Este trabalho foi financiado pela CDMRP, Grants USUHS G188LE, G188MG, e G188QC (para HL), e do Centro USUHS para o Estudo do Estresse Traumático.

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Não há conflitos de interesse declarados.

O diagnóstico de TEPT é baseado em auto-relatos de sintomas psiquiátricos (DSM IV) por potenciais sujeitos. Não biomarcador bem definido está atualmente disponível para acessar o estado fisiopatológico do potencial pacientes com TEPT. TEPT é um transtorno provocado por risco de vida eventos traumáticos e os principais sintomas psiquiátricos continuam presentes na vida do sobrevivente por meses e até mesmo anos após os eventos iniciais. Os sintomas mais proeminentes e persistentes revelados em pacientes com TEPT são hipervigilância, atraso resposta exagerada sobressalto 14, 15, 16 e um compromisso aparente do eixo HPA. Nos humanos os sintomas permanecem, ou aparecer com um atraso de três meses, após a cessação do estressor traumático 24. Nosso atual modelo para estudos de biomarcadores de PTSD emprega contenção e choques cauda repetidas por três dias consecutivos (2-h sessões de 40, 2 tailshocks MA) - o modelo rabo-choque inescapável (ITS) em ratos com peso de 150 gramas. Este modelo tem SEUFoi demonstrado que imitam de forma substancial a fisiopatologia da PTSD 17, 7, 4, 10. Nosso laboratório e outros laboratórios têm verificado que o seu modelo de estresse em ratos induz alterações comportamentais e neurobiológicos que são semelhantes aos encontrados no TEPT assuntos 17, 7, 10, 9. Especificamente, estes ratos estressados ​​apresentam (1) uma resposta atrasada e exagerada de sobressalto aparecimento de vários dias após a cessação do estressor, que dada a escala de tempo comprimido de vida dos ratos em comparação a um ser humano, corresponde ao atraso meses 1-3 de sintomas em pacientes com TEPT (DSM-IV-TR PTSD Criterian D / E 13), (2) aumento da corticosterona no plasma (CORT) para vários (10) dias, indicando o comprometimento do eixo hypothalamopituitary (HPA), e (3) o ganho de peso corporal após retardado stressor cessação, correspondente à disfunção da regulação metabólica de PTSD. Não há evidências na literatura que a Fox odor, a exposição predador, ou temem resposta sobressalto potencializado, expopouco esses fenótipos comportamentais e persistentes neuroendocrinologic associado com PTSD. Ratos expostos a uma sessão de tensão simples (1DS) exibiram transiente, mas não as anormalidades persistentes exibidos por 3DS ratos 17 A presente experiência comparou a resposta de sobressalto de 3DS e 1DS ratos 4, 7, e 10 dias após a cessação do estressor. Consistente com trabalhos anteriores, salientou ratos apresentaram elevados níveis plasmáticos basais de CORT o estresse pós primeiro dia 17. Estes níveis de CORT foram sensíveis ao número de exposições estressores com níveis mais elevados em ratos CORT 3DS que em 1DS ratos. Quanto à resposta de sobressalto, os ratos 1DS exibem uma resposta de sobressalto exagerada 7 dias pós estressor, enquanto sensibilização sobressalto só se torna aparente 10 dias após o estressor em ratos 3DS. Assim, o aparecimento de uma resposta exagerada de sobressalto após cessação estressor parece estar relacionado com o número de posições de sessão de stress. O modelo 3DS estressado pareceser útil para obter insights sobre a expressão alterada de biomarcadores associados com os sintomas da PTSD e os principais mensuráveis ​​fenótipos comportamentais associadas com o tempo após a cessação do sofrimento. Genômicas resultados apresentados fornecem prova de princípio para a aplicação de ferramentas de bioinformática e rMNChip para identificar vias de diferenciais, e biomarcadores de genes e proteínas, o que vai facilitar muito sistemas biológicos estudo e compreensão dos mecanismos moleculares subjacentes multifatoriais e complexos distúrbios neurológicos, incluindo TEPT. Enquanto o nosso paradigma não mergulhar no cognitivo e mais complexos aspectos comportamentais do TEPT, notamos que os padrões de sono alterados no modelo STI 1 corresponde à dificuldade e manter o sono e os pesadelos de 11 pacientes com TEPT (DSM-IV-TR TEPT Critérios D 13), e as deficiências na fuga / esquiva de aprendizagem e de aprendizagem de uma tarefa apetitiva no seu modelo <sup&gt; 12 corresponde às concentrações de pobres e déficits de memória de TEPT 5 (DSM-IV-TR TEPT Critério C 13). O modelo atual se correlaciona bem com os principais sintomas característicos do TEPT e fornece um bom modelo para correlacionar biomarcadores periféricos de TEPT, que podem ser coletados de pacientes, com centrais, biomarcadores mecanicistas, que não pode 6.

biomarkers in an animal model for revealing neural, hematologic, and behavioral correlates of ptsd

Identificação de biomarcadores que representam a evolução da fisiopatologia do Transtorno de Estresse Pós-Traumático (TEPT) é de vital importância, não só para o diagnóstico objetivo, mas também para a avaliação da eficácia terapêutica e resistência ao trauma. A pesquisa em curso é dirigido a identificar biomarcadores de TEPT, incluindo as proteínas de stress traumático, induzidas transcriptomas genômicas, variâncias e moduladores genéticos, utilizando amostras biológicas a partir de sangue dos sujeitos, a saliva, a urina, e tecidos cerebrais post-mortem. No entanto, a correlação destas moléculas de biomarcadores em amostras de periféricos ou post-mortem para funções cerebrais alteradas associados a sintomas psiquiátricos em TEPT continua por resolver. Aqui, nós apresentamos um modelo animal de PTSD em que tanto no sangue periférico e biomarcadores cerebrais centrais, bem como fenótipo comportamental, pode ser recolhido e medido, fornecendo assim a correlação necessária dos biomarcadores centrais de PTSD, que são mecânica e pathognomônico, mas não podem ser coletadas de pessoas, com os biomarcadores periféricos e fenótipos comportamentais, que podem. Nosso modelo animal de TEPT emprega contenção e choques cauda repetidas por três dias consecutivos - o modelo rabo-choque inescapável (ITS) em ratos. Este modelo ITS imita a patofisiologia da PTSD 17, 7, 4, 10. Nós e outros autores verificaram que o modelo ITS induz alterações comportamentais e neurobiológicos semelhantes aos encontrados em indivíduos PTSD 17, 7, 10, 9. Especificamente, estes ratos estressados ​​apresentam (1) uma resposta atrasada e exagerada de sobressalto aparecimento de vários dias após a cessação do estressor, que dada a escala de tempo comprimido de vida dos ratos em comparação a um ser humano, corresponde ao atraso meses 1-3 de sintomas em pacientes com TEPT (DSM-IV-TR PTSD Criterian D / E 13), (2) aumento da corticosterona no plasma (CORT) por vários dias, o que indica o comprometimento do eixo hypothalamopituitary (HPA), e (3) bo retardadoganho de peso após a cessação dy estressor, indicando disfunção de regulação metabólica. Os paradigmas experimentais utilizados para este modelo são: (1) um paradigma desamparo aprendido em ratos ensaiadas por medição da resposta de sobressalto acústico (ASR) e um gráfico de massa corporal, (2) microdissecção do cérebro de rato em regiões e núcleos, ( 3) ensaio de imunossorvente ligado a enzima (ELISA) para os níveis sanguíneos de CORT, (4) um microarray de expressão de genes, mais relacionada com as ferramentas da bioinformática 18. Este microarray, apelidado rMNChip, centra-se na mitocôndria mitocondriais e nucleares relacionadas com genes em ratos, de modo a abordar especificamente as bioenergética neuronais hipótese de estar envolvido no TEPT.

Watch this Scientific Journal Video about Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD at JoVE.com

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

Nós descrevemos um modelo de rato do transtorno de estresse pós-traumático (TEPT), que revela as alterações persistentes na função neuroendócrina eo atraso de longo prazo, resposta de medo exagerado, característico de pacientes com TEPT. O modelo animal e os métodos descritos aqui são úteis para correlacionar biomarcadores em núcleos do cérebro, que são mecanicista, mas não pode ser medido em pacientes com biomarcadores em células brancas do sangue periférico, os quais podem.

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 ...

biomarkers-an-animal-model-for-revealing-neural-hematologic

Research

JoVE Journal

Medicine

15.9K visualizações.  Uniformed Services University of the Health Sciences, Bethesda, Maryland. Nós descrevemos um modelo de rato do transtorno de estresse pós-traumático (TEPT), que revela as alterações persistentes na função neuroendócrina eo atraso de longo prazo, resposta de medo exagerado, característico de pacientes com TEPT. O modelo animal e os métodos descritos aqui são úteis para correlacionar biomarcadores em núcleos do cérebro, que são mecanicista, mas não pode ser medido em pacientes com biomarcadores em células brancas do sangue perif?...

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

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several techniques, including infusion of exogenous bacterial toxin (endotoxemia) or bacteria (bacteremia), as well as surgical perforation of the cecum by cecal ligation and puncture (CLP)1-3. CLP allows bacteria spillage and fecal contamination of the peritoneal cavity, mimicking the human clinical disease of perforated appendicitis or diverticulitis. The severity of sepsis, as reflected by the eventual mortality rates, can be controlled surgically by varying the size of the needle used for cecal puncture2. In animals, CLP induces similar, biphasic hemodynamic cardiovascular, metabolic, and immunological responses as observed during the clinical course of human sepsis3. Thus, the CLP model is considered as one of the most clinically relevant models for experimental sepsis1-3.
Various animal models have been used to elucidate the intricate mechanisms underlying the pathogenesis of experimental sepsis. The lethal consequence of sepsis is attributable partly to an excessive accumulation of early cytokines (such as TNF, IL-1 and IFN-&gamma;)4-6 and late proinflammatory mediators (e.g., HMGB1)7. Compared with early proinflammatory cytokines, late-acting mediators have a wider therapeutic window for clinical applications. For instance, delayed administration of HMGB1-neutralizing antibodies beginning 24 hours after CLP, still rescued mice from lethality8,9, establishing HMGB1 as a late mediator of lethal sepsis. The discovery of HMGB1 as a late-acting mediator has initiated a new field of investigation for the development of sepsis therapies using Traditional Chinese Herbal Medicine. In this paper, we describe a procedure of CLP-induced sepsis, and its usage in screening herbal medicine for HMGB1-targeting therapies.

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection, and can be simulated by a surgical technique termed cecal ligation and puncture (CLP). Here we describe a method to use CLP-induced animal model to screen medicinal herbs for therapeutic agents.

Uso de Modelo Animal de Sepse para avaliar novas terapias à base de plantas

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several ...

Medicina

An Isolated Working Heart System for Large Animal Models

Since its introduction in the late 19th century, the Langendorff isolated heart perfusion apparatus, and the subsequent development of the working heart model, have been invaluable tools for studying cardiovascular function and disease1-15. Although the Langendorff heart preparation can be used for any mammalian heart, most studies involving this apparatus use small animal models (e.g., mouse, rat, and rabbit) due to the increased complexity of systems for larger mammals1,3,11. One major difficulty is ensuring a constant coronary perfusion pressure over a range of different heart sizes &#8211; a key component of any experiment utilizing this device1,11. By replacing the classic hydrostatic afterload column with a centrifugal pump, the Langendorff working heart apparatus described below allows for easy adjustment and tight regulation of perfusion pressures, meaning the same set-up can be used for various species or heart sizes. Furthermore, this configuration can also seamlessly switch between constant pressure or constant flow during reperfusion, depending on the user&#8217;s preferences. The open nature of this setup, despite making temperature regulation more difficult than other designs, allows for easy collection of effluent and ventricular pressure-volume data.

Most studies involving the Langendorff apparatus use small animal models due to the increased complexity of systems for larger mammals. We describe a Langendorff system for large animal models that allows for use across a range of species, including humans, and relatively easy data acquisition.

Um Sistema de coração isolado para Grandes Modelos Animais

Since its introduction in the late 19th century, the Langendorff isolated heart perfusion apparatus, and the subsequent development of the working heart ...

Bioluminescence Imaging of an Immunocompetent Animal Model for Glioblastoma

In contrast to commonly reported human glioma xenograft animal models, GL261 murine glioma xenografts recapitulate nearly all relevant clinical and histopathologic features of the human disease. When GL261 cells are implanted intracranially in syngeneic C57BL/6 mice, the model has the added advantage of maintaining an intact immune microenvironment. Stable expression of luciferase in GL261 cells allows non-invasive cost effective bioluminescence monitoring of intracranial tumor growth. We have recently demonstrated that luciferase expression in GL261 cells does not affect the tumor growth properties, tumor cell immunomodulatory cytokine expression, infiltration of immune cells into the tumor, or overall survival of animals bearing the intracranial tumor. Therefore, it appears that the GL261 luciferase glioma model can be useful in the study of novel chemotherapeutic and immunotherapeutic modalities. Here we report the technique for generating stable luciferase expression in GL261 cells and how to study the in vitro and in vivo growth of the tumor cells by bioluminescence imaging.

GL261 glioma cells provide a useful immunocompetent animal model of glioblastoma. The goals of this protocol are to demonstrate proper techniques for monitoring intracranial tumor growth using in vivo bioluminescence imaging, and to verify the utility of luciferase-modified GL261 cells for studying tumor immunology and immunotherapeutic approaches for treating glioblastoma.

Bioluminescência imagem de um modelo animal para imunocompetentes Glioblastoma

In contrast to commonly reported human glioma xenograft animal models, GL261 murine glioma xenografts recapitulate nearly all relevant clinical and ...

A Repetitive Concussive Head Injury Model in Mice

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of knowledge on the injury and its effects. To develop a better understanding of concussions, animals are often used because they provide a controlled, rigorous, and efficient model. Studies have adapted traditional animal models to perform mTBI to stimulate mild injury severity by changing the injury parameters. These models have been used because they can produce morphologically similar brain injuries to the clinical condition and provide a spectrum of injury severities. However, they are limited in their ability to present the identical features of injuries in patients. Using a traditional impact system, a repetitive concussive injury (rCHI) model can induce mild to moderate human-like concussion. The injury degree can be determined by measuring the period of loss of consciousness (LOC) with a sign of a transient termination of breathing. The rCHI model is beneficial to use for its accuracy and simplicity in determining mTBI effects and potential treatments.

Concussion presents the most common type of traumatic brain injury. Therefore, a repetitive concussive animal model, which replicates the important features of an injury in patients, may provide a means to study concussion in a rigorous, controlled, and efficient manner.

A principal modelo lesões repetitivas Concussive em Ratos

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of ...

An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model

Pedicle screw fixation is the gold standard for the treatment of spinal diseases. However, many studies have reported the issue of loosening pedicle screws after spinal surgery, which is a serious concern. To address this problem, diverse types of pedicle screws have been examined to identify those with good fixation strength and osseointegration in spine bone. The porcine spine is a good alternative for the human spine in the evaluation of pedicle screws due to the anatomical size, mechanical characteristics, and cost. Although several studies have reported that pedicle screws are efficient in the porcine model, no study has described detailed protocols for the evaluation of a pedicle screw using the porcine model. Here, we describe a detailed method for evaluating transpedicular screws using an in vivo porcine lumbar spine model. The technical details for anesthesia, spine surgery, and harvest provided here will facilitate with the evaluation of the transpedicular screw fixation model.

An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model

Here, we introduce a method to evaluate transpedicular screws by using an in vivo porcine lumbar spine model.

Método de anestesia, cirurgia e colheita para avaliação de parafusos transpediculares usando um In Vivo Modelo de espinha lombar porcina

Pedicle screw fixation is the gold standard for the treatment of spinal diseases. However, many studies have reported the issue of loosening pedicle ...

A Chronic Cardiac Ischemia Model in Swine Using an Ameroid Constrictor

Cardiovascular disease remains the number one cause of mortality in the United States. There are numerous approaches to treating these diseases, but regardless of the approach, an in vivo model is needed to test each treatment. The pig is one of the most used large animal models for cardiovascular disease. Its heart is very similar in anatomy and function to that of a human. The ameroid placement technique creates an ischemic area of the heart, which has many useful applications in studying myocardial infarction. This model has been used for surgical research, pharmaceutical studies, imaging techniques, and cell therapies.
There are several ways of inducing an ischemic area in the heart. Each has its advantages and disadvantages, but the placement of an ameroid constrictor remains the most widely used technique. The main advantages to using the ameroid are its prevalence in existing research, its availability in various sizes to accommodate the anatomy and size of the vessel to be constricted, the surgery is a relatively simple procedure, and the post-operative monitoring is minimal, since there are no external devices to maintain. This paper provides a detailed overview of the proper technique for the placement of the ameroid constrictor.

The purpose of this protocol is to demonstrate the placement of a delayed constricting device (an ameroid constrictor) around a coronary artery in a swine model. This device creates an ischemic area of the heart that is useful for studying new diagnostic imaging techniques and new methods of treatment.

Um modelo de isquemia cardíaca crônica em suínos, usando um Ameroid Constrictor

Cardiovascular disease remains the number one cause of mortality in the United States. There are numerous approaches to treating these diseases, but ...

Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo

Plasma coagulation as a form of electrocautery is used in liver surgery for decades to seal the large liver cut surface after major hepatectomy to prevent hemorrhages at a later stage. The exact effects of plasma coagulation on liver tissue are only poorly examined. In our porcine model, the coagulation effects can be examined close to the clinical application. A combined laser Doppler flowmeter and spectrophotometer documents microcirculation changes during coagulation at 8 mm tissue depth noninvasively, providing quantifiable information about hemostasis beyond the subjective clinical impression. The temperature at coagulation site is assessed with an infrared thermometer prior and post coagulation and with a thermographic camera during coagulation, a measurement of the gas beam temperature is not possible due to the upper threshold of the devices. The depth of coagulation is measured microscopically on hematoxylin/eosin stained sections after calibration with an object micrometer and gives an exact information about the power setting-coagulation depth-relation. The sealing effect is examined on the bile ducts as it is not possible for a plasma coagulator to seal larger blood vessels. Burst pressure experiments are carried out on explanted organs to rule out blood pressure related effects.

Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo

Here we present a protocol to experimentally assess plasma coagulation in liver tissue in vivo. In a porcine model, microcirculation is examined by laser Doppler, coagulation depth is measured histologically, temperature at coagulation site by infrared thermometer and thermographic camera, and duct sealing effect is documented by burst pressure experiments.

Avaliação da coagulação do Plasma no tecido do fígado em um modelo Animal grande na Vivo

Plasma coagulation as a form of electrocautery is used in liver surgery for decades to seal the large liver cut surface after major hepatectomy to prevent ...

Intra-Operative Neural Monitoring of Thyroid Surgery in a Porcine Model

Intraoperative injury to the recurrent laryngeal nerve (RLN) can cause vocal cord paralysis, which interferes with speech and can potentially interfere with breathing. In recent years, intraoperative neural monitoring (IONM) has been widely adapted as an adjunct technique to localize the RLN, detect RLN injury, and predict vocal cord function during the operations. Many studies have also used animal models to investigate new applications of IONM technology and to develop reliable strategies for preventing intraoperative RLN injury. The aim of this article is to introduce a standard protocol for using a porcine model in IONM research. The article demonstrates the procedures for inducing general anesthesia, performing tracheal intubation, and experimental design to investigate the electrophysiological characteristics of RLN injuries. Applications of this protocol can improve overall efficacy in implementing the 3R principle (replacement, reduction and refinement) in porcine IONM studies.

This study aims to develop a standard protocol of intra-operative neural monitoring of thyroid surgery in a porcine model. Here, we present a protocol to demonstrate general anesthesia, to compare different types of electrodes, and to investigate the electrophysiological characteristics of the normal and injured recurrent laryngeal nerves.

Monitoramento Neural intra-operatória de cirurgia de tireoide em um modelo de suínos

Intraoperative injury to the recurrent laryngeal nerve (RLN) can cause vocal cord paralysis, which interferes with speech and can potentially interfere ...

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 ...

Manufacture of a Multi-Purpose Low-Cost Animal Bench-Model for Teaching Tracheostomy

Tracheostomy is one of the most frequent procedures, performed through various techniques in the intensive care unit and emergency situations. Despite this, there is a lack of training on this procedure that affects its outcome, which is also dependent on operator's dexterity. Here, we take the specific training and simulation into consideration. This article aims to describe every step of the manufacture of a new multi-purpose low-cost animal bench-model, with the support of video and images, and to obtain an opinion about the quality of this model by administering a questionnaire to professionals with experience in the procedures. 
Ten experts in the technique were enrolled. The model scored an average of 3.45/5 for its anatomical realism; 4.75/5 for its usefulness as a training tool for simulation courses and assessments. The time necessary to build the model was 15 minutes, and the cost amounted to 10&#8364;. The animal bench-model was considered a very useful simulator for tracheostomy training and assessments. Therefore, it could be used as a tool for medical courses and residencies.

This article illustrates every step of the manufacture of a new multi-purpose low-cost animal bench-model for subglottic airway access management. All the procedures are shown in the video. The model's realism and its suitability for training the given clinical maneuvers were assessed by independent senior otolaryngologists and anesthesiologists.

Fabricação de um modelo de banco de animais de baixo custo de múltiplos propósitos para o ensino de traqueostomia

Tracheostomy is one of the most frequent procedures, performed through various techniques in the intensive care unit and emergency situations. Despite ...