We thank all the midwives and anesthesiologists who have collaborated on this project. David Sim&#243; has specially collaborated with this project recording the procedure.

This study was approved by the Clinic Research Committee of the &#34;Virgen de la Arrixaca&#34; at the University Clinical Hospital. Written informed consent was obtained from all participants.
1. Offer external cephalic version at consult (36 weeks)
<ol>
	<li>Identify the fetal position with an ultrasound.
	<ol>
		<li>Apply ultrasound gel on the patient&#8217;s abdomen.</li>
		<li>Place the abdominal probe on the hypogastric region.</li>
		<li>Identify the fetal position.</li>
	</ol>
	</li>
	<li>Offer external cephalic version.</li>
	<li>Have the patient sign informed consent.</li>
</ol>
2. Admit in the obstetric emergency room (&#8805;37 weeks)
<ol>
	<li>Identify the fetal position with an ultrasound.
	<ol>
		<li>Apply ultrasound gel on the patient&#8217;s abdomen.</li>
		<li>Place the abdominal probe on the hypogastric region.</li>
		<li>Identify the fetal position.</li>
	</ol>
	</li>
	<li>Prepare the patient and check requirements (blood test and informed consent).</li>
</ol>
3. Admit in the obstetric delivery room
<ol>
	<li>Perform cardiotocography assessment for fetal well-being.</li>
	<li>Add 10 mL of ritodrine in 500 mL of glucose solution.</li>
	<li>30 minutes before the procedure, administer 6 mg of ritodrine at 60 mL/h.</li>
	<li>Invite the patient to empty her bladder.</li>
	<li>Stop ritodrine perfusion before moving to the obstetric operating room.</li>
</ol>
4. External cephalic version procedure in obstetric operating room
<ol>
	<li>Move to the obstetric operating room.</li>
	<li>Monitor maternal vital signs (heartrate, EKG, temperature, noninvasive blood pressure, oxygen saturation).</li>
	<li>Identify the fetal position with an ultrasound.
	<ol>
		<li>Apply ultrasound gel on the patient&#8217;s abdomen.</li>
		<li>Place the abdominal probe on the hypogastric region.</li>
		<li>Identify the fetal position.</li>
	</ol>
	</li>
	<li>Position the patient in Trendelenburg (15&#176;).</li>
	<li>Perform analgesia.
	<ol>
		<li>Sedate the patient with 1-1.5 mg/kg propofol IV.</li>
		<li>Alternatively: Provide spinal anesthesia with 10 mL of 0.1% bupivacaine.</li>
	</ol>
	</li>
	<li>External cephalic version procedure (2 attempts)
	<ol>
		<li>Apply an abundant quantity of ultrasound gel on the patient&#8217;s abdomen.</li>
		<li>Place hands in the hypogastric region to identify the fetal buttocks (Obstetrician A).</li>
		<li>Elevate fetal buttocks (Obstetrician A).</li>
		<li>Place hands in the patient&#8217;s abdomen to locate the fetal head (Obstetrician B).</li>
		<li>Guide the fetal buttocks to the fundus (Obstetrician A).</li>
		<li>Consecutively direct the fetal head to the pelvis (Obstetrician B).</li>
	</ol>
	</li>
	<li>Identify the fetal well-being with ultrasound.
	<ol>
		<li>Place the abdominal probe on the abdomen.</li>
		<li>Identify the fetal heart.</li>
		<li>Observe the fetal heart rate for at least a minute.</li>
	</ol>
	</li>
	<li>Check for vaginal bleeding.</li>
	<li>Identify the fetal position with ultrasound.
	<ol>
		<li>Place the abdominal probe on the hypogastric region.</li>
		<li>Identify the fetal position.</li>
	</ol>
	</li>
</ol>
5. Move to obstetric delivery room
<ol>
	<li>Perform cardiotocography assessment for fetal well-being for at least 4 hours.</li>
	<li>Discharge the patient.</li>
</ol>
6. Admit in obstetric emergency room (24 h post-procedure)
<ol>
	<li>Perform cardiotocography assessment for fetal well-being for 30 minutes.</li>
</ol>

<ol>
	<li>Melo, P., Georgiou, E. X., Hedditch, A., Ellaway, P., Impey, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=External+cephalic+version+at+term:+a+cohort+study+of+18+years'+experience.">External cephalic version at term: a cohort study of 18 years' experience.</a> British Journal of Obstetrics and Gynecology. 126, 493-499 (2019).</li><li>Hofmeyr, G. J., Kulier, R., West, H. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=External+cephalic+version+for+breech+presentation+at+term.">External cephalic version for breech presentation at term.</a> Cochrane Database of Systematic Reviews. 4, 83 (2015).</li><li>Isakov, O., 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=Prediction+of+Success+in+External+Cephalic+Version+for+Breech+Presentation+at+Term.">Prediction of Success in External Cephalic Version for Breech Presentation at Term.</a> Obstetrics and Gynecology. 133 (5), 857-866 (2019).</li><li>Ebner, F., 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=Predictors+for+a+successful+external+cephalic+version:+a+single+centre+experience.">Predictors for a successful external cephalic version: a single centre experience.</a> Archives of Gynecology and Obstetrics. 293 (4), 749-755 (2016).</li><li>Castan&#771;er-Ma&#769;rmol, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Results of external cephalic version in our area without rutinary tocolysis. Review and implementation strategy. , (2015).</li><li>Burgos, J., 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=Probability+of+cesarean+delivery+after+successful+external+cephalic+version.">Probability of cesarean delivery after successful external cephalic version.</a> International Journal of Gynecology &amp; Obstetrics. 131 (2), 192-195 (2015).</li><li>Mu&#241;oz, M., 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=External+cephalic+version+at+term:+accumulated+experience.">External cephalic version at term: accumulated experience.</a> Progresos de Obstetricia y Ginecolog&#237;a. 48 (12), 574-580 (2005).</li><li>Velzel, J., 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=Atosiban+versus+fenoterol+as+a+uterine+relaxant+for+external+cephalic+version:+randomised+controlled+trial.">Atosiban versus fenoterol as a uterine relaxant for external cephalic version: randomised controlled trial.</a> British Medical Journal. 356 (26), 6773 (2017).</li><li>Thissen, D., Swinkels, P., Dullemond, R. C., van der Steeg, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Introduction+of+a+dedicated+team+increases+the+success+rate+of+external+cephalic+version:+A+prospective+cohort+study.">Introduction of a dedicated team increases the success rate of external cephalic version: A prospective cohort study.</a> European Journal of Obstetrics &amp; Gynecology and Reproductive Biology. 236 (3), 193-197 (2019).</li><li>Hindawi, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Value+and+pregnancy+outcome+of+external+cephalic+versi&#243;n.">Value and pregnancy outcome of external cephalic versi&#243;n.</a> Eastern Mediterranean Health Journal. 11 (4), 633-639 (2005).</li><li>Levin, G., Rottenstreich, A., Weill, Y., Pollack, R. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+bladder+volume+in+the+success+of+external+cephalic+version.">The role of bladder volume in the success of external cephalic version.</a> European Journal of Obstetrics &amp; Gynecology and Reproductive Biology. 230, 178-181 (2018).</li><li>Vani, S., Lau, S. Y., Lim, B. K., Omar, S. Z., Tan, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intravenous+salbutamol+for+external+cephalic+version.">Intravenous salbutamol for external cephalic version.</a> International Journal of Gynecology &amp; Obstetrics. 104, 28-31 (2009).</li><li>Sullivan, J. T., 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=A+randomized+controlled+trial+of+the+effect+of+combined+spinal-epidural+analgesia+on+the+success+of+external+cephalic+version+for+breech+presentation.">A randomized controlled trial of the effect of combined spinal-epidural analgesia on the success of external cephalic version for breech presentation.</a> International Journal of Obstetric Anesthesia. 18, 328-334 (2009).</li><li>Weiniger, C. F., 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=Randomized+controlled+trial+of+external+cephalic+version+in+term+multiparae+with+or+without+spinal+analgesia.">Randomized controlled trial of external cephalic version in term multiparae with or without spinal analgesia.</a> British Journal of Anaesthesia. 104 (5), 613-618 (2010).</li><li>Grootscholten, K., Kok, M., Oei, S. G., Mol, B. W., van der Post, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=External+cephalic+version-related+risks:+a+meta-analysis.">External cephalic version-related risks: a meta-analysis.</a> Obstetrics and Gynecology. 112 (5), 1143-1151 (2008).</li><li>Sociedad Espa&#241;ola de Ginecolog&#237;a y Obstetricia. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=External+cephalic+version+(updated+March+2014).">External cephalic version (updated March 2014).</a> Progresos de Obstetricia y Ginecolog&#237;a. 58 (7), 337-340 (2015).</li><li>Chaudhary, S., Contag, S., Yao, 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+impact+of+maternal+body+mass+index+on+external+cephalic+version+success.">The impact of maternal body mass index on external cephalic version success.</a> Journal of Maternal-Fetal and Neonatal Medicine. 32, 21-59 (2019).</li><li>de Hundt, M., Velzel, J., de Groot, C. J., Mol, B. W., Kok, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mode+of+delivery+after+successful+external+cephalic+version:+a+systematic+review+and+meta-analysis.">Mode of delivery after successful external cephalic version: a systematic review and meta-analysis.</a> Obstetrics and Gynecology. 123 (6), 1327-1334 (2014).</li><li>Krueger, S., 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=Labour+Outcomes+After+Successful+External+Cephalic+Version+Compared+With+Spontaneous+Cephalic+Version.">Labour Outcomes After Successful External Cephalic Version Compared With Spontaneous Cephalic Version.</a> Journal of Obstetrics and Gynecology Canada. 40, 61-67 (2018).</li><li>Kean, L. H., Suwanrath, C., Gargari, S. S., Sahota, D. S., James, D. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparison+of+fetal+behaviour+in+breech+and+cephalic+presentations+at+term.">A comparison of fetal behaviour in breech and cephalic presentations at term.</a> British Journal of Obstetrics and Gynecology. 106, 1209-1213 (1999).</li></ol>

The authors have nothing to disclose.

This article shows the procedure to carry out the ECV. The ECV procedure success rate in this study is 82.5% (78.1-86.4%), which is higher than the success rate found in international literature 49.0% (47-50.9%)1,2,3,4 or Spanish literature 53.49% (42.9-64.0%)5,6,7. This difference may be due to the us...

An erratum was issued for:&#160;<a href="https://www.jove.com/t/60636">External Cephalic Version: Is it an Effective and Safe Procedure?</a>. Figure 1 was updated from:
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/60636/60636fig1.jpg" /> 
Figure 1: Comparison of type of delivery: General pregnant population in 2018, ECV cohort between 2014-2018, Successful ECV cohort between 2014-2018. * Chi-squared test: p&#60;0.05. <a href="https://www.jove.com/files/ftp_upload/60636/60636fig1large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
to:
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/60636/60636fig1v3.jpg" /> 
Figure 1: Comparison of type of delivery: General pregnant population in 2018, ECV cohort between 2014-2018, Successful ECV cohort between 2014-2018. * Chi-squared test: p&#60;0.05. <a href="https://www.jove.com/files/ftp_upload/60636/60636fig1v2large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>

An erratum was issued for:&#160;<a href="https://www.jove.com/t/60636">External Cephalic Version: Is it an Effective and Safe Procedure?</a>. Figure 1 was updated.

Erratum: External Cephalic Version: Is it an Effective and Safe Procedure?

External cephalic version (ECV) is a procedure for modifying the fetal position and achieving a cephalic presentation. The objective of the ECV is to offer an opportunity for cephalic delivery to occur, which is widely known to be safer than breech or cesarean section. ECV is usually performed before the active labor period begins. Factors associated with a higher ECV success rate include1,4: multiparity, a transverse presentation, black race, posterior placenta, amniotic fluid index higher than 10 cm.
However, ECV is not an innocuous procedure and may present7,11 intraversion complications such as premature rupture of membranes, vaginal bleeding, transitory changes of fetal heart rate, cord prolapse, abruptio placentae, even stillbirth.
In this article, we analyzed ECVs performed under analgesia and tocolysis. To date, no video report has been published showing how to perform this procedure with analgesia and tocolysis. The main objective of this study is to show how ECV is performed. We also describe some key actions that could improve the procedural success. As a secondary objective, we analyzed the results of ECV obtained following the specific protocol with tocolysis and analgesia and compared the results with the literature. We also analyzed factors associated with ECV success rate, type of delivery and a comparison of the type of delivery in ECV pregnant women with non-ECV pregnant women were also included.

<table border="0" cellpadding="0" cellspacing="0" style="width:1081px;" width="1081">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="56">
			<td height="56" style="height:56px;width:267px;">Avalon FM20 Fetal monitor</td>
			<td style="width:197px;">Koninklijke Philips N.V</td>
			<td style="width:107px;"></td>
			<td style="width:511px;">https://www.usa.philips.com/healthcare/product/HC862198/avalon-fm20-fetal-monitor</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">Convex Array Probe 4C-RS</td>
			<td style="width:197px;">General Electric Health Care Company</td>
			<td style="width:107px;"></td>
			<td style="width:511px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:23px;width:267px;">Aquasonic&#174; 100 Ultrasound Transmission Gel</td>
			<td style="width:197px;">Parker Laboratories, INC</td>
			<td style="width:107px;"></td>
			<td style="width:511px;">https://www.parkerlabs.com/aquasonic-100.asp</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">Propofol Lipuro (10 mg/mL Inject 20 mL)</td>
			<td style="width:197px;">B. Braun Medical, SA</td>
			<td style="width:107px;"></td>
			<td style="width:511px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">Ritodrine Pre-par (10 mg/mL)</td>
			<td style="width:197px;">Laboratorio Reig Jofr&#233;, S.A</td>
			<td style="width:107px;"></td>
			<td style="width:511px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:23px;width:267px;">Viridia series 50 XM Fetal Monitor</td>
			<td style="width:197px;">Koninklijke Philips N.V</td>
			<td style="width:107px;"></td>
			<td style="width:511px;">https://www.usa.philips.com/healthcare/product/HC865071/avalon-fm50-fetal-monitor</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:267px;">Voluson P6</td>
			<td style="width:197px;">General Electric Health Care Company</td>
			<td style="width:107px;"></td>
			<td style="width:511px;">https://www.ge-sonostore.com/en/voluson/p6</td>
		</tr>
	</tbody>
</table>

external cephalic version: is it an effective and safe procedure?

External cephalic version (ECV) is an effective procedure for reducing the number of cesarean sections. To date, there is no video publication showing the methodology of this procedure. The main objective is to show how to perform ECV with a specific protocol with tocolysis before the procedure and analgesia. Moreover, we describe and analyze the factors associated with successful ECV, and also compare to deliveries in the general pregnant population.
A retrospective and descriptive analysis of ECV carried out at the Hospital Clinico Universitario Virgen de la Arrixaca in Murcia (Spain) between 1/1/2014 and 12/31/2018 was assessed. The latest data available of labor deliveries in the local center, which is the biggest maternity department in Spain, were from 2018.
320 patients were recruited and 3 pregnant women were lost during the study. ECV was carried out at 37&#177;3 weeks gestation. ECV was successful in 82.5% (N=264). 19 complications were reported (5.9%): 8 vaginal bleeding (2.5%), 9 fetal bradycardia (2.8%), 1 preterm rupture of membranes (0.3%) and 1 cord prolapse (0.3%). A previous vaginal delivery increases the success rate of ECV ORadjusted=3.03 (1.62-5.68). Maternal Body Mass Index (BMI) affects the success of ECV ORadjusted=0.94 (0.89-0.99). Patients with BMI&#62;40 kg/m2 have an ORadjusted=0.09 (0.009-0.89) compared with those with BMI &#60;25 kg/m2. If ECV was successful, the cesarean delivery index is 22.2% (17.5-27.6%), the eutocic delivery index is 52.1% (46.1-58.1%) and the instrumented vaginal delivery index is 25.7% (20.7-31.2%). There are no differences in cesarean and eutocic delivery indexes after successful ECV. However, a successful ECV is associated with a 6.29% increase in the instrumented delivery rate (OR=1.63).
ECV is an effective procedure to reduce the number of cesarean sections for breech presentations. Maternal BMI and previous vaginal delivery are associated with ECV success. Successful ECV does not modify the usual delivery pattern.

Three hundred and twenty patients were recruited between January 1, 2014 and December 31, 2018. Three patients were lost during the follow-up after the ECV because delivery was not carried out in our hospital.
Statistics were derived from the raw data. To study the differences between groups, unpaired Student&#39;s t-tests were used for quantitative variables and chi-square tests for dichotomous variables. All tests were two-tailed at a 0.05 significance level.
The ...

Watch this Scientific Journal Video about External Cephalic Version: Is it an Effective and Safe Procedure? at JoVE.com

External Cephalic Version: Is it an Effective and Safe Procedure?

This article shows how to perform the external cephalic version (ECV) by two experienced obstetricians in the obstetric operating room in the presence of an anesthesiologist and a midwife. The ECV is carried out with analgesia and tocolysis. Two attempts are made under ultrasonography control.

External cephalic version (ECV) is an effective procedure for reducing the number of cesarean sections. To date, there is no video publication showing the ...

external-cephalic-version-is-it-an-effective-and-safe-procedure

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14.3K Views.  University Clinical Hospital. This article shows how to perform the external cephalic version (ECV) by two experienced obstetricians in the obstetric operating room in the presence of an anesthesiologist and a midwife. The ECV is carried out with analgesia and tocolysis. Two attempts are made under ultrasonography control.

Video: External Cephalic Version: Is it an Effective and Safe Procedure?

Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model creating an artificial fistula between the common carotid artery (CCA) and the external jugular vein (EJV) has been widely described and proved technically feasible. This construct provokes a consistent cerebral venous hypertension (CVH), and therefore has helped studying the contribution of venous hypertension to formation, clinical symptoms, and prognosis of brain AVMs and dural AVFs. Equivalent mice models have been only scarcely described and have shown trouble with stenosis of the fistula. An established murine model would allow the study of not only pathophysiology but also potential genetic therapies for these cerebrovascular diseases.
We present a model of arteriovenous fistula that produces a durable intracranial venous hypertension in the mouse. Microsurgical anastomosis of the murine CCA and EJV can be difficult due to diminutive anatomy and frequently result in a non-patent fistula. In this step-by-step protocol we address all the important challenges encountered during this procedure. Avoiding excessive retraction of the vein during the exposure, using 11-0 sutures instead of 10-0, and making a carefully planned end-to-side anastomosis are some of the critical steps. Although this method requires advanced microsurgical skills and a longer learning curve that the equivalent in the rat, it can be consistently developed.
This novel model has been designed to integrate transgenic mouse techniques with a previously well-established experimental system that has proved useful to study brain AVMs and dural AVFs. By opening the possibility of using transgenic mice, a broader spectrum of valid models can be achieved and genetic treatments can also be tested. The experimental construct could also be further adapted to the study of other cerebrovascular diseases related with venous hypertension such as migraine, transient global amnesia, transient monocular blindness, etc.

We describe a method for creating a reliable model of cerebral venous hypertension in the adult mouse. This model has been widely described and tested in the rat. This new counterpart in the mice opens the possibility of using genetic modified animals and thereby broadens the applications of the model.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model ...

A Protocol for Genetic Induction and Visualization of Benign and Invasive Tumors in Cephalic Complexes of Drosophila melanogaster

Drosophila has illuminated our understanding of the genetic basis of normal development and disease for the past several decades and today it continues to contribute immensely to our understanding of complex diseases 1-7. Progression of tumors from a benign to a metastatic state is a complex event 8 and has been modeled in Drosophila to help us better understand the genetic basis of this disease 9. Here I present a simple protocol to genetically induce, observe and then analyze the progression of tumors in Drosophila larvae. The tumor induction technique is based on the MARCM system 10 and exploits the cooperation between an activated oncogene, RasV12 and loss of cell polarity genes (scribbled, discs large and lethal giant larvae) to generate invasive tumors 9. I demonstrate how these tumors can be visualized in the intact larvae and then how these can be dissected out for further analysis. The simplified protocol presented here should make it possible for this technique to be utilized by investigators interested in understanding the role of a gene in tumor invasion.

A Protocol for Genetic Induction and Visualization of Benign and Invasive Tumors in Cephalic Complexes of Drosophila melanogaster

Cooperation between an activated oncogene like RASV12 and mutations in cell polarity genes like scribbled, result in tumor growth in Drosophila where tumor cells also display invasive behaviors. Here a simple protocol for the induction and observation of the benign and invasive tumors is presented.

Drosophila has illuminated our  understanding of the genetic basis of normal development and disease for the  past several decades and today it continues ...

Adjustable Stiffness, External Fixator for the Rat Femur Osteotomy and Segmental Bone Defect Models

The mechanical environment around the healing of broken bone is very important as it determines the way the fracture will heal. Over the past decade there has been great clinical interest in improving bone healing by altering the mechanical environment through the fixation stability around the lesion. One constraint of preclinical animal research in this area is the lack of experimental control over the local mechanical environment within a large segmental defect as well as osteotomies as they heal. In this paper we report on the design and use of an external fixator to study the healing of large segmental bone defects or osteotomies. This device not only allows for controlled axial stiffness on the bone lesion as it heals, but it also enables the change of stiffness during the healing process in vivo. The conducted experiments have shown that the fixators were able to maintain a 5 mm femoral defect gap in rats in vivo during unrestricted cage activity for at least 8 weeks. Likewise, we observed no distortion or infections, including pin infections during the entire healing period. These results demonstrate that our newly developed external fixator was able to achieve reproducible and standardized stabilization, and the alteration of the mechanical environment of in vivo rat large bone defects and various size osteotomies. This confirms that the external fixation device is well suited for preclinical research investigations using a rat model in the field of bone regeneration and repair.

One constraint of preclinical research in the field of bone repair is the lack of experimental control over the local mechanical environment within a healing bone lesion. We report the design and use of an external fixator for bone repair with the ability to change fixator stiffness in vivo.

The mechanical environment around the healing of broken bone is very important as it determines the way the fracture will heal. Over the past decade there ...

Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement

The mainstay of contemporary therapies for extensive occlusive arterial disease is venous bypass graft. However, its durability is threatened by intimal hyperplasia (IH) that eventually leads to vessel occlusion and graft failure. Mechanical forces, particularly low shear stress and high wall tension, are thought to initiate and to sustain these cellular and molecular changes, but their exact contribution remains to be unraveled. To selectively evaluate the role of pressure and shear stress on the biology of IH, an ex&#160;vivo perfusion system (EVPS) was created to perfuse segments of human saphenous veins under arterial regimen (high shear stress and high pressure). Further technical innovations allowed the simultaneous perfusion of two segments from the same vein, one reinforced with an external mesh. Veins were harvested using a no-touch technique and immediately transferred to the laboratory for assembly in the EVPS. One segment of the freshly isolated vein was not perfused (control, day 0). The two others segments were perfused for up to 7 days, one being completely sheltered with a 4 mm (diameter) external mesh. The pressure, flow velocity, and pulse rate were continuously monitored and adjusted to mimic the hemodynamic conditions prevailing in the femoral artery. Upon completion of the perfusion, veins were dismounted and used for histological and molecular analysis. Under ex&#160;vivo conditions, high pressure perfusion (arterial, mean = 100 mm Hg) is sufficient to generate IH and remodeling of human veins. These alterations are reduced in the presence of an external polyester mesh.

Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement

The mechanisms leading to the development of intimal hyperplasia (IH) and vein graft failure are still poorly understood. This study describes an ex&#160;vivo system to perfuse human veins under controlled flow and pressure. Furthermore the efficiency of external mesh reinforcement to limit the development of IH was evaluated.

The mainstay of contemporary therapies for extensive occlusive arterial disease is venous bypass graft. However, its durability is threatened by intimal ...

Glaucoma-inducing Procedure in an In Vivo Rat Model and Whole-mount Retina Preparation

Glaucoma is a disease of the central nervous system affecting retinal ganglion cells (RGCs). RGC axons making up the optic nerve carry visual input to the brain for visual perception. Damage to RGCs and their axons leads to vision loss and/or blindness. Although the specific cause of glaucoma is unknown, the primary risk factor for the disease is an elevated intraocular pressure. Glaucoma-inducing procedures in animal models are a valuable tool to researchers studying the mechanism of RGC death. Such information can lead to the development of effective neuroprotective treatments that could aid in the prevention of vision loss. The protocol in this paper describes a method of inducing glaucoma - like conditions in an in vivo rat model where 50 &#181;l of 2 M hypertonic saline is injected into the episcleral venous plexus. Blanching of the vessels indicates successful injection. This procedure causes loss of RGCs to simulate glaucoma. One month following injection, animals are sacrificed and eyes are removed. Next, the cornea, lens, and vitreous are removed to make an eyecup. The retina is then peeled from the back of the eye and pinned onto sylgard dishes using cactus needles. At this point, neurons in the retina can be stained for analysis. Results from this lab show that approximately 25% of RGCs are lost within one month of the procedure when compared to internal controls. This procedure allows for quantitative analysis of retinal ganglion cell death in an in vivo rat glaucoma model.

Glaucoma-inducing Procedure in an In Vivo Rat Model and Whole-mount Retina Preparation

Glaucoma is characterized by damage to retinal ganglion cells. Inducing glaucoma in animal models can provide insight into the study of this disease. Here, we outline a procedure that induces loss of RGCs in an in vivo rat model and demonstrates the preparation of whole-mount retinas for analysis.

Glaucoma is a disease of the central nervous system affecting retinal ganglion cells (RGCs). RGC axons making up the optic nerve carry visual input to the ...

Pancreatic Duct Infusion: An Effective and Selective Method of Drug and Viral Delivery

The pancreas is a bifunctional organ with both endocrine and exocrine components. A number of pathologies can afflict the pancreas, including diabetes, pancreatitis, and pancreatic cancer. All three of these diseases mark active areas of study, not only to develop immediate therapy, but also to better understand their pathophysiology. There are few tools to further these areas of study. Pancreatic duct infusion is an important technique that can allow for lineage tracing, gene introduction, and cell line-specific targeting. The technique requires the intricate dissection of the second portion of the duodenum and ampulla, followed by the occlusion of the bile duct and the cannulation of the pancreatic duct. Although the technique is technically challenging at first, the applications are myriad. Ambiguity in the specifics of the procedure between groups highlighted the need for a standard protocol. This work describes the expression of a green fluorescent protein (GFP) within the pancreas after the pancreatic duct infusion of a viral vector expressing GFP versus a sham surgery. The infusion and therefore expression is specific to the pancreas, without expression present in any other tissue type.

Pancreatic duct infusion is an important technique that can allow for lineage tracing, gene introduction, and cell line-specific targeting. A pancreatic duct infusion technique for drug and viral delivery to pancreatic cells is presented here.

The pancreas is a bifunctional organ with both endocrine and exocrine components. A number of pathologies can afflict the pancreas, including diabetes, ...

3D Ultrasound Imaging: Fast and Cost-effective Morphometry of Musculoskeletal Tissue

The developmental goal of 3D ultrasound imaging (3DUS) is to engineer a modality to perform 3D morphological ultrasound analysis of human muscles. 3DUS images are constructed from calibrated freehand 2D B-mode ultrasound images, which are positioned into a voxel array. Ultrasound (US) imaging allows quantification of muscle size, fascicle length, and angle of pennation. These morphological variables are important determinants of muscle force and length range of force exertion. The presented protocol describes an approach to determine volume and fascicle length of m.&#160;vastus lateralis and m.&#160;gastrocnemius medialis. 3DUS facilitates standardization using 3D anatomical references. This approach provides a fast and cost-effective approach for quantifying 3D morphology in skeletal muscles. In healthcare and sports, information on the morphometry of muscles is very valuable in diagnostics and/or follow-up evaluations after treatment or training.

3D ultrasound imaging (3DUS) allows fast and cost-effective morphometry of musculoskeletal tissues. We present a protocol to measure muscle volume and fascicle length using 3DUS.

The developmental goal of 3D ultrasound imaging (3DUS) is to engineer a modality to perform 3D morphological ultrasound analysis of human muscles. 3DUS ...

Optimized Griess Reaction for UV-Vis and Naked-eye Determination of Anti-malarial Primaquine

Primaquine (PMQ), an important anti-malarial drug, has been recommended by the World Health Organization (WHO) for the treatment of life-threatening infections caused by P. vivax and ovale. However, PMQ has unwanted adverse effects that lead&#160;to acute hemolysis in patients with glucose-6-phosphate dehydrogenase (G6PD) deficiency. There is a need to develop simple and reliable methods for PMQ determination with the purpose of dosage monitoring. In early 2019, we have reported an UV-Vis and naked-eye based approach for PMQ colorimetric quantification. The detection was based on a Griess-like reaction between PMQ and anilines, which can generate colored azo products. The detection limit for direct measurement of PMQ in synthetic urine is in the nanomolar range. Moreover, this method has shown great potential for PMQ quantification from human serum samples at clinically relevant concentrations. In this protocol, we will describe the technical details regarding the syntheses and characterization of colored azo products, the reagent preparation, and the procedures for PMQ determination.

This protocol describes a novel colorimetric method for antimalarial primaquine (PMQ) detection in synthetic urines and human serums.

Primaquine (PMQ), an important anti-malarial drug, has been recommended by the World Health Organization (WHO) for the treatment of life-threatening ...

Determining and Controlling External Power Output During Regular Handrim Wheelchair Propulsion

The use of a manual wheelchair is critical to 1% of the world&#39;s population. Human powered wheeled mobility research has considerably matured, which has led to improved research techniques becoming available over the last decades. To increase the understanding of wheeled mobility performance, monitoring, training, skill acquisition, and optimization of the wheelchair-user interface in rehabilitation, daily life, and sports, further standardization of measurement set-ups and analyses is required. A crucial stepping-stone is the accurate measurement and standardization of external power output (measured in Watts), which is pivotal for the interpretation and comparison of experiments aiming to improve rehabilitation practice, activities of daily living, and adaptive sports. The different methodologies and advantages of accurate power output determination during overground, treadmill, and ergometer-based testing are presented and discussed in detail. Overground propulsion provides the most externally valid mode for testing, but standardization can be troublesome. Treadmill propulsion is mechanically similar to overground propulsion, but turning and accelerating is not possible. An ergometer is the most constrained and standardization is relatively easy. The goal is to stimulate good practice and standardization to facilitate the further development of theory and its application among research facilities and applied clinical and sports sciences around the world.

Accurate and standardized assessment of external power output is crucial in the evaluation of physiological, biomechanical, and perceived stress, strain, and capacity in manual wheelchair propulsion. The current article presents various methods to determine and control power output during wheelchair propulsion studies in the laboratory and beyond.

The use of a manual wheelchair is critical to 1% of the world&#39;s population. Human powered wheeled mobility research has considerably matured, which ...

TBase - an Integrated Electronic Health Record and Research Database for Kidney Transplant Recipients

TBase is an electronic health record (EHR) for kidney transplant recipients (KTR) combining automated data entry of key clinical data (e.g., laboratory values, medical reports, radiology and pathology data) via standardized interfaces with manual data entry during routine treatment (e.g., clinical notes, medication list, and transplantation data). By this means, a comprehensive database for KTR is created with benefits for routine clinical care and research. It enables both easy everyday clinical use and quick access for research questions with highest data quality. This is achieved by the concept of data validation in clinical routine in which clinical users and patients have to rely on correct data for treatment and medication plans and thereby validate and correct the clinical data in their daily practice. This EHR is tailored for the needs of transplant outpatient care and proved its clinical utility for more than 20 years at Charit&#233; - Universit&#228;tsmedizin Berlin. It facilitates efficient routine work with well-structured, comprehensive long-term data and allows their easy use for clinical research. To this point, its functionality covers automated transmission of routine data via standardized interfaces from different hospital information systems, availability of transplant-specific data, a medication list with an integrated check for drug-drug interactions, and semi-automated generation of medical reports among others. Key elements of the latest reengineering are a robust privacy-by-design concept, modularity, and hence portability into other clinical contexts as well as usability and platform independence enabled by HTML5 (Hypertext Markup Language) based responsive web design. This allows fast and easy scalability into other disease areas and other university hospitals. The comprehensive long-term datasets are the basis for the investigation of Machine Learning algorithms, and the modular structure allows to rapidly implement these into clinical care. Patient reported data and telemedicine services are integrated into TBase in order to meet future needs of the patients. These novel features aim to improve clinical care as well as to create new research options and therapeutic interventions.

TBase combines an electronic health record with an innovative research database for kidney transplant recipients. TBase is built upon an in-memory database platform, connected to different hospital systems, and used for regular outpatient care. It automatically integrates all relevant clinical data including transplantation-specific data creating a unique research database.

TBase is an electronic health record (EHR) for kidney transplant recipients (KTR) combining automated data entry of key clinical data (e.g., laboratory ...