<ol>
	<li>Harding, R., Hooper, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+lung+expansion+and+lung+growth+before+birth.">Regulation of lung expansion and lung growth before birth.</a> J Appl Physiol (1985). 81 (1), 209-224 (1996).</li><li>Olver, R. E., Strang, L. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ion+fluxes+across+the+pulmonary+epithelium+and+the+secretion+of+lung+liquid+in+the+foetal+lamb.">Ion fluxes across the pulmonary epithelium and the secretion of lung liquid in the foetal lamb.</a> J Physiol. 241 (2), 327-357 (1974).</li><li>Schittny, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+the+lung.">Development of the lung.</a> Cell Tissue Res. 367 (3), 427-444 (2017).</li><li>Vilos, G. A., Liggins, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intrathoracic+pressures+in+fetal+sheep.">Intrathoracic pressures in fetal sheep.</a> J Dev Physiol. 4 (4), 247-256 (1982).</li><li>Dawes, G. S., Fox, H. E., Leduc, B. M., Liggins, G. C., Richards, R. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+movements+and+rapid+eye+movement+sleep+in+the+foetal+lamb.">Respiratory movements and rapid eye movement sleep in the foetal lamb.</a> J Physiol. 220 (1), 119-143 (1972).</li><li>Bowes, G., 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=Development+of+patterns+of+respiratory+activity+in+unanesthetized+fetal+sheep+in+utero.">Development of patterns of respiratory activity in unanesthetized fetal sheep in utero.</a> J Appl Physiol Respir Environ Exerc Physiol. 50 (4), 693-700 (1981).</li><li>Clewlow, F., Dawes, G., Johnston, B., Walker, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+breathing,+electrocortical+and+muscle+activity+in+unanaesthetized+fetal+lambs+with+age.">Changes in breathing, electrocortical and muscle activity in unanaesthetized fetal lambs with age.</a> J Physiol. 341, 463-476 (1983).</li><li>Chapman, R. L., Dawes, G. S., Rurak, D. W., Wilds, P. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Breathing+movements+in+fetal+lambs+and+the+effect+of+hypercapnia.">Breathing movements in fetal lambs and the effect of hypercapnia.</a> J Physiol. 302, 19-29 (1980).</li><li>Badalian, S. S., Fox, H. E., Chao, C. R., Timor-Tritsch, I. E., Stolar, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fetal+breathing+characteristics+and+postnatal+outcome+in+cases+of+congenital+diaphragmatic+hernia.">Fetal breathing characteristics and postnatal outcome in cases of congenital diaphragmatic hernia.</a> Am J Obstet Gynecol. 171, 970-976 (1994).</li><li>Badalian, S. S., Fox, H. E., Zimmer, E. Z., Fifer, W. P., Stark, R. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterns+of+perinasal+fluid+flow+and+contractions+of+the+diaphragm+in+the+human+fetus.">Patterns of perinasal fluid flow and contractions of the diaphragm in the human fetus.</a> Ultrasound Obstet Gynecol. 8 (2), 109-113 (1996).</li><li>Fox, H. E., Moessinger, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fetal+breathing+movements+and+lung+hypoplasia:+preliminary+human+observations.">Fetal breathing movements and lung hypoplasia: preliminary human observations.</a> Am J Obstet Gynecol. 151 (4), 531-533 (1985).</li><li>Fox, H. E., Badalian, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasound+prediction+of+fetal+pulmonary+hypoplasia+in+pregnancies+complicated+by+oligohydramnios+and+in+cases+of+congenital+diaphragmatic+hernia:+A+review.">Ultrasound prediction of fetal pulmonary hypoplasia in pregnancies complicated by oligohydramnios and in cases of congenital diaphragmatic hernia: A review.</a> Am J Perinatol. 11 (2), 104-108 (1994).</li><li>Fox, H. E., Badalian, S. S., Fifer, W. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterns+of+fetal+perinasal+fluid+flow+in+cases+of+congenital+diaphragmatic+hernia.">Patterns of fetal perinasal fluid flow in cases of congenital diaphragmatic hernia.</a> Am J Obstet Gynecol. 176 (4), 807-813 (1997).</li><li>Carmel, J. A., Friedman, F., Adams, F. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fetal+tracheal+ligation+and+lung+development.">Fetal tracheal ligation and lung development.</a> Am J Dis Child. 109, 452-456 (1965).</li><li>Deprest, J. 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=Randomized+trial+of+fetal+surgery+for+moderate+left+diaphragmatic+hernia.">Randomized trial of fetal surgery for moderate left diaphragmatic hernia.</a> N Engl J Med. 385 (2), 119-129 (2021).</li><li>Deprest, J. 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=Randomized+trial+of+fetal+surgery+for+severe+left+diaphragmatic+hernia.">Randomized trial of fetal surgery for severe left diaphragmatic hernia.</a> N Engl J Med. 385 (2), 107-118 (2021).</li><li>Van Calster, B., 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=The+randomized+TOTAL-trials+on+fetal+surgery+for+congenital+diaphragmatic+hernia:+re-analysis+using+pooled+data.">The randomized TOTAL-trials on fetal surgery for congenital diaphragmatic hernia: re-analysis using pooled data.</a> Am J Obstet Gynecol. 226 (4), 560.e1-560.e24 (2021).</li><li>NHMRC. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> National Health and Medical Research Council (2013) Australian Code for the Care and Use of Animals for Scientific Purposes. , (2021).</li><li>Robbiani, 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=An+implantable+electronic+device+for+monitoring+fetal+lung+pressure+in+a+lamb+model+of+Congenital+Diaphragmatic+Hernia.">An implantable electronic device for monitoring fetal lung pressure in a lamb model of Congenital Diaphragmatic Hernia.</a> IEEE Transactions on Instrumentation and Measurement. 71, 1-10 (2022).</li><li>Moessinger, A. C., Harding, R., Adamson, T. M., Singh, M., Kiu, G. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+lung+fluid+volume+in+growth+and+maturation+of+the+fetal+sheep+lung.">Role of lung fluid volume in growth and maturation of the fetal sheep lung.</a> J Clin Invest. 86 (4), 1270-1277 (1990).</li><li>Dickson, K. A., Harding, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fetal+breathing+and+pressures+in+the+trachea+and+amniotic+sac+during+oligohydramnios+in+sheep.">Fetal breathing and pressures in the trachea and amniotic sac during oligohydramnios in sheep.</a> J Appl Physiol (1985). 70 (1), 293-299 (1991).</li><li>Alcorn, D., 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=Morphological+effects+of+chronic+tracheal+ligation+and+drainage+in+the+fetal+lamb+lung.">Morphological effects of chronic tracheal ligation and drainage in the fetal lamb lung.</a> J Anatomy. 123 (3), 649-660 (1977).</li><li>Wigglesworth, J. S., Desai, R., Hislop, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fetal+lung+growth+in+congenital+laryngeal+atresia.">Fetal lung growth in congenital laryngeal atresia.</a> Pediatr Pathol. 7 (5-6), 515-525 (1987).</li><li>Khan, P. A., Cloutier, M., Piedboeuf, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tracheal+occlusion:+a+review+of+obstructing+fetal+lungs+to+make+them+grow+and+mature.">Tracheal occlusion: a review of obstructing fetal lungs to make them grow and mature.</a> Am J Med Genet C Semin Med Genet. 145C (2), 125-138 (2007).</li><li>De Paepe, M. E., Johnson, B. D., Papadakis, K., Luks, F. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lung+growth+response+after+tracheal+occlusion+in+fetal+rabbits+is+gestational+age-dependent.">Lung growth response after tracheal occlusion in fetal rabbits is gestational age-dependent.</a> Am J Respir Cell Mol Biol. 21 (1), 65-76 (1999).</li><li>Probyn, M. E., Wallace, M. J., Hooper, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+increased+lung+expansion+on+lung+growth+and+development+near+midgestation+in+fetal+sheep.">Effect of increased lung expansion on lung growth and development near midgestation in fetal sheep.</a> Pediatr Res. 47 (6), 806-812 (2000).</li><li>Hashim, E., Laberge, J. M., Chen, M. F., Quillen, E. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reversible+tracheal+obstruction+in+the+fetal+sheep:+effects+on+tracheal+fluid+pressure+and+lung+growth.">Reversible tracheal obstruction in the fetal sheep: effects on tracheal fluid pressure and lung growth.</a> J Pediatr Surg. 30 (8), 1172-1177 (1995).</li><li>Hellmeyer, L., 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=Telemetric+monitoring+of+tracheal+pressure+after+tracheal+occlusion+for+treatment+of+severe+congenital+diaphragmatic+hernia.">Telemetric monitoring of tracheal pressure after tracheal occlusion for treatment of severe congenital diaphragmatic hernia.</a> Arch Gynecol Obstet. 275 (4), 245-248 (2007).</li></ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1.59 mm (outside diameter, OD) to 3.18 mm (OD) connector</td>
			<td>Qosina</td>
			<td>11913</td>
			<td></td>
		</tr>
		<tr>
			<td>3.18 mm (OD) to 3.18 mm (OD) connector</td>
			<td>Qosina</td>
			<td>11684</td>
			<td></td>
		</tr>
		<tr>
			<td>70 % Alcohol</td>
			<td>Henry Schein</td>
			<td>1127067</td>
			<td></td>
		</tr>
		<tr>
			<td>Absorbable poliglecaprone 25 monofilament 3-0</td>
			<td>Riverpoint Medical</td>
			<td>Q316</td>
			<td></td>
		</tr>
		<tr>
			<td>Absorbable polydioxanone monofilament 1</td>
			<td>Riverpoint Medical</td>
			<td>D879</td>
			<td></td>
		</tr>
		<tr>
			<td>Absorbable polyglactin 910 braided 2-0</td>
			<td>Riverpoint Medical</td>
			<td>V317</td>
			<td></td>
		</tr>
		<tr>
			<td>Absorbable polydioxanone monofilament 5-0</td>
			<td>Riverpoint Medical</td>
			<td>D303</td>
			<td></td>
		</tr>
		<tr>
			<td>Acepromazine</td>
			<td>Ceva Animal Health</td>
			<td>APVMA No: 36680</td>
			<td></td>
		</tr>
		<tr>
			<td>Babcock, uterine forceps 6.25 inch</td>
			<td>Roboz</td>
			<td>RS-8022</td>
			<td></td>
		</tr>
		<tr>
			<td>Betamethasone</td>
			<td>Merck Sharp &#38; Dohme</td>
			<td>Aust R 18777</td>
			<td></td>
		</tr>
		<tr>
			<td>Blade, size 10</td>
			<td>Becton Dickinson</td>
			<td>371110</td>
			<td></td>
		</tr>
		<tr>
			<td>Blade, size 15</td>
			<td>Becton Dickinson</td>
			<td>371115</td>
			<td></td>
		</tr>
		<tr>
			<td>Bupivacaine</td>
			<td>Pfizer Australia Pty Ltd</td>
			<td>AUST R 11312</td>
			<td></td>
		</tr>
		<tr>
			<td>Cefazolin</td>
			<td>AFT pharmaceuticals</td>
			<td>AUST R 171582</td>
			<td></td>
		</tr>
		<tr>
			<td>Chlorhexidine</td>
			<td>Henry Schein</td>
			<td>0404-0175-02</td>
			<td></td>
		</tr>
		<tr>
			<td>Endotracheal tube (size 8.0)</td>
			<td>&#160;Jorgen Kruuse</td>
			<td>272411</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps, Potts-Smith</td>
			<td>Roboz</td>
			<td>RS-5314</td>
			<td></td>
		</tr>
		<tr>
			<td>Iodine solution (10 %)</td>
			<td>Henry Schein</td>
			<td>6907281</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Piramal Critical Care&#160;</td>
			<td>APVMA No: 53120/112272 
			M. L. No.:220/AP/MD/96/B&#38;F/R</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Ceva Animal Health</td>
			<td>APVMA 37711/58317 
			KETALAB04</td>
			<td></td>
		</tr>
		<tr>
			<td>Hartmanns Solution</td>
			<td>Baxter&#160;</td>
			<td>AUST R 48510</td>
			<td></td>
		</tr>
		<tr>
			<td>Hemostats, Mosquito forceps curved delicate</td>
			<td>Roboz</td>
			<td>RS-7271</td>
			<td></td>
		</tr>
		<tr>
			<td>Medroxyprogesterone acetate</td>
			<td>Pfizer Australia Pty Ltd</td>
			<td>AUST R 12300</td>
			<td></td>
		</tr>
		<tr>
			<td>Meloxicam</td>
			<td>Ilium</td>
			<td>APVMA Approval No.: 62535/127884 
			LI0119V1</td>
			<td></td>
		</tr>
		<tr>
			<td>Methocel</td>
			<td>Colorcon&#160;</td>
			<td>ID34435</td>
			<td></td>
		</tr>
		<tr>
			<td>Microcuff endotracheal tube (3.0)</td>
			<td>Halyard</td>
			<td>35111</td>
			<td></td>
		</tr>
		<tr>
			<td>Midazolam</td>
			<td>Mylan</td>
			<td>AUST R 160205</td>
			<td></td>
		</tr>
		<tr>
			<td>Morphine</td>
			<td>Pfizer Australia Pty Ltd</td>
			<td>AUST R 101240</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle, 22 G</td>
			<td>Becton Dickinson</td>
			<td>305155</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle, 27 G</td>
			<td>Becton Dickinson</td>
			<td>305109</td>
			<td></td>
		</tr>
		<tr>
			<td>Nonabsorbable nylon monofilament 4-0</td>
			<td>Riverpoint Medical</td>
			<td>662BL</td>
			<td></td>
		</tr>
		<tr>
			<td>Nonabsorbable polypropylene monofilament 2-0</td>
			<td>Riverpoint Medical</td>
			<td>P8411</td>
			<td></td>
		</tr>
		<tr>
			<td>OpSite Transparent Film</td>
			<td>Smith and Nephew</td>
			<td>66000040</td>
			<td></td>
		</tr>
		<tr>
			<td>Oxytetracycline</td>
			<td>Norbrook</td>
			<td>APVMA Approval No: 53087/49616</td>
			<td></td>
		</tr>
		<tr>
			<td>Pentobarbitone sodium 300 mg/mL</td>
			<td>Jurox</td>
			<td>APVMA 36208</td>
			<td></td>
		</tr>
		<tr>
			<td>Peracetic acid/hydrogen peroxide</td>
			<td>Medivators Inc&#160;</td>
			<td>ref: 78401-649</td>
			<td></td>
		</tr>
		<tr>
			<td>Piperacillin/Tazobactam&#160;</td>
			<td>Sandoz Pty Ltd</td>
			<td>AUST R 140840</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors, Metzenbaum Surgical 7 inch straight</td>
			<td>Roboz</td>
			<td>RS-6955SC</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors, Vannas 0.15 mm tip width</td>
			<td>Roboz</td>
			<td>RS-5618</td>
			<td></td>
		</tr>
		<tr>
			<td>Silicone tubing (1.59 mm inside diameter)</td>
			<td>Qosina</td>
			<td>T2013</td>
			<td></td>
		</tr>
		<tr>
			<td>Suction catheter (5 French)</td>
			<td>Covidien</td>
			<td>30500</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe, 1 mL</td>
			<td>Becton Dickinson</td>
			<td>309659</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe, 10 mL</td>
			<td>Becton Dickinson</td>
			<td>309604</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe, 60 mL</td>
			<td>Becton Dickinson</td>
			<td>309654</td>
			<td></td>
		</tr>
		<tr>
			<td>Telemetry device</td>
			<td>Politecnico di Milano</td>
			<td>-</td>
			<td>Not commercially available</td>
		</tr>
		<tr>
			<td>Thiopentone sodium</td>
			<td>Jurox Pty Ltd</td>
			<td>APVMA No. 51520/5g/0809</td>
			<td></td>
		</tr>
		<tr>
			<td>Transdermal fentanyl patch</td>
			<td>Janssen-Cilag Pty Ltd</td>
			<td>AUST R 112371</td>
			<td></td>
		</tr>
	</tbody>
</table>

continuous telemetric in utero tracheal pressure measurements in fetal lambs

Read this Scientific Article Protocol about Continuous Telemetric In Utero Tracheal Pressure Measurements in Fetal Lambs at JoVE.com

<meta charset="utf8"></head><body>Medidas Telemétricas Contínuas de Pressão Traqueal In Utero em Cordeiros Fetais

Medidas Telemétricas Contínuas de Pressão Traqueal In Utero em Cordeiros Fetais

Normal in utero lung development and growth rely upon the expansion of airspaces and the controlled efflux of lung liquid into the amniotic space. Infants ...

continuous-telemetric-utero-tracheal-pressure-measurements-fetal

Research

JoVE Journal

Medicine

Continuous Telemetric In Utero Tracheal Pressure Measurements in Fetal Lambs

239 visualizações.  The University of Western Australia. A otimização da pressão de oclusão e da duração necessária para melhorar a hipoplasia pulmonar com oclusão traqueal é essencial para melhorar os tratamentos corretivos in utero para hérnia diafragmática congênita (HDC). Este estudo relata um novo método para medição contínua da pressão traqueal em um modelo cirúrgico de CDH de cordeiro fetal ocluído e não ocluído.

Vídeo: Medidas Telemétricas Contínuas de Pressão Traqueal In Utero em Cordeiros Fetais

A Mouse Model of in Utero Transplantation

The transplantation of stem cells and viruses in utero has tremendous potential for treating congenital disorders in the human fetus. For example, in utero transplantation (IUT) of hematopoietic stem cells has been used to successfully treat patients with severe combined immunodeficiency.1,2 In several other conditions, however, IUT has been attempted without success.3 Given these mixed results, the availability of an efficient non-human model to study the biological sequelae of stem cell transplantation and gene therapy is critical to advance this field. We and others have used the mouse model of IUT to study factors affecting successful engraftment of in utero transplanted hematopoietic stem cells in both wild-type mice4-7 and those with genetic diseases.8,9 The fetal environment also offers considerable advantages for the success of in utero gene therapy. For example, the delivery of adenoviral10, adeno-associated viral10, retroviral11, and lentiviral vectors12,13 into the fetus has resulted in the transduction of multiple organs distant from the site of injection with long-term gene expression. in utero gene therapy may therefore be considered as a possible treatment strategy for single gene disorders such as muscular dystrophy or cystic fibrosis. Another potential advantage of IUT is the ability to induce immune tolerance to a specific antigen. As seen in mice with hemophilia, the introduction of Factor IX early in development results in tolerance to this protein.14 
In addition to its use in investigating potential human therapies, the mouse model of IUT can be a powerful tool to study basic questions in developmental and stem cell biology. For example, one can deliver various small molecules to induce or inhibit specific gene expression at defined gestational stages and manipulate developmental pathways. The impact of these alterations can be assessed at various timepoints after the initial transplantation. Furthermore, one can transplant pluripotent or lineage specific progenitor cells into the fetal environment to study stem cell differentiation in a non-irradiated and unperturbed host environment.
The mouse model of IUT has already provided numerous insights within the fields of immunology, and developmental and stem cell biology. In this video-based protocol, we describe a step-by-step approach to performing IUT in mouse fetuses and outline the critical steps and potential pitfalls of this technique.

A Mouse Model of in Utero Transplantation

The mouse model of in utero transplantation is a versatile tool that can be used to study the potential clinical applications of stem cell transplantation and gene therapy in the fetus. In this protocol, we present a general approach to performing this technique

Um modelo do rato de No Utero Transplante

The transplantation of stem cells and viruses in utero has tremendous potential for treating congenital disorders in the human fetus.  For example, in ...

Medicina

In vitro Measurements of Tracheal Constriction Using Mice

Transgenic and knockout mice have been powerful tools for the investigation of the physiology and pathophysiology of airways1,2. In vitro tensometry of isolated tracheal preparations has proven to be a useful assay of airway smooth muscle (ASM) contractile response in genetically modified mice. These in vitro tracheal preparations are relatively simple, provide a robust response, and retain both functional cholinergic nerve endings and muscle responses, even after long incubations.
Tracheal tensometry also provides a functional assay to study a variety of second messenger signaling pathways that affect contraction of smooth muscle. Contraction in trachea is primarily mediated by parasympathetic, cholinergic nerves that release acetylcholine onto ASM (Figure 1). The major ASM acetylcholine receptors are muscarinic M2 and M3 which are Gi/o and Gq coupled receptors, respectively3,4,5. M3 receptors evoke contraction by coupling to Gq to activate phospholipase C, increase IP3 production and IP3-mediated calcium release from the sarcoplasmic reticulum3,6,7. M2/Gi/o signaling is believed to enhance contractions by inhibition of adenylate cyclase leading to a decrease in cAMP levels5,8,9,10. These pathways constitute the so called "pharmaco-contraction coupling" of airway smooth muscle11. In addition, cholinergic signaling through M2 receptors (and modulated by M3 signaling) involves pathways that depolarize the ASM which in turn activate L-type, voltage-dependent calcium channels (Figure 1) and calcium influx (so called "excitation-contraction coupling")4,7. More detailed reviews on signaling pathways controlling airway constriction can be found4,12. The above pathways appear to be conserved between mice and other species. However, mouse tracheas differ from other species in some signaling pathways. Most prominent is their lack of contractile response to histamine and adenosine13,14, both well-known ASM modulators in humans and other species5,15.
Here we present protocols for the isolation of murine tracheal rings and the in vitro measurement of their contractile output. Included are descriptions of the equipment configuration, trachea ring isolation and contractile measurements. Examples are given for evoking contractions indirectly using high potassium stimulation of nerves and directly by depolarization of ASM muscle to activate voltage-dependent calcium influx (1. high K+, Figure 1). In addition, methods are presented for stimulations of nerves alone using electric field stimulation (2. EFS, Figure 1), or for direct stimulation of ASM muscle using exogenous neurotransmitter applied to the bath (3. exogenous ACH, Figure 1). This flexibility and ease of preparation renders the isolated trachea ring model a robust and functional assay for a number of signaling cascades involved in airway smooth muscle contraction.

In vitro Measurements of Tracheal Constriction Using Mice

Transgenic mice have been extremely useful in ascribing physiological function to genes. As such, research in general, and functional studies of airway, in particular, have undergone a remarkable shift toward murine models. Here we provide protocols for in vitro trachea constriction studies to evaluate smooth muscle function in murine airway.

Em medições in vitro de constrição traqueal Utilizando camundongos

Transgenic and knockout mice have been powerful tools for the investigation of the physiology and pathophysiology of airways1,2. In vitro tensometry of ...

Assessing Teratogenic Changes in a Zebrafish Model of Fetal Alcohol Exposure

Fetal alcohol syndrome (FAS) is a severe manifestation of embryonic exposure to ethanol. It presents with characteristic defects to the face and organs, including mental retardation due to disordered and damaged brain development. Fetal alcohol spectrum disorder (FASD) is a term used to cover a continuum of birth defects that occur due to maternal alcohol consumption, and occurs in approximately 4% of children born in the United States. With 50% of child-bearing age women reporting consumption of alcohol, and half of all pregnancies being unplanned, unintentional exposure is a continuing issue2. In order to best understand the damage produced by ethanol, plus produce a model with which to test potential interventions, we developed a model of developmental ethanol exposure using the zebrafish embryo. Zebrafish are ideal for this kind of teratogen study3-8. Each pair lays hundreds of eggs, which can then be collected without harming the adult fish. The zebrafish embryo is transparent and can be readily imaged with any number of stains. Analysis of these embryos after exposure to ethanol at different doses and times of duration and application shows that the gross developmental defects produced by ethanol are consistent with the human birth defect. Described here are the basic techniques used to study and manipulate the zebrafish FAS model.

In order to understand the molecular mechanisms of the ethanol-induced developmental damage, we have developed a zebrafish model of ethanol exposure and are exploring the physical, cellular, and genetic alterations that occur after ethanol exposure1. We then seek to find potential interventions and rapidly test them in this animal model.

Avaliar as alterações teratogênicos em um modelo de Zebrafish de exposição fetal ao álcool

Fetal alcohol syndrome (FAS) is a severe manifestation of embryonic exposure to ethanol. It presents with characteristic defects to the face and organs, ...

Fetal Echocardiography and Pulsed-wave Doppler Ultrasound in a Rabbit Model of Intrauterine Growth Restriction

Fetal intrauterine growth restriction (IUGR) results in abnormal cardiac function that is apparent antenatally due to advances in fetoplacental Doppler ultrasound and fetal echocardiography. Increasingly, these imaging modalities are being employed clinically to examine cardiac function and assess wellbeing in utero, thereby guiding timing of birth decisions. Here, we used a rabbit model of IUGR that allows analysis of cardiac function in a clinically relevant way. Using isoflurane induced anesthesia, IUGR is surgically created at gestational age day 25 by performing a laparotomy, exposing the bicornuate uterus and then ligating 40-50% of uteroplacental vessels supplying each gestational sac in a single uterine horn. The other horn in the rabbit bicornuate uterus serves as internal control fetuses. Then, after recovery at gestational age day 30 (full term), the same rabbit undergoes examination of fetal cardiac function. Anesthesia is induced with ketamine and xylazine intramuscularly, then maintained by a continuous intravenous infusion of ketamine and xylazine to minimize iatrogenic effects on fetal cardiac function. A repeat laparotomy is performed to expose each gestational sac and a microultrasound examination (VisualSonics VEVO 2100) of fetal cardiac function is performed. Placental insufficiency is evident by a raised pulsatility index or an absent or reversed end diastolic flow of the umbilical artery Doppler waveform. The ductus venosus and middle cerebral artery Doppler is then examined. Fetal echocardiography is performed by recording B mode, M mode and flow velocity waveforms in lateral and apical views. Offline calculations determine standard M-mode cardiac variables, tricuspid and mitral annular plane systolic excursion, speckle tracking and strain analysis, modified myocardial performance index and vascular flow velocity waveforms of interest. This small animal model of IUGR therefore affords examination of in utero cardiac function that is consistent with current clinical practice and is therefore useful in a translational research setting.

We describe examination of fetal cardiac function with contemporary functional fetal echocardiography and fetoplacental Doppler ultrasound using the VisualSonics VEVO 2100 microultrasound in a surgically induced model of intrauterine fetal growth restriction in a rabbit.

Ecocardiografia fetal e ultra-som Doppler pulsátil em um modelo de coelho de restrição do crescimento intra-uterino

Fetal intrauterine growth restriction (IUGR) results in abnormal cardiac function that is apparent antenatally due to advances in fetoplacental Doppler ...

In utero Measurement of Heart Rate in Mouse by Noninvasive M-mode Echocardiography

Congenital heart disease (CHD) is the most frequent noninfectious cause of death at birth. The incidence of CHD ranges from 4 to 50/1,000 births (Disease and injury regional estimates, World Health Organization, 2004). Surgeries that often compromise the quality of life are required to correct heart defects, reminding us of the importance of finding the causes of CHD. Mutant mouse models and live imaging technology have become essential tools to study the etiology of this disease. Although advanced methods allow live imaging of abnormal hearts in embryos, the physiological and hemodynamic states of the latter are often compromised due to surgical and/or lengthy procedures. Noninvasive ultrasound imaging, however, can be used without surgically exposing the embryos, thereby maintaining their physiology. Herein, we use simple M-mode ultrasound to assess heart rates of embryos at E18.5 in utero. The detection of abnormal heart rates is indeed a good indicator of dysfunction of the heart and thus constitutes a first step in the identification of developmental defects that may lead to heart failure.

In utero Measurement of Heart Rate in Mouse by Noninvasive M-mode Echocardiography

Ultra-high frequency ultrasound is a powerful live imaging tool to examine cardiac abnormalities in small animals. Its noninvasiveness allows maintaining the physiologic state of embryos. Herein, we show the use of M-mode ultrasound to measure the heart rates of embryos at E18.5 in utero.

Na medição utero da frequência cardíaca em rato por não invasiva em modo M Ecocardiografia

Congenital heart disease (CHD) is the most frequent noninfectious cause of death at birth. The incidence of CHD ranges from 4 to 50/1,000 births (Disease ...

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. Despite the discovery of key regulators of uterine contractility, this process is still not fully understood. Transgenic mice provide an ideal model in which to study parturition. Previously, the only method to study uterine contractility in the mouse was ex vivo isometric tension recordings, which are suboptimal for several reasons. The uterus must be removed from its physiological environment, a limited time course of investigation is possible, and the mice must be sacrificed. The recent development of radiometric telemetry has allowed for longitudinal, real-time measurements of in vivo intrauterine pressure in mice. Here, the implantation of an intrauterine telemeter to measure pressure changes in the mouse uterus from mid-pregnancy until delivery is described. By comparing differences in pressures between wild type and transgenic mice, the physiological impact of a gene of interest can be elucidated. This technique should expedite the development of therapeutics used to treat myometrial disorders during pregnancy, including preterm labor.

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

This manuscript provides a protocol for implanting telemeters in the mouse for the purpose of measuring intrauterine pressures during pregnancy.

Telemetria Intrauterine medir Rato contrátil Pressão<em&gt; In Vivo</em

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. ...

Spatial Measurements of Perfusion, Interstitial Fluid Pressure and Liposomes Accumulation in Solid Tumors

The heterogeneous intra-tumoral accumulation of liposomes is a critical determinant of their efficacy. Both the chaotic tumor microcirculation and elevated IFP are linked to the heterogeneous intra-tumoral distribution of nanotechnology-based drug delivery systems such as liposomes. In the present study, the relationship between tumor microcirculation, elevated IFP, and accumulation of nanoparticles was investigated through in vivo experimentation. This was accomplished by evaluation of the tumor microcirculation using dynamic contrast enhanced computed tomography (DCE-CT) and measurement of tumor IFP using a novel image-guided robotic needle placement system connected to the micro-CT scanner. The intra-tumoral accumulation of liposomes was determined by CT image-based assessment of a nanoparticle liposomal formulation that stably encapsulate the contrast agent iohexol (CT-liposomes). CT imaging allowed for co-localization of the spatial distribution of tumor hemodynamics, IFP and CT-liposome accumulation in an individual subcutaneous xenograft mouse model of breast cancer. Measurements led to the discovery that perfusion and plasma volume fraction are strong mediators of the intra-tumoral distribution of liposomes. Furthermore, the results suggest that IFP plays an indirect role in mediating liposome distribution through modulating blood flow.

The heterogeneous intra-tumoral accumulation of liposomes has been linked to an abnormal tumor microenvironment. Herein methods are presented to measure tumor microcirculation by perfusion imaging and elevated interstitial fluid pressure (IFP) using an image-guided robotic system. Measurements are compared to the intra-tumoral accumulation of liposomes, determined using volumetric micro-CT imaging.

Medidas espaciais de Perfusão, intersticial pressão do produto e lipossomas Acumulação em tumores sólidos

The heterogeneous intra-tumoral accumulation of liposomes is a critical determinant of their efficacy. Both the chaotic tumor microcirculation and ...

A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the luminal surface area in arteries thereby reducing adequate blood flow to organs and distal tissues. Clinically, revascularization procedures such as balloon angioplasty with or without stent placement aim to restore blood flow. Although these procedures reestablish blood flow by reducing plaque burden, they damage the vessel wall, which initiates the arterial healing response. The prolonged healing response causes arterial restenosis, or re-narrowing, ultimately limiting the long-term success of these revascularization procedures. Therefore, preclinical animal models are integral for analyzing the pathophysiological mechanisms driving restenosis, and provide the opportunity to test novel therapeutic strategies. Murine models are cheaper and easier to operate on than large animal models. Balloon or wire injury are the two commonly accepted injury modalities used in murine models. Balloon injury models in particular mimic the clinical angioplasty procedure and cause adequate damage to the artery for the development of restenosis. Herein we describe the surgical details for performing and histologically analyzing the modified, pressure-controlled rat carotid artery balloon injury model. Additionally, this protocol highlights how local periadventitial application of therapeutics can be used to inhibit neointimal hyperplasia. Lastly, we present light sheet fluorescence microscopy as a novel approach for imaging and visualizing the arterial injury in three-dimensions.

The rat carotid artery balloon injury mimics the clinical angioplasty procedure performed to restore blood flow in atherosclerotic vessels. This model induces the arterial injury response by distending the arterial wall, and denuding the intimal layer of endothelial cells, ultimately causing remodeling and an intimal hyperplastic response.

Uma lesão de balão segmental controlada por pressão da artéria de rato com aplicação terapêutica periadventicional

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the ...

Using a Chemical Biopsy for Graft Quality Assessment

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

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

Usando uma biópsia química para avaliação da qualidade do enxerto

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

On-Site Sampling and Extraction of Brain Tumors for Metabolomics and Lipidomics Analysis

Despite the variety of tools available for cancer diagnosis and classification, methods that enable fast and simple characterization of tumors are still in need. In recent years, mass spectrometry has become a method of choice for untargeted profiling of discriminatory compound as potential biomarkers of a disease. Biofluids are generally considered as preferable matrices given their accessibility and easier sample processing while direct tissue profiling provides more selective information about a given cancer. Preparation of tissues for the analysis via traditional methods is much more complex and time-consuming, and, therefore, not suitable for fast on-site analysis. The current work presents a protocol combining sample preparation and extraction of small molecules on-site, immediately after tumor resection. The sampling device, which is of the size of an acupuncture needle, can be inserted directly into the tissue and then transported to the nearby laboratory for instrumental analysis. The results of metabolomics and lipidomics analyses demonstrate the capability of the approach for the establishment of phenotypes of tumors related to the histological origin of the tumor, malignancy, and genetic mutations, as well as for the selection of discriminating compounds or potential biomarkers. The non-destructive nature of the technique permits subsequent performance of routinely used tests e.g., histological tests, on the same samples used for SPME analysis, thus enabling attainment of more comprehensive information to support personalized diagnostics.

The manuscript presents on-site sampling of human brain tumors with solid phase microextraction followed by their biochemical profiling towards the discovery of biomarkers.

Amostragem e Extração de Tumores Cerebrais no local para análise metabolômica e lipidômica

Despite the variety of tools available for cancer diagnosis and classification, methods that enable fast and simple characterization of tumors are still ...