Os autores têm sem agradecimentos.

todas as experiências em animais foram realizadas em conformidade com a lei alemã de proteção Animal (TierSchG) e foram aprovadas pelo Comitê de bem-estar dos animais locais (Baixa Saxônia Secretaria de defesa do consumidor e da segurança alimentar, protocolo TSA 14/1556). O peso mínimo de mouse apropriado para este modelo é de 25 g.
 1. preparativos pré-operatório Nota: todos os procedimentos são realizados sob condições limpas, não estéril, com instrumentos esterilizados.
<ol><li>Lugar 50-60 8 cm longo propileno oco fibras em paralelo em um tubo e se conectar com um adaptador T em ambos os lados. Preencher o espaço entre as fibras ocas e o ramo externo do adaptador em T com cola.
	<ol><li>Permitir pelo menos 24 h para a cola endurecer. Cortar as fibras ocas que se estendem do T-adaptador usando um micrótomo padrão e encostar o tubo do silicone para os sites de conexão.</li>
	</ol></li> <li>Agulha 27G metal (a partir de um branule de 26G) Introduza uma cânula de poliuretano 2 Fr. Use uma lâmina cirúrgica e microtesoura sob um microscópio (ampliação de X 8-12), para fazer três ou quatro fenestrações de cerca de 0,15 cm cada dentro do terço distal da cânula para assegurar ideal venoso retorna fluxo.
	<ol><li>Remover o fio quando concluído. Use cotonetes para dissecção romba e retração de estruturas. Usar compressas de gaze (2 de 5 x 5 cm) para absorver o excesso de líquido para evitar a desidratação do tecido.</li>
	</ol></li> <li>Preparar o volume de escorva. Conclusão, adicione 30 UI de heparina por mL de solução e 2,5% v/v de uma solução de 8,4% de NaHCO 3 para o buffer de escorva. Armazene esta solução a 4 ° C até usado.</li>
	<li>Encher o circuito de CEC (ou seja,, bomba, caçador de ar, tubulação e ambas as cânulas) com solução de escorva µ l 850 através da cânula venosa. Uma vez preenchido, mantenha o CPB circular até que o animal está pronto para canulação.</li>
</ol> 2. Anestesia animal <ol><li>para administrar anestesia, coloque o animal em uma câmara de indução abaixo dos 2,5% mistura de isoflurano/oxigênio v/v. Confirme a anestesia adequada avaliando o reflexo de retirada de pedal, reflexo de pitada de cauda e reflexo de piscar de olhos. Aplicar a pomada veterinária para evitar ressecamento do olho.</li>
	<li>Colocar o animal sobre uma almofada de aquecimento com função de regulação de temperatura. Medir a temperatura do corpo com uma sonda inserida no reto e conectado ao sistema de aquisição de dados.</li>
	<li>Após anestesia completa é alcançada, entubar o animal orotracheally usando um braunule de plástico de 20G, inseri-lo oralmente e empurrá-lo para a traqueia sob controle visual. Inicie ventilação mecânica usando um vaporizador de isoflurano, ligado a um ventilador de rato. Dependendo do peso do animal, ajustar a ventilação para que seja alcançado um volume tidal de 250-350 µ l.</li>
	<li>Para garantir a completa analgesia, injetar carprofeno animais de peso corporal de 5 mg/kg por via subcutânea. É recomendável que a concentração de isoflurano ser mantido entre 1,3-2,5% de oxigênio. Concentração de isoflurano pode ser ajustada manualmente dependendo da fase do procedimento.</li>
</ol> 3. Procedimentos cirúrgicos <ol><li>após intubação e anestesia completa é alcançada, realizar uma incisão de pele no pescoço com uma tesoura bem afiada, separe os músculos de forma contundente e expor o direito de veia jugular e carótida esquerda. Durante a preparação, coagular pequenos vasos usando um Coagulador veterinário ligado a pinça bipolar para garantir a perda de sangue mínima.</li>
	<li>Após a preparação dos vasos jugular, cranialmente ligate do segmento distal da artéria carótida comum esquerda até sua bifurcação usando suturas de seda de 8-0.</li>
	<li>Conectar-se a extremidade distal de uma cânula de 27 G para a tubagem de influxo arterial usando um conector de silicone (ID de 0,5 mm, 1 mm OD), deslizamento de sutura seda lugar 8-0 nós no segmento proximal da artéria e inserir a ponta da cânula na artéria carótida.</li>
	<li>Após o correto posicionamento da ponta da cânula, avançar a ponta da cânula para que ela põe de 3 a 4 mm proximal para o arco aórtico. Fixar a ponta da cânula fixando com nodos seda 8-0.</li>
	<li>Usando microcirúrgico fórceps e a tesoura, faz dissecação romba e afiada, expor a veia jugular direita por uma preparação sem corte de tecido lateralmente ao músculo esternocleidomastoideo, perto da clavícula e coloque um nó de seda de 8-0 na extremidade distal e um 8-0 laço na extremidade proximal.</li>
	<li>Depois da ligadura da extremidade distal da veia jugular, coloque a ponta da cânula venosa na veia jugular certa e progresso para o átrio direito e fixá-lo com o nó de seda. Para retorno venoso ideal, pode ser necessário avançar a ponta venosa no ventrículo direito.</li>
	<li>Uma vez que a posição correta da cânula é alcançada, realizar heparinização sistémica adicionando 2,5 UI de heparina por grama do peso do corpo animal através de injeção intravenosa direta na veia jugular.</li>
	<li>Para monitorização de pressão invasiva em tempo real, Canule a artéria femoral esquerda. Expor a zona da virilha e suavemente separar a artéria femoral comum a veia femoral e o nervo femoral sob 25 ampliação X.
	<ol><li>Ligam a parte distal da artéria femoral. Temporariamente occlude a parte proximal com um deslizamento-nó e faça uma pequena incisão na parede anterior usando microfórceps.</li>
		<li>Depois, introduza uma cânula de poliuretano 1 Fr na artéria femoral e fixe-o com uma seda sutura de 10-0. Esta cânula é usada para extrair amostras de sangue para gasometria arterial.</li>
	</ol></li> <li>Após a colocação do sucesso das cânulas arteriais e venosas, iniciar extracorpórea por ligar a bomba. O tempo de intubação para arranque do CPB é aproximadamente 45 min. iniciar usando uma taxa de fluxo de 0,5 mL/min, dependendo as medições da pressão sistêmica e aumento do fluxo sanguíneo dentro de 2 min para o fluxo total (4-6 mL/min), aumentando a velocidade da bomba.</li>
	<li>Sob monitoramento completo, para executar uma esternotomia superior usando a tesoura afiada, a partir do manúbrio e indo 5 mm na direção caudal. Coagular as bordas esternais imediatamente para evitar sangramento. Expor o arco aórtico, puxando a artéria carótida direita em direção cranial.</li>
	<li>Controle ótimo, circunferencialmente gratuitamente o arco aórtico do Timo e do tecido circundante, para facilitar a fixação. Coloque um laço de seda de 8-0 em torno da aorta ascendente. Para colocar a braçadeira transversal para cardioplegia, puxe o laço de seda na direção cranial para expor melhor a aorta ascendente.</li>
</ol> 4. Bypass cardiopulmonar e gasometria arterial <ol><li>para gasometria arterial (BGA), usar tubos capilares de vidro para coletar sangue arterial de 60-90 µ l através do cateter de artéria femoral.
	<ol><li>Usar uma pinça pequena do tubo do silicone e retire o tubo do cateter. Liberar a pressão lentamente sobre a mordaça para permitir o enchimento controlado do tubo capilar.</li>
		<li>Reanexar o tubo do silicone para o cateter. Inserir os tubos capilares o analisador de gás do sangue para medições nos seguintes pontos tempo: 10 min após início da CEC com ventilação (BGA1) após 25 min de CEC e a 15 min de parada respiratória (BGA2) após 40 min de CEC e 30 min de parada respiratória (BGA3) após 55 min de CEC, 45 min de respiratória e 20 min de parada cardíaca (BGA4)</li></ol></li> <li>sob fluxo estável do CPB, initia parada respiratória te parando ventilação.</li>
	<li>Após o término da ventilação, conjunto a almofada de aquecimento para 28 ° c e começar a refrigeração topicamente o animal à temperatura de 28 º c dentro o primeiro 20 min de parada respiratória, usando gaze embebida em soro fisiológico frio.</li>
	<li>Uma vez atingida uma temperatura de 28 º c e depois de um total de 30 min de parada respiratória, administrar 0,3 mL de solução de KCl de 7,45% no circuito CPB para iniciar cardioplégica.</li>
	<li>Para cruzar o aperto da aorta, puxe o laço de seda previamente colocado (etapa 3.10) na direção cranial para melhor exposição e coloque uma pinça de micro-serrefine na parte ascendente da aorta.</li>
	<li>Realizar um total de 60 min de parada respiratória e 30 min de parada cardíaca. Retire o grampo de micro-serrefine para iniciar a reperfusão do coração, a aorta ascendente. Simultaneamente, re-ventilar e aquecer o animal a 37 ° C.</li>
	<li>Após a reperfusão é concluída e o animal chegou a normotermia, encerrar o CPB, desligando a bomba e continuar a monitorar medidas fisiológicas (por exemplo, a temperatura do corpo, ECG e pressão arterial invasiva) por 20 min.</li>
	<li>No final do experimento, extrair sangue do animal sob anestesia completa (5% de isoflurano) e coletar sangue e órgãos para ainda mais análise 14.</li>
</ol>

<ol>
	<li>Edmunds, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiopulmonary+Bypass+after+50+Years.">Cardiopulmonary Bypass after 50 Years.</a> N. Engl. J. Med. 351 (16), 1601-1603 (2004).</li><li>Goto, T., Maekawa, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cerebral+dysfunction+after+coronary+artery+bypass+surgery.">Cerebral dysfunction after coronary artery bypass surgery.</a> J. Anesth. 28 (2), 242-248 (2014).</li><li>Uysal, S., Reich, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurocognitive+outcomes+of+cardiac+surgery.">Neurocognitive outcomes of cardiac surgery.</a> J. Cardiothorac. Vasc. Anesth. 27 (5), 958-971 (2013).</li><li>Ballaux, P. K., Gourlay, T., Ratnatunga, C. P., Taylor, K. 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Nós desenvolvemos um modelo clinicamente relevante pleno funcionamento da CEC em um rato. Com mais de 30 cepas de ratos a ter doenças cardiovasculares, nosso modelo poderia ser um ponto de partida para o desenvolvimento de novos protocolos potenciais relacionados à CEC. Além disso, devido a infinidade de reagentes específicos de rato e camundongos nocaute-para fora, este modelo não pode apenas substituir o actual modelo de rato da CEC, mas irá facilitar a dissecação dos mecanismos moleculares envolvidos em danos...

Desde a introdução do extracorpórea (CEC), para a clínica, ele tem desempenhado um papel essencial na cirurgia cardíaca1. Na moderna cirurgia cardíaca, prolongado tempo de CEC é essencial para realizar procedimentos combinados e extensas reconstruções da aorta. Embora os avanços tecnológicos têm sido tremendos, o uso de circulação extracorporais é associado com intra e pós-operatória sistêmica e órgão local danificar2,3.
Grandes modelos animais foram desenvolvidos para investigar o papel da CEC em processos fisiológicos4,5. Embora estes modelos forneceu insights sobre alguns do CPB associada a complicações, eles são extremamente caros e ferramentas moleculares (por exemplo, anticorpos) são muito limitadas. Desenvolveu-se uma alternativa mais custo-eficiente em pequenos animais. Desde o seu desenvolvimento, vários estudos têm sido realizados para otimizar um modelo CPB em ratos e coelhos5,6,7,8,9. Esses modelos fornecem uma boa base para medições de processos fisiopatológicos da doença; no entanto, eles são ainda insuficientes para investigar Imunologia celular e humoral, devido à falta de reagentes e anticorpos pertinentes. Isto prejudica o seu papel neste campo de pesquisa.
Recentemente desenvolvemos um modelo do rato do CPB. Devido a uma grande variedade de reagentes específicos de rato e camundongos geneticamente modificados, modelos de mouse são, em geral, o modelo de escolha para a pesquisa fisiológica, molecular e imunológico10,11. Portanto, nosso modelo facilitará o estudo da CEC em relação a várias comorbidades, como existem muitas cepas de ratos disponíveis com doenças clinicamente relevante12,13. Nesse sentido, este paper descreve, em detalhes, como executar CPB em camundongos. Oxigênio e parâmetros hemodinâmicos são monitorados de perto após a detenção de fundo respiratória e circulatória.

<table border="0" cellpadding="0" cellspacing="0" width="1191">
	<tbody>
		<tr height="23">
			<td height="23" style="height:23px;width:297px;">Sterofundin</td>
			<td style="width:279px;">B.Braun Petzold GmbH</td>
			<td style="width:181px;">PZN:8609189</td>
			<td style="width:435px;">priming volume, 1:1 with Tetraspan</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Tetraspan 6% HES Solution</td>
			<td style="width:279px;">B. Braun Melsungen AG</td>
			<td style="width:181px;">PZN: 05565416</td>
			<td>priming volume, 1:1 with Sterofundin</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Heparin Natrium 25.000</td>
			<td style="width:279px;">Ratiopharm GmbH</td>
			<td style="width:181px;">PZN: 3029843</td>
			<td>2.5 IU per ml of priming solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">NaHCO3 8,4% Solution</td>
			<td style="width:279px;">B. Braun Melsungen AG</td>
			<td style="width:181px;">PZN: 1579775</td>
			<td>3% in priming solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">KCL 7,45 % Solution</td>
			<td style="width:279px;">B. Braun Melsungen AG</td>
			<td style="width:181px;">PZN: 2418577</td>
			<td>0.1 ml for cardioplegia</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Carprofen</td>
			<td style="width:279px;">Zoetis Inc., USA</td>
			<td>PZN:00289615 08859153</td>
			<td>5 mg/kg/BW</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">1 Fr PU Catheter</td>
			<td style="width:279px;">Instechlabs INC., USA</td>
			<td style="width:181px;">C10PU-MCA1301</td>
			<td>carotid artery</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">2 Fr PU Catheter</td>
			<td style="width:279px;">Instechlabs INC., USA</td>
			<td style="width:181px;">C20PU-MJV1302</td>
			<td>jugular vein</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vasofix Safety catheter 20G</td>
			<td style="width:279px;">B.Braun Medical</td>
			<td style="width:181px;">4268113S-01</td>
			<td>orotracheal intubation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">8-0 Silk suture braided</td>
			<td>Ashaway Line &#38; Twine Mfg. Co., USA</td>
			<td style="width:181px;">75290</td>
			<td>ligature</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Isoflurane</td>
			<td style="width:279px;">Piramal Critical Care Deutschland GmbH</td>
			<td style="width:181px;">PZN:9714675</td>
			<td>narcosis</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">CLINITUBES blood capillaries</td>
			<td style="width:279px;">Radiomed GmbH</td>
			<td style="width:181px;">51750132</td>
			<td>blood sampling 60 - 95 microliter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Spring Scissors - 6mm Blades</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">15020-15</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Spring Scissors - 2mm Blades</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">15000-03</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Halsted-Mosquito Hemostat</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">13009-12</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dumont #55 Forceps</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">11295-51</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Castroviejo Micro Needle Holder - 9cm</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">12060-02</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro Serrefines</td>
			<td>Fine Science Tools GmbH</td>
			<td>18555-01</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bulldog Serrefine</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">18050-28</td>
			<td>instruments</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:297px;">MiniVent Ventilator for Mice (Model 845)</td>
			<td style="width:279px;">Harvard Apparatus</td>
			<td style="width:181px;">73-0044</td>
			<td>mechanical ventilation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Isoflurane Vaporizer Drager 19.1</td>
			<td style="width:279px;">Dr&#228;gerwerk AG &#38; Co. KGaA</td>
			<td style="width:181px;"></td>
			<td>anesthesia 1.3 -2.5%</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PowerLab data acquisition device 4/35</td>
			<td>ADInstruments Ltd, New Zealand</td>
			<td style="width:181px;">PL3504</td>
			<td>invasive pressure, ECG, temperature</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ABL 800 Flex</td>
			<td>Radiometer GmbH</td>
			<td style="width:181px;"></td>
			<td>blood gas analysis</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">NMRI mice</td>
			<td style="width:279px;">Charles River Laboratories</td>
			<td style="width:181px;">Crl:NMRI(Han)</td>
			<td>male, 30-35 g, 12 weeks old, housed at least 1 week before the experiment</td>
		</tr>
	</tbody>
</table>

cardiopulmonary bypass in a mouse model: a novel approach

Como extracorpórea prolongada torna-se mais essencial durante intervenções cardíacas, uma crescente demanda clínica surge para otimização do processo e para minimizar danos nos órgãos resultantes de circulação extracorporais prolongada. O objetivo deste trabalho foi demonstrar um modelo totalmente funcional e clinicamente relevante de bypass cardiopulmonar em um rato. Relatamos o dispositivo design, otimização do circuito de perfusão e técnicas microcirúrgicos. Este modelo é um modelo agudo, que não é compatível com a sobrevivência, devido à necessidade de vários desenhos de sangue. Devido a gama de ferramentas disponíveis para ratos (por exemplo, marcadores, nocautes, etc), este modelo facilitará a investigação sobre os mecanismos moleculares de danos nos órgãos e o efeito de bypass cardiopulmonar em relação a outros comorbidades.

Este protocolo descreve o circuito de perfusão, procedimentos cirúrgicos e monitoramento de parâmetros fisiológicos durante a CEC de um rato. Quando realizada por um micro-cirurgião adequadamente qualificado, os resultados são consistentemente e reproducibly obtidos.
Para manter a perfusão adequada do tecido, a pressão arterial média é sempre mantida entre 40 e 60 mmHg, ajustando o fluxo de sangue do CPB e adicionando ...

Watch this Scientific Journal Video about Cardiopulmonary Bypass in a Mouse Model: A Novel Approach at JoVE.com

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

Este artigo descreve como realizar bypass cardiopulmonar em ratos. Este novo modelo facilitará a investigação dos mecanismos moleculares envolvidos na lesão de órgão.

Bypass cardiopulmonar em um modelo do rato: uma nova abordagem

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization ...

cardiopulmonary-bypass-in-a-mouse-model-a-novel-approach

Research

JoVE Journal

Medicine

10.8K visualizações.  Hannover Medical School. Este artigo descreve como realizar bypass cardiopulmonar em ratos. Este novo modelo facilitará a investigação dos mecanismos moleculares envolvidos na lesão de órgão.

Vídeo: Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

Vascular Occlusion Training for Inclusion Body Myositis: A Novel Therapeutic Approach

Inclusion body myositis (IBM) is a rare idiopathic inflammatory myopathy. It is known to produces remarkable muscle weakness and to greatly compromise function and quality of life. Moreover, clinical practice suggests that, unlike other inflammatory myopathies, the majority of IBM patients are not responsive to treatment with immunosuppressive or immunomodulatory drugs to counteract disease progression1. Additionally, conventional resistance training programs have been proven ineffective in restoring muscle function and muscle mass in these patients2,3. Nevertheless, we have recently observed that restricting muscle blood flow using tourniquet cuffs in association with moderate intensity resistance training in an IBM patient produced a significant gain in muscle mass and function, along with substantial benefits in quality of life4. Thus, a new non-pharmacological approach for IBM patients has been proposed. Herein, we describe the details of a proposed protocol for vascular occlusion associated with a resistance training program for this population.

The present article presents the details pertaining to the application of resistance training associated to vascular occlusion in IBM patients.

Oclusão Vascular Formação para a Inclusão Miosite Corpo: uma nova abordagem terapêutica

Inclusion body myositis (IBM) is a rare idiopathic inflammatory myopathy. It is known to produces remarkable muscle weakness and to greatly compromise ...

Medicina

Bronchial Thermoplasty:  A Novel Therapeutic Approach to Severe Asthma

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled manner to reduce excessive airway smooth muscle. Reducing airway smooth muscle decreases the ability of the airways to constrict, thereby reducing the frequency of asthma attacks. Bronchial thermoplasty is delivered by the Alair System and is performed in three outpatient procedure visits, each scheduled approximately three weeks apart. The first procedure treats the airways of the right lower lobe, the second treats the airways of the left lower lobe and the third and final procedure treats the airways in both upper lobes. After all three procedures are performed the bronchial thermoplasty treatment is complete.
Bronchial thermoplasty is performed during bronchoscopy with the patient under moderate sedation. All accessible airways distal to the mainstem bronchi between 3 and 10 mm in diameter, with the exception of the right middle lobe, are treated under bronchoscopic visualization. Contiguous and non-overlapping activations of the device are used, moving from distal to proximal along the length of the airway, and systematically from airway to airway as described previously. Although conceptually straightforward, the actual execution of bronchial thermoplasty is quite intricate and procedural duration for the treatment of a single lobe is often substantially longer than encountered during routine bronchoscopy. As such, bronchial thermoplasty should be considered a complex interventional bronchoscopy and is intended for the experienced bronchoscopist. Optimal patient management is critical in any such complex and longer duration bronchoscopic procedure. This article discusses the importance of careful patient selection, patient preparation, patient management, procedure duration, postoperative care and follow-up to ensure that bronchial thermoplasty is performed safely.
Bronchial thermoplasty is expected to complement asthma maintenance medications by providing long-lasting asthma control and improving asthma-related quality of life of patients with severe asthma. In addition, bronchial thermoplasty has been demonstrated to reduce severe exacerbations (asthma attacks) emergency rooms visits for respiratory symptoms, and time lost from work, school and other daily activities due to asthma.

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled manner to reduce excessive airway smooth muscle. Reducing airway smooth muscle decreases the ability of the airways to constrict, thereby reducing the frequency of asthma attacks.

Termoplastia brônquica: uma nova abordagem terapêutica para Asma Grave

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled ...

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient management strategies. Orthotopic mouse models of oral cancer have been developed to facilitate the study of factors that impact invasion and serve as model system for evaluating anti-tumor therapeutics. In these systems, visualization of disseminated tumor cells within oral cavity tissues has typically been conducted by either conventional histology or with in vivo bioluminescent methods. A primary drawback of these techniques is the inherent inability to accurately visualize and quantify early tumor cell invasion arising from the primary site in three dimensions. Here we describe a protocol that combines an established model for squamous cell carcinoma of the tongue (SCOT) with two-photon imaging to allow multi-vectorial visualization of lingual tumor spread. The OSC-19 head and neck tumor cell line was stably engineered to express the F-actin binding peptide LifeAct fused to the mCherry fluorescent protein (LifeAct-mCherry). Fox1nu/nu mice injected with these cells reliably form tumors that allow the tongue to be visualized by ex-vivo application of two-photon microscopy. This technique allows for the orthotopic visualization of the tumor mass and locally invading cells in excised tongues without disruption of the regional tumor microenvironment. In addition, this system allows for the quantification of tumor cell invasion by calculating distances that invaded cells move from the primary tumor site. Overall this procedure provides an enhanced model system for analyzing factors that contribute to SCOT invasion and therapeutic treatments tailored to prevent local invasion and distant metastatic spread. This method also has the potential to be ultimately combined with other imaging modalities in an in vivo setting.

A comprehensive overview of the techniques involved in generating a mouse model of oral cancer and quantitative monitoring of tumor invasion within the tongue through multi-photon microscopy of labeled cells is presented. This system can serve as a useful platform for the molecular assessment and drug efficacy of anti-invasive compounds.

Multi-fotão imagem de invasão das células tumorais em um modelo de camundongo ortotópico de carcinoma epidermóide oral

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused by whole-body hypoperfusion.5,6 Successful reproduction of whole-body hypoperfusion in rodent models has been fraught with 
difficulty.7-9,9,10 Models which employ focal ischemia have repeatedly demonstrated results which do not translate to the clinical setting, and larger 
animal models which allow for whole body hypoperfusion lack access to the full toolset of genetic manipulation possible in the mouse.11,12 However, in recent 
years a mouse model of cardiac arrest and cardiopulmonary resuscitation has emerged which can be adapted to model AKI.13 This model reliably reproduces 
physiologic, functional, anatomic, and histologic outcomes seen in clinical AKI, is rapidly repeatable, and offers all of the significant advantages of a murine surgical 
model, including access to genetic manipulative techniques, low cost relative to large animals, and ease of use. Our group has developed extensive experience 
with use of this model to assess a number of organ-specific outcomes in AKI.14,15

A powerful model for perioperative and critical care related acute kidney injury is presented. Using whole body hypoperfusion induced by cardiac arrest it is possible to nearly replicate the histologic and functional changes of clinical AKI.

Cardiac Arrest normotérmica e Ressuscitação Cardiopulmonar: um modelo do rato de isquemia-reperfusão

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused ...

Roux-en-Y Gastric Bypass Operation in Rats

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality benefit is bariatric surgery1,2. Consequently, there has been a steady rise in the number of bariatric operations done worldwide in recent years with the Roux-en-Y gastric bypass (gastric bypass) being the most commonly performed operation3. Against this background, it is important to understand the physiological mechanisms by which gastric bypass induces and maintains body weight loss. These mechanisms are yet not fully understood, but may include reduced hunger and increased satiation4,5, increased energy expenditure6,7, altered preference for food high in fat and sugar8,9, altered salt and water handling of the kidney10 as well as alterations in gut microbiota11. Such changes seen after gastric bypass may at least partly stem from how the surgery alters the hormonal milieu because gastric bypass increases the postprandial release of peptide-YY (PYY) and glucagon-like-peptide-1 (GLP-1), hormones that are released by the gut in the presence of nutrients and that reduce eating12.
During the last two decades numerous studies using rats have been carried out to further investigate physiological changes after gastric bypass. The gastric bypass rat model has proven to be a valuable experimental tool not least as it closely mimics the time profile and magnitude of human weight loss, but also allows researchers to control and manipulate critical anatomic and physiologic factors including the use of appropriate controls. Consequently, there is a wide array of rat gastric bypass models available in the literature reviewed elsewhere in more detail 13-15. The description of the exact surgical technique of these models varies widely and differs e.g. in terms of pouch size, limb lengths, and the preservation of the vagal nerve. If reported, mortality rates seem to range from 0 to 35%15. Furthermore, surgery has been carried out almost exclusively in male rats of different strains and ages. Pre- and postoperative diets also varied significantly.
Technical and experimental variations in published gastric bypass rat models complicate the comparison and identification of potential physiological mechanisms involved in gastric bypass. There is no clear evidence that any of these models is superior, but there is an emerging need for standardization of the procedure to achieve consistent and comparable data. This article therefore aims to summarize and discuss technical and experimental details of our previously validated and published gastric bypass rat model.

Numerous studies using gastric bypass rat models have been recently conducted to uncover the underlying physiological mechanisms of Roux-en-Y gastric bypass operations. This article aims to demonstrate and discuss the technical and experimental details of our published gastric bypass rat model to understand advantages and limitations of this experimental tool.

Roux-en-Y operação de bypass gástrico em ratos

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality ...

A Novel Surgical Approach for Intratracheal Administration of Bioactive Agents in a Fetal Mouse Model

Prenatal pulmonary delivery of cells, genes or pharmacologic agents could provide the basis for new therapeutic strategies for a variety of genetic and acquired diseases. Apart from congenital or inherited abnormalities with the requirement for long-term expression of the delivered gene, several non-inherited perinatal conditions, where short-term gene expression or pharmacological intervention is sufficient to achieve therapeutic effects, are considered as potential future indications for this kind of approach. Candidate diseases for the application of short-term prenatal therapy could be the transient neonatal deficiency of surfactant protein B causing neonatal respiratory distress syndrome1,2 or hyperoxic injuries of the neonatal lung3. Candidate diseases for permanent therapeutic correction are Cystic Fibrosis (CF)4, genetic variants of surfactant deficiencies5 and &alpha;1-antitrypsin deficiency6.
Generally, an important advantage of prenatal gene therapy is the ability to start therapeutic intervention early in development, at or even prior to clinical manifestations in the patient, thus preventing irreparable damage to the individual. In addition, fetal organs have an increased cell proliferation rate as compared to adult organs, which could allow a more efficient gene or stem cell transfer into the fetus. Furthermore, in utero gene delivery is performed when the individual's immune system is not completely mature. Therefore, transplantation of heterologous cells or supplementation of a non-functional or absent protein with a correct version should not cause immune sensitization to the cell, vector or transgene product, which has recently been proven to be the case with both cellular and genetic therapies7.
In the present study, we investigated the potential to directly target the fetal trachea in a mouse model. This procedure is in use in larger animal models such as rabbits and sheep8, and even in a clinical setting9, but has to date not been performed before in a mouse model. When studying the potential of fetal gene therapy for genetic diseases such as CF, the mouse model is very useful as a first proof-of-concept because of the wide availability of different transgenic mouse strains, the well documented embryogenesis and fetal development, less stringent ethical regulations, short gestation and the large litter size.
Different access routes have been described to target the fetal rodent lung, including intra-amniotic injection10-12, (ultrasound-guided) intrapulmonary injection13,14 and intravenous administration into the yolk sac vessels15,16 or umbilical vein17. Our novel surgical procedure enables researchers to inject the agent of choice directly into the fetal mouse trachea which allows for a more efficient delivery to the airways than existing techniques18.

We developed a novel surgical approach for intratracheal administration of bioactive agents into the mouse fetus. The delivery route is more efficient in targeting the fetal mouse lungs than the commonly used intra-amniotic injection. This procedure has to date not been described in a mouse model.

Uma nova abordagem cirúrgica para administração intratraqueal de agentes bioativos em um modelo de camundongo Fetal

Prenatal pulmonary delivery of cells, genes or pharmacologic agents could provide the basis for new therapeutic strategies for a variety of genetic and ...

Evaluation of a Novel Laser-assisted Coronary Anastomotic Connector - the Trinity Clip - in a Porcine Off-pump Bypass Model

To simplify and facilitate beating heart (i.e., off-pump), minimally invasive coronary artery bypass surgery, a new coronary anastomotic connector, the Trinity Clip, is developed based on the excimer laser-assisted nonocclusive anastomosis technique. The Trinity Clip connector enables simplified, sutureless, and nonocclusive connection of the graft to the coronary artery, and an excimer laser catheter laser-punches the opening of the anastomosis. Consequently, owing to the complete nonocclusive anastomosis construction, coronary conditioning (i.e., occluding or shunting) is not necessary, in contrast to the conventional anastomotic technique, hence simplifying the off-pump bypass procedure. Prior to clinical application in coronary artery bypass grafting, the safety and quality of this novel connector will be evaluated in a long-term experimental porcine off-pump coronary artery bypass (OPCAB) study. In this paper, we describe how to evaluate the coronary anastomosis in the porcine OPCAB model using various techniques to assess its quality. Representative results are summarized and visually demonstrated.

This paper describes a novel nonocclusive coronary anastomotic connector in a porcine off-pump coronary artery bypass (OPCAB) model. This easy-to-use coronary connector has intrinsic potential to facilitate minimally invasive OPCAB surgery.

Avaliação de uma nova Laser-assistida Coronary Anastomotic Connector - Trindade Clip - em um modelo de Bypass Porcina Off-bomba

To simplify and facilitate beating heart (i.e., off-pump), minimally invasive coronary artery bypass surgery, a new coronary anastomotic connector, the ...

A Novel Approach for the Administration of Medications and Fluids in Emergency Scenarios and Settings

The available routes of administration commonly used for medications and fluids in the acute care setting are generally limited to oral, intravenous, or intraosseous routes, but in many patients, particularly in the emergency or critical care settings, these routes are often unavailable or time-consuming to access. A novel device is now available that offers an easy route for administration of medications or fluids via rectal mucosal absorption (also referred to as proctoclysis in the case of fluid administration and subsequent absorption). Although originally intended for the palliative care market, the utility of this device in the emergency setting has recently been described. Specifically, reports of patients being treated for dehydration, alcohol withdrawal, vomiting, fever, myocardial infarction, hyperthyroidism, and cardiac arrest have shown success with administration of a wide variety of medications or fluids (including water, aspirin, lorazepam, ondansetron, acetaminophen, methimazole, and buspirone). Device placement is straightforward, and based on the observation of expected effects from the medication administrations, absorption is rapid. The rapidity of absorption kinetics are further demonstrated in a recent report of the measurement of phenobarbital pharmacokinetics. We describe here the placement and use of this device, and demonstrate methods of pharmacokinetic measurements of medications administered by this method.

This study presents a novel device that offers an easy route to quickly provide medications and fluids in patients with limited or difficult IV access. This small-diameter device is placed in the distal third of the rectum, allowing for ongoing medication and fluids administration.

Uma nova abordagem para a administração de medicamentos e fluídos em cenários de emergência e Configurações

The available routes of administration commonly used for medications and fluids in the acute care setting are generally limited to oral, intravenous, or ...

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. Research models for bypass graft failure are essential for a better understanding of pathobiological and pathophysiological processes during graft patency loss. Large animal models, such as pigs or sheep, resemble human anatomical structures but require special facilities and equipment. This video describes a rat vein interposition model to investigate vein graft patency loss. Rats are inexpensive and easy to handle. Compared to mouse models, the convenient size of rats permits better operability and enables a sufficient amount of material to be obtained for further diverse analysis. In brief, the inferior epigastric vein of a donor rat is harvested and used to replace a segment of the femoral artery. Anastomosis is conducted via single stitches and sealed with fibrin glue. Graft patency can be monitored non-invasively using duplex sonography. Myointimal hyperplasia, which is the main cause for graft patency loss, develops progressively over time and can be calculated from histological cross sections.

This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats.

Veia Interposição Modelo: Um modelo adequado para estudar Bypass Graft Perviedade

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Medição de contratilidade isométrica da artéria mesentérica Mouse usando fio miografia

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...