Gli autori desiderano ringraziare pascal Lejeune per il suo supporto tecnico durante tutte le procedure.

Il protocollo presentato viene utilizzato per le fattrici presentate nell'ospedale equino e segue le linee guida istituzionali per la cura degli animali.
NOTA: Eseguire la fotoablazione delle cisti endometriali prima dell'estro: i riscaldamenti osservati a fine luglio o agosto sembrano quindi più affidabili. Non eseguire questa procedura troppo tardi nella stagione riproduttiva in quanto può limitare il numero di estro disponibili per il trattamento di una potenziale endometrite.
1. Durante l'estro precedente
<ol><li>Eseguire un esame completo della solidità riproduttiva della cavalla, compresa una biopsia endometriale1 (non descritta qui) per garantire che la cavalla possa potenzialmente rimanere incinta dopo la procedura.</li>
	<li>In caso di scarsa stadiazione della biopsia endometriale secondo la classificazione di Kenney1,dare al programma di trasferimento dell'embrione o iniezione intracitoplasmatica di spermatozoi (ICSI) la preferenza rispetto alla fotoabalazione delle cisti endometriali.</li>
</ol>2. Durante il diestro precedente
<ol><li>Una volta che il retto è vuoto, contare, misurare e mappare le cisti endometriali della cavalla nell'utero utilizzando l'ecografia trans-rettale guidata dalla palpazione rettale con una sonda rettale lineare a 7,5 o 5 MHz e pinze sullo schermo (Figura 3) in un ceppo di contesa.</li>
	<li>Somministrare 250 μg di cloprostenolo per via intramuscolare per indurre l'estro entro 2 o 3 giorni dopo il giorno 6\u20127 post-ovulazione.</li>
	<li>Su base giornaliera, eseguire l'ecografia di routine comune e la palpazione della cervice per osservare i segni dell'estro, tra cui edema endometriale significativo, follicolo antrale di 35 mm e cervice ammorbidita e aperta.</li>
</ol>3. Preparazione quando la cavalla è in calore
<ol><li>Limitare l'accesso alla sala operatoria alle persone che indossano occhiali di protezione per gli occhi.</li>
	<li>Dopo lo svuotamento del retto, avvolgere la coda e pulire la vulva in modo asettico usando iodio-povidone.</li>
	<li>Per garantire una facile manipolazione e il comfort della cavalla, somministrare detomidina (10 μg/kg) e butorfanolo (0,1 mg/kg) per via endovenosa nella vena giugulare con un ago da 21 G.</li>
	<li>Per vagina, installare il catetere di raccolta degli embrioni attraverso la cervice e insufflare il palloncino con 40 ml di aria.</li>
	<li>Infondere l'utero con 1,5 L di soluzione salina sterile (0,9% NaCl) per gonfiare l'utero.</li>
	<li>Sgonfiare il palloncino e rimuovere il catetere.</li>
	<li>Passare l'endoscopio in modo asettico attraverso la cervice all'interno di una manica sterile.</li>
</ol>4. Tecnica
<ol><li>Collegare la fibra di quarzo da 600 μm a un laser a diodi funzionante a 980 nm. Impostare la potenza a 20\u201225 W in modalità continua.</li>
	<li>Passare la fibra di quarzo flessibile attraverso il canale biopsia di un video-endoscopio di 10 mm di diametro fino a quando 3\u20124 cm di fibra libera possono essere identificati sullo schermo. Un minimo di 10 mm dall'estremità della fibra dovrebbe essere visibile.</li>
	<li>Attivare il laser per forare la cisti idealmente al suo apice: sul dispositivo è disponibile un pedale dedicato per tenere le mani libere. In alcuni casi, sono necessarie più forature in varie posizioni per ottenere lo svuotamento completo della cisti. Il liquido cistico viene drenato passivamente nell'utero.</li>
	<li>Dopo che il rivestimento della cisti è collassato intorno alla fibra, distribuire il laser fino a quando la membrana non si restringe.</li>
	<li>Eseguire l'ecografia transrettale dell'utero tutte le volte che è necessario durante la procedura per confermare che le cisti endometriali sono scomparse.</li>
</ol>5. Trattamenti post-operatori
<ol><li>Somministrare un trattamento antibiotico generale: 22.000 UI/kg di sospensione di penicillina-procaina, per via intramuscolare, due volte al giorno, per 3 giorni.</li>
	<li>Subito dopo l'intervento, eseguire il lavaggio uterino come descritto sopra (vedere i passaggi 3.4-3.6).<ol>
<li>Raccogliere il liquido infuso nell'utero prima della procedura endoscopica attraverso un catetere sterile ammanettato.</li>
		<li>Lavare l'utero con successive instillazioni di soluzione isotonica sterile da 1 L (0,9% NaCl) fino a quando non viene raccolto un liquido limpido.</li>
		<li>Alla fine della procedura, somministrare 20 UI di ossitocina per via endovenosa.</li>
		<li>Eseguire l'ecografia transrettale dell'utero di cavalla il giorno dopo: si osserva frequentemente liquido libero intraluminale (vedere passo 2.1).</li>
	</ol>
</li>
	<li>Finché l'ecografia intra-luminale libera viene osservata con fluido transrettale e fino alla fine del calore, eseguire lavaggi uterini ogni giorno con la procedura sopra descritta (passaggi 3.4-3.6). A volte, il liquido simile a un siero, limpido e colorato di giallo viene raccolto per diversi giorni. NOTA: Apporto ripetute iniezioni intramuscolari di ossitocina (20 UI) ogni 4 ore o meno quando la cavalla è in estro può evitare l'accumulo di liquidi e ridurre il rischio di contaminazione batterica nell'utero.</li>
	<li>Eseguire l'ecografia uterina trans-rettale durante il successivo diestro per confermare la scomparsa delle cisti endometriali e l'assenza di liquido libero intraluminale. NOTA: Se si osserva liquido intraluminale libero, si raccomanda l'iniezione di prostaglandine per indurre un nuovo estro, per diagnosticare un'eventuale endometrite successiva e per trattarla.</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>

Tutti gli autori sono pienamente impiegati dalla Clinica Equina dell'Università di Liegi e non hanno conflitti di interesse con alcuna azienda che commercia uno dei prodotti sopra menzionati.

L'isteroscopia e la fotoablazione laser delle cisti endometriali è una procedura semplice per ridurre le grandi cisti intra-luminali nella cavalla, come precedentemente suggerito14. Tecnicamente, questo metodo è migliorato se eseguito su una cavalla in estro. Gonfiare l'utero con una soluzione salina sterile ha migliorato la visibilità rispetto al riempimento dell'aria e all'aspirazione continua durante l'esecuzione della procedura.  Le piccole cisti intra-parietali (<10 mm) sono più difficil...

Le cisti endometriali nelle fattrici sono state descritte istopatologicamente negli anni settanta da Kenney1. Alla fine degli anni Ottanta, l'uso dell'isteroscopia e dell'ecografia ha portato alla loro descrizione clinica e ha contribuito a capire le loro conseguenze sulla fertilità2,3,4,5.
Nelle fasi iniziali, le cisti endometriali sono state associate a una degenerazione dell'endometrio ora chiamata endometrosi1,3,4,6,7,8 ed è attualmente accettato che piccole cisti (<1 cm) sono correlate alla fibrosi delle ghiandole endometriali quando le cisti più grandi sono correlate alla dilatazione linfatica6 . Queste grandi cisti hanno maggiori probabilità di disturbare la migrazione dell'embrione equino tra il giorno 6.5 e il giorno 17, diminuendo così la fertilità compromettendo il riconoscimento materno1,3,4,6,8. Le cisti endometriali possono anche interferire con la diagnosi precoce della gravidanza, specialmente se sono rotonde e hanno una dimensione simile a quella di una vescicola embrionale3,4,6,9 (vedi Figura 1). Pertanto, possono potenzialmente aumentare l'intervallo tra due inseminazioni ritardando la prima ecografia embrionale e la valutazione della gestazione gemellare (vedi Figura 2). Le cisti endometriali possono anche interferire meccanicamente con la raccolta di liquidi durante il lavaggio uterino eseguito per trattare l'endometrite o per raccogliere l'embrione. Di conseguenza, le cisti endometriali hanno effetti sulla fertilità della cavalla osservata, con potenziali conseguenze economiche negli allevamenti.
Oltre ai tentativi di trattare l'endometrosisottostante 10,sono stati proposti trattamenti focali endoscopicamente guidati delle cisti. È stata descritta la cauterizzazione elettrochirurgica, ma l'esposizione al calore potrebbe danneggiare l'endometrio intorno alla cisti11. L'aspirazione della cisti è efficiente solo se la struttura secernente non è più attiva al momento dellapuntura 12. L'irradiazione laser con un laser a diodi a 980 nm con una fibra di contatto sembra mostrare cicatrici endometriali successive più piccole6,13. Il miglioramento dei dispositivi e delle procedure laser6 ci consente ora di proporre questa tecnica sicura ed efficiente nella pratica clinica equina. Tuttavia, i proprietari di cavalle devono essere informati sui limiti di questa opzione. La fotoablazione delle cisti aumenterà il riconoscimento embrionale in alcune fattrici, farà avanzare la diagnosi di gravidanza in alcuni casi e potrebbe limitare l'effetto meccanico negativo delle cisti endometriali sul lavaggio uterino eseguito per trattare l'endometrite o per raccogliere embrioni. Tuttavia, non tratterà mai l'endometriosi o l'endometrite sottostante concomitante.
Questo rapporto mira a descrivere una procedura efficiente per eseguire l'ablazione della cisti laser a diodi guidata da isteroscopia nelle fattrici. La preparazione e le procedure saranno descritte praticamente per un'applicazione clinica.

<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

Nelle fattrici, le cisti endometriali sono associate all'endometriosi e possono causare il fallimento del riconoscimento materno o compromettere e ritardare le diagnosi di gravidanza. I trattamenti storici erano invasivi e avevano effetti avversi sull'endometrio. La terapia laser a guida isteroscopica è facile ed efficace per la resezione delle cisti endometriali, senza effetti deleteri per l'endometrio.
Un endoscopio lungo 110 cm e largo 1,0 cm viene introdotto sterilemente nell'utero attraverso la cervice aperta di una cavalla estrale dopo la pulizia vulvare. L'utero viene lentamente infuso con meno di 1 L di soluzione fisiologica e la fibra laser viene inserita nel canale biopsia dell'endoscopio. Le cisti vengono quindi cauterizzate con il laser a diodi da 980 nm con una fibra di contatto impostata a 20\u20122 5W in modalità continua. Ogni cisti viene perforata fino al completo svuotamento della cisti e al restringimento della parete della cisti attorno alla fibra. I lavaggi uterini con soluzione salina sterile vengono eseguiti direttamente dopo l'intervento chirurgico e per uno o due giorni poiché si può osservare fluido non infiammatorio.
Questa procedura è facile e rapidamente eseguita, senza evidenti effetti deleteri. La resezione delle cisti rende più facile la diagnosi di gravidanza ad ultrasuoni e, in alcuni casi, potrebbe ripristinare la corretta migrazione dell'embrione nelle corna uterine tra il giorno 6,5 e il giorno 17. Tuttavia, questo trattamento non migliora le lesioni istologiche sottostanti correlate all'endometriosi. Queste considerazioni dovrebbero essere chiaramente espresse all'allevatore prima di questa procedura.

Gli interventi chirurgici sono stati eseguiti su fattrici con cisti con diametri superiori a 15 mm. Tuttavia, se erano presenti anche altre cisti più piccole, è stata tentata la fotoablazione delle altre cisti. Generalmente, queste grandi cisti erano alla biforcazione o alla base delle corna. Tuttavia, alcuni di loro erano presenti nel corpo dell'utero e sono stati trattati allo stesso modo.
Nella nostra esperienza, le cisti intraluminali sono molto facili da localizzare con l'endoscopio. In...

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

Lo scopo di questo rapporto è quello di descrivere la fotoablazione delle cisti endometriali nella cavalla utilizzando un dispositivo laser endoscopico e di discutere l'interesse e i limiti di questo nuovo approccio per affrontare i problemi di bassa fertilità.

Fotoablazione guidata da endoscopia di cisti endometriali utilizzando un laser a 980 nm con una fibra di contatto in 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

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Medicine

4.2K Views.  Liège University. Lo scopo di questo rapporto è quello di descrivere la fotoablazione delle cisti endometriali nella cavalla utilizzando un dispositivo laser endoscopico e di discutere l'interesse e i limiti di questo nuovo approccio per affrontare i problemi di bassa fertilità.

Video: 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.

MRI-guidato Interruzione della barriera emato-encefalica transcranica con ultrasuoni focalizzati in un modello murino

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

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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.

Epidurale Misurazione della pressione intracranica in ratto utilizzando una fibra ottica trasduttore di pressione

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.

Riproducibile Paraplegia da Toracica aortica occlusione in un modello murino di Spinal Cord ischemia-riperfusione

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

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.

Un possibile modello Zebrafish della malattia del rene policistico: Knockdown di<em&gt; Wnt5a</em&gt; Provoca cisti in Zebrafish Reni

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.

Analisi semi-automatica del Mouse del muscolo scheletrico morfologia e composizione di 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.

Una Mobile esterno-in tecnica di endoscopia lombare transforaminale per ernie del disco lombare

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.

Enucleazione della prostata per il trattamento dell'iperplasia prostatica benigna mediante laser a diodi da 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.

Valutazione dell'autoregolazione del flusso sanguigno cerebrale nel ratto utilizzando 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 ...

Fertility Sparing Procedure using Carbon Dioxide Fiber Laser Vaporization of Ovarian Endometrioma

The surgical management of endometrioma is still a matter of debate. Cystectomy, which is recognized as the standard technique, seems to be associated with a potential reduction in the ovarian reserve due to the inadvertent removal and thermal damage of healthy ovarian tissue. New ablative techniques with reduced tissue penetration depth and less thermal spread to the surrounding parenchyma may represent a viable alternative to cystectomy. For these reasons, the aim of this manuscript is to demonstrate the ablation of the endometrioma capsule using a CO2 fiber laser technique and discuss the clinical outcomes. Once the cyst has been drained and washed, a biopsy is taken. After cyst eversion, vaporization of the inner surface of the cyst is performed using a CO2 fiber laser. The technique is simple and reproducible as even young surgeons without any surgical experience were more confident in performing laser CO2 vaporization instead of cystectomy. The positive effects of CO2 technology are reported in a randomized controlled trial, where the postoperative changes in the antral follicular count (AFC) and antimullerian hormone (AMH) levels were compared between patients who had their endometrioma excised (cystectomy) and those who had undergone endometrioma vaporization with CO2 laser. The patients treated with CO2 laser showed significantly increased AFC without a reduction in serum AMH levels as compared to the cystectomy group, in which both parameters were significantly reduced. The postoperative pregnancy rate was also assessed, and comparable pregnancy rates were found after both treatments. On the contrary, patients treated with the CO2 fiber laser technique had more favorable in-vitro fertilization (IVF) outcomes compared to cystectomy.
In conclusion, the CO2 fiber laser technique may represent a viable alternative to cystectomy in the surgical treatment of endometrioma in terms of ovarian preservation, pregnancy rates, and IVF outcomes. Moreover, it has the advantage of being independent of the surgeon's skills and personal experience.

In this protocol, CO2 fiber laser technique is demonstrated for the surgical treatment of ovarian endometriosis, which represents a viable alternative in terms of fertility preservation with the major advantage of not being dependent on the surgeon's skills and personal experience.

Procedura di risparmio della fertilità utilizzando la vaporizzazione laser a fibra di anidride carbonica dell'endometrioma ovarico

The surgical management of endometrioma is still a matter of debate. Cystectomy, which is recognized as the standard technique, seems to be associated ...

Doppio approccio posteromediale artroscopico per l'escissione di cisti del LCP

Arthroscopic Excision of Posterior Cruciate Ligament Cysts Using a Double Posteromedial Approach

Cruciate ligament cysts of the knee are a rare condition. Posterior cruciate ligament cysts of the knee are less common than anterior cruciate ligament cysts. In patients with asymptomatic isolated cruciate ligament cysts of the knee, conservative treatment is recommended. Symptomatic cruciate ligament cysts of the knee are mostly manifested as knee hyperflexion pain, straightening pain, knee discomfort after standing for a long time or walking for a long time, etc., which seriously affects the quality of life, surgical treatment can be performed. The surgical treatments can be divided into ultrasound-guided cyst puncture and fluid extraction procedure and arthroscopic cystectomy. Cysts are mostly lobulated with a multi-layer cyst wall, cyst fluid extraction does not remove the cyst wall completely but simply extracts cyst fluid, leading to a high recurrence rate. Arthroscopic surgery can completely remove the cyst wall with little trauma, a low recurrence rate, and fast postoperative recovery, so arthroscopic resection is the most common and preferred method of treatment. Since posterior cruciate ligament cysts mostly occur posterior to the ligament, we remove the cyst wall by adding a double posteromedial approach to the knee joint, and the cyst wall is removed under direct vision, which is simple to operate, the cyst wall is completely cleared, the trauma is small, the postoperative recovery is fast, and there is no recurrence. Here, 8 posterior cruciate ligament cysts were removed with complete postoperative symptom relief, no surgical complications, and no recurrence at 1-year follow-up.

Autore in primo piano: Doppio approccio posteromediale per il trattamento delle cisti del legamento crociato posteriore

We present a surgical approach to treat posterior cruciate ligament cysts by an arthroscopic double posteromedial approach.

Escissione artroscopica delle cisti del legamento crociato posteriore utilizzando un doppio approccio posteromediale

Cruciate ligament cysts of the knee are a rare condition. Posterior cruciate ligament cysts of the knee are less common than anterior cruciate ligament ...