Name: endoscopy guided photoablation of endometrial cysts using a 980 nm laser with a contact fiber in mares
Uploaded: 2020-07-16T14:30:00
Description: 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

Authors would like to thank Mr. Pascal Lejeune for his technical support during all procedures.

The presented protocol is used for mares presented in the equine hospital and follows institutional animal care guidelines.
NOTE: Perform endometrial cysts photoablation before anestrus: heats observed in late July or August thus seem most reliable. Do not perform this procedure too late in the breeding season as it can limit the number of estruses available to treat a potential endometritis.
1. During the preceding estrus
<ol>
	<li>Perform a complete breeding sou...

<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. 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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. 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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. 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Hysteroscopy and laser photoablation of endometrial cysts is an easy procedure to reduce large intra-luminal cysts in the mare, as previously suggested14. Technically, this method is improved when performed on a mare in estrus. Inflating the uterus with sterile saline solution improved visibility when compared to air filling and continuous aspiration while performing the procedure.&#160; Small intra-parietal cysts (&#60;10 mm) are more difficult to reach and to treat. Moreover, treating these smal...

Endometrial cysts in mares were histopathologically described in the seventies by Kenney1. In the late eighties, the use of hysteroscopy and ultrasonography led to their clinical description and helped to understand their consequences on fertility2,3,4,5.
In the early stages, endometrial cysts have been associated with a degeneration of the endometrium now called endometrosis1,3,4,6,7,8 and it is currently accepted that small (&#60;1 cm) cysts are related to endometrial glands fibrosis when larger cysts are related to lymphatic dilatation6. These large cysts are the most likely to disturb equine embryo migration between day 6.5 and day 17, thus decreasing the fertility by impairing maternal recognition1,3,4,6,8. Endometrial cysts may also interfere with early pregnancy diagnosis, especially if they are round and have a size similar to that of an embryonic vesicle3,4,6,9 (see Figure 1). Thus, they can potentially increase the interval between two inseminations by delaying first embryo ultrasonography and twin gestation assessment (see Figure 2). Endometrial cysts may also interfere mechanically with fluid collection during uterine lavage performed to treat endometritis or to collect the embryo. Consequently, endometrial cysts have effects on observed mare&#8217;s fertility, with potential economic consequences in breeding farms.
Besides attempts to treat the underlying endometrosis10, endoscopically-guided focal treatments of cysts have been proposed. Electrosurgical cauterization has been described, but heat exposure could injure the endometrium around the cyst11. Aspiration of the cyst is only efficient if the secreting structure is no more active at the time of the puncture12. Laser irradiation with a 980 nm diode laser with a contact fiber seems to show smaller subsequent endometrial scars6,13. Improvement of laser devices and procedures6 now allows us to propose this safe and efficient technique in equine clinical practice. However, mare owners must be informed about the limits of this option. Photoablation of the cysts will increase embryo recognition in some mares, will advance the pregnancy diagnosis in some cases and could limit the negative mechanical effect of endometrial cysts on uterine lavage performed to treat endometritis or to collect embryos. However, it will never treat the concomitant underlying endometriosis or endometritis.
This report aims to describe an efficient procedure to perform hysteroscopy-guided diode laser cyst ablation in mares. Preparation and procedures will be practically described for a clinical application.

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	<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

In mares, endometrial cysts are associated with endometriosis and can cause maternal recognition failure or compromise and delay pregnancy diagnoses. Historical treatments were invasive and had adverse effects on the endometrium. Hysteroscopically guided laser therapy is easy and effective for endometrial cysts resection, with no deleterious effects for the endometrium.
A 110 cm long and 1.0 cm wide endoscope is sterilely introduced in the uterus through the open cervix of an estrous mare after vulvar cleaning. The uterus is slowly infused with less than 1 L of physiologic solution and the laser fiber is inserted in the biopsy canal of the endoscope. Cysts are then cauterized with the 980 nm diode laser with a contact fiber set at 20&#8210;2 5W in continuous mode. Each cyst is punctured&#160;until complete voiding of the cyst and shrinking of the cyst wall around the fiber. Uterine lavages with sterile saline solution are performed directly after the surgery and for one or two days as non-inflammatory fluid can be observed.
This procedure is easy and quickly performed, with no obvious deleterious effects. Cysts resection makes ultrasound pregnancy diagnosis easier and, in some cases, could restore proper embryo migration in the uterine horns between day 6.5 and 17. However, this treatment does not improve the underlying histological lesions related to endometriosis. These considerations should be clearly expressed to the breeder before this procedure.

Surgeries were performed on mares having cysts with diameters over 15 mm. However, if other smaller cysts were also present, photoablation of the other cysts was attempted. Generally, these large cysts were at the bifurcation or the basis of the horns. However, some of them were present in the body of the uterus and were equally treated.&#160;
In our experience, intraluminal cysts are very easy to localize with the endoscope. Moreover, performing this procedure in saline solution increases the...

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

The aim of this report is to describe the photoablation of endometrial cysts in the mare using an endoscopic laser device, and to discuss the interest and limitations of this new approach to deal with low fertility issues.

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

Reducing the number of Endomentrial Cysts with laser and a hysteroscopy control can improve fertility by reducing the delay for pregnancy confirmation and improving embryo migration between day six and half and seventeen. Laser irradiation with a contact fiber, results in smaller subsequent endometrial scars. Performing this procedure in liquid environment, avoids smoke emission or foam production inside the uterus during the procedure.Diode laser is mainly used in overseas to treat upper airways pathologies. It could also be used for other pathologies, such as some skin disorders or for facilitated ankylosis. Before cyst ablation by laser, perform breeding soundness examination, including endometrial biopsy.To establish the prognosis and treat the underlying pathologies. It is impossible for placebo embryo transfer or exit. Demonstrating the procedure will be Pascal Lejeune and endoscopy specialized technician from our clinic.During the preceding Estrus, perform a complete breeding soundness examination of the mare, including an endometrial biopsy to ensure that the mare can potentially get pregnant after the procedure. During the preceding Diestrous, count, measure and map endometriosis in the uterus using transrectal ultrasonography, guided by palpation with a 7.5 or 5 MHz linear rectal probe and calipers on the screen in a contention stock. Intramuscularly administer 250 micrograms of Cloprostenol to induce estrus within two to three days, at day six to seven, post ovulation.Perform routine daily ultrasonography and cervix palpation to observe estrous signs. Including significant endometrial edema, antral follicle of 35 mm, or a softened and open cervix. Make sure that anyone entering the operating room is wearing eye protection.After emptying of the mare's rectum, wrap the tail and clean the vulva aseptically using Iodine-Povidone. To ensure easy manipulation and comfort of the mare, intravenously administer 10 mg/kg Detomidine and 0.1 mg/kg Butorphanol in the juggler vein with a 21 gauge needle. Install the embryo collection catheter through the cervix and inflate the balloon with 40 ml of air.Infuse the uterus 1.5 liters of sterile saline solution to inflate it, then deflate the balloon and remove the catheter. Pass the endoscope aseptically through the cervix within a sterile sleeve. Connect the 600 m quartz fiber to a diode laser operating at 980 nm.And set the power at 20 to 25 Watts in continuous mode. Pass the flexible quartz fiber via the biopsy channel of a 10 mm diameter video endoscope. Until 3 to 4 cm of free fiber can be identified on the screen.A minimum of 10 mm from the end of the fibers should be visible. Activate the laser to puncture the cyst. Ideally at its apex.In some cases, multiple punctures at various locations are necessary to achieve complete voiding. The cystic fluid is then passively drained out into the uterus. After the cyst lining has collapsed around the fiber, deploy the laser until the membrane shrinks.When acting in liquid medium, be sure that the fiber is in close contact with the cyst and verify the potential in the wound of cyst out punctured Perform transrectal ultrasonography of the uterus as many times as necessary during the procedure, to confirm that the endometrial cysts have disappeared. Collect the liquid infused in the uterus before the endoscopic procedure through a sterile cuffed catheter. Then flush the uterus with successive installations of one liter of sterile isotonic solution, until a clear liquid is collected.After the procedure, administer general antibiotic treatment intramuscularly twice a day for three days. Administer 20 IU of oxytocin intravenously at the end of the procedure. On the next day, perform transrectal ultrasonography of the mare uterus.Intraluminal free liquid is frequently observed. As long as free intraluminal fluid is observed by transrectal ultrasonography, and until the end of the heat, perform uterine lavage daily. Sometimes serum like limpid and yellow stained liquid is collected for several days.Perform transrectal uterine ultrasonography, during the following diestrus to confirm disappearance of endometrial cysts and the absence of intraluminal free fluid. This protocol was used to treat cyst with diameters over 15 millimeters. Photoablation of the smaller cyst was also attempted, if they were present.Generally large cysts were at the bifurcation or the bases of the horns. However, some of them were present in the body of the uterus and were also treated. Parietal cysts were more difficult to identify because of their location and their smaller size.Only the smallest parietal cyst were impossible to treat, but they do not interfere with the embryonic vesicles movements and are not easily confused with day 14 embryos. Never forget the associated underlying causes of infertility. Perform uterine swaps on biopsies, then treat and prevent these fertility dealing pathologies with Verve surgeries or uterine allergies.Multicentric studies about fertility observed before and after laser cyst ablation should be performed. However, semen quality, intimination timings and other external factors may induce extrinsic causes of variation.

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Medicine

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.

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

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.

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.

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.

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.

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.

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.

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.

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.

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

Arthroscopic Double Posteromedial Approach for PCL Cyst Excision

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.

Author Spotlight: Double Posteromedial Approach for Treating Posterior Cruciate Ligament Cysts

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