Este trabalho foi financiado pelo NIH NINDS conceder R01 NS074388 581 (CL Limoli), Instituto da Califórnia de Medicina Regenerativa (CIRM) Grant RS1-00413 (CLL) e Grant CIRM para JOVE em apoio à documentação em vídeo. Gostaríamos também de agradecer a Rudy Limburg e Edward Lau, da Universidade da Califórnia, Irvine Loja da Máquina de Ciências Biológicas para os seus conselhos, apoio e excelente capacidade técnica na preparação de todos os materiais feitos sob medida usada para testes comportamentais.

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	<li>Abayomi, O. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogenesis+of+irradiation-induced+cognitive+dysfunction.">Pathogenesis of irradiation-induced cognitive dysfunction.</a> Acta. Oncol. 35, 659-663 (1996).</li><li>Anderson, V. A., Godber, T., Smibert, E., Weiskop, S., Ekert, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cognitive+and+academic+outcome+following+cranial+irradiation+and+chemotherapy+in+children:+a+longitudinal+study.">Cognitive and academic outcome following cranial irradiation and chemotherapy in children: a longitudinal study.</a> Br. J. Cancer. 82, 255-262 (2000).</li><li>Acharya, M. 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M., Harris, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Memory+consolidation+for+contextual+and+auditory+fear+conditioning+is+dependent+on+protein+synthesis,+PKA,+and+MAP+kinase.">Memory consolidation for contextual and auditory fear conditioning is dependent on protein synthesis, PKA, and MAP kinase.</a> Learn. Mem. 6, 97-110 (1999).</li><li>Winocur, G., Wojtowicz, J. M., Sekeres, M., Snyder, J. S., Wang, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+neurogenesis+interferes+with+hippocampus-dependent+memory+function.">Inhibition of neurogenesis interferes with hippocampus-dependent memory function.</a> Hippocampus. 16, 296-304 (2006).</li><li>Phillips, R. G., LeDoux, J. 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B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cerebral+hyporesponsiveness+and+cognitive+impairment+10+years+after+chemotherapy+for+breast+cancer.">Cerebral hyporesponsiveness and cognitive impairment 10 years after chemotherapy for breast cancer.</a> Hum. Brain. Mapp. , (2010).</li><li>Hess, D. C., Borlongan, C. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell-based+therapy+in+ischemic+stroke.">Cell-based therapy in ischemic stroke.</a> Expert. Rev. Neurother. 8, 1193-1201 (2008).</li></ol>

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Há uma necessidade urgente de estratégias de longo prazo de forma a prevenir o desenvolvimento de disfunção cognitiva que está se tornando cada vez mais comum em sobreviventes de câncer 12,13. Terapias com células-tronco pode fornecer os meios para amenizar uma série de seqüelas tecido normal no sistema nervoso central, como eles fornecem a oportunidade para substituir as células perdidas ou danificadas de tratamentos de câncer citotóxicas 14. Para acelerar a realização destes esforços, temos realizado uma série de experimentos da prova principal que confirmaram a capacidade de hNSCs cranialmente enxertada para restaurar funcionalmente cognição dentro do leito do tecido irradiado três CNS. Crítico para este esforço foi a capacidade de forma confiável realizar uma série de testes cognitivos cuidadosamente controlado de uma forma reprodutível. Temos detalhada desses procedimentos crítico nos esforços para acelerar o potencial de translação de terapias com células-tronco na clínica. Outras considerações que sãosusceptível de ter um impacto significativo da qualidade dos dados são destacadas a seguir. Cognitiva testes exige atenção cuidadosa aos detalhes e aderir a certas práticas padrão podem ajudar a minimizar os artefatos que dificultam a interpretação dos resultados. O uso de salas dedicadas testes comportamentais separadas das salas de animais exploração e experimentadores é preferível. Também é prudente usar &quot;ruído branco&quot; que bloqueia ruídos externos nas instalações de animais que podem distrair os animais da tarefa em mãos. Para minimizar o stress para os animais destinados a testes cognitivos, é importante para eles para se familiarizar com os experimentadores que irão realizar testes de comportamento, antes da administração de tarefas. Em geral, se o teste fêmeas e machos, é recomendável testar as fêmeas primeiro como eles tendem a ser mais ansiosos durante os testes, se do sexo masculino pistas odor permanecer no arenas testes. Ao administrar uma bateria de série de tarefas, é importante para expor animais para least tarefas estressantes primeiro (como EPM, NPR) e mais tarefas estressantes última (como FC, MWM).

quantifying cognitive decrements caused by cranial radiotherapy

Com a exceção de sobrevivência, comprometimento cognitivo decorrente do manejo clínico do cancro é um dos principais fatores que ditam os resultados terapêuticos. Para muitos pacientes aflitos com CNS e não-malignas do SNC, radioterapia e quimioterapia oferecer as melhores opções para controle da doença. Estes tratamentos no entanto têm um custo, e sobreviventes de câncer de quase todos (~ 11 milhões em os EUA sozinhos em 2006) incorrer algum risco para o desenvolvimento de disfunção cognitiva, com os casos mais graves encontrados em pacientes submetidos à radioterapia craniana (~ 200 mil / ano ) para o controle de tumores cerebrais primários e metastáticos 1. Particularmente problemático são os casos pediátricos, cuja sobrevivência a longo prazo atormentado com marcada resultados cognitivos decréscimos significativos nos encargos socioeconômico 2. Até à data, ainda não existem soluções satisfatórias para este problema clínico significativo. Temos abordado este grave problema de saúde utilizando-tronco transplantadas cells para combater a radiação induzida por declínio cognitivo em ratos atímicos submetido a irradiação craniana 3. Detalhes da irradiação estereotáxica ea cultura em vitro e transplante de células-tronco neurais (hNSCs) pode ser encontrado em nosso papel de companheiro (Acharya et al., Jove de referência). Após irradiação e cirurgia de transplante, os ratos são então avaliadas para alterações na cognição, a sobrevivência de células enxertadas e expressão de marcadores de diferenciação específicos 1 e 4 meses após a irradiação. Avaliar criticamente o sucesso ou fracasso de qualquer intervenção projetada para melhorar o potencial induzido pela radiação cognitiva sequelas, uma série rigorosa de tarefas cognitivas quantitativa deve ser realizada. Para conseguir isso, submetemos nossos animais a um conjunto de paradigmas testes cognitivos, incluindo o reconhecimento romance lugar, labirinto aquático, labirinto elevado e condicionamento do medo, a fim de quantificar a aprendizagem do hipocampo e não-hipocampal e memória. Temos demonstrado a utilidade daestes testes para quantificar tipos específicos de decréscimos cognitivos em animais irradiados, e os usou para mostrar que os animais enxertados com hNSCs apresentam melhoras significativas na função cognitiva 3. Os benefícios cognitivos derivados de células-tronco humanas enxertados sugerem que estratégias semelhantes podem proporcionar um dia tão necessário o recurso clínico para sobreviventes de câncer que sofrem de disfunção cognitiva. Consequentemente, nós fornecemos a documentação escrita e visual dos passos críticos usados ​​em nossos paradigmas testes cognitivos para facilitar a tradução de nossos resultados promissores para a clínica.

Watch this Scientific Journal Video about Quantifying Cognitive Decrements Caused by Cranial Radiotherapy at JoVE.com

Quantifying Cognitive Decrements Caused by Cranial Radiotherapy

Comprometimento cognitivo decorrentes da gestão radioterápico de tumores cerebrais representa uma condição clinicamente intratáveis ​​que impacta negativamente a qualidade de vida. A capacidade de avaliar criticamente potenciais intervenções para amenizar induzida por radiação decrementa cognitivo depende em última instância a capacidade de realizar rigorosa avaliações quantitativas da cognição.

Diminui quantificar cognitivos causados ​​pela radioterapia craniana

With the exception of survival, cognitive impairment stemming from the clinical management of cancer is a major factor dictating therapeutic outcome. For ...

quantifying-cognitive-decrements-caused-by-cranial-radiotherapy

Research

JoVE Journal

Medicine

12.5K visualizações.  University of California Irvine. Comprometimento cognitivo decorrentes da gestão radioterápico de tumores cerebrais representa uma condição clinicamente intratáveis ​​que impacta negativamente a qualidade de vida. A capacidade de avaliar criticamente potenciais intervenções para amenizar induzida por radiação decrementa cognitivo depende em última instância a capacidade de realizar rigorosa avaliações quantitativas da cognição.

Vídeo: Quantifying Cognitive Decrements Caused by Cranial Radiotherapy

Stem Cell Transplantation Strategies for the Restoration of Cognitive Dysfunction Caused by Cranial Radiotherapy

Radiotherapy often provides the only clinical recourse for those afflicted with primary or metastatic brain tumors. While beneficial, cranial irradiation can induce a progressive and debilitating decline in cognition that may, in part, be caused by the depletion of neural stem cells. Given the increased survival of patients diagnosed with brain cancer, quality of life in terms of cognitive health has become an increasing concern, especially in the absence of any satisfactory long-term treatments.
To address this serious health concern we have used stem cell replacement as a strategy to combat radiation-induced cognitive decline. Our model utilizes athymic nude rats subjected to cranial irradiation. The ionizing radiation is delivered as either whole brain or as a highly focused beam to the hippocampus via linear accelerator (LINAC) based stereotaxic radiosurgery. Two days following irradiation, human neural stem cells (hNSCs) were stereotaxically transplanted into the hippocampus. Rats were then assessed for changes in cognition, grafted cell survival and for the expression of differentiation-specific markers 1 and 4-months after irradiation. Our cognitive testing paradigms have demonstrated that animals engrafted with hNSCs exhibit significant improvements in cognitive function. Unbiased stereology reveals significant survival (10-40%) of the engrafted cells at 1 and 4-months after transplantation, dependent on the amount and type of cells grafted. Engrafted cells migrate extensively, differentiate along glial and neuronal lineages, and express a range of immature and mature phenotypic markers.
Our data demonstrate direct cognitive benefits derived from engrafted human stem cells, suggesting that this procedure may one day afford a promising strategy for the long-term functional restoration of cognition in individuals subjected to cranial radiotherapy. To promote the dissemination of the critical procedures necessary to replicate and extend our studies, we have provided written and visual documentation of several key steps in our experimental plan, with an emphasis on stereotaxic radiosurgey and transplantation.

Brain tumor patients routinely undergo cranial radiotherapy, and while beneficial, this treatment often results in debilitating cognitive dysfunction. This serious unresolved problem has at present, no clinical recourse, and has driven our efforts to devise stem cell based therapies for the recovery of radiation-induced cognitive decrements.

Transplante de Células-tronco Estratégias para a Restauração da disfunção cognitiva causada pela radioterapia craniana

Radiotherapy often provides the only clinical recourse for those afflicted with primary or metastatic brain tumors. While beneficial, cranial irradiation ...

Medicina

Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model

Glioblastoma multiforme (GBM) are the most common and aggressive adult primary brain tumors1. In recent years there has been substantial progress in the understanding of the mechanics of tumor invasion, and direct intracerebral inoculation of tumor provides the opportunity of observing the invasive process in a physiologically appropriate environment2. As far as human brain tumors are concerned, the orthotopic models currently available are established either by stereotaxic injection of cell suspensions or implantation of a solid piece of tumor through a complicated craniotomy procedure3. In our technique we harvest cells from tissue culture to create a cell suspension used to implant directly into the brain. The duration of the surgery is approximately 30 minutes, and as the mouse needs to be in a constant surgical plane, an injectable anesthetic is used. The mouse is placed in a stereotaxic jig made by Stoetling (figure 1). After the surgical area is cleaned and prepared, an incision is made; and the bregma is located to determine the location of the craniotomy. The location of the craniotomy is 2 mm to the right and 1 mm rostral to the bregma. The depth is 3 mm from the surface of the skull, and cells are injected at a rate of 2 &mu;l every 2 minutes. The skin is sutured with 5-0 PDS, and the mouse is allowed to wake up on a heating pad. From our experience, depending on the cell line, treatment can take place from 7-10 days after surgery. Drug delivery is dependent on the drug composition. For radiation treatment the mice are anesthetized, and put into a custom made jig. Lead covers the mouse's body and exposes only the brain of the mouse. The study of tumorigenesis and the evaluation of new therapies for GBM require accurate and reproducible brain tumor animal models. Thus we use this orthotopic brain model to study the interaction of the microenvironment of the brain and the tumor, to test the effectiveness of different therapeutic agents with and without radiation.

The purpose of this article is to describe the use of an orthotopic glioblastoma model for chemoradiation studies. This article will go though cell processing, implanting, and radiotherapy of the mouse using an intracranial model.

Radioterapia em uma combinação ortotópico modelo de tumor de cérebro de camundongos

Glioblastoma multiforme (GBM) are the most common and aggressive adult primary brain tumors1. In recent years there has been substantial progress in the ...

Voluntary Breath-hold Technique for Reducing Heart Dose in Left Breast Radiotherapy

Breath-holding techniques reduce the amount of radiation received by cardiac structures during tangential-field left breast radiotherapy. With these techniques, patients hold their breath while radiotherapy is delivered, pushing the heart down and away from the radiotherapy field. Despite clear dosimetric benefits, these techniques are not yet in widespread use. One reason for this is that commercially available solutions require specialist equipment, necessitating not only significant capital investment, but often also incurring ongoing costs such as a need for daily disposable mouthpieces. The voluntary breath-hold technique described here does not require any additional specialist equipment. All breath-holding techniques require a surrogate to monitor breath-hold consistency and whether breath-hold is maintained. Voluntary breath-hold uses the distance moved by the anterior and lateral reference marks (tattoos) away from the treatment room lasers in breath-hold to monitor consistency at CT-planning and treatment setup. Light fields are then used to monitor breath-hold consistency prior to and during radiotherapy delivery.

The current priority in breast cancer radiotherapy is to reduce cardiac doses without compromising target tissue coverage. The voluntary breath-hold technique described here is a simple, inexpensive solution to this problem and capable of being instituted widely without the need for specialized equipment.

Voluntário Técnica de apneia para a Redução Coração Dose em Radioterapia mama esquerda

Breath-holding techniques reduce the amount of radiation received by cardiac structures during tangential-field left breast radiotherapy. With these ...

State of the Art Cranial Ultrasound Imaging in Neonates

Cranial ultrasound (CUS) is a reputable tool for brain imaging in critically ill neonates. It is safe, relatively cheap and easy to use, even when a patient is unstable. In addition it is radiation-free and allows serial imaging. CUS possibilities have steadily expanded. However, in many neonatal intensive care units, these possibilities are not optimally used. We present a comprehensive approach for neonatal CUS, focusing on optimal settings, different probes, multiple acoustic windows and Doppler techniques. This approach is suited for both routine clinical practice and research purposes. In a live demonstration, we show how this technique is performed in the neonatal intensive care unit. Using optimal settings and probes allows for better imaging quality and improves the diagnostic value of CUS in experienced hands. Traditionally, images are obtained through the anterior fontanel. Use of supplemental acoustic windows (lambdoid, mastoid, and lateral fontanels) improves detection of brain injury. Adding Doppler studies allows screening of patency of large intracranial arteries and veins. Flow velocities and indices can be obtained. Doppler CUS offers the possibility of detecting cerebral sinovenous thrombosis at an early stage, creating a window for therapeutic intervention prior to thrombosis-induced tissue damage. Equipment, data storage and safety aspects are also addressed.

Cranial ultrasound (CUS) is a valuable tool for brain imaging in critically ill neonates. This video shows a comprehensive approach for neonatal (Doppler) CUS for both clinical and research purposes, including a bedside demonstration of the technique.

Estado da Arte Cranial Ecografia em recém-nascidos

Cranial ultrasound (CUS) is a reputable tool for brain imaging in critically ill neonates. It is safe, relatively cheap and easy to use, even when a ...

Treatment of Liver Metastases Using an Internal Target Volume Method for Stereotactic Body Radiotherapy

The prognosis of patients with metastatic cancers has improved in the past decades due to effective chemotherapy and oligometastatic surgery. For inoperable patients, local ablation therapies, such as stereotactic body radiotherapy (SBRT), can provide effective local tumor control with minimal toxicity. Because of its high precision and accuracy, SBRT delivers a higher radiation dose per fraction, is more effective, and targets smaller irradiation volumes than does conventional radiotherapy. In addition, steep dose gradients from target lesions to surrounding normal tissues are achieved using SBRT; thus, SBRT provides more effective tumor control and exhibits fewer side effects than conventional radiotherapy. The use of SBRT is prevalent for treating intracranial lesions (known as stereotactic radiosurgery); however, it is now also used for treating spinal and adrenal metastases. Because of advancements in image-guided assistance and respiratory motion management, several studies have investigated the use of SBRT for treating lung or liver tumors, which move as a patient breathes. The results of these studies have suggested that SBRT favorably controls tumors in the case of moving lesions.
Four-dimensional computed tomography (4D-CT) with an abdominal compressor (AC) is clinically convenient for effective respiratory motion management. Because this method is noninvasive and allows free breathing, its use reduces complications. Furthermore, patients consider this method convenient. Moreover, it is considered more efficient than other methods of respiratory motion management by physicians and therapists. The use of 4D-CT with an AC for treating pulmonary lesions has also been widely investigated, and the technique is gaining acceptance for treating hepatic lesions. However, the protocols for using 4D-CT with an AC for treating hepatic lesions are different from those used for treating pulmonary lesions. In this article, we describe a new protocol for SBRT with 4D-CT and an AC for treating liver metastases.

Stereotactic body radiotherapy (SBRT) requires rigorous accuracy and precision for delivering high radiation doses per fraction to small treatment volumes to improve tumor control and simultaneously reduce toxicity. Herein, we present a noninvasive and clinically convenient respiratory motion management protocol for SBRT for liver metastases.

Tratamento de metástases hepáticas, usando um método do Volume alvo interno para radioterapia estereotáxica corpo

The prognosis of patients with metastatic cancers has improved in the past decades due to effective chemotherapy and oligometastatic surgery. For ...

Radiation Planning Assistant - A Streamlined, Fully Automated Radiotherapy Treatment Planning System

The Radiation Planning Assistant (RPA) is a system developed for the fully automated creation of radiotherapy treatment plans, including volume-modulated arc therapy (VMAT) plans for patients with head/neck cancer and 4-field box plans for patients with cervical cancer. It is a combination of specially developed in-house software that uses an application programming interface to communicate with a commercial radiotherapy treatment planning system. It also interfaces with a commercial secondary dose verification software. The necessary inputs to the system are a Treatment Plan Order, approved by the radiation oncologist, and a simulation computed tomography (CT) image, approved by the radiographer. The RPA then generates a complete radiotherapy treatment plan. For the cervical cancer treatment plans, no additional user intervention is necessary until the plan is complete. For head/neck treatment plans, after the normal tissue and some of the target structures are automatically delineated on the CT image, the radiation oncologist must review the contours, making edits if necessary. They also delineate the gross tumor volume. The RPA then completes the treatment planning process, creating a VMAT plan. Finally, the completed plan must be reviewed by qualified clinical staff.

Radiation therapy is a highly complex cancer treatment that requires multiple specialists to create a treatment plan and provide quality assurance (QA) prior to delivery to a patient. This protocol describes the use of a fully automated system, the Radiation Planning Assistant (RPA), to create high-quality radiation treatment plans.

Radiação Planejamento Assistente - uma simplificada, totalmente automatizado sistema de planeamento de tratamento de radioterapia

The Radiation Planning Assistant (RPA) is a system developed for the fully automated creation of radiotherapy treatment plans, including volume-modulated ...

Digital Handwriting Analysis of Characters in Chinese Patients with Mild Cognitive Impairment

An increasing amount of evidence shows that cognitive deficits and movement dysfunctions are not separated. Patients with mild cognitive impairment (MCI) can manifest fine motor disorders of the upper extremities. Handwriting is a complex and unique human activity involving both motor and cognitive coordination. Researchers from western countries have discovered that patients with MCI have abnormal handwriting features. However, no relevant studies have been conducted in the Chinese population. Owing to the cross-culture phenomenon of handwriting, the aim of this study is to find new handwriting tasks to demonstrate the differences in handwriting features between elderly patients with MCI and age-matched healthy individuals.

This study proposed a digital handwriting analysis of characters in individuals with mild cognitive impairment to find more information than is revealed by traditional pencil&#8211;paper handwriting analysis.

Análise de Caligrafia Digital de Personagens em Pacientes Chineses com Prejuízo Cognitivo Leve

An increasing amount of evidence shows that cognitive deficits and movement dysfunctions are not separated. Patients with mild cognitive impairment (MCI) ...

Aprimorando a radioterapia com o assistente de planejamento de radiação baseado na Web

Radiation Planning Assistant - A Web-based Tool to Support High-quality Radiotherapy in Clinics with Limited Resources

Access to radiotherapy worldwide is limited. The Radiation Planning Assistant (RPA) is a fully automated, web-based tool that is being developed to offer fully automated radiotherapy treatment planning tools to clinics with limited resources. The goal is to help clinical teams scale their efforts, thus reaching more patients with cancer. The user connects to the RPA via a webpage, completes a Service Request (prescription and information about the radiotherapy targets), and uploads the patient&#39;s CT image set. The RPA offers two approaches to automated planning. In one-step planning, the system uses the Service Request and CT scan to automatically generate the necessary contours and treatment plan. In two-step planning, the user reviews and edits the automatically generated contours before the RPA continues to generate a volume-modulated arc therapy plan. The final plan is downloaded from the RPA website and imported into the user&#39;s local treatment planning system, where the dose is recalculated for the locally commissioned linac; if necessary, the plan is edited prior to approval for clinical use.

Autor em destaque: Melhorando o acesso à radioterapia com o Assistente de Planejamento de Radiação

This protocol describes a series of automated tools designed for high-quality radiotherapy autocontouring and autoplanning that are being packaged into a web-based service to maximize robustness and scalability while minimizing operational costs.

Assistente de Planejamento de Radiação - Uma Ferramenta Baseada na Web para Suporte à Radioterapia de Alta Qualidade em Clínicas com Recursos Limitados

Access to radiotherapy worldwide is limited. The Radiation Planning Assistant (RPA) is a fully automated, web-based tool that is being developed to offer ...

Diminui quantificar cognitivos causados ​​pela radioterapia craniana

Diminui quantificar cognitivos causados pela radioterapia craniana