OcrV1, PascalFrancis, Curation, bibRecord, 000660

Assessment of Tissue-Engineered Islet Graft Viability by Fluorine Magnetic Resonance Spectroscopy

Identifieur interne : 000660 ( PascalFrancis/Curation ); précédent : 000659; suivant : 000661

Assessment of Tissue-Engineered Islet Graft Viability by Fluorine Magnetic Resonance Spectroscopy

Auteurs : T. M. Suszynski [États-Unis] ; E. S. Avgoustiniatos [États-Unis] ; S. A. Stein [États-Unis] ; E. J. Falde [États-Unis] ; B. E. Hammer [États-Unis] ; K. K. Papas [États-Unis]

Source :

Transplantation proceedings [ 0041-1345 ] ; 2011.

RBID : Pascal:12-0017463

Descripteurs français

Pascal (Inist)
- Greffe, Culture tissu, Génie tissulaire, Spectrométrie RMN, Ilot Langerhans, Viabilité, Fluor, Médecine, Transplantation, Traitement.
Wicri :
- topic : Fluor, Médecine.

English descriptors

KwdEn :
- Fluorine, Graft, Langerhans islet, Medicine, NMR spectrometry, Tissue culture, Tissue engineering, Transplantation, Treatment, Viability.

Abstract

Introduction. Despite significant progress in the last decade, islet transplantation remains an experimental therapy for a limited number of patients with type 1 diabetes. Tissue-engineered approaches may provide promising alternatives to the current clinical protocol and would benefit greatly from concurrent development of graft quality assessment techniques. This study was designed to evaluate whether viability of tissue-engineered islet grafts can be assessed using fluorine magnetic resonance spectroscopy (¹⁹F-MRS), by the noninvasive measurement of oxygen partial pressure (pO₂) and the subsequent calculation of islet oxygen consumption rate (OCR). Methods. Scaffolds composed of porcine plasma were seeded with human islets and perfluorodecalin. Each graft was covered with the same volume of culture media in a Petri dish. Four scaffolds were seeded with various numbers (0-8000) of islet equivalents (IE) aliquoted from the same preparation. After randomizing run order, grafts were examined by ¹⁹F-MRS at 37°C using a 5T spectrometer and a single-loop surface coil placed underneath. A standard inversion recovery sequence was used to obtain characteristic ¹⁹F spin-lattice relaxation times (T1), which were converted to steady-state average pO₂ estimates using a previously determined linear calibration (R² = 1.000). Each condition was assessed using replicate ¹⁹F-MRS measurements (n = 6-8). Results. Grafts exhibited IE dose-dependent increases in T1 and decreases in pO₂ estimates. From the difference between scaffold pO₂ estimates and ambient pO₂, the islet preparation OCR was calculated to be 95 ± 12 (mean ± standard error of the mean) nmol/(min . mg DNA) using theoretical modeling. This value compared well with OCR values measured using established methods for human islet preparations. Conclusions. ¹⁹F-MRS can be used for noninvasive pre- and possibly posttransplant assessment of tissue-engineered islet graft viability by estimating the amount of viable, oxygen-consuming tissue in a scaffold.

A01	`01`	`1`		`@0 0041-1345`
A02	`01`			`@0 TRPPA8`
A03		`1`		`@0 Transplant. proc.`
A05				`@2 43`
A06				`@2 9`
A08	`01`	`1`	`ENG`	`@1 Assessment of Tissue-Engineered Islet Graft Viability by Fluorine Magnetic Resonance Spectroscopy`
A11	`01`	`1`		`@1 SUSZYNSKI (T. M.)`
A11	`02`	`1`		`@1 AVGOUSTINIATOS (E. S.)`
A11	`03`	`1`		`@1 STEIN (S. A.)`
A11	`04`	`1`		`@1 FALDE (E. J.)`
A11	`05`	`1`		`@1 HAMMER (B. E.)`
A11	`06`	`1`		`@1 PAPAS (K. K)`
A14	`01`			`@1 Schulze Diabetes Institute, Department of Surgery, University of Minnesota @2 Minneapolis, Minnesota @3 USA @Z 1 aut. @Z 2 aut. @Z 3 aut. @Z 4 aut. @Z 6 aut.`
A14	`02`			`@1 Center for Interdisciplinary Applications in Magnetic Resonance, Department of Radiology, University of Minnesota @2 Minneapolis, Minnesota @3 USA @Z 5 aut.`
A20				`@1 3221-3225`
A21				`@1 2011`
A23	`01`			`@0 ENG`
A43	`01`			`@1 INIST @2 14765 @5 354000507348890180`
A44				`@0 0000 @1 © 2012 INIST-CNRS. All rights reserved.`
A45				`@0 38 ref.`
A47	`01`	`1`		`@0 12-0017463`
A60				`@1 P @2 C`
A61				`@0 A`
A64	`01`	`1`		`@0 Transplantation proceedings`
A66	`01`			`@0 NLD`
C01	`01`		`ENG`	@0 Introduction. Despite significant progress in the last decade, islet transplantation remains an experimental therapy for a limited number of patients with type 1 diabetes. Tissue-engineered approaches may provide promising alternatives to the current clinical protocol and would benefit greatly from concurrent development of graft quality assessment techniques. This study was designed to evaluate whether viability of tissue-engineered islet grafts can be assessed using fluorine magnetic resonance spectroscopy (¹⁹F-MRS), by the noninvasive measurement of oxygen partial pressure (pO₂) and the subsequent calculation of islet oxygen consumption rate (OCR). Methods. Scaffolds composed of porcine plasma were seeded with human islets and perfluorodecalin. Each graft was covered with the same volume of culture media in a Petri dish. Four scaffolds were seeded with various numbers (0-8000) of islet equivalents (IE) aliquoted from the same preparation. After randomizing run order, grafts were examined by ¹⁹F-MRS at 37°C using a 5T spectrometer and a single-loop surface coil placed underneath. A standard inversion recovery sequence was used to obtain characteristic ¹⁹F spin-lattice relaxation times (T1), which were converted to steady-state average pO₂ estimates using a previously determined linear calibration (R² = 1.000). Each condition was assessed using replicate ¹⁹F-MRS measurements (n = 6-8). Results. Grafts exhibited IE dose-dependent increases in T1 and decreases in pO₂ estimates. From the difference between scaffold pO₂ estimates and ambient pO₂, the islet preparation OCR was calculated to be 95 ± 12 (mean ± standard error of the mean) nmol/(min . mg DNA) using theoretical modeling. This value compared well with OCR values measured using established methods for human islet preparations. Conclusions. ¹⁹F-MRS can be used for noninvasive pre- and possibly posttransplant assessment of tissue-engineered islet graft viability by estimating the amount of viable, oxygen-consuming tissue in a scaffold.
C02	`01`	`X`		`@0 002B25`
C02	`02`	`X`		`@0 002A06F`
C03	`01`	`X`	`FRE`	`@0 Greffe @5 01`
C03	`01`	`X`	`ENG`	`@0 Graft @5 01`
C03	`01`	`X`	`SPA`	`@0 Injerto @5 01`
C03	`02`	`X`	`FRE`	`@0 Culture tissu @5 02`
C03	`02`	`X`	`ENG`	`@0 Tissue culture @5 02`
C03	`02`	`X`	`SPA`	`@0 Cultivo tejido @5 02`
C03	`03`	`X`	`FRE`	`@0 Génie tissulaire @5 03`
C03	`03`	`X`	`ENG`	`@0 Tissue engineering @5 03`
C03	`03`	`X`	`SPA`	`@0 Ingeniería de tejidos @5 03`
C03	`04`	`X`	`FRE`	`@0 Spectrométrie RMN @5 04`
C03	`04`	`X`	`ENG`	`@0 NMR spectrometry @5 04`
C03	`04`	`X`	`SPA`	`@0 Espectrometría RMN @5 04`
C03	`05`	`X`	`FRE`	`@0 Ilot Langerhans @5 05`
C03	`05`	`X`	`ENG`	`@0 Langerhans islet @5 05`
C03	`05`	`X`	`SPA`	`@0 Isla Langerhans @5 05`
C03	`06`	`X`	`FRE`	`@0 Viabilité @5 06`
C03	`06`	`X`	`ENG`	`@0 Viability @5 06`
C03	`06`	`X`	`SPA`	`@0 Viabilidad @5 06`
C03	`07`	`X`	`FRE`	`@0 Fluor @2 NC @2 FX @5 08`
C03	`07`	`X`	`ENG`	`@0 Fluorine @2 NC @2 FX @5 08`
C03	`07`	`X`	`SPA`	`@0 Fluor @2 NC @2 FX @5 08`
C03	`08`	`X`	`FRE`	`@0 Médecine @5 09`
C03	`08`	`X`	`ENG`	`@0 Medicine @5 09`
C03	`08`	`X`	`SPA`	`@0 Medicina @5 09`
C03	`09`	`X`	`FRE`	`@0 Transplantation @5 11`
C03	`09`	`X`	`ENG`	`@0 Transplantation @5 11`
C03	`09`	`X`	`SPA`	`@0 Trasplantación @5 11`
C03	`10`	`X`	`FRE`	`@0 Traitement @5 25`
C03	`10`	`X`	`ENG`	`@0 Treatment @5 25`
C03	`10`	`X`	`SPA`	`@0 Tratamiento @5 25`
C07	`01`	`X`	`FRE`	`@0 Pancréas endocrine @5 37`
C07	`01`	`X`	`ENG`	`@0 Endocrine pancreas @5 37`
C07	`01`	`X`	`SPA`	`@0 Páncreas endocrino @5 37`
C07	`02`	`X`	`FRE`	`@0 Chirurgie @5 38`
C07	`02`	`X`	`ENG`	`@0 Surgery @5 38`
C07	`02`	`X`	`SPA`	`@0 Cirugía @5 38`
N21				`@1 002`
N44	`01`			`@1 OTO`
N82				`@1 OTO`

A30	`01`	`1`	`ENG`	`@1 World Congress of the International Pancreas and Islet Transplant Association (IPITA) @3 CZE @4 2011-06-01`

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Le document en format XML

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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Assessment of Tissue-Engineered Islet Graft Viability by Fluorine Magnetic Resonance Spectroscopy</title>
<author><name sortKey="Suszynski, T M" sort="Suszynski, T M" uniqKey="Suszynski T" first="T. M." last="Suszynski">T. M. Suszynski</name>
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<country>États-Unis</country>
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<author><name sortKey="Avgoustiniatos, E S" sort="Avgoustiniatos, E S" uniqKey="Avgoustiniatos E" first="E. S." last="Avgoustiniatos">E. S. Avgoustiniatos</name>
<affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Schulze Diabetes Institute, Department of Surgery, University of Minnesota</s1>
<s2>Minneapolis, Minnesota</s2>
<s3>USA</s3>
<sZ>1 aut.</sZ>
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<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
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<country>États-Unis</country>
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<author><name sortKey="Stein, S A" sort="Stein, S A" uniqKey="Stein S" first="S. A." last="Stein">S. A. Stein</name>
<affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Schulze Diabetes Institute, Department of Surgery, University of Minnesota</s1>
<s2>Minneapolis, Minnesota</s2>
<s3>USA</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>6 aut.</sZ>
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<country>États-Unis</country>
</affiliation>
</author>
<author><name sortKey="Falde, E J" sort="Falde, E J" uniqKey="Falde E" first="E. J." last="Falde">E. J. Falde</name>
<affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Schulze Diabetes Institute, Department of Surgery, University of Minnesota</s1>
<s2>Minneapolis, Minnesota</s2>
<s3>USA</s3>
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<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
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<s2>Minneapolis, Minnesota</s2>
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<author><name sortKey="Papas, K K" sort="Papas, K K" uniqKey="Papas K" first="K. K" last="Papas">K. K. Papas</name>
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<s3>USA</s3>
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<sZ>6 aut.</sZ>
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</affiliation>
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<series><title level="j" type="main">Transplantation proceedings</title>
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<idno type="ISSN">0041-1345</idno>
<imprint><date when="2011">2011</date>
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<idno type="ISSN">0041-1345</idno>
</seriesStmt>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Fluorine</term>
<term>Graft</term>
<term>Langerhans islet</term>
<term>Medicine</term>
<term>NMR spectrometry</term>
<term>Tissue culture</term>
<term>Tissue engineering</term>
<term>Transplantation</term>
<term>Treatment</term>
<term>Viability</term>
</keywords>
<keywords scheme="Pascal" xml:lang="fr"><term>Greffe</term>
<term>Culture tissu</term>
<term>Génie tissulaire</term>
<term>Spectrométrie RMN</term>
<term>Ilot Langerhans</term>
<term>Viabilité</term>
<term>Fluor</term>
<term>Médecine</term>
<term>Transplantation</term>
<term>Traitement</term>
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<term>Médecine</term>
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<front><div type="abstract" xml:lang="en">Introduction. Despite significant progress in the last decade, islet transplantation remains an experimental therapy for a limited number of patients with type 1 diabetes. Tissue-engineered approaches may provide promising alternatives to the current clinical protocol and would benefit greatly from concurrent development of graft quality assessment techniques. This study was designed to evaluate whether viability of tissue-engineered islet grafts can be assessed using fluorine magnetic resonance spectroscopy (<sup>19</sup>
F-MRS), by the noninvasive measurement of oxygen partial pressure (pO<sub>2</sub>
) and the subsequent calculation of islet oxygen consumption rate (OCR). Methods. Scaffolds composed of porcine plasma were seeded with human islets and perfluorodecalin. Each graft was covered with the same volume of culture media in a Petri dish. Four scaffolds were seeded with various numbers (0-8000) of islet equivalents (IE) aliquoted from the same preparation. After randomizing run order, grafts were examined by <sup>19</sup>
F-MRS at 37°C using a 5T spectrometer and a single-loop surface coil placed underneath. A standard inversion recovery sequence was used to obtain characteristic <sup>19</sup>
F spin-lattice relaxation times (T1), which were converted to steady-state average pO<sub>2</sub>
 estimates using a previously determined linear calibration (R<sup>2</sup>
 = 1.000). Each condition was assessed using replicate <sup>19</sup>
F-MRS measurements (n = 6-8). Results. Grafts exhibited IE dose-dependent increases in T1 and decreases in pO<sub>2</sub>
 estimates. From the difference between scaffold pO<sub>2</sub>
 estimates and ambient pO<sub>2</sub>
, the islet preparation OCR was calculated to be 95 ± 12 (mean ± standard error of the mean) nmol/(min . mg DNA) using theoretical modeling. This value compared well with OCR values measured using established methods for human islet preparations. Conclusions. <sup>19</sup>
F-MRS can be used for noninvasive pre- and possibly posttransplant assessment of tissue-engineered islet graft viability by estimating the amount of viable, oxygen-consuming tissue in a scaffold.</div>
</front>
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<fA08 i1="01" i2="1" l="ENG"><s1>Assessment of Tissue-Engineered Islet Graft Viability by Fluorine Magnetic Resonance Spectroscopy</s1>
</fA08>
<fA11 i1="01" i2="1"><s1>SUSZYNSKI (T. M.)</s1>
</fA11>
<fA11 i1="02" i2="1"><s1>AVGOUSTINIATOS (E. S.)</s1>
</fA11>
<fA11 i1="03" i2="1"><s1>STEIN (S. A.)</s1>
</fA11>
<fA11 i1="04" i2="1"><s1>FALDE (E. J.)</s1>
</fA11>
<fA11 i1="05" i2="1"><s1>HAMMER (B. E.)</s1>
</fA11>
<fA11 i1="06" i2="1"><s1>PAPAS (K. K)</s1>
</fA11>
<fA14 i1="01"><s1>Schulze Diabetes Institute, Department of Surgery, University of Minnesota</s1>
<s2>Minneapolis, Minnesota</s2>
<s3>USA</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>6 aut.</sZ>
</fA14>
<fA14 i1="02"><s1>Center for Interdisciplinary Applications in Magnetic Resonance, Department of Radiology, University of Minnesota</s1>
<s2>Minneapolis, Minnesota</s2>
<s3>USA</s3>
<sZ>5 aut.</sZ>
</fA14>
<fA20><s1>3221-3225</s1>
</fA20>
<fA21><s1>2011</s1>
</fA21>
<fA23 i1="01"><s0>ENG</s0>
</fA23>
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<s2>14765</s2>
<s5>354000507348890180</s5>
</fA43>
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<s1>© 2012 INIST-CNRS. All rights reserved.</s1>
</fA44>
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</fA45>
<fA47 i1="01" i2="1"><s0>12-0017463</s0>
</fA47>
<fA60><s1>P</s1>
<s2>C</s2>
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<fC01 i1="01" l="ENG"><s0>Introduction. Despite significant progress in the last decade, islet transplantation remains an experimental therapy for a limited number of patients with type 1 diabetes. Tissue-engineered approaches may provide promising alternatives to the current clinical protocol and would benefit greatly from concurrent development of graft quality assessment techniques. This study was designed to evaluate whether viability of tissue-engineered islet grafts can be assessed using fluorine magnetic resonance spectroscopy (<sup>19</sup>
F-MRS), by the noninvasive measurement of oxygen partial pressure (pO<sub>2</sub>
) and the subsequent calculation of islet oxygen consumption rate (OCR). Methods. Scaffolds composed of porcine plasma were seeded with human islets and perfluorodecalin. Each graft was covered with the same volume of culture media in a Petri dish. Four scaffolds were seeded with various numbers (0-8000) of islet equivalents (IE) aliquoted from the same preparation. After randomizing run order, grafts were examined by <sup>19</sup>
F-MRS at 37°C using a 5T spectrometer and a single-loop surface coil placed underneath. A standard inversion recovery sequence was used to obtain characteristic <sup>19</sup>
F spin-lattice relaxation times (T1), which were converted to steady-state average pO<sub>2</sub>
 estimates using a previously determined linear calibration (R<sup>2</sup>
 = 1.000). Each condition was assessed using replicate <sup>19</sup>
F-MRS measurements (n = 6-8). Results. Grafts exhibited IE dose-dependent increases in T1 and decreases in pO<sub>2</sub>
 estimates. From the difference between scaffold pO<sub>2</sub>
 estimates and ambient pO<sub>2</sub>
, the islet preparation OCR was calculated to be 95 ± 12 (mean ± standard error of the mean) nmol/(min . mg DNA) using theoretical modeling. This value compared well with OCR values measured using established methods for human islet preparations. Conclusions. <sup>19</sup>
F-MRS can be used for noninvasive pre- and possibly posttransplant assessment of tissue-engineered islet graft viability by estimating the amount of viable, oxygen-consuming tissue in a scaffold.</s0>
</fC01>
<fC02 i1="01" i2="X"><s0>002B25</s0>
</fC02>
<fC02 i1="02" i2="X"><s0>002A06F</s0>
</fC02>
<fC03 i1="01" i2="X" l="FRE"><s0>Greffe</s0>
<s5>01</s5>
</fC03>
<fC03 i1="01" i2="X" l="ENG"><s0>Graft</s0>
<s5>01</s5>
</fC03>
<fC03 i1="01" i2="X" l="SPA"><s0>Injerto</s0>
<s5>01</s5>
</fC03>
<fC03 i1="02" i2="X" l="FRE"><s0>Culture tissu</s0>
<s5>02</s5>
</fC03>
<fC03 i1="02" i2="X" l="ENG"><s0>Tissue culture</s0>
<s5>02</s5>
</fC03>
<fC03 i1="02" i2="X" l="SPA"><s0>Cultivo tejido</s0>
<s5>02</s5>
</fC03>
<fC03 i1="03" i2="X" l="FRE"><s0>Génie tissulaire</s0>
<s5>03</s5>
</fC03>
<fC03 i1="03" i2="X" l="ENG"><s0>Tissue engineering</s0>
<s5>03</s5>
</fC03>
<fC03 i1="03" i2="X" l="SPA"><s0>Ingeniería de tejidos</s0>
<s5>03</s5>
</fC03>
<fC03 i1="04" i2="X" l="FRE"><s0>Spectrométrie RMN</s0>
<s5>04</s5>
</fC03>
<fC03 i1="04" i2="X" l="ENG"><s0>NMR spectrometry</s0>
<s5>04</s5>
</fC03>
<fC03 i1="04" i2="X" l="SPA"><s0>Espectrometría RMN</s0>
<s5>04</s5>
</fC03>
<fC03 i1="05" i2="X" l="FRE"><s0>Ilot Langerhans</s0>
<s5>05</s5>
</fC03>
<fC03 i1="05" i2="X" l="ENG"><s0>Langerhans islet</s0>
<s5>05</s5>
</fC03>
<fC03 i1="05" i2="X" l="SPA"><s0>Isla Langerhans</s0>
<s5>05</s5>
</fC03>
<fC03 i1="06" i2="X" l="FRE"><s0>Viabilité</s0>
<s5>06</s5>
</fC03>
<fC03 i1="06" i2="X" l="ENG"><s0>Viability</s0>
<s5>06</s5>
</fC03>
<fC03 i1="06" i2="X" l="SPA"><s0>Viabilidad</s0>
<s5>06</s5>
</fC03>
<fC03 i1="07" i2="X" l="FRE"><s0>Fluor</s0>
<s2>NC</s2>
<s2>FX</s2>
<s5>08</s5>
</fC03>
<fC03 i1="07" i2="X" l="ENG"><s0>Fluorine</s0>
<s2>NC</s2>
<s2>FX</s2>
<s5>08</s5>
</fC03>
<fC03 i1="07" i2="X" l="SPA"><s0>Fluor</s0>
<s2>NC</s2>
<s2>FX</s2>
<s5>08</s5>
</fC03>
<fC03 i1="08" i2="X" l="FRE"><s0>Médecine</s0>
<s5>09</s5>
</fC03>
<fC03 i1="08" i2="X" l="ENG"><s0>Medicine</s0>
<s5>09</s5>
</fC03>
<fC03 i1="08" i2="X" l="SPA"><s0>Medicina</s0>
<s5>09</s5>
</fC03>
<fC03 i1="09" i2="X" l="FRE"><s0>Transplantation</s0>
<s5>11</s5>
</fC03>
<fC03 i1="09" i2="X" l="ENG"><s0>Transplantation</s0>
<s5>11</s5>
</fC03>
<fC03 i1="09" i2="X" l="SPA"><s0>Trasplantación</s0>
<s5>11</s5>
</fC03>
<fC03 i1="10" i2="X" l="FRE"><s0>Traitement</s0>
<s5>25</s5>
</fC03>
<fC03 i1="10" i2="X" l="ENG"><s0>Treatment</s0>
<s5>25</s5>
</fC03>
<fC03 i1="10" i2="X" l="SPA"><s0>Tratamiento</s0>
<s5>25</s5>
</fC03>
<fC07 i1="01" i2="X" l="FRE"><s0>Pancréas endocrine</s0>
<s5>37</s5>
</fC07>
<fC07 i1="01" i2="X" l="ENG"><s0>Endocrine pancreas</s0>
<s5>37</s5>
</fC07>
<fC07 i1="01" i2="X" l="SPA"><s0>Páncreas endocrino</s0>
<s5>37</s5>
</fC07>
<fC07 i1="02" i2="X" l="FRE"><s0>Chirurgie</s0>
<s5>38</s5>
</fC07>
<fC07 i1="02" i2="X" l="ENG"><s0>Surgery</s0>
<s5>38</s5>
</fC07>
<fC07 i1="02" i2="X" l="SPA"><s0>Cirugía</s0>
<s5>38</s5>
</fC07>
<fN21><s1>002</s1>
</fN21>
<fN44 i1="01"><s1>OTO</s1>
</fN44>
<fN82><s1>OTO</s1>
</fN82>
</pA>
<pR><fA30 i1="01" i2="1" l="ENG"><s1>World Congress of the International Pancreas and Islet Transplant Association (IPITA)</s1>
<s3>CZE</s3>
<s4>2011-06-01</s4>
</fA30>
</pR>
</standard>
</inist>
</record>

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Assessment of Tissue-Engineered Islet Graft Viability by Fluorine Magnetic Resonance Spectroscopy

Assessment of Tissue-Engineered Islet Graft Viability by Fluorine Magnetic Resonance Spectroscopy

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