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1. Preparação da placenta no momento do parto <ol><li> Rotular o cordão umbilical dos gêmeos com um (para o primeiro-nascido) ou dois (para o segundo-nascido) grampos. </li><li> Inspecionar a superfície materna e fetal da placenta para a completude ou interrupção. </li><li> Registrar os seguintes dados: tipo de inserção do cordão umbilical (central, excêntrico, marginal ou vilamentosa), número de vasos sanguíneos no cordão umbilical (geralmente uma veia e duas artérias, às vezes apenas uma artéria) e diferença de cor entre as duas partes da placenta. A seção das membranas dividindo podem ser enviadas para Patologia para confirmar o tipo de corionicidade. </li><li> A placenta pode ser colocado em uma bacia plástica e refrigerado até o exame final (melhor dentro de uma semana) e injeção de corante de cor. </li><li> A placenta não deve ser congelado ou fixo (não use formol). </li></ol> 2. Cateterismo dos vasos umbilicais <ol><li> Lavar a placenta com água morna ou soro fisiológico. </li><li> Aparar as membranas periférica, remover a membrana inter-twin dividindo e descolar as âmnions (para melhor visualização das anastomoses vascular e melhor qualidade das imagens da placenta). </li><li> Transecto cada cordão umbilical com aproximadamente 5 cm de distância a partir da inserção do cordão umbilical. </li><li> Delicadamente esprema coágulos de sangue dos vasos umbilicais e vasos placentários. </li><li> A veia umbilical é geralmente fácil de identificar devido ao seu maior diâmetro, em comparação com o menor diâmetro das duas artérias umbilical. </li><li> Canular a veia umbilical com um cateter de tamanho adequado. Evite passagens falsas. </li><li> Canular uma artéria umbilical com um pequeno cateter. Use uma pinça para alargar a luz da artéria umbilical. Evite passagens falsas. Apenas uma das duas artérias umbilicais precisa ser cateterizada desde uma anastomose (de Hyrtl) conecta as 2 artérias perto da inserção do cordão umbilical. </li><li> Repita os passos para o cordão umbilical outros. </li><li> Colocação do cateter pode ser facilitada pela volta suave e massagear diante dos vasos umbilicais. Qualquer tipo de cateter pode ser usado para este procedimento. Nós escolhemos usar (e reciclar) os cateteres usados ​​em nossa ala de neonatologia para cateterismo umbilical em recém-nascidos. </li><li> Amarre um pedaço de fita adesiva em torno de ambos os cabos para evitar o refluxo do corante de cor durante a injeção de corante. </li></ol> 3. Injeção com corante de cor <ol><li> Conectar uma seringa de 20 ml cheia com um corante de cor para cada cateter. </li><li> Toda a tintura viscoso colorido pode ser usado para visualizar a placenta angio-arquitetura. Use cores contrastantes para permitir uma boa visualização das anastomoses (cores escuras para as artérias, cores brilhantes para as veias). </li><li> Suavemente injetar (com baixa pressão) o corante de cor na veia, enquanto um assistente delicadamente empurra o corante para permitir que a tinta colorida para preencher todos os vasos placentários, também os mais pequenos. </li><li> Preste atenção especial aos vasos pequenos perto do equador vascular (o equador vascular é o lugar onde as anastomoses de ambos os gêmeos se conectar uns com os outros). </li><li> Repita os passos anteriores para injetar tinta colorida na artéria. De nota: as artérias podem ser mais difíceis de injetar e exigem mais paciência. </li><li> Repita os passos para o cordão umbilical outros. </li></ol> 4. Avaliação e documentação da placenta após a injeção de corante de cor <ol><li> Examinar cuidadosamente o equador vascular e gravar o número e tipos de anastomoses. </li><li> Coloque uma fita métrica na placenta para medir os diâmetros e partes da placenta na imagem digital. </li><li> Use uma câmera digital de alta resolução e tirar fotos da placenta injetado. Certifique-se que as fotos são tiradas perpendicular à placenta. </li></ol> 5. Resultados representativos: A placenta angio-arquitetura em gêmeos monocoriônicos varia de acordo com o tipo de gravidez gemelar monocoriônicas. Estudos têm demonstrado que a injeção de anastomoses AA, AV e VV estão presentes em 80%, respectivamente, 95% e 20% de gestações gemelares monocoriônicas simples 1 (Figura 1). Anastomoses AA são considerados para proteger contra o desenvolvimento da STT e TAPS 6. Estudos têm demonstrado que injeções de anastomoses AA ocorrem em apenas 20% da STT placentas e 10% das placentas TAPS 1,2,7 (Figura 2). Em placentas STT tratados com cirurgia a laser fetoscopic, as cicatrizes causadas por coagulação a laser das anastomoses vasculares podem ser vistas na superfície da placenta (Figura 3 e 4). TAPS placentas são caracterizados pela presença de apenas alguns anastomoses minúscula AV 2 (Figura 5). A parcela da placenta do destinatário TAPS é muitas vezes pletórica, enquanto a parte da placenta do doador é pálida (Figura 6). Em gestações gemelares monocoriônicas discordância com peso de nascimento, o crescimento restrito fetomuitas vezes tem uma inserção do cordão umbilical vilamentosa e uma parcela muito menor da placenta (Figura 7). Anastomoses AA estão freqüentemente presentes em placentas monocoriônicas de gêmeos com discordância peso 8. Monocoriônicas-monoamniotic placentas têm uma característica angio-arquitetura com uma pequena distância entre as duas inserções do cordão umbilical (Figura 8). A incidência de anastomoses AA em monoamniotic-monocoriônicas placentas é praticamente 100% e impede o desenvolvimento da STT em gêmeos monoamniotic 9. Os números neste artigo mostrar o quadro característico de placentas monocoriônicas injetada no nosso centro com corante de cor. Rotineiramente usar cores mais escuras (azul ou verde) para as artérias e cores mais claras (amarelo, rosa ou laranja) para as veias. <img alt="Figura 1" src="/files/ftp_upload/3208/3208fig1.jpg" /> Figura 1. Placenta monocoriônicas de uma normal, a gravidez uncomplicated monocoriônicas gêmeos mostrando várias anastomoses AV das artérias às veias verde-rosa (estrelas brancas), várias anastomoses VA das artérias às veias azuis amarelo (estrelas verdes) e um grande AA anastomose (identificado pelo mistura de corante azul e verde; estrela azul). <img alt="Figura 2" src="/files/ftp_upload/3208/3208fig2.jpg" /> Figura 2. Placenta monocoriônicas em uma gravidez STT tratados com amnioreduction série mostrando a presença de apenas AV (estrelas brancas) e VA (estrelas verdes) anastomoses sem anastomose AA. <img alt="Figura 3" src="/files/ftp_upload/3208/3208fig3.jpg" /> Figura 3. STT placenta após a coagulação a laser fetoscopic das anastomoses vascular utilizando a técnica a laser seletiva em que um pequeno residual anastomose foi inadvertidamente deixou patente (estrela branca). Com a técnica a laser seletiva, as anastomoses vasculares são identificado pela primeira vez e, posteriormente, coagulado, um por um. <img alt="Figura 4" src="/files/ftp_upload/3208/3208fig4.jpg" /> Figura 4. STT placenta após a coagulação a laser fetoscopic utilizando a técnica de Salomão, em que, após identificação e de coagulação de cada anastomose individual, o equador vascular completa é coagulado de uma margem placentária para o outro. <img alt="Figura 5" src="/files/ftp_upload/3208/3208fig5.jpg" /> Figura 5. TAPS placenta em que apenas alguns, anastomoses minúscula VA (estrelas brancas) são visíveis ao longo do equador vascular. <img alt="Figura 6" src="/files/ftp_upload/3208/3208fig6.jpg" /> Figura 6. O lado materno da placenta TAPS (mostrado na Figura 5) demonstra a diferença de cor entre as duas partes característica da placenta. <img alt="Figura 7" src="/files/ftp_upload/3208/3208fig7.jpg" /> Figura 7. Placenta monocoriônicas de uma gravidez de gêmeos com restrição de crescimento intra-uterino seletiva de um dos gêmeos. O feto tem um crescimento restrito inserção do cordão umbilical vilamentosa e uma parcela muito menor da placenta. A estrela branca indica uma anastomose AA. <img alt="Figura 8" src="/files/ftp_upload/3208/3208fig8.jpg" /> Figura 8 placenta Monoamniotic:. Observe o AV diversas (estrela verde), VA (blue star) e 2 AA anastomoses (branco estrelas), ea curta distância entre as duas inserções do cordão umbilical.

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STT e TAPS são duas doenças graves que podem ocorrer em gêmeos monocoriônicos. Ambos os distúrbios são devidos a anastomoses AV causando o fluxo de sangue imbalanced intertwin. Nos últimos 2 décadas, a cirurgia a laser fetoscopic tornou-se disponível e mostrou ser o melhor método para o tratamento da STT 10. O objetivo do tratamento a laser fetoscopic é interromper a circulação inter-twin através da coagulação do anastomoses vascular na superfície placentária. No entanto, o tratamento a lase...

<table border="1"><tbody><tr><th> Nome do reagente </th><th> Companhia </th><th> Número de catálogo </th><th> Comentários </th></tr><tr><td> Cateteres umbilicais (várias dimensões, comprimento 40 cm) </td><td> Vygon </td><td> 1.270,08 (8 F) 1.270,08 (5 F) 1.270,04 (4 F) 1.270,03 (3,5 F) 1.270,02 (2,5 F) </td><td> Qualquer tipo de cateter pode ser usado </td></tr><tr><td> 20 ml seringas </td><td> BD Plastipak </td><td> 300613 </td><td></td></tr><tr><td> Cor da tintura (cores diferentes) </td><td> Real Talens Schoolverf </td><td> 36716010 (azul) 36715010 (verde) 36712350 (amarelo) 36713570 (rosa) </td><td> Qualquer corante de cor viscoso pode ser utilizado </td></tr></tbody></table>

accurate and simple evaluation of vascular anastomoses in monochorionic placenta using colored dye

A presença de anastomoses vascular placentária é uma conditio sine qua non para o desenvolvimento de twin-to-twin síndrome de transfusão (STT) e seqüência de policitemia twin anemia (TAPS) 1,2. Estudos injeção de placentas de gêmeos têm mostrado que tais anastomoses são quase invariavelmente presentes em gêmeos monocoriônicos e extremamente rara em gêmeos dicoriônicas 1. Três tipos de anastomoses foram documentados: de esquerda para artéria, da veia a veia e da artéria para a veia. Artério-venosa (AV) anastomoses são unidirecionais e são referidos como &quot;profunda&quot; anastomoses, uma vez que proceda através de um cotilédone da placenta partilhada, enquanto arterio-arterial (AA) e veno-venoso (VV) anastomoses são bi-direcional e são referidos como &quot;superficial&quot;, uma vez que se encontram na placa coriônica. Ambos STT e TAPS são causados ​​por desequilíbrio líquido do fluxo de sangue entre os gêmeos, devido à anastomoses AV. Sangue de um gêmeo (o doador) é bombeado através de uma artéria no cotilédone compartilhada placentária e, em seguida, drenados através de uma veia para a circulação do outro gêmeo (o destinatário). A menos que o sangue é bombeado de volta do destinatário para o doador através direção oposta profunda anastomoses AV ou através de anastomoses superficial, um desequilíbrio de volumes de sangue ocorre, gradativamente, levando ao desenvolvimento da STT ou TAPS. A presença de uma anastomose AA foi mostrado para proteger contra o desenvolvimento da STT e TAPS, compensando o desequilíbrio circulatório causado pela uni-direcional 1,2 anastomoses AV. Injeção de placentas monocoriônicas logo após o nascimento é um meio útil para entender a etiologia de várias (hematológicas) complicações em gêmeos monocoriônicos e é um teste necessário para atingir o diagnóstico de TAPS 2. Além disso, a injeção de placentas STT tratados com cirurgia a laser fetoscopic permite a identificação de possíveis anastomoses residual 3-5. Esta informação adicional é de suma importância para todos os perinatologists envolvidos na gestão e cuidado dos gêmeos monocoriônicos com STT ou TAPS. Várias técnicas de injeção da placenta estão sendo usados ​​atualmente. Nós fornecemos um protocolo simples para avaliar com precisão a presença de anastomoses (residual) vascular usando injeção de tinta colorida.

Watch this Scientific Journal Video about Accurate and Simple Evaluation of Vascular Anastomoses in Monochorionic Placenta using Colored Dye at JoVE.com

Accurate and Simple Evaluation of Vascular Anastomoses in Monochorionic Placenta using Colored Dye

Twin-to-twin síndrome de transfusão e seqüência policitemia twin anemia são dois problemas potencialmente devastadores na medicina perinatal. Ambos os distúrbios ocorrem apenas em gêmeos monocoriônicos e resultam de fluxo sanguíneo através da placenta desequilibrada anastomoses vascular. Nós fornecemos um protocolo simples para avaliar com precisão a presença de anastomoses vascular usando injeção de tinta colorida dos vasos da placenta após o nascimento.

Avaliação precisas e simples de anastomoses vasculares em Placenta monocoriônicas usando Dye Colorido

The presence of placental vascular anastomoses is a conditio sine qua non for the development of twin-to-twin transfusion syndrome (TTTS) and twin anemia ...

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26.6K visualizações.  Leiden University Medical Center. Twin-to-twin síndrome de transfusão e seqüência policitemia twin anemia são dois problemas potencialmente devastadores na medicina perinatal. Ambos os distúrbios ocorrem apenas em gêmeos monocoriônicos e resultam de fluxo sanguíneo através da placenta desequilibrada anastomoses vascular. Nós fornecemos um protocolo simples para avaliar com precisão a presença de anastomoses vascular usando injeção de tinta colorida dos vasos da placenta após o nascimento.

Vídeo: Accurate and Simple Evaluation of Vascular Anastomoses in Monochorionic Placenta using Colored Dye

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&beta; using a Whole Blood Stimulation Assay

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues as well as altered cytokine homeostasis. The innate immune system plays a crucial role in recognizing pathogens and other endogenous danger stimuli. One of the major cytokines released by innate immune cells is Interleukin (IL)-1. Therefore, we utilize a whole blood stimulation assay in order to measure the secretion of inflammatory cytokines and specifically of the pro-inflammatory cytokine IL-1&beta; 1, 2, 3.
Patients with genetic dysfunctions of the innate immune system causing autoinflammatory syndromes show an exaggerated release of mature IL-1&beta; upon stimulation with LPS alone. In order to evaluate the innate immune component of patients who present with inflammatory-associated pathologies, we use a specific immunoassay to detect cellular immune responses to pathogen-associated molecular patterns (PAMPs), such as the gram-negative bacterial endotoxin, lipopolysaccharide (LPS). These PAMPs are recognized by pathogen recognition receptors (PRRs), which are found on the cells of the innate immune system 4, 5, 6, 7. A primary signal, LPS, in conjunction with a secondary signal, ATP, is necessary for the activation of the inflammasome, a multiprotein complex that processes pro-IL-1&beta; to its mature, bioactive form 4, 5, 6, 8, 9, 10.
The whole blood assay requires minimal sample manipulation to assess cytokine production when compared to other methods that require labor intensive isolation and culturing of specific cell populations. This method differs from other whole blood stimulation assays; rather than diluting samples with a ratio of RPMI media, we perform a white blood cell count directly from diluted whole blood and therefore, stimulate a known number of white blood cells in culture 2. The results of this particular whole blood assay demonstrate a novel technique useful in elucidating patient cohorts presenting with autoinflammatory pathophysiologies.

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&amp;#946; using a Whole Blood Stimulation Assay

We describe a simple immunoassay to measure the production of pro-inflammatory cytokines, such as IL-1 beta production, in patients presenting with autoinflammatory phenotypes. By activating cells in whole blood cultures with pathogen-associated molecular patterns, specifically with lipopolysaccharide, cytokine secretion can be conveniently evaluated in whole blood supernatants.

Medição precisa e simples do pró-inflamatórias IL-1β citocinas usando um ensaio Estimulação Sangue Total

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues ...

Medicina

Guide Wire Assisted Catheterization and Colored Dye Injection for Vascular Mapping of Monochorionic Twin Placentas

Monochorionic (MC) twin pregnancies are associated with significantly higher morbidity and mortality rates than dichorionic twins. Approximately 50% of MC twin pregnancies develop complications arising from the shared placenta and associated vascular connections1. Severe twin-to-twin syndrome (TTTS) is reported to account for approximately 20% of these complications2,3. Inter-twin vascular connections occur in almost all MC placentas and are related to the prognosis and outcome of these high-risk twin pregnancies. The number, size and type of connections have been implicated in the development of TTTS and other MC twin conditions. Three types of inter-twin vascular connections occur: 1) artery to vein connections (AVs) in which a branch artery carrying deoxygenated blood from one twin courses along the fetal surface of the placenta and dives into a placental cotyledon. Blood flows via a deep intraparenchymal capillary network into a draining vein that emerges at the fetal surface of the placenta and brings oxygenated blood toward the other twin. There is unidirectional flow from the twin supplying the afferent artery toward the twin receiving the efferent vein; 2) artery to artery connections (AAs) in which a branch artery from each twin meets directly on the superficial placental surface resulting in a vessel with pulsatile bidirectional flow, and 3) vein to vein connections (VVs) in which a branch vein from each twin meets directly on the superficial placental surface allowing low pressure bidirectional flow. In utero obstetric sonography with targeted Doppler interrogation has been used to identify the presence of AV and AA connections4. Prenatally detected AAs that have been confirmed by postnatal placental injection studies have been shown to be associated with an improved prognosis for both twins5. Furthermore, fetoscopic laser ablation of inter-twin vascular connections on the fetal surface of the shared placenta is now the preferred treatment for early, severe TTTS. 
Postnatal placental injection studies provide a valuable method to confirm the accuracy of prenatal Doppler ultrasound findings and the efficacy of fetal laser therapy6. Using colored dyes separately hand-injected into the arterial and venous circulations of each twin, the technique highlights and delineates AVs, AAs, and VVs. This definitive demonstration of MC placental vascular anatomy may then be correlated with Doppler ultrasound findings and neonatal outcome to enhance our understanding of the pathophysiology of MC twinning and its sequelae. Here we demonstrate our placental injection technique.

Vascular mapping of monochorionic (MC) twin placentas after birth provides a means for detailed demonstration of vascular connections between the twins&#x2019; circulations. 
Imbalance of these connections is thought to play a pivotal role in the development of complications of MC twinning including twin-to-twin transfusion syndrome.

Guia de Cateterismo Fio Assistida e injeção de corante colorido para o Mapeamento de Vascular monocoriônicas placentas gêmeas

Monochorionic (MC) twin pregnancies are associated with significantly higher morbidity and mortality rates than dichorionic twins. Approximately 50% of MC ...

A Simple Method of Mouse Lung Intubation

A simple procedure to intubate mice for pulmonary function measurements would have several advantages in longitudinal studies with limited numbers or expensive animal. One of the reasons that this is not done more routinely is that it is relatively difficult, despite there being several published studies that describe ways to achieve it. In this paper we demonstrate a procedure that eliminates one of the major hurdles associated with this intubation, that of visualizing the trachea during the entire time of intubation. The approach uses a 0.5 mm fiberoptic light source that serves as an introducer to direct the intubation cannula into the mouse trachea. We show that it is possible to use this procedure to measure lung mechanics in individual mice over a time course of at least several weeks. The technique can be set up with relatively little expense and expertise, and it can be routinely accomplished with relatively little training. This should make it possible for any laboratory to routinely carry out this intubation, thereby allowing longitudinal studies in individual mice, thereby minimizing the number of mice needed and increasing the statistical power by using each mouse as its own control.

This paper describes a striaghforward and efficient method of intubating mice for pulmonary function measurements or pulmonary instillation, that allows the mice to recover and be studied at later times. The procedure involves an inexpensive fiberoptic light source that directly illuminates the trachea.

Um método simples de intubação Lung Rato

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

Aspectos técnicos da artocava Fístula Rato

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

Fast and Accurate Exhaled Breath Ammonia Measurement

This exhaled breath ammonia method uses a fast and highly sensitive spectroscopic method known as quartz enhanced photoacoustic spectroscopy (QEPAS) that uses a quantum cascade based laser. The monitor is coupled to a sampler that measures mouth pressure and carbon dioxide. The system is temperature controlled and specifically designed to address the reactivity of this compound. The sampler provides immediate feedback to the subject and the technician on the quality of the breath effort. Together with the quick response time of the monitor, this system is capable of accurately measuring exhaled breath ammonia representative of deep lung systemic levels. 
Because the system is easy to use and produces real time results, it has enabled experiments to identify factors that influence measurements. For example, mouth rinse and oral pH reproducibly and significantly affect results and therefore must be controlled. Temperature and mode of breathing are other examples. As our understanding of these factors evolves, error is reduced, and clinical studies become more meaningful. This system is very reliable and individual measurements are inexpensive. 
The sampler is relatively inexpensive and quite portable, but the monitor is neither. This limits options for some clinical studies and provides rational for future innovations.

Ammonia is an important physiologic metabolite relevant to various disease and wellness states. It is also a difficult molecule to measure in breath, which demands particular precautions be taken to obtain accurate results. Not all factors influencing ammonia are known, but progress can be difficult without accounting for these factors.

Medição Amônia ar exalado Rápido e Preciso

This exhaled breath ammonia method uses a fast and highly sensitive spectroscopic method known as quartz enhanced photoacoustic spectroscopy (QEPAS) that ...

Evaluation of Tumor-infiltrating Leukocyte Subsets in a Subcutaneous Tumor Model

Specialized immune cells that infiltrate the tumor microenvironment regulate the growth and survival of neoplasia.&#160; Malignant cells must elude or subvert anti-tumor immune responses in order to survive and flourish. Tumors take advantage of a number of different mechanisms of immune &#8220;escape,&#8221; including the recruitment of tolerogenic DC, immunosuppressive regulatory T cells (Tregs), and myeloid-derived suppressor cells (MDSC) that inhibit cytotoxic anti-tumor responses. Conversely, anti-tumor effector immune cells can slow the growth and expansion of malignancies: immunostimulatory dendritic cells, natural killer cells which harbor innate anti-tumor immunity, and cytotoxic T cells all can participate in tumor suppression. The balance between pro- and anti-tumor leukocytes ultimately determines the behavior and fate of transformed cells; a multitude of human clinical studies have borne this out. Thus, detailed analysis of leukocyte subsets within the tumor microenvironment has become increasingly important. Here, we describe a method for analyzing infiltrating leukocyte subsets present in the tumor microenvironment in a mouse tumor model. Mouse B16 melanoma tumor cells were inoculated subcutaneously in C57BL/6 mice. At a specified time, tumors and surrounding skin were resected en bloc and processed into single cell suspensions, which were then stained for multi-color flow cytometry. Using a variety of leukocyte subset markers, we were able to compare the relative percentages of infiltrating leukocyte subsets between control and chemerin-expressing tumors. Investigators may use such a tool to study the immune presence in the tumor microenvironment and when combined with traditional caliper size measurements of tumor growth, will potentially allow them to elucidate the impact of changes in immune composition on tumor growth. Such a technique can be applied to any tumor model in which the tumor and its microenvironment can be resected and processed.

This protocol describes a method for the detailed evaluation of leukocyte subsets within the tumor microenvironment in a mouse tumor model. Chemerin-expressing B16 melanoma cells were implanted subcutaneously into syngeneic mice. Cells from the tumor microenvironment were then stained and analyzed by flow cytometry, allowing for detailed leukocyte subset analyses.

Avaliação do tumor infiltrantes Leucócitos Subconjuntos em um modelo de tumor subcutâneo

Specialized immune cells that infiltrate the tumor microenvironment regulate the growth and survival of neoplasia.&#160; Malignant cells must elude or ...

Using the Sleeve Technique in a Mouse Model of Aortic Transplantation - An Instructional Video

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the aorta in mice using the sleeve method was short and easy averaging 20 min, permitting studies of iso/allo grafts. The following article describes the aortic transplantation procedure used in our laboratory. The mice were anesthetized with a mixture of 1.5% volume isoflurane and 100% oxygen through a face mask. At this point, the segment of the aorta between the renal arteries and its bifurcation was separated from the vena cava, freely prepared and clampedat the proximal and distal segments with a single silk suture. Prior to the removal of the aorta, a saline solution containing heparin was injected into the inferior vena cava. Then the aorta was cut between the clamps and a saline heparin solution was used to flush the lumen. The sleeve technique with monofilament sutures was used in order to transplant the abdominal aorta in the orthotopic position.

We present an orthotopic aortic transplantation model using the sleeve technique in mice. It is a very rapid anastomosis method, which can be employed in studies of vascular disease.

Usando a técnica de manga em um modelo do rato da transplantação da aorta - um vídeo instrutivo

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the ...

Model Surgical Training: Skills Acquisition in Fetoscopic Laser Photocoagulation of Monochorionic Diamniotic Twin Placenta Using Realistic Simulators

Fetoscopic laser coagulation of arterio-venous anastomoses (AVA) in a monochorionic placenta is the standard of care for twin-twin transfusion syndrome (TTTS), but is technically challenging and can lead to significant complications. Acquiring and maintaining the necessary surgical skills require consistent practice, a critical caseload, and time. Training on realistic surgical simulators can potentially shorten this steep learning curve and enables several proceduralists to acquire procedure-specific skills simultaneously. Here we describe realistic simulators designed to allow the user familiarity with the equipment and specific steps required in the surgical treatment of TTTS, including fetoscopic handling, approaches to anterior and posterior placenta, recognition of anastomoses, and efficient coagulation of vessels. We describe the skills that are especially important in conducting placental laser coagulation that the surgeon can practice on the model and apply in a clinical case. These models can be adapted easily depending on the availability of materials and require standard fetoscopy equipment. Such training systems are complementary to traditional surgical apprenticeships and can be useful aids for fetal medicine units that provide this clinical service.

Practicing the specific skills required for fetoscopic laser coagulation of monochorionic placental anastomoses on realistic models can aid less experienced surgeons in overcoming the steep learning curve associated with this procedure that is now regarded as the standard of care for twin-twin transfusion syndrome.

Modelo de Treinamento Cirúrgico: Aquisição de Competências em Fotocoagulação Fetoscópica a Laser de Placenta Gêmea Diamniótica Monocoriônica Utilizando Simuladores Realistas

Fetoscopic laser coagulation of arterio-venous anastomoses (AVA) in a monochorionic placenta is the standard of care for twin-twin transfusion syndrome ...

Noninvasive Determination of Vortex Formation Time Using Transesophageal Echocardiography During Cardiac Surgery

Trans-mitral blood flow produces a three-dimensional rotational body of fluid, known as a vortex ring, that enhances the efficiency of left ventricular (LV) filling compared with a continuous linear jet. Vortex ring development is most often quantified with vortex formation time (VFT), a dimensionless parameter based on fluid ejection from a rigid tube. Our group is interested in factors that affect LV filling efficiency during cardiac surgery. In this report, we describe how to use standard two-dimensional (2D) and Doppler transesophageal echocardiography (TEE) to noninvasively derive the variables needed to calculate VFT. We calculate atrial filling fraction (&#946;) from velocity-time integrals of trans-mitral early LV filling and atrial systole blood flow velocity waveforms measured in the mid-esophageal four-chamber TEE view. Stroke volume (SV) is calculated as the product of the diameter of the LV outflow track measured in the mid-esophageal long axis TEE view and the velocity-time integral of blood flow through the outflow track determined in the deep transgastric view using pulse-wave Doppler. Finally, mitral valve diameter (D) is determined as the average of major and minor axis lengths measured in orthogonal mid-esophageal bicommissural and long axis imaging planes, respectively. VFT is then calculated as 4 &#215; (1-&#946;) &#215; SV/(&#960;D3). We have used this technique to analyze VFT in several groups of patients with differing cardiac abnormalities. We discuss our application of this technique and its potential limitations and also review our results to date. Noninvasive measurement of VFT using TEE is straightforward in anesthetized patients undergoing cardiac surgery. The technique may allow cardiac anesthesiologists and surgeons to assess the impact of pathological conditions and surgical interventions on LV filling efficiency in real time.

We describe a protocol to measure vortex formation time, an index of left ventricular filling efficiency, using standard transesophageal echocardiography techniques in patients undergoing cardiac surgery. We apply this technique to analyze vortex formation time in several groups of patients with differing cardiac pathologies.

Determinação não-invasiva de tempo de formação do vórtice usando ecocardiografia transesofágica durante cirurgia cardíaca

Trans-mitral blood flow produces a three-dimensional rotational body of fluid, known as a vortex ring, that enhances the efficiency of left ventricular ...

Echocardiographic Evaluation of Atrial Communications before Transcatheter Closure

Transthoracic (TTE) and transesophageal echocardiography (TEE) is the standard imaging method for atrial septal defect (ASD) and patent foramen ovale (PFO) detection, for patient selection for transcatheter ASD/PFO closure,&#160;for intraoperative guidance and for long-term follow-up. The size, shape, location and the number of the atrial communications schould be determined. The accuracy of PFO detection can be improved by using agitated saline together with maneuvers to transiently increase the right atrial (RA) pressure. The appearance of microbubbles in the left atrium (LA) within 3 cardiac cycles after opacification of the RA is considered positive for the presence of an intracardiac shunt. Three dimensional TEE identifies further septal fenestrations and describes the dynamic morphology of ASD/PFO and atrial septal aneurysm. Follow-up evaluations with TTE is recommended at 1, 6, and 12 months after the procedure, with a subsequent evaluation every year. Previous studies showed an increased incidence of atrial arrhythmias early after device closure. Speckle tracking analysis may help to understand functional left atrial remodeling following percutaneous closure and its impact on atrial arrhythmias.

Transthoracic (TTE) and transesophageal (TEE) echocardiography represent the basic imaging tools for interatrial septum examination. Three dimensional (3D) TEE provides incremental information in the assessment of the interatrial septum. Further advanced echocardiography techniques using speckle tracking echocardiography are applied for sensitive volumetric and functional assessment of the heart chambers.&#160;

Avaliação Ecocardiográfica das Comunicações Atrial antes do fechamento do Transcateter

Transthoracic (TTE) and transesophageal echocardiography (TEE) is the standard imaging method for atrial septal defect (ASD) and patent foramen ovale ...