Name: surgical placement of catheters for long-term cardiovascular exercise testing in swine
Uploaded: 2016-02-09T15:00:00
Description: Watch this Scientific Journal Video about Surgical Placement of Catheters for Long-term Cardiovascular Exercise Testing in Swine at JoVE.com

This study was supported by Netherlands Heart Foundation grant 2000T038 (to D.J. Duncker) grant 2000T042 (to D. Merkus), European Commission FP7-Health-2010 grant MEDIA-261409 (to D.J. Duncker and D. Merkus), Netherlands CardioVascular Research Initiative: the Dutch Heart Foundation, the Dutch Federation for University Medical Centers, the Netherlands Organisation for Health Research and Development and the Royal Netherlands Academy of Sciences &#160;CVON- ARENA CVON 2011-11 (to D.J. Duncker), CVON-PHAEDRA CVON2012-08 ( to D.&#160;Merkus) &#160;and CVON-RECONNECT CVON 2014-11 (to D.J. Duncker and D. Merkus), Sophia Foundation (to D. de Wijs-Meijler, D. Merkus and I.K.M. Reiss).

Procedures involving animal subjects have been approved by the Animal Care Committee at Erasmus Medical Center Rotterdam (NL). Swine with weights between 6 and 80 kg have been successfully instrumented using this protocol.
1. Adaptation of the Animals to Human Handling
<ol>
	<li>After arrival in the facility, house the animals solitarily but enable them to interact with each other.</li>
	<li>Accustomize swine to human handling and transportation from the animal facility to the experimental laboratory, by handlin...

<ol>
	<li>Laughlin, M. H., 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=Peripheral+circulation.">Peripheral circulation.</a> Compr Physiol. 2, 321-447 (2012).</li><li>Datta, D., Normandin, E., ZuWallack, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiopulmonary+exercise+testing+in+the+assessment+of+exertional+dyspnea.">Cardiopulmonary exercise testing in the assessment of exertional dyspnea.</a> Ann Thorac Med. 10, 77-86 (2015).</li><li>Vatner, S. F., Braunwald, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiovascular+control+mechanisms+in+the+conscious+state.">Cardiovascular control mechanisms in the conscious state.</a> N Engl J Med. 293, 970-976 (1975).</li><li>Duncker, D. J., Bache, R. J. <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+coronary+blood+flow+during+exercise.">Regulation of coronary blood flow during exercise.</a> Physiol Rev. 88, 1009-1086 (2008).</li><li>Tune, J. D., Gorman, M. W., Feigl, E. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Matching+coronary+blood+flow+to+myocardial+oxygen+consumption.">Matching coronary blood flow to myocardial oxygen consumption.</a> J Appl Physiol. 97 (1985), 404-415 (2004).</li><li>van den Heuvel, 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=Coronary+microvascular+dysfunction+in+a+porcine+model+of+early+atherosclerosis+and+diabetes.">Coronary microvascular dysfunction in a porcine model of early atherosclerosis and diabetes.</a> Am J Physiol Heart Circ Physiol. 302, H85-H94 (2012).</li><li>Zhou, Z., 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=Pulmonary+vasoconstrictor+influence+of+endothelin+in+exercising+swine+depends+critically+on+phosphodiesterase+5+activity.">Pulmonary vasoconstrictor influence of endothelin in exercising swine depends critically on phosphodiesterase 5 activity.</a> Am J Physiol Lung Cell Mol Physiol. 306, L442-L452 (2014).</li><li>Pereda, 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=Swine+model+of+chronic+postcapillary+pulmonary+hypertension+with+right+ventricular+remodeling:+long-term+characterization+by+cardiac+catheterization,+magnetic+resonance,+and+pathology.">Swine model of chronic postcapillary pulmonary hypertension with right ventricular remodeling: long-term characterization by cardiac catheterization, magnetic resonance, and pathology.</a> J Cardiovasc Transl Res. 7, 494-506 (2014).</li><li>Mercier, 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=Endothelin+A+receptor+blockade+improves+regression+of+flow-induced+pulmonary+vasculopathy+in+piglets.">Endothelin A receptor blockade improves regression of flow-induced pulmonary vasculopathy in piglets.</a> J Thorac Cardiovasc Surg. 140, 677-683 (2010).</li><li>Spinale, F. 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=Chronic+supraventricular+tachycardia+causes+ventricular+dysfunction+and+subendocardial+injury+in+swine.">Chronic supraventricular tachycardia causes ventricular dysfunction and subendocardial injury in swine.</a> Am J Physiol. 259, H218-H229 (1990).</li><li>Yarbrough, W. M., Spinale, F. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Large+animal+models+of+congestive+heart+failure:+a+critical+step+in+translating+basic+observations+into+clinical+applications.">Large animal models of congestive heart failure: a critical step in translating basic observations into clinical applications.</a> J Nucl Cardiol. 10, 77-86 (2003).</li><li>Duncker, D. J., Stubenitsky, R., Verdouw, P. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autonomic+control+of+vasomotion+in+the+porcine+coronary+circulation+during+treadmill+exercise:+evidence+for+feed-forward+beta-adrenergic+control.">Autonomic control of vasomotion in the porcine coronary circulation during treadmill exercise: evidence for feed-forward beta-adrenergic control.</a> Circ Res. 82, 1312-1322 (1998).</li><li>Stubenitsky, R., Verdouw, P. D., Duncker, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autonomic+control+of+cardiovascular+performance+and+whole+body+O2+delivery+and+utilization+in+swine+during+treadmill+exercise.">Autonomic control of cardiovascular performance and whole body O2 delivery and utilization in swine during treadmill exercise.</a> Cardiovasc Res. 39, 459-474 (1998).</li><li>Zhou, Z., 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=Phosphodiesterase-5+activity+exerts+a+coronary+vasoconstrictor+influence+in+awake+swine+that+is+mediated+in+part+via+an+increase+in+endothelin+production.">Phosphodiesterase-5 activity exerts a coronary vasoconstrictor influence in awake swine that is mediated in part via an increase in endothelin production.</a> Am J Physiol Heart Circ Physiol. 306, H918-H927 (2014).</li><li>Gross, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Animal Models in Cardiovascular Research. , (2009).</li><li>Merkus, D., Duncker, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perspectives:+Coronary+microvascular+dysfunction+in+post-infarct+remodelled+myocardium.">Perspectives: Coronary microvascular dysfunction in post-infarct remodelled myocardium.</a> Eur Heart J Suppl. 16, A74-A79 (2014).</li><li>de Beer, V. J., de Graaff, H. J., Hoekstra, M., Duncker, D. J., Merkus, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrated+control+of+pulmonary+vascular+tone+by+endothelin+and+angiotensin+II+in+exercising+swine+depends+on+gender.">Integrated control of pulmonary vascular tone by endothelin and angiotensin II in exercising swine depends on gender.</a> Am J Physiol Heart Circ Physiol. 298, H1976-H1985 (2010).</li><li>Lautt, W. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Resistance+or+conductance+for+expression+of+arterial+vascular+tone.">Resistance or conductance for expression of arterial vascular tone.</a> Microvasc Res. 37, 230-236 (1989).</li><li>Merkus, 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=Phosphodiesterase+5+inhibition-induced+coronary+vasodilation+is+reduced+after+myocardial+infarction.">Phosphodiesterase 5 inhibition-induced coronary vasodilation is reduced after myocardial infarction.</a> Am J Physiol Heart Circ Physiol. 304, H1370-H1381 (2013).</li><li>Heusch, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+paradox+of+alpha-adrenergic+coronary+vasoconstriction+revisited.">The paradox of alpha-adrenergic coronary vasoconstriction revisited.</a> J Mol Cell Card. 51, 16-23 (2011).</li><li>Merkus, D., Houweling, B., van den Meiracker, A. H., Boomsma, F., Duncker, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+endothelin+to+coronary+vasomotor+tone+is+abolished+after+myocardial+infarction.">Contribution of endothelin to coronary vasomotor tone is abolished after myocardial infarction.</a> Am J Physiol Heart Circ Physiol. 288, H871-H880 (2005).</li><li>Haitsma, D. 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=Minimal+impairment+of+myocardial+blood+flow+responses+to+exercise+in+the+remodeled+left+ventricle+early+after+myocardial+infarction,+despite+significant+hemodynamic+and+neurohumoral+alterations.">Minimal impairment of myocardial blood flow responses to exercise in the remodeled left ventricle early after myocardial infarction, despite significant hemodynamic and neurohumoral alterations.</a> Cardiovasc Res. 52, 417-428 (2001).</li><li>Bender, S. B., van Houwelingen, M. J., Merkus, D., Duncker, D. J., Laughlin, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+analysis+of+exercise-induced+enhancement+of+early-+and+late-systolic+retrograde+coronary+blood+flow.">Quantitative analysis of exercise-induced enhancement of early- and late-systolic retrograde coronary blood flow.</a> J Appl Physiol. 108 (3), 507-514 (2010).</li></ol>

The present study describes the surgery for chronical instrumentation of swine as well as the protocol for exercising the instrumented swine on a motor-driven treadmill while measuring hemodynamics and taking blood samples for measurement of oxygen content in arterial, mixed venous and coronary venous blood.
Critical Steps within the Protocol 
There are several critical steps within the protocol that start already during the intubation procedure. Thiopental (2.1.5) is a respiratory depres...

Adequate cardiopulmonary function is essential to supply the body with oxygen and nutrients, particularly during conditions of increased metabolic demand such as during exercise1. The cardiopulmonary response to exercise is characterized by a number of adaptations in cardiac function, i.e., an increase in heart rate, contractility and stroke volume, and microvascular function, i.e., vasodilation in the vascular beds supplying exercising muscles as well as in the pulmonary vasculature, and vasoconstriction in the vascular beds supplying the gastrointestinal system as well as inactive muscles1. Impaired exercise capacity is an early hallmark of cardiopulmonary dysfunction, and cardiopulmonary exercise testing is used as an effective method to delineate between cardiac dysfunction, vascular dysfunction and/ or pulmonary dysfunction in patients with impaired exercise capacity2. Early cardiopulmonary dysfunction is difficult to diagnose, particularly in animal models, as cardiopulmonary function is often measured invasively, requiring anesthesia, with many anesthetic agents possessing cardiodepressive properties3.
Chronic instrumentation allows for measurement of cardiopulmonary function in the awake state, and when the animals are fully adjusted to the laboratory conditions measurements can be obtained under quiet resting conditions without the effects of anesthesia and acute surgical trauma. Furthermore, when the animals are appropriately trained, measurements can also be obtained during graded treadmill exercise4,5. More specifically, left and right ventricular function can be assessed and related to myocardial perfusion, while regulation of vasomotor tone in the coronary, systemic and pulmonary microcirculation can be determined. The use of fluid-filled catheters allows measurement of pressure as well as taking blood samples without imposing additional stress on the animals. Another advantage of studying chronically instrumented animals is that cardiopulmonary exercise testing can be repeated allowing the use of an animal as its own control, either during disease development or during chronic treatment, thereby increasing statistical power and hence limiting the number of animals required for a study.
Cardiopulmonary anatomy of swine closely resembles that of humans and it is possible to induce various forms of cardiopulmonary disease, such as diabetes 6, myocardial infarction 7, pulmonary hypertension 8,9 and pacing-induced heart failure10,11. Moreover, the size of swine allows chronic instrumentation, and repeated blood sampling of sufficient quantity to analyze not only blood gases, but also to perform neurohumoral measurements and/or to search for biomarkers of disease.
This protocol describes the surgery used to chronically instrument swine as well as the protocol for exercising the swine on a motor-driven treadmill.

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			<td height="21" style="height:21px;width:496px;">3-way stopcocks</td>
			<td style="width:212px;">B. Braun</td>
			<td align="right" style="width:179px;">16496</td>
			<td style="width:167px;"></td>
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			<td height="21" style="height:21px;">Perfusor lines PVC (DEHP-free) 150cm/2.6ml&#160;</td>
			<td>B. Braun</td>
			<td align="right" style="width:179px;">8722960</td>
			<td>Used for fluid filled catheters</td>
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			<td height="21" style="height:21px;">&#8220;python &#8220; silicontubing</td>
			<td>Rubber BV</td>
			<td>1757 ID 1mm, OD 2mm</td>
			<td>Used for fluid filled catheters</td>
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			<td height="21" style="height:21px;width:496px;">Sodium Chloride 0.9%</td>
			<td style="width:212px;">Baxter</td>
			<td style="width:179px;">TKF7124</td>
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			<td height="21" style="height:21px;width:496px;">Suction device</td>
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			<td></td>
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			<td height="21" style="height:21px;">Slim-Line electrosurgical pencil with 2 buttons</td>
			<td>ERBE ELEKTROMEDIZIN GMBH</td>
			<td>20190-066</td>
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			<td>Vererinary Technics Int.</td>
			<td>11.02.47</td>
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			<td style="width:212px;">3M</td>
			<td style="width:179px;">1818FS</td>
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			<td height="21" style="height:21px;width:496px;">surgical hat</td>
			<td style="width:212px;">Klinidrape</td>
			<td align="right" style="width:179px;">621301</td>
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			<td>Molnlycke Health Care</td>
			<td align="right" style="width:179px;">97027809</td>
			<td>Surgical drape, gauze pads, syringes, beaker etc</td>
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			<td style="width:212px;">Alcon</td>
			<td style="width:179px;">288-28282-01</td>
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			<td>Meda Pharma BV</td>
			<td>RVG08939</td>
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			<td>Meda Pharma BV</td>
			<td>RVG01331</td>
			<td></td>
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			<td height="21" style="height:21px;width:496px;">Cuffed Endotracheal tube</td>
			<td style="width:212px;">Emdamed</td>
			<td style="width:179px;"></td>
			<td>size depends on animal size</td>
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			<td height="21" style="height:21px;width:496px;">Breathing filter Hyrdo therm 3HME</td>
			<td style="width:212px;">Intersurgical</td>
			<td align="right" style="width:179px;">1560000</td>
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			<td style="width:212px;">Kawe Germany</td>
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			<td height="21" style="height:21px;width:496px;">Manual resuscitator- Combibag</td>
			<td style="width:212px;">Weinmann</td>
			<td>6515-12-313-5596</td>
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			<td height="21" style="height:21px;width:496px;">Perivascular flow probe 3PS</td>
			<td style="width:212px;">Transonic</td>
			<td style="width:179px;"></td>
			<td>For coronary artery; Size 2.5-4 mm depending on animal size</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Confidence flow probe</td>
			<td style="width:212px;">Transonic</td>
			<td style="width:179px;"></td>
			<td>For aorta/pulmonary artery, 16-20 mm; size depends on animal size</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Venflon-Venisystem 20Gx 32 mm</td>
			<td style="width:212px;">BD</td>
			<td align="right" style="width:179px;">393224</td>
			<td>For coronary venous catheter</td>
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			<td height="21" style="height:21px;width:496px;">Blunt Needle 18G</td>
			<td style="width:212px;"></td>
			<td style="width:179px;"></td>
			<td>For coronary venous catheter</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Tygon Tubing</td>
			<td style="width:212px;">Rubber BV</td>
			<td>2802 ID 0.8mm (1/32&#8217;&#8217;), OD 2.4mm (3/32&#8217;&#8217;)</td>
			<td>For coronary venous catheter</td>
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		<tr height="21">
			<td height="21" style="height:21px;">Suction Handle 17 cm 6 6/8 &#34; Coupland 18/8 martinit with tube connector</td>
			<td>KLS Martin Group</td>
			<td>18-575-24</td>
			<td></td>
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			<td height="21" style="height:21px;width:496px;">Scalple blade&#160;</td>
			<td style="width:212px;"></td>
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			<td></td>
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			<td height="21" style="height:21px;">Scalpel Handle 13.5 cm 5 3/8 &#34; Stainless Steel solid</td>
			<td>KLS Martin Group</td>
			<td>10-100-04</td>
			<td></td>
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			<td height="21" style="height:21px;">Vascular Forceps 20.2 cm 8 &#34; De Bakey Stainless Stee</td>
			<td>KLS Martin Group</td>
			<td>24-388-20</td>
			<td>&#177; 14 cm</td>
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			<td height="21" style="height:21px;">Dressing Forceps 17 cm 6 6/8 &#34; Cushing Stainless Steel</td>
			<td>KLS Martin Group</td>
			<td>12-189-17</td>
			<td>&#177; 18 cm</td>
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			<td height="21" style="height:21px;">halsted-musquito straight 12.5cm - 5&#34;</td>
			<td>Rudolf Medical</td>
			<td>RU-3100-13</td>
			<td>&#177; 12 cm</td>
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			<td height="21" style="height:21px;">halsted-musquito curved 12.5cm - 5&#34;</td>
			<td>Rudolf Medical</td>
			<td>RU-3101-12</td>
			<td>&#177; 12 cm</td>
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			<td height="21" style="height:21px;">Dissecting and Ligature Forceps 13 cm 5 1/8 &#34; Gemini Stainless Steel</td>
			<td>KLS Martin Group</td>
			<td>13-451-13</td>
			<td>&#177; 12 cm</td>
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			<td height="21" style="height:21px;">Dissecting and Ligature Forceps 18.5 cm 7 2/8 &#34; Schnidt Stainless Steel</td>
			<td>KLS Martin Group</td>
			<td>13-363-18</td>
			<td></td>
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			<td height="21" style="height:21px;">Rib Retractor Finochietto, Baby Aluminium -</td>
			<td>KLS Martin Group</td>
			<td>24-162-01</td>
			<td></td>
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			<td height="21" style="height:21px;">suture forceps Mayo-Hegar 3mm 18cm - 7&#34;</td>
			<td>Rudolf Medical</td>
			<td>RU-6050-18</td>
			<td></td>
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			<td height="21" style="height:21px;">Metchenbaum blunt curved 14,5cm - 5(3/4)&#34;</td>
			<td>Rudolf Medical</td>
			<td>RU-1311-14M</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Retrector farabeuf 12cm - 4 (3/4)&#34;</td>
			<td>Rudolf Medical</td>
			<td>RU-4497-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Towel forceps schr&#228;del curved 9cm - 3,5&#34;</td>
			<td>Rudolf Medical</td>
			<td>RU-3550-09</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">surgical scissors blunt 13cm - 5&#34;</td>
			<td>Rudolf Medical</td>
			<td>RU-1001-13</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Gauzes Cutisoft 10 x 10 cm 4-ply</td>
			<td>BSN Medical</td>
			<td>45846-00</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Gauzes Cutisoft 5 x 5 cm 4-ply</td>
			<td>BSN Medical</td>
			<td>45844-00</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:496px;">Flowmeter -CM2 / SF2 - 2gas (O2 and Air)</td>
			<td>UNO BV</td>
			<td align="right" style="width:179px;">180000008</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tec 7 Vaporizer</td>
			<td>Datex-Ohmeda</td>
			<td style="width:179px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Acederm wound spay</td>
			<td style="width:212px;">Ecuphar NV</td>
			<td style="width:179px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Vaseline Album</td>
			<td style="width:212px;">Bufa</td>
			<td align="right" style="width:179px;">165313</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">silkam 3-0 Natural silk, non-absorbable</td>
			<td>B. Braun</td>
			<td>F 1134043</td>
			<td>sutures for placement of catheters</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">silkam 2-0 Natural silk, non-absorbable</td>
			<td>B. Braun</td>
			<td>F 1134051</td>
			<td>sutures for muscular approximation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">dagrofil 3-0 Polyester, non-absorbable</td>
			<td>B. Braun</td>
			<td>C 0842478</td>
			<td>sutures for fluid fille catheters after tunneling</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vicryl rapide 3-0, 1x45 cm FS2, V2930G</td>
			<td>Daxtrio medische producten</td>
			<td>15560</td>
			<td>sutures for electrical catheters after tunneling</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Vitafil 6 USP</td>
			<td style="width:212px;">SMI</td>
			<td align="right" style="width:179px;">6080</td>
			<td>Ties</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Syringes</td>
			<td style="width:212px;"></td>
			<td style="width:179px;"></td>
			<td>10 ml and 2.5 ml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Heparin LEO (heparin sodium)&#160;</td>
			<td>LEO Pharma A/S</td>
			<td style="width:179px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Zoletil</td>
			<td style="width:212px;">Virbac</td>
			<td style="width:179px;"></td>
			<td>tiletamine / zolazepam</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Sedazine</td>
			<td style="width:212px;">AST farma</td>
			<td align="right" style="width:179px;">108855</td>
			<td>xylazine</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Temgesic</td>
			<td style="width:212px;">RB Pharmaceuticals</td>
			<td align="right" style="width:179px;">5429</td>
			<td>buprenorphine</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:496px;">Tensogrip</td>
			<td style="width:212px;">BSN Medical</td>
			<td style="width:179px;">71522-00</td>
			<td>elastic vest</td>
		</tr>
	</tbody>
</table>

surgical placement of catheters for long-term cardiovascular exercise testing in swine

This protocol describes the surgical procedure to chronically instrument swine and the procedure to exercise swine on a motor-driven treadmill. Early cardiopulmonary dysfunction is difficult to diagnose, particularly in animal models, as cardiopulmonary function is often measured invasively, requiring anesthesia. As many anesthetic agents are cardiodepressive, subtle changes in cardiovascular function may be masked. In contrast, chronic instrumentation allows for measurement of cardiopulmonary function in the awake state, so that measurements can be obtained under quiet resting conditions, without the effects of anesthesia and acute surgical trauma. Furthermore, when animals are properly trained, measurements can also be obtained during graded treadmill exercise.
Flow probes are placed around the aorta or pulmonary artery for measurement of cardiac output and around the left anterior descending coronary artery for measurement of coronary blood flow. Fluid-filled catheters are implanted in the aorta, pulmonary artery, left atrium, left ventricle and right ventricle for pressure measurement and blood sampling. In addition, a 20 G&#160;catheter is positioned in the anterior interventricular vein to allow coronary venous blood sampling.
After a week of recovery, swine are placed on a motor-driven treadmill, the catheters are connected to pressure and flow meters, and swine are subjected to a five-stage progressive exercise protocol, with each stage lasting 3 min. Hemodynamic signals are continuously recorded and blood samples are taken during the last 30 sec of each exercise stage.
The major advantage of studying chronically instrumented animals is that it allows serial assessment of cardiopulmonary function, not only at rest but also during physical stress such as exercise. Moreover, cardiopulmonary function can be assessed repeatedly during disease development and during chronic treatment, thereby increasing statistical power and hence limiting the number of animals required for a study.

Exercise up to 5 km/hr resulted in a doubling of cardiac output from 4.3 &#177; 0.3 to 8.5 &#177; 0.7 L/min which was principally accomplished by an increase in heart rate from 137 &#177; 7 to 256 &#177; 8 beats per min in combination with a small increase in stroke volume from 32 &#177; 2 to 36 &#177; 3 ml (Figure 3). The increase in stroke volume was facilitated by an increase in left ventricular contractility, as evidenced by an increase in the maximum of the first der...

Watch this Scientific Journal Video about Surgical Placement of Catheters for Long-term Cardiovascular Exercise Testing in Swine at JoVE.com

Surgical Placement of Catheters for Long-term Cardiovascular Exercise Testing in Swine

Here we present a protocol to assess cardiopulmonary function in awake swine, at rest and during graded treadmill exercise. Chronic instrumentation allows for repeated hemodynamic measurements uninfluenced by cardiodepressive anesthetic agents.

This protocol describes the surgical procedure to chronically instrument swine and the procedure to exercise swine on a motor-driven treadmill. Early ...

The overall goal of this surgery is to chronically instrument animals to allow for hemodynamic measurements during cardiopulmonary stress testing in alert animals with and without cardiovascular disease. This method can help answer key questions in the cardiovascular field, such as how metabolic syndrome affects left ventricular function and perfusion therby contributing to the development of heart failure with preserved ejection fraction. Two key advantages of this technique are that hemodynamic measurements and blood samples are obtained from awake animals and the technique can be performed at rest and during cardiovascular stress such as exercise.The implications of this technique can extend towards early diagnosis of pulmonary hypertension because pulmonary microvascular function, as well as right ventricular function and perfusion can be measured at well defined stages after introduction of the disease. Visual demonstration of this technique is essential. It is difficult to learn how to place a flow probe around a coronary artery due to vasospasm of this vessel.Also, placing the flow probe around aorta is difficult due to location of this vessel with respect to the thoracotomy. Demonstrating the procedure will be Annemarie Verzijl, a technician from our laboratory. After preparing the animal for surgery, make an incision in the skin, starting one centimeter caudal to the left inferior angle of the scapula and cut downward to the left axilla.Following the incision, use diathermy to cauterize blood vessels in the skin. Next, using the cutting modality of the diathermy, cut though the serratus muscle and pectoralis major muscle. Continue using diathermy to cauterize blood vessels in the muscle layer.Now, using blunt dissection, carefully divide the intercostal muscle of the 4th left intercostal space. Finish this using a mosquito clamp. Then expose the costal surface of the left lung, which is covered with visceral and parietal pleura.Next, enter the pleural cavity by carefully piercing both layers of the pleura and tearing them open. Improve the exposure with a thoracic retractor by separting the edges of the wound and ribs then forcefully pushing the tissues apart. To expose the heart and great vessels, push the left lung caudally and wedge it in place with wet gauze.Begin this portion of the surgery with a blunt dissection to remove about 2 square centimeters of connective tissue surrounding the descending thoracic aorta. Next, place a three stitch purse string suture in the aortic wall using non-absorbable USP 3-0 braided silk suture. Then penetrate the aortic vessel wall through the suture with a stainless steel, 16-gauge needle.Following the needle, insert the tip of the fluid-filled catheter into the aorta. Then pull the purse string suture firmly together and tie it off. To secure the catheter, wind the excess suture around the catheter above the ring three times, tie it off, and add a new stitch about 1 centimeter cranial to its insertion site.Now connect the fluid-filled catheter to the calibrated pressure transducer connected to a computer. Thus, monitor the mean arterial pressure during the subsequent steps. Next, without damaging the phrenic nerve, open the pericardium with a crossed cut.Beneath the cut, expose the ascending aorta and aortic arch with a Farabeuf retractor by identifying the pulmonary artery and gently pulling it caudally. With the aorta exposed, make a 1 centimeter cut in the connective tissue between the ascending aorta and the pulmonary artery using Metzenbaum scissors. Now attach the flow probe.first get the rubber band around the vessel by using a suture lead. And secondly, attach the flow probe measurement device to the rubber band. Thirdly, connect the flow probe to the computer and fourthly, check the cardiac output signal to confirm the probe is properly positioned.The next step is to expose and dissect the proximal part of the left anterior descending coronary artery. First, lift the tissue with forceps. And then, using Metzenbaum scissors, make a small cut in the tissue.Then carefully tease away tissues from the artery using a cotton swab. And to ensure the coronary artery is completely dissected, pass a small, straight-angled mosquito clamp underneath it. When vasospasm of the coronary artery occurs, spray with 10%lidocane to relax the vessel.Using the same technique, place fluid-filled catheters in the pulmonary artery, right ventricle, left ventricle, and left atrium. All the catheters should now be connected to the computer. Then make a stitch parallel to the anterior interventricular coronary vein with a suture connected to the coronary venous catheter.Following the stitch, puncture the vein with the 20-gauge needle of the coronary venous catheter and insert the cannula of the catheter intravenously. Then secure the catheter with the existing stitch. Remove the needle and connect to the extension line.Now place a coronary flow probe around the previously dissected left anterior descending coronary artery. With the probe placed, check the signal of the coronary flow on the computer to confirm that the flow probe is correctly placed. The shape of the coronary flow signal should be similar to that shown here.Begin this phase of the surgery by making a 1.5 centimeter incision about 8 centimeters caudal and parallel to the first incision. Then using a large, curved mosquito clamp, subcutaneously lead the drain from the pleural cavity through the 6th intercostal space to this incision. Connect the drain to the suction device to remove remaining fluid from the thorax.Next, use a large, curved mosquito clamp to tunnel the flow probes individually through the 3rd left intercostal space and the muscle above the rib. Tunnel the fluid-filled catheters through either the 3rd or the 5th left intercostal space. Firstly, clamp off the fluid-filled catheters.Next, remove the three-way stopcock to minimize the piercing area. And thirdly, pierce the intercostal muscle. Now, fix the flow probes and the fluid-filled catheters with non-absorbable USP 2-0 braided silk.Use a purse string suture on the intercostal muscle. This sutre will also prevent air leakage after reinstating negative intrathoracic pressure. Next, make three incisions in the skin about 2 centimeters to the left and parallel to the vertebral column.Make these three incisions 3 centimeters long and 3 centimeters apart from each other. Now pierce a trocar beneath the left latissimus dorsi muscle from the rostral incision site to the incisions on the back. Then using the trocar, tunnel the flow probes and fluid catheters to the back.Once tunneling is complete, place stopcocks on the fluid-filled catheters and remove the clamp. Withdraw some blood to remove the clots and air bubbles. Then fill the fluid-filled catheters with heparin at 1, 000 units per milliliter, and fill the coronary venous catheters with heparin at 5, 000 units per milliliter.Close the thorax by pulling together the ribs of the 4th intercostal space at two separate sites. Use non-absorbable USP 6 braided polyester. Then close the serratus muscle and pectoralis major muscle with the running stitch.Close the skin with the running subcuticular suture using non-absorbable USP 2-0 braided silk. Then sutre the incisions on the dorsal side between the catheters. First, tie a knot directly onto the skin to close the incision.Then fixate the catheters to the suture with another knot about 1 centimeter from the skin. Suture the flow probes with polyglactin suture so that the flow probe wire is not cut by the suture. Then carefully remove the drain while applying pressure to the cranial side of the incsion so as to reinstate negative pressure in the pleural cavity.Now complete the surgery by closing the incision with a purse string suture and sealing the wound with petroleum jelly. Follow the text protocol on how to recover the animal and how to conduct the treadmill experiment. Exercising swine to move up to 5 kilometers per hour resulted in a doubling of cardiac output.A tripling of body oxygen consumption was observed. This was due to the increase in cardiac output coupled with an increase in hemoglobin concentration and an increase in body oxygen extraction. This was principally accomplished by an increase in heart rate with a small increase in stroke volume.The increase in stroke volume was facilitated by an increase in left ventricular contractility as evidenced by an increase in the maximum of the 1st derivative of left ventricular pressure together with an increased rate of relaxation of the left ventricle and an increase in left atrial pressure, which is the filling pressure of the left ventricle. Systemic vasodilation occurred as evidenced by an increase in systemic vascular conductance and a decrease in systemic vascular resistance. This accommodated the increase in cardiac output almost completely so that mean aortic pressure increased only slightly.Further interpretations of several other parameters are provided in the text protocol. After watching this video, you should have a good understanding of how to chronically instrument animals to make hemodynamic measurements in subsequent cardiovascular experiments.

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