Name: external cephalic version: is it an effective and safe procedure?
Uploaded: 2020-06-06T13:00:00
Description: Watch this Scientific Journal Video about External Cephalic Version: Is it an Effective and Safe Procedure? at JoVE.com

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>...

<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>

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.

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	<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>
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		<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? External cephalic version is a procedure for modifying fetal position and achieving a cephalic presentation.The aim of ECV is to offer an opportunity to have a cephalic delivery with less risk than a breech or cesarean delivery. ECV is usually done before active labor period begins. The aim of this study was to show how to do the ECV and describe key actions that could contribute to improve the success of this procedure.Our secondary objective was to analyze the results of ECV with our specific protocol with tocolysis and analgesia and compare with those in the literature. This study was approved by the Clinical Research Committee of the Virgen de la Arrixaca at the University Clinical Hospital. Written informed consent was obtained from all participants.A descriptive retrospective analysis of ECV performed in Hospital Clinico Universitario Virgen de la Arrixaca in Murcia between 1st of January of 2014 and 31st of December of 2018 has been carried out. Procedure was performed by two of the four experienced obstetricians of Maternal-Fetal Unit. A midwife and anesthesiologist were present during the ECV.During third trimester ultrasound scan fetal presentation is checked. If non-cephalic presentation is assessed, the patient is seated the next week. During that consult, ECV is offered to every pregnant with non-cephalic presentation and no absolute contraindication for vaginal delivery.Then, procedure is scheduled for the following week. Offer external cephalic version at consult. Identify fetal position with ultrasound offer the external cephalic version, sign informed consent.Admit in obstetric emergency room. Identify fetal position with ultrasound. Prepare the patient and take requirements with test and informed consent.Admit in obstetric delivery room. Perform cardiotocography assessment for fetal well-being. Prepare ritodrine solution.Administer six milligrams of ritodrine at 60 milliliter per hour during 30 minutes before the procedure. Invite the patient to empty her bladder. Stop ritodrine perfusion before moving to obstetric operating room.External cephalic version procedure in obstetric operating room. Move to obstetric operating room. Monitor maternal vital signs.Position the patient in Trendelenburg. There are two options for performing analgesia. Sedate patient with propofol or alternatively spinal anesthesia.External cephalic version procedure following National guidelines, two attempts are done. No more. Obstetrician A elevate fetal buttocks.Then when it is achieved, obstetrician A guide fetal buttocks to the fundus. And when it is achieved, obstetrician B direct fetal head to the pelvis. Identify fetal well-being with ultrasound.Check vaginal bleeding and perform cardiotocography assessment for fetal well-being. Later moved to obstetric delivery room. Perform cardiotocography assessment for fetal well-being during at least four hours.And discharge the patient. Cardiotocography assessment for fetal well-being should be repeated in 24 hours. 320 patients were recruited between 1st of January of 2014 and 31st of December of 2018.Three patients were lost. The ECV were performed in 37 plus three weeks of gestation as an average. ECV was successful in 82.5%Intraversion complication occurred in almost 6%Nine fetal bradycardia more than six minutes, eight vaginal bleeding, one cord prolapse and one preterm rupture of the membranes during the following 24 hours.No newborns were hospitalization in neonatal unit neither neonatal intensive care unit. These are the results and the characteristic of ECV. It should be noted that nulliparity was more prevalent in patient in fail group.It should be highlighted that there was no difference between BMI. It should be emphasized that sedation was the most frequent method for analgesia during ECV and only in three patients spinal anesthesia was performed for ECV. These are the results and delivery depending on ECV result.It should be remarked that almost 78%of patients with successful ECV had vaginal delivery. 52%have eutocic delivery and 26 instrumented delivery. This is the multivariate logistic regression model for predicting ECV result.It should be highlighted that previous vaginal delivery improved the possibilities of success with an adjusted odds ratio of three. And maternal body mass index reduced the possibilities of success with an adjusted odds ratio of 0.94. This is the multivariate logistic regression model for predicting ECV results depending on the BMI.It should be noted that patients with BMI above 40 had far less possibilities of success with an odds ratio of 0.09. This is the comparisons of the type of delivery. It should be note that there were no difference in cesarean rate and eutocic delivery rate between patients with a successful ECV and general pregnant population.However, patients with successful ECV had the higher instrumented delivery rate than general pregnant population. After a successful ECV, nulliparity was the only factor statistically associated with instrumented delivery with an adjusted odds ratio of nine. After a successful ECV, BMI was the only factor statistically associated with urgent cesarean section with an adjusted odds ratio of 1.11.After a successful ECV, instrumented delivery rate increased in 8.56%compared with general population what means an odds ratio of 1.63. ECV procedure rate is higher than literature and Spain. It should be noted that some steps considered as crucial and that may differ from others such as empty the bladder before the procedure, the use of tocolysis only before the ECV, or analgesic with propofol or spinal anesthesia.Bladder volume plays an important role in the success of ECV. The high procedure success rate maybe explained because of the use of this protocol with tocolysis and analgesia, the experienced team and the presence of a midwife and anesthesiologist. ECV complication rate is 5.94%which is similar to literature.It should be remarked that the use of tocolysis and analgesia increase the risk of vaginal bleeding and placental abruption. Successful ECV does not modify eutocic delivery rate. Successful ECV does not modify cesarean delivery rate.Successful ECV increase instrumented delivery rate with an odds ratio 1.63 which is similar to literature.

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

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Medicine

Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules that regulate HSCs. In these transplant model systems, the function of HSCs is determined by the ability of these cells to engraft and reconstitute lethally irradiated recipient mice. Commonly, the donor cell contribution/engraftment is measured by antibodies to donor- specific cell surface proteins using flow cytometry. However, this method heavily depends on the specificity and the ability of the cell surface marker to differentiate donor-derived cells from recipient-originated cells, which may not be available for all mouse strains. Considering the various backgrounds of genetically modified mouse strains in the market, this cell surface/ flow cytometry-based method has significant limitations especially in mouse strains that lack well-defined surface markers to separate donor cells from congenic recipient cells. Here, we reported a PCR-based technique to determine donor cell engraftment/contribution in transplant recipient mice. We transplanted male donor bone marrow HSCs to lethally irradiated congenic female mice. Peripheral blood samples were collected at different time points post transplantation. Bone marrow samples were obtained at the end of the experiments. Genomic DNA was isolated and the Y chromosome specific gene, Zfy1, was amplified using quantitative Real time PCR. The engraftment of male donor-derived cells in the female recipient mice was calculated against standard curve with known percentage of male vs. female DNAs. Bcl2 was used as a reference gene to normalize the total DNA amount. Our data suggested that this approach reliably determines donor cell engraftment and provides a useful, yet simple method in measuring hematopoietic cell reconstitution in murine bone marrow transplantation models. Our method can be routinely performed in most laboratories because no costly equipment such as flow cytometry is required.

Determining donor cell engraftment presents a challenge in mouse bone marrow transplant models that lack well-defined phenotypical markers. We described a methodology to quantify male donor cell engraftment in female transplant recipient mice. This method can be used in all mouse strains for the study of HSC functions.

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules ...

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 ...

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, ...

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 ...

Focal Laser Ablation of Prostate Cancer: An Office Procedure

In this article, we describe and illustrate an outpatient procedure for focal laser ablation (FLA) of prostate cancer (PCa). The procedure is conceptually similar to a fusion biopsy and is performed under local anesthesia in a clinic setting; treatment time is usually less than one hour. Laser insertion is guided by ultrasound; lesion targeting is via magnetic resonance imaging-ultrasound (MRI/US) fusion, as in targeted prostate biopsy. Real-time ablation monitoring is achieved utilizing a thermal probe adjacent to the laser fiber. The video demonstrates procedure planning, patient preparation, various steps during the procedure, and treatment monitoring. Safety, feasibility, and efficacy of this approach have been established during a previous trial. Outpatient FLA under local anesthesia is an option for management of intermediate risk prostate cancer.

This article presents and describes an outpatient treatment for prostate cancer using focal laser ablation. Laser catheter placement is guided by MRI-ultrasound fusion imaging in a fashion similar to prostate needle biopsy. Treatment is monitored in real-time with a thermal probe, placed adjacent to the laser fiber.

In this article, we describe and illustrate an outpatient procedure for focal laser ablation (FLA) of prostate cancer (PCa). The procedure is conceptually ...

An Effective Mouse Model of Unilateral Renal Ischemia-Reperfusion Injury

Ischemia-reperfusion injury (IRI) is the leading cause of acute renal failure and is a significant contributor to delayed graft function. Animal models are the only available resources that mimic the complexities of the IRI-associated damage encountered in vivo. This paper describes an effective mouse model of unilateral renal IRI that delivers highly reproducible data. Ischemia is induced by occluding the right renal pedicle for 30 min followed by reperfusion. In addition to the surgical procedure, a sequential overview of the expected physiological and histopathological changes following renal IRI will be provided by comparing data from seven different reperfusion times (4 h, 8 h, 16 h, 1 day, 2 days, 4 days, and 7 days). Critical data for planning experiments ahead, such as mean surgical time, average anesthetic consumption, and body weight changes over time, will be shared. This work will help researchers implement a reliable renal IRI model and select the appropriate reperfusion time that aligns with their intended investigative goals.

Renal ischemia-reperfusion injury is associated with high morbidity and mortality in hospitalized patients. Here, we present a simple and effective mouse model of unilateral renal ischemia-reperfusion injury and provide a sequential overview of representative pathological changes observed in the kidney.

Ischemia-reperfusion injury (IRI) is the leading cause of acute renal failure and is a significant contributor to delayed graft function. Animal models ...