We would like to thank Dr. Harry Orr at the University of Minnesota for his generous gift of transgenic mice. We would also like to thank the following Skidmore College alum for their work performing the preceding experiments: Monica Villegas, Porter Hall, Mitchell Spring, Nicholas Toker, Jenny Zhang, Chloe Larson and Cheyanne Slocum. Furthermore, we would like to thank Skidmore College for funding the development of these methods.

<ol>
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Os autores não têm nada para revelar.

Se estes métodos são usados, conforme descrito, eles são capazes de detectar e aliviar a disfunção mitocondrial mediada por fosforilação oxidativa em modelos de ratos doenças neurodegenerativas cerebelares. Os ensaios bioquímicos e comportamentais combinados são métodos multifacetadas para determinar a extensão da contribuição mitocondrial para cerebelar patologia doença neurodegenerativa. Ao tratar ratos com ácido succínico para estimular o metabolismo e aumentar a função mitocondrial, somos capazes ...

As mitocôndrias são os principais produtores de trifosfato de adenosina (ATP), uma co-enzima essencial para a energia celular, com a maioria do ATP mitocondrial produzido através da fosforilação oxidativa (FOX) usando a cadeia de transporte de electrões. O cérebro, dadas as suas elevadas exigências metabólicas e sua dependência de fosforilação oxidativa para alimentar a atividade neural, é altamente suscetível a disfunção mitocondrial. Como resultado, disfunção mitocondrial é accionado durante o processo de envelhecimento 1 e está implicada na patogénese de várias doenças neurodegenerativas, 2, 3, 4. Portanto, segue-se que as mitocôndrias são alvos terapêuticos atractivos para a neurodegeneração. Neste protocolo, adotamos o uso de ataxia espinocerebelar tipo 1 (SCA1) como uma doença neurodegenerativa modelo para o estudo das mitocôndriasl disfunção e ao desenvolvimento de terapias alvo-mitocondriais. SCA1 é causada por uma mutação de poliglutamina (poliQ) expansão repetida no produto do gene ataxin-1 que desencadeia a progressiva degeneração dos neurónios de Purkinje do cerebelo e neurónios de outras regiões do cérebro. A linha transgénica rato usado aqui (designado como o rato SCA1), que expressa uma ataxin-1 transgene poliQ-mutante sob o controlo de um promotor específico de células de Purkinje, permite a análise orientada de o componente das células de Purkinje de SCA1 5. Ratos SCA1 sofrer degeneração das células de Purkinje gradual e desenvolver marcha atáxica 6. disfunção complexo mitocondrial e eficácia do tratamento alvejado-mitocondrial pode ser avaliada com uma bateria de ensaios moleculares e comportamentais. Disfunção complexo mitocondrial é medida ex vivo através de ensaios de respiração que detectam o consumo de oxigénio no tecido cerebelar alterada ema presença de substratos cadeia de transporte de elétrons e inibidores 7. Os ensaios de respiração foram previamente utilizadas com tecido permeabilizada, isolados mitocondriais, e tecido inteiro 7, 8, 9. Eles permitem a avaliação direta da função mitocondrial ao contrário dos métodos de recolha de dados morfológicos, como a microscopia eletrônica de transmissão ou imunofluorescência. O uso de tecido todo em vez de isolado mitocôndrias impede a seleção tendenciosa de mitocôndrias saudáveis que podem ocorrer durante o processo de isolamento 7. Quando adaptadas ao protocolo como mostrado, o ensaio a respiração é um valioso método para a detecção de disfunção mitocondrial em estados de doenças neurodegenerativas do cerebelo. activadores não específica de metabolismo pode ser usada para inferir a disfunção mitocondrial em modelos de ratinhos transgénicos de diseas neurodegenerativasE e auxílio no desenvolvimento de novas terapias. A quercetina, coenzima Q10 e creatina têm sido mostrados para melhorar a patologia neurodegenerativa doença em doentes e em modelos animais de doença neurodegenerativa 10, 11, 12, 13, 14, 15, 16. Aqui nós apresentamos um novo activador metabólico, ácido succínico, para estimular o metabolismo e aumentar a função mitocondrial na doença neurodegenerativa. Para assegurar que o activador é de atravessar a barreira hemato-encefálica, a HPLC foi utilizada para detectar a entrega ao tecido neural em ratinhos tratados 17. Para avaliar os efeitos terapêuticos dos compostos solúveis em água metabolicamente alvo, tais como ácido succínico, uma bateria de paradigmas comportamentais e estudos imunopatológicos pode ser usado. due para os défices de coordenação motora encontradas na doença neurodegenerativa cerebelar, o ensaio de pista pegada, ensaio de feixe e ensaio da haste rotativa aceleração são usados para detectar salvamento de patologia comportamental 6, 18, 19. Estas medidas são completadas com a avaliação imunopatológico da citoarquitetura cerebelar, avaliando a espessura molecular camada (definido como o comprimento do mandril Purkinje dendrítica celular) e contagens de células de Purkinje soma dentro de um lóbulo definida de tecido cerebelar 6, 20, 21. Aqui apresentamos vários métodos neuropatológicas e comportamentais para a detecção e tratamento da disfunção mitocondrial com compostos solúveis em água metabolicamente alvo. Nós usamos a análise ex vivo da respiração mitocondrial para analisar a disfunção mitocondrial na tran SCA1rato sgenic. Além disso, mostra-se que os sintomas da doença e patologia são melhoradas pelo impulsionador ácido succínico mitocondrial solúvel em água, implicando ainda a disfunção mitocondrial na progressão da doença SCA1.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Adenosine diphosphate</td>
			<td>Sigma Aldrich</td>
			<td>A2754</td>
			<td>ADP</td>
		</tr>
		<tr>
			<td>Ascorbate</td>
			<td>Sigma Aldrich</td>
			<td>A7631</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine serum albumin</td>
			<td>Sigma Aldrich</td>
			<td>A2153</td>
			<td>BSA</td>
		</tr>
		<tr>
			<td>4&#39;,6-Diamidino-2-phenylindole</td>
			<td>Sigma Aldrich</td>
			<td>D9542</td>
			<td>DAPI</td>
		</tr>
		<tr>
			<td>Digitonin</td>
			<td>Sigma Aldrich</td>
			<td>D141</td>
			<td></td>
		</tr>
		<tr>
			<td>Dithiothreitol</td>
			<td>Sigma Aldrich</td>
			<td>D0632</td>
			<td>DTT</td>
		</tr>
		<tr>
			<td>Donkey serum</td>
			<td>Sigma Aldrich</td>
			<td>D9663</td>
			<td></td>
		</tr>
		<tr>
			<td>Glutamate</td>
			<td>Sigma Aldrich</td>
			<td>1446600</td>
			<td></td>
		</tr>
		<tr>
			<td>Malate</td>
			<td>Sigma Aldrich</td>
			<td>6994</td>
			<td></td>
		</tr>
		<tr>
			<td>Mannitol</td>
			<td>Sigma Aldrich</td>
			<td>M4125</td>
			<td></td>
		</tr>
		<tr>
			<td>Paraformaldehyde</td>
			<td>Sigma Aldrich</td>
			<td>P6148</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium-lactobionate</td>
			<td>Bio-Sugars</td>
			<td>69313-67-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Rotenone</td>
			<td>Sigma Aldrich</td>
			<td>R8875</td>
			<td></td>
		</tr>
		<tr>
			<td>Saponin</td>
			<td>Sigma Aldrich</td>
			<td>47036</td>
			<td></td>
		</tr>
		<tr>
			<td>Succinic Acid</td>
			<td>Sigma Aldrich</td>
			<td>S3674</td>
			<td></td>
		</tr>
		<tr>
			<td>N,N,N&#8242;,N&#8242;-Tetramethyl-p-phenylenediamine</td>
			<td>Sigma Aldrich</td>
			<td>T7394</td>
			<td>TMPD</td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>Sigma Aldrich</td>
			<td>T9284</td>
			<td></td>
		</tr>
		<tr>
			<td>Urea</td>
			<td>Sigma Aldrich</td>
			<td>U0631</td>
			<td></td>
		</tr>
		<tr>
			<td>Vectashield mounting medium</td>
			<td>Vector Labs</td>
			<td>H-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibodies</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>11NQ antibody (anti-ataxin-1 )</td>
			<td></td>
			<td></td>
			<td>Servadio, et al. 1995, PMID: 7647801</td>
		</tr>
		<tr>
			<td>Alexa Fluor 488 anti-mouse secondary antibody</td>
			<td>Life Technologies</td>
			<td>A-11015</td>
			<td></td>
		</tr>
		<tr>
			<td>Alexa Fluor 594 anti-rabbit secondary antibody</td>
			<td>Life Technologies</td>
			<td>A-11012</td>
			<td></td>
		</tr>
		<tr>
			<td>Calbindin antibody (goat)</td>
			<td>Santa Cruz</td>
			<td>C-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Animals</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Control transgenic mice</td>
			<td>Harry Orr, Ph.D.</td>
			<td>A02</td>
			<td>Burright, et al. 1997, PMID: 9217978</td>
		</tr>
		<tr>
			<td>SCA1 mice</td>
			<td>Harry Orr, Ph.D.</td>
			<td>B05</td>
			<td>Burright, et al. 1997, PMID: 9217978</td>
		</tr>
		<tr>
			<td>Wildtype mice</td>
			<td>The Jackson Laboratory</td>
			<td>001800</td>
			<td></td>
		</tr>
		<tr>
			<td>Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>ESM-100L microtome</td>
			<td>ERMA</td>
			<td></td>
			<td>Sledge microtome</td>
		</tr>
		<tr>
			<td>Fluoview FV1200 Confocal Microscope</td>
			<td>Olympus</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Glycerol-gelatin slides</td>
			<td>FD Neuro Technologies</td>
			<td>PO101</td>
			<td></td>
		</tr>
		<tr>
			<td>Hamilton syringe</td>
			<td>Sigma Aldrich</td>
			<td>VCAT 80465</td>
			<td></td>
		</tr>
		<tr>
			<td>OXYT1 Oxytherm Electrode Control Unit</td>
			<td>Hansatech Instruments</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>P.T.F.E. paper</td>
			<td>Cole-Parmer</td>
			<td>UX-08277-15</td>
			<td></td>
		</tr>
		<tr>
			<td>Rotallion Rotarod</td>
			<td>PPP&#38;G</td>
			<td></td>
			<td>contact corresponding author for information</td>
		</tr>
		<tr>
			<td>Ultimate 3000 HPLC</td>
			<td>Dionex</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Software</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>ImageJ</td>
			<td>National Institute of Health</td>
			<td></td>
			<td><a href="http://imagej.nih.gov/ij/">http://imagej.nih.gov/ij/</a></td>
		</tr>
		<tr>
			<td>Cell counter plugin (for ImageJ)</td>
			<td>National Institute of Health</td>
			<td></td>
			<td><a href="http://rsb.info.nih.gov/ij/plugins/cell-counter.html">http://rsb.info.nih.gov/ij/plugins/cell-counter.html</a></td>
		</tr>
		<tr>
			<td>3P&#38;G Rota-Rod v3.3.3 (rotarod software)</td>
			<td>PPP&#38;G</td>
			<td></td>
			<td>contact corresponding author for information</td>
		</tr>
		<tr>
			<td>Phidget21.dll (required for rotarod software)</td>
			<td>DLL-Files.com</td>
			<td></td>
			<td><a href="https://www.dll-files.com/phidget21.dll.html">https://www.dll-files.com/phidget21.dll.html</a></td>
		</tr>
	</tbody>
</table>

treating sca1 mice with water-soluble compounds to non-specifically boost mitochondrial function

disfunção mitocondrial desempenha um papel significativo no processo de envelhecimento e em várias doenças neurodegenerativas, incluindo as ataxias espinocerebelares hereditárias e outras perturbações do movimento marcadas pela degeneração progressiva do cerebelo. O objetivo deste protocolo é avaliar disfunção mitocondrial em ataxia espinocerebelar tipo 1 (SCA1) e avaliar a eficácia da segmentação farmacológico da respiração metabólica através do ácido succínico composto solúvel em água para retardar a progressão da doença. Esta abordagem é aplicável a outras doenças cerebelares e pode ser adaptado a uma série de terapias solúveis em água. Análise ex vivo da respiração mitocondrial é utilizado para detectar e quantificar as alterações relacionadas com a doença em função mitocondrial. Com a evidência genética (dados não publicados) e as provas proteômica de disfunção mitocondrial no modelo do rato SCA1, podemos avaliar a eficácia do tratamento com o metabólica impulsionador s solúvel em águaácido uccinic por dissolução deste composto directamente na água de beber casa gaiola. A capacidade do fármaco para passar a barreira hemato-encefálica pode ser deduzida utilizando cromatografia líquida de alta eficiência (HPLC). A eficácia destes compostos pode ser testada usando múltiplos paradigmas comportamentais, incluindo o rotarod aceleração, teste trave de equilíbrio e análise da pegada. integridade citoarquitectónica do cerebelo pode ser avaliada através de imunofluorescência que detectam os núcleos das células de Purkinje e dendritos das células de Purkinje e soma. Estes métodos são técnicas robustas para a determinação da disfunção mitocondrial e a eficácia do tratamento com compostos solúveis em água na doença neurodegenerativa cerebelar.

Através de segmentação farmacológico da mitocôndria cerebelar com ácido succínico somos capazes de prevenir a disfunção mitocondrial em um modelo do rato do cerebelar SCA1 doença neurodegenerativa. O doador de electrões canónica de succinato desidrogenase, ácido succínico, foi dissolvido em água potável casa gaiola de ratos SCA1 durante um mês, com início avaliação comportamental durante a segunda semana de tratamento e a avaliação neuropatológico após o tratament...

Watch this Scientific Journal Video about Treating SCA1 Mice with Water-Soluble Compounds to Non-Specifically Boost Mitochondrial Function at JoVE.com

Tratamento de camundongos SCA1 com compostos solúveis em água para aumentar a função mitocondrial de maneira não específica | Medicina | Vídeo

Treating SCA1 Mice with Water-Soluble Compounds to Non-Specifically Boost Mitochondrial Function

We present a biochemical and behavioral protocol to evaluate the efficacy of mitochondria-targeted water-soluble compounds for the treatment of Spinocerebellar ataxia type 1 (SCA1) and other cerebellar neurodegenerative diseases.

Tratar ratos SCA1 com solúvel em água compostos não especificamente impulsionar a função mitocondrial

Mitochondrial dysfunction plays a significant role in the aging process and in neurodegenerative diseases including several hereditary spinocerebellar ...

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10.5K visualizações.  Skidmore College. We present a biochemical and behavioral protocol to evaluate the efficacy of mitochondria-targeted water-soluble compounds for the treatment of Spinocerebellar ataxia type 1 (SCA1) and other cerebellar neurodegenerative diseases. 

Vídeo: Treating SCA1 Mice with Water-Soluble Compounds to Non-Specifically Boost Mitochondrial Function

Assessing Endothelial Vasodilator Function with the Endo-PAT 2000

The endothelium is a delicate monolayer of cells that lines all blood vessels, and which comprises the systemic and lymphatic capillaries. By virtue of the panoply of paracrine factors that it secretes, the endothelium regulates the contractile and proliferative state of the underlying vascular smooth muscle, as well as the interaction of the vessel wall with circulating blood elements. Because of its central role in mediating vessel tone and growth, its position as gateway to circulating immune cells, and its local regulation of hemostasis and coagulation, the the properly functioning endothelium is the key to cardiovascular health. Conversely, the earliest disorder in most vascular diseases is endothelial dysfunction.
In the arterial circulation, the healthy endothelium generally exerts a vasodilator influence on the vascular smooth muscle. There are a number of methods to assess endothelial vasodilator function. The Endo-PAT 2000 is a new device that is used to assess endothelial vasodilator function in a rapid and non-invasive fashion. Unlike the commonly used technique of duplex ultra-sonography to assess flow-mediated vasodilation, it is totally non-operator-dependent, and the equipment is an order of magnitude less expensive. The device records endothelium-mediated changes in the digital pulse waveform known as the PAT ( peripheral Arterial Tone) signal, measured with a pair of novel modified plethysmographic probes situated on the finger index of each hand. Endothelium-mediated changes in the PAT signal are elicited by creating a downstream hyperemic response. Hyperemia is induced by occluding blood flow through the brachial artery for 5 minutes using an inflatable cuff on one hand. The response to reactive hyperemia is calculated automatically by the system. A PAT ratio is created using the post and pre occlusion values. These values are normalized to measurements from the contra-lateral arm, which serves as control for non-endothelial dependent systemic effects. Most notably, this normalization controls for fluctuations in sympathetic nerve outflow that may induce changes in peripheral arterial tone that are superimposed on the hyperemic response.
In this video we demonstrate how to use the Endo-PAT 2000 to perform a clinically relevant assessment of endothelial vasodilator function.

A noninvasive procedure to assess endothelial function is demonstrated using the Endo-PAT 2000.

Avaliação da função endotelial com a Vasodilatador 2000 Endo-PAT

The endothelium is a delicate monolayer of cells that lines all blood vessels, and which comprises the systemic and lymphatic capillaries. By virtue of ...

Medicina

Tumor Treating Field Therapy in Combination with Bevacizumab for the Treatment of Recurrent Glioblastoma

A novel device that employs TTF therapy has recently been developed and is currently in use for the treatment of recurrent glioblastoma (rGBM). It was FDA approved in April 2011 for the treatment of patients 22 years or older with rGBM. The device delivers alternating electric fields and is programmed to ensure maximal tumor cell kill1.
Glioblastoma is the most common type of glioma and has an estimated incidence of approximately 10,000 new cases per year in the United States alone2. This tumor is particularly resistant to treatment and is uniformly fatal especially in the recurrent setting3-5. Prior to the approval of the TTF System, the only FDA approved treatment for rGBM was bevacizumab6. Bevacizumab is a humanized monoclonal antibody targeted against the vascular endothelial growth factor (VEGF) protein that drives tumor angiogenesis7. By blocking the VEGF pathway, bevacizumab can result in a significant radiographic response (pseudoresponse), improve progression free survival and reduce corticosteroid requirements in rGBM patients8,9. Bevacizumab however failed to prolong overall survival in a recent phase III trial26. A pivotal phase III trial (EF-11) demonstrated comparable overall survival between physicians&#8217; choice chemotherapy and TTF Therapy but better quality of life were observed in the TTF arm10.
There is currently an unmet need to develop novel approaches designed to prolong overall survival and/or improve quality of life in this unfortunate patient population. One appealing approach would be to combine the two currently approved treatment modalities namely bevacizumab and TTF Therapy. These two treatments are currently approved as monotherapy11,12, but their combination has never been evaluated in a clinical trial. We have developed an approach for combining those two treatment modalities and treated 2 rGBM patients. Here we describe a detailed methodology outlining this novel treatment protocol and present representative data from one of the treated patients.

A novel methodology that is employed for the treatment of recurrent glioblastomas is described. This treatment approach employs the application of alternating electric tumor treating fields (TTFields), known as TTF Therapy in combination with bevacizumab, a targeted agent that is currently FDA approved as monotherapy.

Tumor Tratar Terapia campo em combinação com bevacizumabe para o tratamento do glioblastoma recorrente

A novel device that employs TTF therapy has recently been developed and is currently in use for the treatment of recurrent glioblastoma (rGBM). It was FDA ...

Using Saccadometry with Deep Brain Stimulation to Study Normal and Pathological Brain Function

The oculomotor system involves a large number of brain areas including parts of the basal ganglia, and various neurodegenerative diseases including Parkinson's and Huntington's can disrupt it. People with Parkinson's disease, for example, tend to have increased saccadic latencies. Consequently, the quantitative measurement of saccadic eye movements has received considerable attention as a potential biomarker for neurodegenerative conditions. A lot more can be learned about the brain in both health and disease by observing what happens to eye movements when the function of specific brain areas is perturbed. Deep brain stimulation is a surgical intervention used for the management of a range of neurological conditions including Parkinson's disease, in which stimulating electrodes are placed in specific brain areas including several sites in the basal ganglia. Eye movement measurements can then be made with the stimulator systems both off and on and the results compared. With suitable experimental design, this approach can be used to study the pathophysiology of the disease being treated, the mechanism by which DBS exerts it beneficial effects, and even aspects of normal neurophysiology.

This paper describes the use of quantitative measurement of eye movements in conjunction with stimulation of focal areas of the deep brain in order to study physiology, pathophysiology, and the mechanisms of deep brain stimulation.

Usando Saccadometry com Estimulação Cerebral Profunda para estudar normal e patológico função cerebral

The oculomotor system involves a large number of brain areas including parts of the basal ganglia, and various neurodegenerative diseases including ...

Assessment of Respiratory Function in Conscious Mice by Double-chamber Plethysmography

Air volume changes created by a conscious subject breathing spontaneously within a body box are at the basis of plethysmography, a technique used to non-invasively assess some features of the respiratory function in humans as well as in laboratory animals. The present article focuses on the application of the double-chamber plethysmography (DCP) in small animals. It provides background information on the methodology as well as a detailed step-by-step procedure to successfully assess respiratory function in conscious, spontaneously breathing animals in a non-invasive manner. The DCP can be used to monitor the respiratory function of multiple animals in parallel, as well as to identify changes induced by aerosolized substances over a chosen time period and in a repeated manner. Experiments on control and allergic mice are used herein to demonstrate the utility of the technique, explain the associated outcome parameters, as well as to discuss the related advantages and shortcomings. Overall, the DCP provides valid and theoretically sound readouts that can be trusted to evaluate the respiratory function of conscious small animals both at baseline and after challenges with aerosolized substances.

The objective of the present article is to provide a detailed description of the recommended procedures to evaluate respiratory function in conscious mice by double-chamber plethysmography.

Avaliação da função respiratória em ratos conscientes por pletismografia dobro-câmara

Air volume changes created by a conscious subject breathing spontaneously within a body box are at the basis of plethysmography, a technique used to ...

Non-invasive Assessment of Dorsiflexor Muscle Function in Mice

Assessment of skeletal muscle contractile function is an important measurement for both clinical and research purposes. Numerous conditions can negatively affect skeletal muscle. This can result in a loss of muscle mass (atrophy) and/or loss of muscle quality (reduced force per unit of muscle mass), both of which are prevalent in chronic disease, muscle-specific disease, immobilization, and aging (sarcopenia). Skeletal muscle function in animals can be evaluated by a range of different tests. All tests have limitations related to the physiological testing environment, and the selection of a specific test often depends on the nature of the experiments. Here, we describe an in vivo, non-invasive technique involving a helpful and easy assessment of force frequency-curve (FFC) in mice that can be performed on the same animal over time. This permits monitoring of disease progression and/or efficacy&#160;of a potential therapeutic treatment.

Measurement of rodent skeletal muscle contractile function is a useful tool that can be used to track disease progression as well as efficacy of therapeutic intervention. We describe here the non-invasive, in vivo assessment of the dorsiflexor muscles that can be repeated over time in the same mouse.

Avaliação não-invasiva da função de músculos levantadores em ratos

Assessment of skeletal muscle contractile function is an important measurement for both clinical and research purposes. Numerous conditions can negatively ...

A Non-Invasive Method for Generating the Cyclic Loading-Induced Intra-Articular Cartilage Lesion Model of the Rat Knee

The pathophysiology of primary osteoarthritis (OA) remains unclear. However, a specific subclassification of OA in relatively younger age groups is likely correlated with a history of articular cartilage damage and ligament avulsion. Surgical animal models of OA of the knee play an important role in understanding the onset and progression of post-traumatic OA and aid in the development of novel therapies for this disease. However, non-surgical models have been recently considered to avoid traumatic inflammation that could affect the evaluation of the intervention. 
In this study, an intra-articular cartilage lesion rat model induced by in vivo cyclic compressive loading was developed, which allowed researchers to (1) determine the optimal magnitude, speed, and duration of load that could cause focal cartilage damage; (2) assess post-traumatic spatiotemporal pathological changes in chondrocyte vitality; and (3) evaluate the histological expression of destructive or protective molecules that are involved in the adaptation and repair mechanisms against joint compressive loads. This report describes the experimental protocol for this novel cartilage lesion in a rat model.

Here, we present the cyclic loading-induced intra-articular cartilage lesion model of the rat knee, generated by 60 cyclic compressions over 20 N, resulting in damage to the femoral condylar cartilage in rats.

Um Método Não Invasivo para Gerar o Modelo de Lesão da Cartilagem Intra-Articular Induzida por Carga Cíclica do Joelho de Rato

The pathophysiology of primary osteoarthritis (OA) remains unclear. However, a specific subclassification of OA in relatively younger age groups is likely ...

Determinando a respiração em espermatozóides humanos por respirometria de alta resolução

High-Resolution Respirometry to Assess Mitochondrial Function in Human Spermatozoa

Semen quality is often studied by routine semen analysis, which is descriptive and often inconclusive. Male infertility is associated with altered sperm mitochondrial activity, so the measurement of sperm mitochondrial function is an indicator of sperm quality. High-resolution respirometry is a method of measuring the oxygen consumption of cells or tissues in a closed-chamber system. This technique can be implemented to measure respiration in human sperm and provides information about the quality and integrity of the sperm mitochondria. High-resolution respirometry allows the cells to move freely, which is an a priori advantage in the case of sperm. This technique can be applied with intact or permeabilized spermatozoa and allows for the study of intact sperm mitochondrial function and the activity of individual respiratory chain complexes. The high-resolution oxygraph instrument uses sensors to measure the oxygen concentration coupled with sensitive software to calculate the oxygen consumption. The data are used to calculate respiratory indices based on the oxygen consumption ratios. Consequently, the indices are the proportions of two oxygen consumption rates and are internally normalized to the cell number or protein mass. The respiratory indices are an indicator of sperm mitochondrial function and dysfunction.

Autor em destaque: Avançando na pesquisa de infertilidade masculina, desvendando o metabolismo do esperma e a função mitocondrial 

The analysis of sperm mitochondrial function by high-resolution respirometry permits the measurement of the oxygen consumption of freely moving spermatozoa in a closed-chamber system. The technique can be applied to measure respiration in human spermatozoa, which provides information on sperm mitochondrial characteristics and integrity.

Respirometria de Alta Resolução para Avaliação da Função Mitocondrial em Espermatozoides Humanos

Semen quality is often studied by routine semen analysis, which is descriptive and often inconclusive. Male infertility is associated with altered sperm ...

Protocolo Tuina Padronizado para Modelo de Compactação DRG de Rato

Analgesic Effect of Tuina on Rat Models with Compression of the Dorsal Root Ganglion Pain

Neuropathic pain is a prevalent condition that affects 6.9%-10% of the population and results from nerve damage due to various etiologies, such as lumbar disc herniation, spinal canal stenosis, and intervertebral foramen stenosis. Although Tuina, a traditional Chinese manual therapy, has shown analgesic effects in clinical practice for the treatment of neuropathic pain, its underlying neurobiological mechanisms remain unclear. Animal models are essential for elucidating the basic principles of Tuina. In this study, we propose a standardized Tuina protocol for rats with compression of the dorsal root ganglion (DRG), which involves inducing DRG compression by inserting a stainless steel rod into the intervertebral foramen, performing Tuina manipulation with specific parameters of location, intensity, and frequency in a controlled environment, and assessing the behavioral and histopathological outcomes of Tuina treatment. This article also discusses the potential clinical implications and limitations of the study and suggests directions for future research on Tuina.

Autor em destaque: Efeito analgésico da Tuina em modelos de ratos com compressão da dor do gânglio da raiz dorsal  

This article presents a manipulation for treating chronic compression of the dorsal root ganglion in rats using Tuina therapy, along with a method for evaluating its effectiveness based on pain behavior and histopathological results.

Efeito Analgésico de Tuina em Modelos de Rato com Compressão da Dor do Gânglio da Raiz Dorsal

Neuropathic pain is a prevalent condition that affects 6.9%-10% of the population and results from nerve damage due to various etiologies, such as lumbar ...