relacionadas com a progressão da aterosclerose, embora as imagens transversais e transversais obtidos podem proporcionar uma aproximação grosseira de elastância carótida. Este método, no entanto, sofre de alguns dos mesmos constrangimentos como os estudos de distensão acima descritos. Como os investigadores continuam a olhar para as peças de rigidez arterial papel em vários estados de doença,Medição PWV provavelmente continuará a fornecer um desfecho substituto quantitativa e reprodutível.

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pulse wave velocity testing in the baltimore longitudinal study of aging

Velocidade da onda de pulso carotídeo-femoral é considerada o padrão ouro para a medida da rigidez arterial central obtidas por métodos não invasivos 1. Os indivíduos são colocados em decúbito dorsal e deixados a descansar durante pelo menos 10 minutos antes do início do exame. O tamanho adequado do manguito é selecionado e uma pressão arterial é obtida através de um dispositivo oscilométrico. Uma vez que uma pressão arterial de repouso foi obtida, ondas de pressão são adquiridos a partir das artérias carótidas comum femorais direita e direita. O sistema calcula automaticamente o tempo de passagem de impulsos entre estes dois locais (utilizando a artéria carótida, como um substituto para a aorta descendente). Medições de superfície corporal são usados ​​para determinar a distância percorrida pela onda de pulso entre os dois locais de amostragem. Esta distância é, então, dividido pelo tempo de passagem do pulso, resultando na velocidade da onda de pulso. As medições são realizadas em triplicata ea média é usada para analysis.

<html><img alt="Tabela 1" fo:content-width="5in" fo:src="/files/ftp_upload/50817/50817table1highres.jpg" src="/files/ftp_upload/50817/50817table1.jpg" /> Tabela 1. Resultados da PWV são produzidos em metros por segundo. Em indivíduos saudáveis, essas velocidades podem variar de um 6,2 m / seg média (em indivíduos com menos de 30) para 10,9 m / seg (em indivíduos de 70 anos ou mais). Estes valores foram estabelecidos no artigo &quot;Determinantes da veloci...

Watch this Scientific Journal Video about Pulse Wave Velocity Testing in the Baltimore Longitudinal Study of Aging at JoVE.com

Pulse Wave Velocity Testing in the Baltimore Longitudinal Study of Aging

Velocidade de onda de pulso (VOP) Medição avalia a rigidez arterial por um sistema baseado em tonometria, que mede a velocidade com que a pressão arterial onda viaja ao longo da árvore arterial, normalmente aproximar o tempo que ele leva essa onda de viajar da aorta descendente (usando a carótida artéria como um substituto) a artéria femoral. Carótida-femoral PWV aumenta duas a três vezes ao longo da vida adulta.

Pulso de onda Velocity Ensaios no Estudo Longitudinal de Envelhecimento de Baltimore

Carotid-femoral pulse wave velocity is considered the gold standard for measurements of central arterial stiffness obtained through noninvasive ...

pulse-wave-velocity-testing-baltimore-longitudinal-study

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Vital Signs: Pulse

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16.9K visualizações.  National Institute of Aging.  Velocidade de onda de pulso (VOP) Medição avalia a rigidez arterial por um sistema baseado em tonometria, que mede a velocidade com que a pressão arterial onda viaja ao longo da árvore arterial, normalmente aproximar o tempo que ele leva essa onda de viajar da aorta descendente (usando a carótida artéria como um substituto) a artéria femoral. Carótida-femoral PWV aumenta duas a três vezes ao longo da vida adulta. 

Vídeo: Pulse Wave Velocity Testing in the Baltimore Longitudinal Study of Aging

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for objective diagnosis but also for the evaluation of therapeutic efficacy and resilience to trauma. Ongoing research is directed at identifying molecular biomarkers for PTSD, including traumatic stress induced proteins, transcriptomes, genomic variances and genetic modulators, using biologic samples from subjects' blood, saliva, urine, and postmortem brain tissues. However, the correlation of these biomarker molecules in peripheral or postmortem samples to altered brain functions associated with psychiatric symptoms in PTSD remains unresolved. Here, we present an animal model of PTSD in which both peripheral blood and central brain biomarkers, as well as behavioral phenotype, can be collected and measured, thus providing the needed correlation of the central biomarkers of PTSD, which are mechanistic and pathognomonic but cannot be collected from people, with the peripheral biomarkers and behavioral phenotypes, which can.
Our animal model of PTSD employs restraint and tail shocks repeated for three continuous days - the inescapable tail-shock model (ITS) in rats. This ITS model mimics the pathophysiology of PTSD 17, 7, 4, 10. We and others have verified that the ITS model induces behavioral and neurobiological alterations similar to those found in PTSD subjects 17, 7, 10, 9. Specifically, these stressed rats exhibit (1) a delayed and exaggerated startle response appearing several days after stressor cessation, which given the compressed time scale of the rat's life compared to a humans, corresponds to the one to three months delay of symptoms in PTSD patients (DSM-IV-TR PTSD Criterian D/E 13), (2) enhanced plasma corticosterone (CORT) for several days, indicating compromise of the hypothalamopituitary axis (HPA), and (3) retarded body weight gain after stressor cessation, indicating dysfunction of metabolic regulation.
The experimental paradigms employed for this model are: (1) a learned helplessness paradigm in the rat assayed by measurement of acoustic startle response (ASR) and a charting of body mass; (2) microdissection of the rat brain into regions and nuclei; (3) enzyme-linked immunosorbent assay (ELISA) for blood levels of CORT; (4) a gene expression microarray plus related bioinformatics tools 18. This microarray, dubbed rMNChip, focuses on mitochondrial and mitochondria-related nuclear genes in the rat so as to specifically address the neuronal bioenergetics hypothesized to be involved in PTSD.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

Biomarcadores em um modelo animal para Revelando correlatos neurais, hematológica e Comportamental do TEPT

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

Medicina

Through the Looking Glass: Time-lapse Microscopy and Longitudinal Tracking of Single Cells to Study Anti-cancer Therapeutics

The response of single cells to anti-cancer drugs contributes significantly in determining the population response, and therefore is a major contributing factor in the overall outcome. Immunoblotting, flow cytometry and fixed cell experiments are often used to study how cells respond to anti-cancer drugs. These methods are important, but they have several shortcomings. Variability in drug responses between cancer and normal cells, and between cells of different cancer origin, and transient and rare responses are difficult to understand using population averaging assays and without being able to directly track and analyze them longitudinally. The microscope is particularly well suited to image live cells. Advancements in technology enable us to routinely image cells at a resolution that enables not only cell tracking, but also the observation of a variety of cellular responses. We describe an approach in detail that allows for the continuous time-lapse imaging of cells during the drug response for essentially as long as desired, typically up to 96 hr. Using variations of the approach, cells can be monitored for weeks. With the employment of genetically encoded fluorescent biosensors numerous processes, pathways and responses can be followed. We show examples that include tracking and quantification of cell growth and cell cycle progression, chromosome dynamics, DNA damage, and cell death. We also discuss variations of the technique and its flexibility, and highlight some common pitfalls.

Here, we describe a method of long-term time-lapse microscopy to longitudinally track single cells in response to anti-cancer therapeutics.

Through the Looking Glass: Microscopia Time-lapse e Longitudinal de rastreamento de células individuais para estudar Therapeutics Anti-câncer

The response of single cells to anti-cancer drugs contributes significantly in determining the population response, and therefore is a major contributing ...

A Phenotyping Regimen for Genetically Modified Mice Used to Study Genes Implicated in Human Diseases of Aging

Age-related diseases are becoming increasingly prevalent and the burden continues to grow as our population ages. Effective treatments are necessary to lessen the impact of debilitating conditions but remain elusive in many cases. Only by understanding the causes and pathology of diseases associated with aging, can scientists begin to identify potential therapeutic targets and develop strategies for intervention. The most common age-related conditions are neurodegenerative disorders such as Parkinson's disease and blindness. Age-related macular degeneration (AMD) is the leading cause of blindness in the elderly. Genome wide association studies have previously identified loci that are associated with increased susceptibility to this disease and identified two regions of interest: complement factor H (CFH) and the 10q26 locus, where the age-related maculopathy susceptibility 2 (ARMS2) and high-temperature requirement factor A1 (HtrA1) genes are located. CFH acts as a negative regulator of the alternative pathway (AP) of the complement system while HtrA1 is an extracellular serine protease. ARMS2 is located upstream of HtrA1 in the primate genome, although the gene is absent in mice. To study the effects of these genes, humanized knock-in mouse lines of Cfh and ARMS2, knockouts of Cfh, HtrA1, HtrA2, HtrA3 and HtrA4 as well as a conditional neural deletion of HtrA2 were generated. Of all the genetically engineered mice produced only mice lacking HtrA2, either systemically or in neural tissues, displayed clear phenotypes. In order to examine these mice thoroughly and systematically, an initial phenotyping schedule was established, consisting of a series of tests related to two main diseases of interest: AMD and Parkinson's. Genetically modified mice can be subjected to appropriate experiments to identify phenotypes that may be related to the associated diseases in humans. A phenotyping regimen with a mitochondrial focus is presented here alongside representative results from the tests of interest.

A reverse-genetics approach to understanding gene families associated with human disease is presented, using mouse as a model system, and the subsequent mouse phenotyping schedule is described. Because mice defective in a gene of interest, HtrA2, manifested Parkinsonian symptoms, the phenotyping regimen is focused on identifying neurological defects.

Um regime de fenotipagem de camundongos geneticamente modificados usados ​​para estudar genes implicados nas doenças humanas do envelhecimento

Age-related diseases are becoming increasingly prevalent and the burden continues to grow as our population ages. Effective treatments are necessary to ...

Ultrasound-based Pulse Wave Velocity Evaluation in Mice

Arterial stiffness can be evaluated by calculating pulse wave velocity (PWV), i.e., the speed with which the pulse wave travels in a conduit vessel. This parameter is being increasingly investigated in small rodent models in which it is used for assessing alterations in vascular function related to particular genotypes/treatments or for characterizing cardiovascular disease progression. This protocol describes an image processing algorithm which leads to non-invasive arterial PWV measurement in mice using ultrasound (US) images only. The proposed technique has been used to assess abdominal aorta PWV in mice and evaluate its age-associated changes.
Abdominal aorta US scans are obtained from mice under gaseous anesthesia using a specific US device equipped with high-frequency US probes. B-mode and Pulse-Wave Doppler (PW-Doppler) images are analyzed in order to obtain diameter and mean velocity instantaneous values, respectively. For this purpose, edge detection and contour tracking techniques are employed. The single-beat mean diameter and velocity waveforms are time aligned and combined in order to achieve the diameter-velocity (lnD-V) loop. PWV values are obtained from the slope of the linear part of the loop, which corresponds to the early systolic phase.
With the present approach, anatomical and functional information about the mouse abdominal aorta can be non-invasively achieved. Requiring the processing of US images only, it may represent a useful tool for the non-invasive characterization of different arterial sites in the mouse in terms of elastic properties. The application of the present technique can be easily extended to other vascular districts, such as the carotid artery, thus providing the possibility to obtain a multi-site arterial stiffness assessment.

Arterial stiffness represents a key factor in cardiovascular disease and pulse wave velocity (PWV) can be considered as a surrogate index for arterial stiffness. This protocol describes an image processing algorithm for calculating PWV in mice based on ultrasound image processing that is applicable at different arterial sites.

O ultra-som à base de pulso de onda Velocity Avaliação em ratos

Arterial stiffness can be evaluated by calculating pulse wave velocity (PWV), i.e., the speed with which the pulse wave travels in a conduit vessel. This ...

The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy (rNAION)

Nonarteritic anterior ischemic optic neuropathy (NAION) is a focal ischemic lesion of the optic nerve that affects 1/700 individuals throughout their lifetime. NAION results in optic nerve edema, selective loss of the retinal ganglion cell neurons (RGCs) and atrophy of the optic nerve. A rodent model of NAION that expresses most NAION features and sequelae has been developed, which is applicable to both rats and mice. This model utilizes a focal laser application of 532 nm wavelength to illuminate a photoactive dye, Rose Bengal (RB), to cause capillary damage and leakage at the targeted anterior optic nerve (the laminar region). After rNAION induction, there is an early optic nerve ischemia, optic nerve edema, and intraneural inflammation, followed by selective RGC and axonal loss. Since the optic nerve is a CNS white matter tract, the rNAION model is applicable to mechanistic studies of selective white matter ischemia, as well as neuroprotective analyses and short and long-term mechanisms of glial and neuronal response to ischemia.

The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy rNAION

The following report describes how to replicate the rodent model of nonarteritic anterior ischemic optic neuropathy (rNAION), using the appropriate dye, contact lens and laser parameters. We also reveal the appropriate steps for evaluating the rNAION lesion in vivo.

O modelo de roedor de nonarterítica óptica isquêmica anterior Neuropatia (rNAION)

Nonarteritic anterior ischemic optic neuropathy (NAION) is a focal ischemic lesion of the optic nerve that affects 1/700 individuals throughout their ...

Longitudinal In Vivo Imaging of the Cerebrovasculature: Relevance to CNS Diseases

Remodeling of the brain vasculature is a common trait of brain pathologies. In vivo imaging techniques are fundamental to detect cerebrovascular plasticity or damage occurring overtime and in relation to neuronal activity or blood flow. In vivo two-photon microscopy allows the study of the structural and functional plasticity of large cellular units in the living brain. In particular, the thinned-skull window preparation allows the visualization of cortical regions of interest (ROI) without inducing significant brain inflammation. Repetitive imaging sessions of cortical ROI are feasible, providing the characterization of disease hallmarks over time during the progression of numerous CNS diseases. This technique accessing the pial structures within 250 &#956;m of the brain relies on the detection of fluorescent probes encoded by genetic cellular markers and/or vital dyes. The latter (e.g., fluorescent dextrans) are used to map the luminal compartment of cerebrovascular structures. Germane to the protocol described herein is the use of an in vivo marker of amyloid deposits, Methoxy-O4, to assess Alzheimer&#39;s disease (AD) progression. We also describe the post-acquisition image processing used to track vascular changes and amyloid depositions. While focusing presently on a model of AD, the described protocol is relevant to other CNS disorders where pathological cerebrovascular changes occur.

Longitudinal In Vivo Imaging of the Cerebrovasculature: Relevance to CNS Diseases

This manuscript describes a procedure to track the remodeling of the cerebrovasculature during amyloid plaque accumulation in vivo using longitudinal two-photon microscopy. A thinned-skull preparation enables the visualization of fluorescent dyes to assess the progression of cerebrovascular damage in a mouse model of Alzheimer&#39;s disease.

Longitudinal<em&gt; In Vivo</em&gt; Imagens do Cerebrovasculature: Relevância para doenças do SNC

Remodeling of the brain vasculature is a common trait of brain pathologies. In vivo imaging techniques are fundamental to detect cerebrovascular ...

Development of a Direct Pulp-capping Model for the Evaluation of Pulpal Wound Healing and Reparative Dentin Formation in Mice

Dental pulp is a vital organ of a tooth fully protected by enamel and dentin. When the pulp is exposed due to cariogenic or iatrogenic injuries, it is often capped with biocompatible materials in order to expedite pulpal wound healing. The ultimate goal is to regenerate reparative dentin, a physical barrier that functions as a "biological seal" and protects the underlying pulp tissue. Although this direct pulp-capping procedure has long been used in dentistry, the underlying molecular mechanism of pulpal wound healing and reparative dentin formation is still poorly understood. To induce reparative dentin, pulp capping has been performed experimentally in large animals, but less so in mice, presumably due to their small sizes and the ensuing technical difficulties. Here, we present a detailed, step-by-step method of performing a pulp-capping procedure in mice, including the preparation of a Class-I-like cavity, the placement of pulp-capping materials, and the restoration procedure using dental composite. Our pulp-capping mouse model will be instrumental in investigating the fundamental molecular mechanisms of pulpal wound healing in the context of reparative dentin in vivo by enabling the use of transgenic or knockout mice that are widely available in the research community.

We describe a step-by-step method of performing direct pulp capping on mice teeth for the evaluation of pulpal wound healing and reparative dentin formation in vivo.

Desenvolvimento de um Modelo-capeamento pulpar direto para a Avaliação dos pulpar cicatrização de feridas e Formação reparadora dentina em Ratos

Dental pulp is a vital organ of a tooth fully protected by enamel and dentin. When the pulp is exposed due to cariogenic or iatrogenic injuries, it is ...

Using Retinal Imaging to Study Dementia

The retina offers a unique &#8220;window&#8221; to study pathophysiological processes of dementia in the brain, as it is an extension of the central nervous system (CNS) and shares prominent similarities with the brain in terms of embryological origin, anatomical features and physiological properties.&#160; The vascular and neuronal structure in the retina can now be visualized easily and non-invasively using retinal imaging techniques, including fundus photography and optical coherence tomography (OCT), and quantified semi-automatically using computer-assisted analysis programs. Studying the associations between vascular and neuronal changes in the retina and dementia could improve our understanding of dementia and, potentially, aid in diagnosis and risk assessment.&#160; This protocol aims to describe a method of quantifying and analyzing retinal vasculature and neuronal structure, which are potentially associated with dementia. This protocol also provides examples of retinal changes in subjects with dementia, and discusses technical issues and current limitations of retinal imaging.

The retina shares prominent similarities with the brain and thus represents a unique window to study vasculature and neuronal structure in the brain non-invasively. This protocol describes a method to study dementia using retinal imaging techniques. This method can potentially aid in diagnosis and risk assessment of dementia.

Utilizando imagens da retina para estudar a demência

The retina offers a unique &#8220;window&#8221; to study pathophysiological processes of dementia in the brain, as it is an extension of the central ...

Measuring the Carotid to Femoral Pulse Wave Velocity (Cf-PWV) to Evaluate Arterial Stiffness

For the elderly, arterial stiffening is a good marker for aging evaluation and it is recommended that the arterial stiffness be determined noninvasively by the measurement of carotid to femoral pulse wave velocity (cf-PWV) (Class I; Level of Evidence A). In literature, numerous community-based or disease-specific studies have reported that higher cf-PWV is associated with increased cardiovascular risk. Here, we discuss strategies to evaluate arterial stiffness with cf-PWV. Following the well-defined steps detailed here, e.g., proper position operator, distance measurement, and tonometer position, we will obtain a standard cf-PWV value to evaluate arterial stiffness. In this paper, a detailed stepwise method to record a good quality PWV and pulse wave analysis (PWA) using a non-invasive tonometry-based device will be discussed.

Measuring the Carotid to Femoral Pulse Wave Velocity Cf-PWV to Evaluate Arterial Stiffness

This protocol describes a method to standardize the measurements of carotid-femoral pulse wave velocity to evaluate arterial stiffness.

Medindo a carótida para velocidade de onda de pulso Femoral (Cf-VOP) para avaliar a rigidez Arterial

For the elderly, arterial stiffening is a good marker for aging evaluation and it is recommended that the arterial stiffness be determined noninvasively ...

Integration of Brain Tissue Saturation Monitoring in Cardiopulmonary Exercise Testing in Patients with Heart Failure

Cerebral hypo-oxygenation during rest or exercise negatively impacts the exercise capacity of patients with heart failure with reduced ejection fraction (HF). However, in clinical cardiopulmonary exercise testing (CPET), cerebral hemodynamics is not assessed. NIRS is used to measure cerebral tissue oxygen saturation (SctO2) in the frontal lobe. This method is reliable and valid and has been utilized in several studies. SctO2 is lower during both rest and peak exercise in patients with HF than in healthy controls (66.3 &#177; 13.3% and 63.4 &#177; 13.8% vs. 73.1 &#177; 2.8% and 72 &#177; 3.2%). SctO2 at rest is significantly linearly correlated with peak VO2 (r = 0.602), oxygen uptake efficiency slope (r = 0.501), and brain natriuretic peptide (r = -0.492), all of which are recognized prognostic and disease severity markers, indicating its potential prognostic value. SctO2 is determined mainly by end-tidal CO2 pressure, mean arterial pressure, and hemoglobin in the HF population. This article demonstrates a protocol that integrates SctO2 using NIRS into incremental CPET on a calibrated bicycle ergometer.

This protocol integrated near-infrared spectroscopy into conventional cardiopulmonary exercise testing to identify the involvement of the cerebral hemodynamic response in exercise intolerance in patients with heart failure.

Integração do monitoramento da saturação do tecido cerebral no teste de exercício cardiopulmonar em pacientes com insuficiência cardíaca

Cerebral hypo-oxygenation during rest or exercise negatively impacts the exercise capacity of patients with heart failure with reduced ejection fraction ...