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See detailEVALUATION OF α-SYNUCLEIN AS BIOMARKER OF PARKINSON'S DISEASE
Napp, Aurore ULg; Garraux, Gaëtan ULg; Servais, Anne-Catherine ULg et al

Conference (2013, October 17)

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See detailRole of RIP3 in PDT-induced glioblastoma Cell Death
Fettweis, Grégory ULg; Coupienne, Isabelle; Fillet, Marianne ULg et al

Poster (2013, September 04)

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See detailRunning races: study of the stress and cardiac biomarkers
LE GOFF, Caroline ULg; Kaux, Jean-François ULg; MELON, Pierre ULg et al

in European Journal of Sports Medicine (2013, September), 1(Supplement 1), 212

Background: Cardiac troponins (cTn) are considered as the best biomarkers for detection of myocardial cell injury and NT-proBNP as the best for the cardiac insufficiency. In this study, cTnT was measured ... [more ▼]

Background: Cardiac troponins (cTn) are considered as the best biomarkers for detection of myocardial cell injury and NT-proBNP as the best for the cardiac insufficiency. In this study, cTnT was measured by new commercially available high-sensitive methods in subjects undergoing a marathon and an ultra-trail. Our aim was to compare cTnT and NT-proBNP levels before and after the stress tests, in sportive subjects. Materials and Methods: Twenty eight subjects (26 men, 42.5 ± 11 yo) ran the Maasmarathon (42.195 kilometers) between Visé (Belgium) and Maastricht (The Netherlands) and 33 subjects (33 men, 45.7±9.3yo) ran the Ultratour of Liège (Belgium ; 67 km). We took blood sample before (T0), just after (T1) and 3 hours after the race (T3). In all the patients, cTnT concentrations were measured by high sensitive methods (hsTnT, Roche Diagnostics) on heparin plasma. The NT-proBNP was also determined with the kit Roche on heparin plasma. The protocol was approved by the Ethics Committee of the University of Liège (Belgium). All subjects gave their informed consent. All statistical analyses were performed using Medcalc version 8.1 for Windows. p-value <0.01 was regarded as statistically significant. Results and discussion: A significant difference between hsTnT concentrations at T0 and T1 (p<0.0001), and between T0 and T3 (p<0.001) for NT-proBNP have been observed, but not between T1 and T3. This observation appeared only after a strenuous exercise. However, up to now this type of exercise is not reproducible easily in a laboratory. Moreover, nobody knows if these observations would have cardiac consequences at long terms. Conclusions: Measurement of cardiac troponins by high sensitive methods allows detecting significant release of biomarkers from the heart during exercise. The value of NT-proBNP are also significant but less than TnThs. We think that the TnThs could be an interesting tool in the future to help sport medicine to detect risk of developing a cardiac problem in the future or a sudden death. [less ▲]

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See detailL’électrophorèse capillaire micellaire pour la détermination simultanée de l’insuline et de ses analogues dans des formulations pharmaceutiques
Lamalle, Caroline ULg; Servais, Anne-Catherine ULg; Crommen, Jacques ULg et al

Poster (2013, June)

L’insuline joue un rôle important dans l’homéostasie de la concentration sanguine en glucose. Un déficit de cette hormone cause le diabète (de type I) qui peut être traité par injection sous-cutanée ... [more ▼]

L’insuline joue un rôle important dans l’homéostasie de la concentration sanguine en glucose. Un déficit de cette hormone cause le diabète (de type I) qui peut être traité par injection sous-cutanée d’insuline synthétique. En plus de l’insuline humaine, plusieurs analogues ont été développés pour accélérer son absorption (Lispro, Aspart, Glulisin) ou la retarder (Glargin, Detemir). La protamine est également parfois utilisée avec l’insuline humaine, Lispro ou Aspart pour former un complexe et augmenter la durée d’action. Le diabète est une des maladies métaboliques les plus courantes dans le monde; la prévalence ne cesse de s’élever. Beaucoup de patients sont donc concernés par le traitement qui est relativement cher et nécessite une ordonnance. Certaines formulations pharmaceutiques peuvent être trouvées sans ordonnance sur le marché parallèle mais la possibilité de contrefaçon existe. Et la mauvaise qualité de ces médicaments peut entrainer des conséquences néfastes pour la santé publique. Il est donc essentiel de développer une méthode permettant l’identification et la quantification de l’insuline humaine et de ses analogues. Ortner et al. [1] ont déjà proposé une méthode par électrophorèse capillaire micellaire (MEKC) pour séparer simultanément l’insuline humaine et ses 5 analogues, mais ils n’analysaient pas les formulations à base de protamine. Le nombre de ces formulations n’étant pas négligeable, nous les avons incluses dans notre étude. La première étape d’optimisation a été la préparation de l’échantillon. Une solution acide (0.01M d’HCl) a finalement été choisie pour solubiliser la protamine et la Glargine. Ensuite, la composition du background electrolyte a été investiguée pour séparer les composants des formulations. Un tampon basique (50mM d’acétate d’ammonium à pH 9) a été choisi, engendrant un flux électroosmotique important et stable, une charge négative à l’insuline et empêchant l’adsorption à la paroi du capillaire. L’addition de dodécyl sulfate de sodium et d’acétonitrile s’est ensuite révélée déterminante pour la sélectivité. Les 6 insulines et les 2 excipients majeurs (le phénol et le m-crésol) ont été complètement séparés endéans 15 minutes. La méthode précitée a ensuite été adaptée pour permettre la séparation de chaque insuline et de ses produits de dégradation. [less ▲]

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See detailDéveloppement de méthodes de quantification en milieu complexe par chromatographie liquide microfluidique couplée à la spectrometrie de masse
Houbart, Virginie ULg; Rozet, Eric ULg; Crommen, Jacques ULg et al

Conference (2013, June)

L’hepcidine est un biomarqueur peptidique dont l’intérêt tant comme outil de diagnostic que de suivi de pathologies est de plus en plus solidement établi. Il existe donc une demande forte d’outils ... [more ▼]

L’hepcidine est un biomarqueur peptidique dont l’intérêt tant comme outil de diagnostic que de suivi de pathologies est de plus en plus solidement établi. Il existe donc une demande forte d’outils sensibles et robustes afin de la doser au sein de milieux biologiques tels que le plasma ou le sérum. Une méthode analytique a été développée à l’aide d’un système chromatographique miniaturisé (nanoLC-chip) couplé à un spectromètre de masse permettant d’assurer un dosage de l’hepcidine à la fois sensible et fiable. Lors du développement de cette méthode, il a été constaté que la composition de l’échantillon avait une influence majeure sur la réponse analytique. Or, l’utilisation de systèmes chromatographiques miniaturisés implique souvent l’injection de volumes proportionnellement très importants par rapport aux dimensions du système, ce qui amplifie encore l’impact de sa composition. C’est pourquoi il est capital d’avoir une bonne compréhension des phénomènes qui ont lieu entre l’injection de l’échantillon dans le système chromatographique et la détection par spectrométrie de masse, et particulièrement lors du développement de méthodes analytiques quantitatives. Dans cette étude, nous avons utilisé la planification expérimentale afin de mieux comprendre le comportement chromatographique des peptides, ainsi que les facteurs qui influencent la sensibilité de la méthode développée. Un mélange de peptides a été sélectionné, varié tant du point de vue du poids moléculaire que du point isoélectrique et de l’hydropathie. Un plan de criblage a permis de délimiter le domaine expérimental ainsi que les facteurs significatifs parmi la composition de la phase mobile (proportion et nature de l’agent de paire d’ions) et de l’échantillon en lui-même (nature de l’agent de paire d’ions et proportion de solvant organique). Ensuite, un plan d’expériences factoriel complet a été mis en œuvre afin d’observer plus finement le rôle de chaque facteur ainsi que les interactions éventuelles qui les lient. Certains facteurs ont montré un effet très marqué tant sur l’intensité de la réponse que sur la rétention. Entre autres, la composition de l’échantillon, facteur parfois négligé lors du développement de méthodes, a démontré son importance capitale, en particulier sur les phénomènes de rétention des peptides. Les données obtenues ont également permis de dégager des conditions optimales d’analyse en termes de rétention et de sensibilité. Enfin, une analyse en composantes principales a également été réalisée sur le grand nombre de données récoltées dans le but de mettre en évidence d’éventuels propriétés physicochimiques des peptides qui pourraient avoir un impact significatif sur les réponses étudiées. [less ▲]

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See detailComparison of the Heart-type fatty acid-binding protein (H-FABP) with the high sensitive cardiac troponin T in healthy runners
LE GOFF, Caroline ULg; MELON, Pierre ULg; BREVERS, Eric ULg et al

in Book of abstracts of 18th Annual Congress of the ECSS (2013, June)

Background: Heart-type fatty acid-binding protein (H-FABP) is a low molecular weight protein involved in the intracellular uptake and buffering of long chain fatty in the myocardium. It is an early marker ... [more ▼]

Background: Heart-type fatty acid-binding protein (H-FABP) is a low molecular weight protein involved in the intracellular uptake and buffering of long chain fatty in the myocardium. It is an early marker for acute coronary syndrome. Troponin T (TnT) is a component of the contractile apparatus of the striated musculature. Cardiac TnT is a cardio-specific, highly sensitive marker for myocardial damage. The aim of our study was to compare the results obtained with the H-FABP and the highly sensitive cardiac troponins (hsTnT) and to test their cardiospecificity in healthy runners. Methods: Twenty three runners (marathon) were enrolled. We drowned samples at three times: just before (T0), just after (T1), and three hours after the end of the race (T3). H-FABP was determined with a Randox immunoturbidimetric assay and hs-TnT with a Roche electrochemiluminescence immunoassay, both on Cobas 6000. A linear regression was calculated to observe if there is any correlation between the two biomarkers. Values above the 95th percentile for H-FABP (2.5ng/mL) and the 99th percentile for hsTnT (14ng/L) were considered as positive. Results: At T0, none of the subjects were positive for hsTnT but 35% were positive for H-FABP; at T1, 83% for hsTnT and 100% for H-FABP; at T3, 83% for hsTnT and 96% for H-FABP. At T0, the regression equation was H-FABP T0 = 3.9454 – 0.1001 x hsTnT T0; at T1: H-FABP T1 = 51.838 – 1.7026 x hsTnT T1; at T3: H-FABP T3 = 47.977 – 1.6193 x hsTnT T3. No correlation was observed between the two biomarkers at the different time. Conclusions: We observed a significant increase of H-FABP and hsTnT in runners. These markers are independent to each other. These values could biologically correspond to a heart ischemia. However, we suggested that exercise-induced cardiac hsTnT and H-FABP release is not a marker of exercise-induced pathology but likely a physiologic response to effort or an exercise-induced cardiac remodelling. [less ▲]

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See detailLes biomarqueurs de la maladie de Parkinson
Napp, Aurore ULg; Fillet, Marianne ULg

Scientific conference (2013, May 23)

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See detailImpact of strenuous exercise on the release of cardiac biomarkers
LE GOFF, Caroline ULg; MELON, Pierre ULg; Kaux, Jean-François ULg et al

in Biochimica Clinica (2013, May), 37(SS), 545

Background: Cardiac troponins (cTn) are considered as the best biomarkers for detection of myocardial cell injury and NT-proBNP as the best for the cardiac insufficiency. In this study, cTnT was measured ... [more ▼]

Background: Cardiac troponins (cTn) are considered as the best biomarkers for detection of myocardial cell injury and NT-proBNP as the best for the cardiac insufficiency. In this study, cTnT was measured by new commercially available high-sensitive methods in subjects un-dergoing the Maasmarathon. Our aim was to compare cTnT and NT-proBNP levels before and after the stress tests, in sportive subjects. Methods: Twenty eight subjects (26 ♂, 42.5±11yo) underwent a race of 42.195 kilometers be-tween Visé (Belgium) and Maastricht (The Netherlands). We drowned blood sample before (T0), just after (T1) and three hours after the race (T3). In all patients, cTnT concentrations were measured by high sensitive methods (hsTnT, Roche Diagnostics) on heparin plasma. The NT-proBNP was also determined with the kit Roche on heparin plasma. The protocol was approved by the ethics committee of the University of Liège (Belgium). All subjects gave their informed consent. All statistical analyses were performed using Medcalc version 8.1 for Windows. p-value <0.01 was regarded as statistically significant. Results: There was a significant difference between hsTnT concentrations at T0 and T1 (p<0.0001), and between T0 and T3 (p<0.001) for NT-proBNP, but not between T1 and T3. This observation appeared only after a strenuous exercise but today this type of exercise is not reproduce easier in a laboratory of sport. Moreover, at this moment, nobody knows if these observations would have cardiac consequences at long terms. Conclusions: Measurement of cardiac troponins by high sensitive methods allows detecting significant release of biomarkers from the heart during exercise. The value of NT-proBNP are also significant but less than TnThs. We think that the TnThs could be an interesting tool in the future to help sport medicine to detect risk of developing a cardiac problem in the future or a sudden death. [less ▲]

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See detailComparison of the Heart-type fatty acid-binding protein (H-FABP) with the high sensitive cardiac troponin T in healthy runners
LE GOFF, Caroline ULg; Kaux, Jean-François ULg; BREVERS, Eric ULg et al

in Biochimica Clinica (2013, May), 37(SS), 544

Background: Heart-type fatty acid-binding protein (H-FABP) is a low molecular weight protein involved in the intracellular uptake and buffering of long chain fatty in the myocardium. It is an early marker ... [more ▼]

Background: Heart-type fatty acid-binding protein (H-FABP) is a low molecular weight protein involved in the intracellular uptake and buffering of long chain fatty in the myocardium. It is an early marker for acute coronary syndrome. Troponin T (TnT) is a component of the contractile apparatus of the striated musculature. Cardiac TnT is a cardio-specific, highly sensitive marker for myocardial damage. The aim of our study was to compare the results obtained with the H-FABP and the highly sensitive cardiac troponins (hsTnT) and to test their cardiospecificity in healthy runners. Methods: Twenty three runners (marathon) were enrolled. We drowned samples at three times: just before (T0), just after (T1), and three hours after the end of the race (T3). H-FABP was determined with a Randox immunoturbidimetric assay and hs-TnT with a Roche electrochemiluminescence immunoassay, both on Cobas 6000. A linear regression was calculated to observe if there is any correlation between the two biomarkers. Values above the 95th percentile for H-FABP (2.5ng/mL) and the 99th percentile for hsTnT (14ng/L) were considered as positive. Results: At T0, none of the subjects were positive for hsTnT but 35% were positive for H-FABP; at T1, 83% for hsTnT and 100% for H-FABP; at T3, 83% for hsTnT and 96% for H-FABP. At T0, the regression equation was H-FABP T0 = 3.9454 – 0.1001 x hsTnT T0; at T1: H-FABP T1 = 51.838 – 1.7026 x hsTnT T1; at T3: H-FABP T3 = 47.977 – 1.6193 x hsTnT T3. No correlation was observed between the two biomarkers at the different time. Conclusions: We observed a significant increase of H-FABP and hsTnT in runners. These markers are independent to each other. These values could biologically correspond to a heart ischemia. However, we suggested that exercise-induced cardiac hsTnT and H-FABP release is not a marker of exercise-induced pathology but likely a physiologic response to effort or an exercise-induced cardiac remodelling. [less ▲]

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See detailRace Cycling: biological evolution
LE GOFF, Caroline ULg; Kaux, Jean-François ULg; MELON, Pierre ULg et al

in Biochimica Clinica (2013, May), 37(SS), 544

Background: The metabolic and cardiac impact of a cycling effort on blood biology is not very well described in the literature. We aimed to measure the concentration of different biomarkers (cardiac and ... [more ▼]

Background: The metabolic and cardiac impact of a cycling effort on blood biology is not very well described in the literature. We aimed to measure the concentration of different biomarkers (cardiac and metabolic) released during an international cycling race. Methods: Venous blood samples of 15 young men (25.1 ± 6.4 y.o.) were collected just before (T1), just after (T2), 3 hours (T3) after an international cycling race of 179.6 kilometers in Belgium for the determination of cardiac and metabolic biomarkers: red blood cell (RBC), haemoglobin (HgB), creatinin (Cr), highly sensitive troponin T (hsTnT), myoglobin (MYO) and NT-proBNP. All automated assays were performed according to the manufacter’s specifications. For the statistical analysis, an Anova calculated with the Statistica Software version 9.1 was used. Results: RBC and HgB levels varied significantly between T0 and T3 (respectively p=0.0026, and p=0.002). Cr concentration also varied significantly between all times (T0-T1:p<0.0001, T1-T3:p=0.0326 and T0-T3 p=0.0001). These changes might be related to renal flow depletion during exercice. MYO increased significantly between T0 and T1 (p<0.0001), but quickly decreased between T1 and T3, however the T3 level stay higher than T0 (p=0.014). The stress delivered from the physical activity performed during the race induced a significant variation of hsTnT which increased significantly between T0 and T1 (p<0.0001) and stayed higher 3 hours after the end of the exercise (T0-T3: p<0.0001) .The intense exercise delivery by the race induced a significant variation of NT-proBNP, that followed the same kinetic of hsTnT but in smaller proportion. We noticed variations statistically significant between T0 and T1 and between T0 and T3 for NT-proBNP. These increases of cardiac biomarkers were significant but reasonable and could not allow us to talk about cellular necrosis or irreversible injury. Conclusion: Our results show that stress generated by a cycling race could be the cause for the different metabolic variations observed. Troponin T stays without a doubt the most specific marker for stress related to myocardial tissue. Its increase can then be considered as being of interest. [less ▲]

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See detailFalsification des médicaments: mythe ou réalité ?
Marini Djang'Eing'A, Roland ULg; Fillet, Marianne ULg; Vancauwenberghe, Roy et al

Conference (2013, April 24)

La santé publique est de nos jours minée par la problématique des médicaments falsifiés ou de qualité inférieure, avec plusieurs conséquences sanitaires, économiques voire professionnelles. On estime à 7 ... [more ▼]

La santé publique est de nos jours minée par la problématique des médicaments falsifiés ou de qualité inférieure, avec plusieurs conséquences sanitaires, économiques voire professionnelles. On estime à 7% la part du marché pharmaceutique mondial que représenterait ce fléau; l’Afrique, l’Asie et de nombreux pays d'Amérique latine étant les régions les plus touchées avec plus de 30% de médicaments falsifiés. D’après l'OMS, plus de 50% des médicaments achetés à partir des sites internet illégaux sont contrefaits, annihilant très fortement les chances de succès thérapeutique. Ces médicaments viennent dans la plupart des cas des pays asiatiques et de l’Eurasie. Le trafic de faux médicaments est un crime contre l'humanité qui représente environ 50 milliards de dollars par an (10-15 % de plus que le marché de la drogue). Au travers de deux leçons, la situation de la falsification des médicaments sera présentée au grand public dans le but de le sensibiliser à ce fléau. La première leçon présentera la situation en Europe avec un accent sur la Belgique. La problématique du droit à la propriété intellectuelle et de l’encadrement législatif sera abordée, ainsi que la falsification des médicaments modernes et des phytomédicaments, ces derniers étant utilisés par plus de 40% de la population en Europe et aux Etats-Unis. Dans la seconde leçon sera abordée la situation vécue en Afrique. L’approvisionnement en médicaments de qualité par le partage de l’information sera présenté ainsi que les moyens analytiques à la disposition de ce continent pour combattre ce fléau. Des membres du Département de Pharmacie de l’Université de Liège, de l’Agence Fédérale des Médicaments et des Produits de Santé ainsi que du programme QUAMED (Quality Medicines for All) feront partager leur expérience sur cette question d’une brûlante actualité.  [less ▲]

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See detailUnusual Amino Acids and Monofluoroacetate from Dichapetalum michelsonii (Umutambasha), a Toxic Plant from Rwanda
Esters, Virginie ULg; Karangwa, Charles; Tits, Monique ULg et al

in Planta Medica (2013), 79

In the course of our investigations on Umutambasha in order to identify its convulsant principles, small quantities of monofluoroacetate were observed in stem bark, leaves, and fruits of this plant newly ... [more ▼]

In the course of our investigations on Umutambasha in order to identify its convulsant principles, small quantities of monofluoroacetate were observed in stem bark, leaves, and fruits of this plant newly identified as Dichapetalum michelsonii Hauman. Conclusive evidence for a monofluoroacetate presence came from its isolation from the freeze-dried extract of stem bark. Three free unusual amino acids, named N-methyl-α-alanine, N-methyl-β-alanine, and 2,7-diaminooctan-1,8-dioic acid, described for the first time in a plant, and known trigonelline were also isolated from the stem bark of D. michelsonii. Structure elucidations were mainly achieved by spectroscopic methods (1H-NMR, 2D-NMR, MS) and by comparison with authentic references. These unusual amino acids were detected by a fast, reliable TLC analysis in all our batches of Umutambasha, suggesting that they could be used for identification purposes in case of human or livestock intoxications. Finally, EEG recordings and behavioural observations performed in mice suggested that the convulsive patterns produced by Umutambasha are the consequence of monofluoroacetate presence in D. michelsonii. [less ▲]

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See detailSimultaneous determination of insulin and its analogues in pharmaceutical formulations by micellar electrokinetic chromatography (MEKC)
Lamalle, Caroline ULg; Servais, Anne-Catherine ULg; Crommen, Jacques ULg et al

Poster (2013)

Insulin plays an important role in the homeostasis of blood glucose concentration. A deficiency of this hormone causes diabetes, which can be treated by subcutaneous injection of synthetic insulin ... [more ▼]

Insulin plays an important role in the homeostasis of blood glucose concentration. A deficiency of this hormone causes diabetes, which can be treated by subcutaneous injection of synthetic insulin. Besides human regular insulin, several modified analogues have been developed to accelerate (Lispro, Aspart, Glulisin) or delay (Glargin, Detemir) its absorption. Moreover, protamine is sometimes associated with human, Lispro or Aspart insulin to give a crystalline form, which delays the action of insulin, providing it with a prolonged absorption profile after injection. Diabetes is one of the most common metabolic diseases in the world; its prevalence increases continuously. A lot of patients are therefore concerned with the treatment, which is relatively expensive and requires a prescription. Some pharmaceutical formulations can sometimes be found without prescription on the parallel market but the risk of drug counterfeiting is then considerably increased. The poor quality of these drugs can lead to harmful consequences for the public health. It is therefore essential to develop a suitable method for the identification and quantification of human insulin and its analogues. Ortner et al. have already proposed micellar electrokinetic chromatography (MEKC) methods to analyse simultaneously human insulin and its five analogues but formulations containing protamine were not tested. Since the number of these formulations is significant, we included them in our study. The first optimisation step involved the sample preparation procedure. An acidic sample solution (0.01 M HCl) was finally selected to solubilise protamine and Glargin. Then the background electrolyte composition was investigated to separate the components present in the formulations. A basic buffer (50 mM ammonium acetate pH 9) was selected, providing an important and stable electroosmotic flow, a negative charge to insulin and related compounds and avoiding any adsorption to the capillary wall. The addition of sodium dodecyl sulfate and acetonitrile were also found crucial for selectivity. The six insulins and the two major excipients (phenol and m-cresol) were fully separated within 15 minutes The aforementioned method was then adapted to permit the separation of each insulin from its degradation products. [less ▲]

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See detailEnantioseparations in nonaqueous capillary electrophoresis using charged cyclodextrins.
Servais, Anne-Catherine ULg; Fillet, Marianne ULg

in Methods in Molecular Biology 970: Chiral Separations (2013)

The enantioseparation of acidic and basic compounds can be successfully achieved in nonaqueous capillary electrophoresis using single-isomer charged beta-cyclodextrin (beta-CD) derivatives of opposite ... [more ▼]

The enantioseparation of acidic and basic compounds can be successfully achieved in nonaqueous capillary electrophoresis using single-isomer charged beta-cyclodextrin (beta-CD) derivatives of opposite charge to that of the analytes. This chapter describes how to separate the enantiomers of three basic substances selected as model compounds, i.e., alprenolol, bupranolol, and terbutaline, using the negatively charged heptakis(2,3-di-O-acetyl-6-O-sulfo)-beta-CD. The enantiomers of three acidic drugs (tiaprofenic acid, suprofen, and flurbiprofen) are resolved using a monosubstituted amino beta-CD derivative, namely, 6-monodeoxy-6-mono(3-hydroxy)propylamino-beta-CD. [less ▲]

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See detailApplication of dual cyclodextrin systems in capillary electrophoresis enantioseparations.
Servais, Anne-Catherine ULg; Fillet, Marianne ULg

in Methods in Molecular biology 970: Chiral Separations (2013)

The enantioseparation of acidic and neutral compounds can be successfully achieved in capillary electrophoresis (CE) using dual cyclodextrin (CD) systems. This chapter describes how to separate the ... [more ▼]

The enantioseparation of acidic and neutral compounds can be successfully achieved in capillary electrophoresis (CE) using dual cyclodextrin (CD) systems. This chapter describes how to separate the enantiomers of acidic or neutral substances using dual CD systems made up of a negatively charged CD derivative, i.e., sulfobutyl-beta-CD (SB-beta-CD) or carboxymethyl-beta-CD (CM-beta-CD), in combination with a neutral one, namely, heptakis(2,3,6-tri-O-methyl)-beta-CD (TM-beta-CD). An acidic compound (carprofen) and a weakly acidic drug (pentobarbital) were selected as model compounds. [less ▲]

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See detailDetermination of inhibitory potency of argatroban toward thrombin by electrophoretically mediated microanalysis
Pochet, Lionel; Servais, Anne-Catherine ULg; Farcas, Elena ULg et al

in Talanta (2013), 116

Developing an EMMA method for enzymatic assay remains a challenge, particularly using UV detection. Indeed, it is necessary to optimize the separation conditions while allowing the enzymatic reaction to ... [more ▼]

Developing an EMMA method for enzymatic assay remains a challenge, particularly using UV detection. Indeed, it is necessary to optimize the separation conditions while allowing the enzymatic reaction to occur within the capillary respecting kinetic constraints and achieving enough sensitivity. In this work, such EMMA methodology was set up to evaluate the inhibitory potency of drugs toward thrombin, a serine protease implicated in the coagulation cascade. To achieve our goal, the separation buffer, the injection sequence, the internal standard and the chromogenic substrate were investigated. The newly developed system was then assessed determining the kinetic Km constant for the selected substrate and compared with the results obtained with a continuous spectrophotometer cell assay. Secondly, the Ki inhibitory constant of the thrombin inhibitor argatroban was determined and found in agreement with the published value. [less ▲]

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See detailIdentification of protein biomarkers associated with cardiac ischemia by a proteomic approach.
Fillet, Marianne ULg; Deroyer, Céline ULg; COBRAIVILLE, G. et al

in Biomarkers : biochemical indicators of exposure, response, and susceptibility to chemicals (2013), 18(7), 614-24

Angina is chest pain induced by ischemia of the heart muscle, generally due to obstruction or spasm of the coronary arteries. People that suffer from average to severe cases of angina have an increased ... [more ▼]

Angina is chest pain induced by ischemia of the heart muscle, generally due to obstruction or spasm of the coronary arteries. People that suffer from average to severe cases of angina have an increased percentage of death before the age of 55, usually around 60%. Therefore, prevention of major complications, optimizing diagnosis, prognosis and therapeutics are of primary importance. The main objective of this study was to uncover biomarkers by comparing serum protein profiles of patients suffering from stable or unstable angina and controls. We identified by non-targeted proteomic approach and confirmed by the means of independent techniques, the differential expression of several proteins indicating significantly increased vascular inflammation response, disturbance in the lipid metabolism and in atherogenic plaques stability. [less ▲]

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See detailKinetics of IL-7 and IL-15 Levels after Allogeneic Peripheral Blood Stem Cell Transplantation following Nonmyeloablative Conditioning
De Bock, Muriel; Fillet, Marianne ULg; Hannon, Muriel ULg et al

in PLoS ONE (2013), 8(2), 55876

Background: We analysed kinetics of IL-7 and IL-15 levels in 70 patients given peripheral blood stem cells after nonmyeloablative conditioning. Methods: EDTA-anticoagulated plasma and serum samples were ... [more ▼]

Background: We analysed kinetics of IL-7 and IL-15 levels in 70 patients given peripheral blood stem cells after nonmyeloablative conditioning. Methods: EDTA-anticoagulated plasma and serum samples were obtained before conditioning and about once per week after transplantation until day 100. Samples were aliquoted and stored at 280uC within 3 hours after collection until measurement of cytokines. IL-7 and IL-15 levels were measured by ELISAs. Results: Median IL-7 plasma levels remained below 6 pg/L throughout the first 100 days, although IL-7 plasma levels were significantly higher on days 7 (5.1 pg/mL, P = 0.002), 14 (5.2 pg/mL, P,0.001), and 28 (5.1 pg/mL, P = 0.03) (but not thereafter) than before transplantation (median value of 3.8 pg/mL). Median IL-15 serum levels were significantly higher on days 7 (12.5 pg/mL, P,0.001), 14 (10.5 pg/mL, P,0.001), and 28 (6.2 pg/mL, P,0.001) than before transplantation (median value of 2.4 pg/mL). Importantly, IL-7 and IL-15 levels on days 7 or 14 after transplantation did not predict grade II–IV acute GVHD. Conclusions: These data suggest that IL-7 and IL-15 levels remain relatively low after nonmyeloablative transplantation, and that IL-7 and IL-15 levels early after nonmyeloablative transplantation do not predict for acute GVHD. [less ▲]

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