Os autores gostariam de agradecer ao Sr. Pascal Lejeune por seu apoio técnico durante todos os procedimentos.

O protocolo apresentado é utilizado para éguas apresentadas no hospital equino e segue as diretrizes institucionais de cuidados com os animais.
NOTA: Realizar fotoablação de cistos endometrial antes do anestrus: os calores observados no final de julho ou agosto parecem, portanto, mais confiáveis. Não realize este procedimento tarde demais na estação de reprodução, pois pode limitar o número de estruses disponíveis para tratar uma endometrite potencial.
1. Durante as estrus anteriores
<ol><li>Realize um exame completo de solidez de reprodução da égua, incluindo uma biópsia endometrial1 (não descrita aqui) para garantir que a égua possa potencialmente engravidar após o procedimento.</li>
	<li>Em caso de má biópsia endometrial de acordo com a classificação1de Kenney, dê a transferência de embrião ou a injeção intra citoplasmática de espermatozoides (ICSI) preferência sobre a fotoabalização dos cistos endometrial.</li>
</ol>2. Durante o diestrus anterior
<ol><li>Uma vez que o reto esteja vazio, conte, meça e mapeie cistos endometrial da égua no útero usando ultrassonografia trans-retal guiada pela palpação retal com uma sonda retatal linear de 7,5 ou 5 MHz e pinças na tela(Figura 3) em um estoque de contenção.</li>
	<li>Administrar 250 μg de cloprostenol intra-muscularmente para induzir estrus dentro de 2 ou 3 dias após o dia 6\u20127 pós-ovulação.</li>
	<li>Diariamente, realizam ultrassonografia de rotina comum e palpação do colo do útero para observar sinais de estrus, incluindo edema endometrial significativo, folículo antral de 35 mm e colo uterino suavizado e aberto.</li>
</ol>3. Preparação quando a égua está no calor
<ol><li>Restringir o acesso à sala de operação para pessoas que usam óculos de proteção ocular.</li>
	<li>Após o esvaziamento do reto, enrole a cauda e limpe a vulva asepticamente usando iodo-povidone.</li>
	<li>Para garantir fácil manipulação e conforto da égua, administre a detomidina (10 μg/kg) e o butorphanol (0,1 mg/kg) por via intravenosa na veia jugular com uma agulha de 21 G.</li>
	<li>Por vagina, instale o cateter de coleta de embriões através do colo uterino e insuflar o balão com 40 mL de ar.</li>
	<li>Infundir o útero com 1,5 L de solução salina estéril (0,9% NaCl) para inflar o útero.</li>
	<li>Esvazie o balão e remova o cateter.</li>
	<li>Passe o endoscópio asepticamente através do colo uterino dentro de uma manga estéril.</li>
</ol>4. Técnica
<ol><li>Conecte a fibra de quartzo de 600 μm a um laser de diodo operando a 980 nm. Defina a potência em 20\u201225 W em modo contínuo.</li>
	<li>Passe a fibra de quartzo flexível através do canal de biópsia de um vídeo-endoscópio de 10 mm de diâmetro até que 3\u20124 cm de fibra livre possam ser identificados na tela. Um mínimo de 10 mm a partir da extremidade da fibra deve ser visível.</li>
	<li>Ative o laser para perfurar o cisto idealmente em seu ápice: um pedal dedicado está disponível no dispositivo para manter as mãos livres. Em alguns casos, múltiplas perfurações em vários locais são necessárias para alcançar a anulação completa do cisto. O fluido cístico é passivamente drenado para dentro do útero.</li>
	<li>Depois que o revestimento do cisto entrou em colapso ao redor da fibra, implante o laser até que a membrana encole.</li>
	<li>Realize a ultrassonografia transretal do útero quantas vezes for necessário durante o procedimento para confirmar que cistos endometrial desapareceram.</li>
</ol>5. Tratamentos pós-operatórios
<ol><li>Administrou o tratamento geral com antibióticos: 22.000 UI/kg penicilina-procaina suspensão, intra-muscular, duas vezes por dia, durante 3 dias.</li>
	<li>Logo após a intervenção, realize lavage uterino conforme descrito acima (ver passos 3.4-3.6).<ol>
<li>Colete o líquido infundido no útero antes do procedimento endoscópico através de um cateter algemado estéril.</li>
		<li>Lave o útero com sucessivas instilações de 1 L solução isotônica estéril (0,9% NaCl) até que um líquido claro seja coletado.</li>
		<li>Ao final do procedimento, administre 20 UI de ocitocina por via intravenosa.</li>
		<li>Realize a ultrassonografia transretal do útero da égua no dia seguinte: o líquido livre intraluminal é frequentemente observado (ver passo 2.1).</li>
	</ol>
</li>
	<li>Desde que o fluido intra-luminal livre seja observado pela ultrassonografia transretal e até o fim do calor, realize lavages uterinos diariamente com o procedimento descrito acima (etapas 3.4-3.6). Às vezes, o líquido manchado de soro, límpido e amarelo é coletado por vários dias. NOTA: Dar repetidas injeções intra-musculares de ocitocina (20 UI) a cada 4h ou menos quando a égua está em estrus pode evitar o acúmulo de fluidos e reduzir o risco de contaminação bacteriana no útero.</li>
	<li>Realizar ultrassonografia uterina trans-retal durante os seguintes diestrus para confirmar o desaparecimento de cistos endometrial e a ausência de fluido livre intraluminal. NOTA: Se for observado fluido intraluminal livre, recomenda-se a injeção de prostaglandina para induzir um novo estrus, diagnosticar uma eventual endometrite subsequente e tratá-la.</li>
</ol>

<ol>
	<li>Kenney, R. M., Ganjam, V. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selected+pathological+changes+of+the+mare+uterus+and+ovary.">Selected pathological changes of the mare uterus and ovary.</a> Journal of Reproduction and Fertility. (23), 335-339 (1975).</li><li>Bracher, V., Mathias, S., Allen, W. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Videoendoscopic+evaluation+of+the+mare's+uterus:+II.+Findings+in+subfertile+mares.">Videoendoscopic evaluation of the mare's uterus: II. Findings in subfertile mares.</a> Equine Veterinary Journal. 24 (4), 279-284 (1992).</li><li>Kaspar, B., Kahn, W., Laging, C., Leidl, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endometrial+cysts+in+the+mare.+1.+Post-mortem+studies:+occurrence+and+morphology.">Endometrial cysts in the mare. 1. Post-mortem studies: occurrence and morphology.</a> Tier&#228;rztliche Praxis. 15 (2), 161-166 (1987).</li><li>Leidl, W., Kaspar, B., Kahn, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endometrial+cysts+in+the+mare.+2.+Clinical+studies:+occurrence+and+significance.">Endometrial cysts in the mare. 2. Clinical studies: occurrence and significance.</a> Tier&#228;rztliche Praxis. 15 (3), 281-289 (1987).</li><li>Mather, E. C., Refsal, K. R., Gustafsson, B. K., Seguin, B. E., Whitmore, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+fibre-optic+techniques+in+clinical+diagnosis+and+visual+assessment+of+experimental+intrauterine+therapy+in+mares.">The use of fibre-optic techniques in clinical diagnosis and visual assessment of experimental intrauterine therapy in mares.</a> Journal of Reproduction and Fertility. (27), 293-297 (1979).</li><li>Stanton, M. E., Mc Kinnon, A. O., Squires, E. L., Vaala, W. E., Varner, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Equine Reproduction Vol. 2. , 2665-2668 (2011).</li><li>Bracher, V., Mathias, S., Allen, W. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+chronic+degenerative+endometritis+(endometrosis)+on+placental+development+in+the+mare.">Influence of chronic degenerative endometritis (endometrosis) on placental development in the mare.</a> Equine Veterinary Journal. 28 (3), 180-188 (1996).</li><li>Tannus, R. J., Thun, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+endometrial+cysts+on+conception+rate+of+mares.">Influence of endometrial cysts on conception rate of mares.</a> Zentralbl Veterinarmed A. 42 (4), 275-283 (1995).</li><li>Pipers, F. S., Zent, W., Holder, R., Asbury, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasonography+as+an+adjunct+to+pregnancy+assessments+in+the+mare.">Ultrasonography as an adjunct to pregnancy assessments in the mare.</a> Journal of the American Veterinary Medical Association. 184 (3), 328-334 (1984).</li><li>LeBlanc, M., Mc Kinnon, A. O., Mc Kinnon, A. O., Squires, E. L., Vaala, W. E., Varner, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Equine Reproduction Vol. 2. , 2620-2642 (2011).</li><li>van Ittersum, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+electrosurgical+treatment+of+endometrial+cysts+in+the+mare.">The electrosurgical treatment of endometrial cysts in the mare.</a> Tijdschr Diergeneeskd. 124 (21), 630-633 (1999).</li><li>Rambags, B. P., Stout, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transcervical+endoscope-guided+emptying+of+a+transmural+uterine+cyst+in+a+mare.">Transcervical endoscope-guided emptying of a transmural uterine cyst in a mare.</a> Vet Record. 156 (21), 679-682 (2005).</li><li>Blikslager, A. T., Tate, L. P., Weinstock, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+neodymium:yttrium+aluminum+garnet+laser+irradiation+on+endometrium+and+on+endometrial+cysts+in+six+mares.">Effects of neodymium:yttrium aluminum garnet laser irradiation on endometrium and on endometrial cysts in six mares.</a> Veterinary Surgery. 22 (5), 351-356 (1993).</li><li>Bartmann, C. P., Stief, B., Schoon, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thermal+injury+and+wound+healing+of+the+endometrium+subsequent+to+minimally+invasive+transendoscopic+use+of+Nd:YAG-laser-and+electrosurgery+in+horses.">Thermal injury and wound healing of the endometrium subsequent to minimally invasive transendoscopic use of Nd:YAG-laser-and electrosurgery in horses.</a> Dtsch Tierarztl Wochenschr. 110 (7), 271-280 (2003).</li><li>Conley, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Review+of+the+reproductive+endocrinology+of+the+pregnant+and+parturient+mare.">Review of the reproductive endocrinology of the pregnant and parturient mare.</a> Theriogenology. 86 (1), 355-365 (2016).</li><li>Guzeloglu, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+enzymes+and+receptors+of+leukotriene+pathway+genes+in+equine+endometrium+during+the+estrous+cycle+and+early+pregnancy.">Expression of enzymes and receptors of leukotriene pathway genes in equine endometrium during the estrous cycle and early pregnancy.</a> Theriogenology. 80 (2), 145-152 (2013).</li><li>Senger, P. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Pathways to pregnancy and parturition, 3rd edn. , (2015).</li></ol>

Todos os autores são totalmente empregados pela Clínica Equina da Universidade liège e não têm conflitos de interesse com qualquer empresa negociando um dos produtos mencionados acima.

A histeroscopia e a fotoablação a laser dos cistos endometrial é um procedimento fácil para reduzir grandes cistos intra-luminais na égua, como sugerido anteriormente14. Tecnicamente, este método é melhorado quando realizado em uma égua em estrus. Inflar o útero com solução salina estéril melhorou a visibilidade quando comparado ao enchimento do ar e à aspiração contínua durante a realização do procedimento.  Pequenos cistos intra parietais (<10 mm) são mais difíceis de alcan?...

Cistos endometrial em éguas foram histopatológicos descritos nos anos 70 por Kenney1. No final dos anos 80, o uso de histeroscopia e ultrassonografia levou à sua descrição clínica e ajudou a entender suas consequências sobre a fertilidade2,3,4,5.
Nos estágios iniciais, os cistos endometrial têm sido associados a uma degeneração do endométrio agora chamado de endometrose1,3,4,6,7,8 e atualmente é aceito que pequenos cistos (<1 cm) estão relacionados à fibrose endometrial das glândulas quando cistos maiores estão relacionados à dilatação linfática6 . Esses cistos grandes são os mais propensos a perturbar a migração de embriões equinos entre o dia 6,5 e o dia 17, diminuindo assim a fertilidade ao prejudicar o reconhecimento materno1,3,4,6,8. Cistos endometrial também podem interferir no diagnóstico precoce da gravidez, especialmente se forem redondos e tiverem um tamanho semelhante ao de uma vesícula embrionária3,4,6,9 (ver Figura 1). Assim, podem potencialmente aumentar o intervalo entre duas inseminações, atrasando a ultrassonografia do primeiro embrião e a avaliação da gestação de gêmeos (ver Figura 2). Cistos endometrial também podem interferir mecanicamente com a coleta de fluidos durante a lavagem uterina realizada para tratar endometrite ou coletar o embrião. Consequentemente, os cistos endometrial têm efeitos sobre a fertilidade observada da égua, com potenciais consequências econômicas nas fazendas de reprodução.
Além das tentativas de tratar a endometrosesubjacente 10,foram propostos tratamentos focais endoscopicamente guiados de cistos. A cauterização eletrocirúrgica foi descrita, mas a exposição ao calor pode ferir o endométrio ao redor do cisto11. A aspiração do cisto só é eficiente se a estrutura de segregação não estiver mais ativa no momento da punção12. Irradiação a laser com um laser de diodo de 980 nm com fibra de contato parece mostrar cicatrizes endometrial subsequentes menores6,13. O aprimoramento de dispositivos e procedimentos a laser6 agora nos permite propor essa técnica segura e eficiente na prática clínica equina. No entanto, os proprietários de éguas devem ser informados sobre os limites desta opção. A fotoablação dos cistos aumentará o reconhecimento de embriões em algumas éguas, avançará no diagnóstico da gravidez em alguns casos e poderá limitar o efeito mecânico negativo dos cistos endometrial sobre o lavage uterino realizado para tratar endometrite ou coletar embriões. No entanto, nunca tratará a endometriose ou endometriose subjacente concomitante.
Este relatório tem como objetivo descrever um procedimento eficiente para realizar a ablação de cisto a laser de diodo guiado por histeroscopia em éguas. A preparação e os procedimentos serão praticamente descritos para uma aplicação clínica.

<table border="0" cellpadding="0" cellspacing="0" style="width:664px;" width="663">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:214px;">Butorphanol</td>
			<td style="width:121px;">Animedica</td>
			<td style="width:161px;">Morphasol 10 mg/mL</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:214px;">Detomidine</td>
			<td style="width:121px;">Dechra</td>
			<td style="width:161px;">Domidine 10 mg/mL</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:214px;">Endoscope</td>
			<td style="width:121px;">Olympus</td>
			<td style="width:161px;">Vet 170-10 SN2012729</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:214px;">Endoscope Vid&#233;o Tower</td>
			<td style="width:121px;">Olympus</td>
			<td style="width:161px;">CLE-E &#38; CV-E</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:214px;">Laser device</td>
			<td style="width:121px;">Wuhan Giga Optronics</td>
			<td style="width:161px;">Velas II-60B GA19-V3186</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:214px;">NaCl 0.9%</td>
			<td style="width:121px;">Baxter</td>
			<td style="width:161px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:214px;">P&#233;nicilline-proca&#239;ne</td>
			<td style="width:121px;">Kela</td>
			<td style="width:161px;">PENI-Kel 300.00 UI/ml</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:214px;">Ultrasonography</td>
			<td style="width:121px;">Aloka</td>
			<td style="width:161px;">Aloka 500 &#38; 5mHz transrectal probe</td>
			<td></td>
		</tr>
	</tbody>
</table>

endoscopy guided photoablation of endometrial cysts using a 980 nm laser with a contact fiber in mares

Nas éguas, os cistos endometrial estão associados à endometriose e podem causar falha de reconhecimento materno ou comprometer e retardar o diagnóstico da gravidez. Os tratamentos históricos foram invasivos e tiveram efeitos adversos no endométrio. A laserterapia histeroscópica guiada é fácil e eficaz para a ressecção de cistos endometrial, sem efeitos deletérios para o endométrio.
Um endoscópio de 110 cm de comprimento e 1,0 cm de largura é introduzido estérilmente no útero através do colo uterino aberto de uma égua estrâmica após a limpeza vulvar. O útero é lentamente infundido com menos de 1 L de solução fisiológica e a fibra laser é inserida no canal de biópsia do endoscópio. Os cistos são então cauterizados com o laser de diodo de 980 nm com um conjunto de fibra de contato a 20\u20122 5W no modo contínuo. Cada cisto é perfurado até a anulação completa do cisto e o encolhimento da parede do cisto ao redor da fibra. Lavages uterinas com solução salina estéril são realizadas logo após a cirurgia e por um ou dois dias como fluido não inflamatório pode ser observado.
Este procedimento é fácil e rapidamente realizado, sem efeitos deletérios óbvios. A ressecção de cistos facilita o diagnóstico de gravidez por ultrassom e, em alguns casos, pode restaurar a migração adequada de embriões nos chifres uterinos entre os dias 6,5 e 17. No entanto, este tratamento não melhora as lesões histológicas subjacentes relacionadas à endometriose. Essas considerações devem ser expressas claramente ao criador antes deste procedimento.

Foram realizadas cirurgias em éguas com cistos com diâmetros acima de 15 mm. No entanto, se outros cistos menores também estivessem presentes, a fotoablação dos outros cistos foi tentada. Geralmente, esses grandes cistos estavam na bifurcação ou na base dos chifres. No entanto, alguns deles estavam presentes no corpo do útero e foram igualmente tratados.
Em nossa experiência, cistos intraluminais são muito fáceis de localização com o endoscópio. Além disso, a realização desse ...

Watch this Scientific Journal Video about Endoscopy Guided Photoablation of Endometrial Cysts using a 980 nm Laser with a Contact Fiber in Mares at JoVE.com

Endoscopy Guided Photoablation of Endometrial Cysts using a 980 nm Laser with a Contact Fiber in Mares

O objetivo deste relatório é descrever a fotoablação de cistos endometrial na égua usando um dispositivo laser endoscópico, e discutir os interesses e limitações dessa nova abordagem para lidar com problemas de baixa fertilidade.

Fotoablação Guiada de Endoscopia de Cistos Endometrial usando um Laser de 980 nm com uma fibra de contato em Mares

In mares, endometrial cysts are associated with endometriosis and can cause maternal recognition failure or compromise and delay pregnancy diagnoses. ...

endoscopy-guided-photoablation-endometrial-cysts-using-980-nm-laser

Research

JoVE Journal

Medicine

4.2K visualizações.  Liège University. O objetivo deste relatório é descrever a fotoablação de cistos endometrial na égua usando um dispositivo laser endoscópico, e discutir os interesses e limitações dessa nova abordagem para lidar com problemas de baixa fertilidade.

Vídeo: Endoscopy Guided Photoablation of Endometrial Cysts using a 980 nm Laser with a Contact Fiber in Mares

MRI-guided Disruption of the Blood-brain Barrier using Transcranial Focused Ultrasound in a Rat Model

Focused ultrasound (FUS) disruption of the blood-brain barrier (BBB) is an increasingly investigated technique for circumventing the BBB1-5. The BBB is a significant obstacle to pharmaceutical treatments of brain disorders as it limits the passage of molecules from the vasculature into the brain tissue to molecules less than approximately 500 Da in size6. FUS induced BBB disruption (BBBD) is temporary and reversible4 and has an advantage over chemical means of inducing BBBD by being highly localized. FUS induced BBBD provides a means for investigating the effects of a wide range of therapeutic agents on the brain, which would not otherwise be deliverable to the tissue in sufficient concentration. While a wide range of ultrasound parameters have proven successful at disrupting the BBB2,5,7, there are several critical steps in the experimental procedure to ensure successful disruption with accurate targeting. This protocol outlines how to achieve MRI-guided FUS induced BBBD in a rat model, with a focus on the critical animal preparation and microbubble handling steps of the experiment.

Microbubble-mediated focused ultrasound disruption of the blood-brain barrier (BBB) is a promising technique for non-invasive targeted drug delivery in the brain1-3. This protocol outlines the experimental procedure for MRI-guided transcranial BBB disruption in a rat model.

Guiada por MRI rompimento da barreira hemato-encefálica utilizando Ultrassom Focalizado transcraniana em um modelo de rato

Focused ultrasound (FUS) disruption of the blood-brain barrier (BBB) is an increasingly investigated technique for circumventing the BBB1-5. The BBB is a ...

Medicina

Epidural Intracranial Pressure Measurement in Rats Using a Fiber-optic Pressure Transducer

Elevated intracranial pressure (ICP) is a significant problem in several forms of ischemic brain injury including stroke, traumatic brain injury and cardiac arrest. This elevation may result in further neurological injury, in the form of transtentorial herniation1,2,3,4, midbrain compression, neurological deficit or increased cerebral infarct2,4. Current therapies are often inadequate to control elevated ICP in the clinical setting5,6,7 . Thus there is a need for accurate methods of ICP measurement in animal models to further our understanding of the basic mechanisms and to develop new treatments for elevated ICP.
In both the clinical and experimental setting ICP cannot be estimated without direct measurement. Several methods of ICP catheter insertion currently exist. Of these the intraventricular catheter has become the clinical 'gold standard' of ICP measurement in humans8. This method involves the partial removal of skull and the instrumentation of the catheter through brain tissue. Consequently, intraventricular catheters have an infection rate of 6-11%9. For this reason, subdural and epidural cannulations have become the preferred methods in animal models of ischemic injury. 
Various ICP measurement techniques have been adapted for animal models, and of these, fluid-filled telemetry catheters10 and solid state catheters are the most frequently used11,12,13,14,15. The fluid-filled systems are prone to developing air bubbles in the line, resulting in false ICP readings. Solid state probes avoid this problem (Figure 1). An additional problem is fitting catheters under the skull or into the ventricles without causing any brain injury that might alter the experimental outcomes. Therefore, we have developed a method that places an ICP catheter contiguous with the epidural space, but avoids the need to insert it between skull and brain. 
An optic fibre pressure catheter (420LP, SAMBA Sensors, Sweden) was used to measure ICP at the epidural location because the location of the pressure sensor (at the very tip of the catheter) was found to produce a high fidelity ICP signal in this model. There are other manufacturers of similar optic fibre technologies13 that may be used with our methodology. Alternative solid state catheters, which have the pressure sensor located at the side of the catheter tip, would not be appropriate for this model as the signal would be dampened by the presence of the monitoring screw. 
Here, we present a relatively simple and accurate method to measure ICP. This method can be used across a wide range of ICP related animal models.

A novel technique to record the pressures within the skull is described. The minimally invasive method uses a fibre-optic pressure sensing system to accurately measure intracranial pressure (ICP) in anaesthetized rats without causing significant brain trauma. The technique may be used in a wide range of experimental models.

Medida da Pressão Intracraniana peridural em ratos, utilizando um transdutor de pressão de fibra óptica

Elevated intracranial pressure (ICP) is a significant problem in several forms of ischemic brain injury including stroke, traumatic brain injury and ...

Reproducable Paraplegia by Thoracic Aortic Occlusion in a Murine Model of Spinal Cord Ischemia-reperfusion

Background 
Lower extremity paralysis continues to complicate aortic interventions. The lack of understanding of the underlying pathology has hindered advancements to decrease the occurrence this injury.&#160;The current model demonstrates reproducible lower&#160;extremity paralysis following thoracic aortic occlusion.
Methods 
Adult male C57BL6 mice were anesthetized with isoflurane. Through a cervicosternal incision the aorta was exposed. The descending thoracic aorta and left subclavian arteries were identified without entrance into pleural space. Skeletonization of these arteries was followed by immediate closure (Sham) or occlusion for 4 min (moderate ischemia) or 8 min (prolonged ischemia). The sternotomy and skin were closed and the mouse was transferred to warming bed for recovery.&#160; Following recovery, functional analysis was obtained at 12 hr intervals until 48 hr.
Results 
Mice that underwent sham surgery showed no observable hind&#160;limb deficit. Mice subjected to moderate ischemia for 4 min had minimal functional deficit at 12 hr followed by progression to complete paralysis at 48 hr. Mice subjected to prolonged ischemia had an immediate paralysis with no observable hind-limb movement at any point in the postoperative period. There was no observed intraoperative or post&#160;operative mortality.
Conclusion 
Reproducible lower extremity paralysis whether immediate or delayed can be achieved in a murine model. Additionally, by using a median sternotomy and careful dissection, high survival rates, and reproducibility can be achieved.

The lack of mechanistic understanding of spinal cord ischemia-reperfusion injury has hindered further adjuncts to prevent paraplegia following high risk aortic operations. Thus, the development of animal models is imperative. This manuscript demonstrates reproducible lower extremity paralysis following thoracic aortic occlusion in a murine model.

Reproducable Paraplegia por Thoracic Aorta Oclusão em um modelo murino de Medula Espinhal A isquemia-reperfusão

Background
Lower extremity paralysis continues to complicate aortic interventions. The lack of understanding of the underlying pathology has hindered ...

Image-guided Convection-enhanced Delivery into Agarose Gel Models of the Brain

Convection-enhanced delivery (CED) has been proposed as a treatment option for a wide range of neurological diseases. Neuroinfusion catheter CED allows for positive pressure bulk flow to deliver greater quantities of therapeutics to an intracranial target than traditional drug delivery methods. The clinical utility of real time MRI guided CED (rCED) lies in the ability to accurately target, monitor therapy, and identify complications. With training, rCED is efficient and complications may be minimized. The agarose gel model of the brain provides an accessible tool for CED testing, research, and training. Simulated brain rCED allows practice of the mock surgery while also providing visual feedback of the infusion. Analysis of infusion allows for calculation of the distribution fraction (Vd/Vi) allowing the trainee to verify the similarity of the model as compared to human brain tissue. This article describes our agarose gel brain phantom and outlines important metrics during a CED infusion and analysis protocols while addressing common pitfalls faced during CED infusion for the treatment of neurological disease.

Convection-enhanced delivery (CED) has been proposed as a treatment option for a wide range of neurological diseases. In order to prepare health care professionals for adoption of CED, accessible training models are needed. We describe the use of agarose gel as such a model of the human brain for testing, research, and training.

Imagem-guiado Entrega reforçada-Convecção em Modelos gel de agarose do Cérebro

Convection-enhanced delivery (CED) has been proposed as a treatment option for a wide range of neurological diseases. Neuroinfusion catheter CED allows ...

A Possible Zebrafish Model of Polycystic Kidney Disease: Knockdown of wnt5a Causes Cysts in Zebrafish Kidneys

Polycystic kidney disease (PKD) is one of the most common causes of end-stage kidney disease, a devastating disease for which there is no cure. The molecular mechanisms leading to cyst formation in PKD remain somewhat unclear, but many genes are thought to be involved. Wnt5a is a non-canonical glycoprotein that regulates a wide range of developmental processes. Wnt5a works through the planar cell polarity (PCP) pathway that regulates oriented cell division during renal tubular cell elongation. Defects of the PCP pathway have been found to cause kidney cyst formation. Our paper describes a method for developing a zebrafish cystic kidney disease model by knockdown of the wnt5a gene with wnt5a antisense morpholino (MO) oligonucleotides. Tg(wt1b:GFP) transgenic zebrafish were used to visualize kidney structure and kidney cysts following wnt5a knockdown. Two distinct antisense MOs (AUG - and splice-site) were used and both resulted in curly tail down phenotype and cyst formation after wnt5a knockdown. Injection of mouse Wnt5a mRNA, resistant to the MOs due to a difference in primary base pair structure, rescued the abnormal phenotype, demonstrating that the phenotype was not due to &#8220;off-target&#8221; effects of the morpholino. This work supports the validity of using a zebrafish model to study wnt5a function in the kidney.

A Possible Zebrafish Model of Polycystic Kidney Disease: Knockdown of wnt5a Causes Cysts in Zebrafish Kidneys

We describe a method of generating a possible zebrafish model of polycystic kidney disease. We used Tg(wt1b:GFP) fish to visualize kidney structure. Knockdown of wnt5a was by morpholino injection. Pronephric cyst formation after wnt5a knockdown was observed in this GFP transgenic zebrafish.

Um Modelo Possível Zebrafish da doença renal policística: Knockdown de<em&gt; Wnt5a</em&gt; Faz com cistos no peixe-zebra Rins

Polycystic kidney disease (PKD) is one of the most common causes of end-stage kidney disease, a devastating disease for which there is no cure. The ...

Semi-automated Analysis of Mouse Skeletal Muscle Morphology and Fiber-type Composition

For years, distinctions between skeletal muscle fiber types were best visualized by myosin-ATPase staining. More recently, immunohistochemical staining of myosin heavy chain (MyHC) isoforms has emerged as a finer discriminator of fiber-type. Type I, type IIA, type IIX and type IIB fibers can now be identified with precision based on their MyHC profile; however, manual analysis of these data can be slow and down-right tedious. In this regard, rapid, accurate assessment of fiber-type composition and morphology is a very desirable tool. Here, we present a protocol for state-of-the-art immunohistochemical staining of MyHCs in frozen sections obtained from mouse hindlimb muscle in concert with a novel semi-automated algorithm that accelerates analysis of fiber-type and fiber morphology. As expected, the soleus muscle displayed staining for type I and type IIA fibers, but not for type IIX or type IIB fibers. On the other hand, the tibialis anterior muscle was composed predominantly of type IIX and type IIB fibers, a small fraction of type IIA fibers and little or no type I fibers. Several image transformations were used to generate probability maps for the purpose of measuring different aspects of fiber morphology (i.e., cross-sectional area (CSA), maximal and minimal Feret diameter). The values obtained for these parameters were then compared with manually-obtained values. No significant differences were observed between either mode of analysis with regards to CSA, maximal or minimal Feret diameter (all p &#62; 0.05), indicating the accuracy of our method. Thus, our immunostaining analysis protocol may be applied to the investigation of effects on muscle composition in many models of aging and myopathy.

Immunohistochemical staining of myosin heavy chain isoforms has emerged as the state-of-the-art discriminator of skeletal muscle fiber-type (i.e., type I, type IIA, type IIX, type IIB). Here, we present a staining protocol along with a novel semi-automated algorithm that facilitates rapid assessment of fiber-type and fiber morphology.

Semi automatizados de análise da morfologia do músculo esquelético de rato e composição de fibra-tipo

For years, distinctions between skeletal muscle fiber types were best visualized by myosin-ATPase staining. More recently, immunohistochemical staining of ...

A Mobile Outside-in Technique of Transforaminal Lumbar Endoscopy for Lumbar Disc Herniations

Percutaneous endoscopic transforaminal lumbar discectomy (PETLD) has now become a standard of care for the management of lumbar disc disease. There are two techniques for the introduction of a working cannula with respect to disc&#8212;outside-in and inside-out. The aim of this prospective study is to describe the technical aspects of a novel mobile outside-in method in dealing with different types of disc prolapse. A total of 184 consecutive patients with unilateral lower limb radiculopathy due to lumbar disc prolapse were operated on with the mobile outside-in technique of PETLD. Their clinical outcomes were evaluated based on the type of disc prolapse they had, a visual analog scale (VAS) leg pain score, the Oswestry Disability Index (ODI), and the Macnab criteria. The completeness of the decompression was documented with a postoperative magnetic resonance imaging. The mean age of the patients was 50 &#177; 16 years and the male/female ratio was 2:1. The mean follow-up was 19 &#177; 6 months. A total of 190 lumbar levels were operated on (L1&#8211;L2: n = 4, L2&#8211;L3: n = 17, L3&#8211;L4: n = 27, L4&#8211;5: n = 123, and L5&#8211;S1: n = 19). Divided into types, the patient distribution was central: n = 14, paracentral: n = 74, foraminal: n = 28, far lateral: n = 13, superior-migrated: n = 8, inferior migrated: n = 38, and high canal compromise: n = 9. The mean operative time was 35 &#177; 12 (25 - 56) min and the mean hospital stay was 1.2 &#177; 0.5 (1&#8211;3) days. The VAS score for leg pain improved from 7.5 &#177; 1 to 1.7 &#177; 0.9. The ODI improved from 70 &#177; 8.3 to 23 &#177; 5. According to the Macnab criteria, 75 patients (40.8%) had excellent results, 104 patients (56.5%) had good results, and 5 patients (2.7%) had fair results. Recurrence (including early and late) was seen in 15 out of the 190 levels that were operated on (7.89%). This article presents a novel outside-in approach that relies on a precise landing within the foramen in a mobile manner and does not solely depend upon the enlargement of the foramen. It is more versatile in application and useful in the management of all types of disc prolapse, even in severe canal compromise and high migration.

Here we present a protocol of a novel outside-in technique of transforaminal endoscopic discectomy for lumbar disc herniations. The technical aspects of the technique, the wide indications of use, and the results of the treatment in 184 patients are described in detail.

Uma técnica de fora para dentro móvel de endoscopia Transforaminal lombar para hérnias de disco lombar

Percutaneous endoscopic transforaminal lumbar discectomy (PETLD) has now become a standard of care for the management of lumbar disc disease. There are ...

Enucleation of the Prostate for the Treatment of Benign Prostatic Hyperplasia Using a 980 nm Diode Laser

In the aging male population, the occurrence of lower urinary tract symptoms (LUTS) caused by benign prostatic hyperplasia (BPH) is a common problem. Here, we introduce a new technique called 980 nm diode laser enucleation (DiLEP) to treat BPH1. Diode lasers can absorb both water and hemoglobin at the same time, so they are good for cutting and hemostasis2. The diode laser was approved by the FDA in 2007, and has been used in the treatment of BPH because of its effective cutting and hemostasis effect3. DiLEP presents several advantages over other techniques, such as TURP, HoLEP, and PVP. During the procedure, we define the boundary of a high-volume prostate and separate it into three lobes with a diode laser by burning two rings and one groove (like a Cupid&#39;s arrow). Compared to other procedures, mDiLEP has fewer intraoperative complications, a shorter learning curve, and achieves more tissue resection.

Here, we present a protocol for modified 980 nm diode laser enucleation to treat large volume benign prostatic hyperplasia.

Enucleação da próstata para o tratamento da hiperplasia benigna da próstata usando um laser de diodo de 980 nm

In the aging male population, the occurrence of lower urinary tract symptoms (LUTS) caused by benign prostatic hyperplasia (BPH) is a common problem. ...

Evaluation of Cerebral Blood Flow Autoregulation in the Rat Using Laser Doppler Flowmetry

When investigating the body&#39;s mechanisms for regulating cerebral blood flow, a relative measurement of microcirculatory blood flow can be obtained using laser Doppler flowmetry (LDF). This paper demonstrates a closed skull preparation that allows cerebral blood flow to be assessed without penetrating the skull or installing a chamber or cerebral window. To evaluate autoregulatory mechanisms, a model of controlled blood pressure reduction via graded hemorrhage can be utilized while simultaneously employing LDF. This enables the real time tracking of the relative changes in the blood flow in response to reductions in arterial blood pressure produced by the withdrawal of circulating blood volume. This paradigm is a valuable approach to study cerebral blood flow autoregulation during reductions in arterial blood pressure and, with minor modifications in the protocol, is also valuable as an experimental model of hemorrhagic shock. In addition to evaluating autoregulatory responses, LDF can be used to monitor the cortical blood flow when investigating metabolic, myogenic, endothelial, humoral, or neural mechanisms that regulate cerebral blood flow and the impact of various experimental interventions and pathological conditions on cerebral blood flow.

This article demonstrates the use of laser Doppler flowmetry to evaluate the ability of the cerebral circulation to autoregulate its blood flow during reductions in arterial blood pressure.

Avaliação da autoregulação do fluxo sanguíneo cerebral no rato usando fluimetria de laser

When investigating the body&#39;s mechanisms for regulating cerebral blood flow, a relative measurement of microcirculatory blood flow can be obtained ...

Using a Chemical Biopsy for Graft Quality Assessment

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated with an increased survival rate and greater quality of patient&#8217;s life when compared to conventional dialysis. Regrettably, transplantology suffers from a lack of reliable methods for organ quality assessment. Standard diagnostic techniques are limited to macroscopic appearance inspection or invasive tissue biopsy, which do not provide comprehensive information about the graft. The proposed protocol aims to introduce solid phase microextraction (SPME) as an ideal analytical method for comprehensive metabolomics and lipidomic analysis of all low molecular compounds present in kidneys allocated for transplantation. The small size of the SPME probe enables performance of a chemical biopsy, which enables extraction of metabolites directly from the organ without any tissue collection. The minimum invasiveness of the method permits execution of multiple analyses over time: directly after organ harvesting, during its preservation, and immediately after revascularization at the recipient&#8217;s body. It is hypothesized that the combination of this novel sampling method with a high-resolution mass spectrometer will allow for discrimination of a set of characteristic compounds that could serve as biological markers of graft quality and indicators of possible development of organ dysfunction.

The protocol presents utilization of the chemical biopsy approach followed by comprehensive metabolomic and lipidomic analysis for quality assessment of kidney grafts allocated for transplantation.

Usando uma biópsia química para avaliação da qualidade do enxerto

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated ...