References of "De Pauw, Edwin"
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See detailAdvances in proteomics for the FP7 Venomics project
Degueldre, Michel ULg; Quinton, Loïc ULg; De Pauw, Edwin ULg

Scientific conference (2012)

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See detailRaman spectroscopy and laser desorption mass spectrometry for minimal destructive forensic analysis of black and color inkjet printed documents
Heudt, Laetitia ULg; Debois, Delphine ULg; Zimmerman, Tyler ULg et al

in Forensic Science International (2012), 219

Inkjet ink analysis is the best way to discriminate between printed documents, or even though more difficult, to connect an inkjet printed document with a brand or model of printers. Raman spectroscopy ... [more ▼]

Inkjet ink analysis is the best way to discriminate between printed documents, or even though more difficult, to connect an inkjet printed document with a brand or model of printers. Raman spectroscopy and laser desorption mass spectrometry (LDMS) have been demonstrated as powerful tools for dyes and pigments analysis, which are ink components. The aim of this work is to evaluate the aforementioned techniques for inkjet inks analysis in terms of discriminating power, information quality, and nondestructive capability. So, we investigated 10 different inkjet ink cartridges (primary colors and black), 7 from the HP manufacturer and one each from Epson, Canon and Lexmark. This paper demonstrates the capabilities of three methods: Raman spectroscopy, LDMS and MALDI-MS. Raman spectroscopy, as it is preferable to try the nondestructive approach first, is successfully adapted to the analysis of color printed documents in most cases. For analysis of color inkjet inks by LDMS, we show that a MALDI matrix (9- aminoacridine, 9AA) is needed to desorb and to ionize dyes from most inkjet inks (except Epson inks). Therefore, a method was developed to apply the 9AA MALDI matrix directly onto the piece of paper while avoiding analyte spreading. The obtained mass spectra are very discriminating and lead to information about ink additives and paper compositions. Discrimination of black inkjet printed documents is more difficult because of the common use of carbon black as the principal pigment. We show for the first time the possibility to discriminate between two black-printed documents coming from different, as well as from the same, manufacturers. Mass spectra recorded from black inks in positive ion mode LDMS detect polyethylene glycol polymers which have characteristic mass distributions and end groups. Moreover, software has been developed for rapid and objective comparison of the low mass range of these positive mode LDMS spectra which have characteristic unknown peaks. [less ▲]

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See detailSparc-like protein 1 is a new marker of human glioma progression.
Turtoi, Andrei ULg; Musmeci, Davide; Naccarato, Antonio Giuseppe et al

in Journal of Proteome Research (2012), 11(10), 5011-21

High-grade gliomas (glioblastomas) are the most common and deadly brain tumors in adults, currently with no satisfactory treatment available. Apart from de novo glioblastoma, it is currently accepted that ... [more ▼]

High-grade gliomas (glioblastomas) are the most common and deadly brain tumors in adults, currently with no satisfactory treatment available. Apart from de novo glioblastoma, it is currently accepted that these malignancies mainly progress from lower grade glial tumors. However, the molecular entities governing the progression of gliomas are poorly understood. Extracellular and membrane proteins are key biomolecules found at the cell-to-cell communication interface and hence are a promising proteome subpopulation that could help understand the development of glioma. Accordingly, the current study aims at identifying new protein markers of human glioma progression. For this purpose, we used glial tumors generated orthotopically with T98G and U373 human glioma cells in nude mice. This setup allowed also to discriminate the protein origin, namely, human (tumor) or mouse (host). Extracellular and membrane proteins were selectively purified using biotinylation followed by streptavidin affinity chromatography. Isolated proteins were digested and then identified and quantified employing 2D-nano-HPLC-MS/MS analysis. A total of 23 and 27 up-regulated extracellular and membrane proteins were identified in the T98G and U373 models, respectively. Approximately two-thirds of these were predominantly produced by the tumor, whereas the remaining proteins appeared to be mainly overexpressed by the host tissue. Following extensive validation, we have focused our attention on sparc-like protein 1. This protein was further investigated using immunohistochemistry in a large collection of human glioma samples of different grades. The results showed that sparc-like protein 1 expression correlates with glioma grade, suggesting the possible role for this protein in the progression of this malignancy. [less ▲]

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See detailThe angiogenesis suppressor gene AKAP12 is under the epigenetic control of HDAC7 in endothelial cells.
Turtoi, Andrei ULg; Mottet, Denis ULg; Matheus, Nicolas ULg et al

in Angiogenesis (2012)

Histone deacetylases (HDACs) are a family of 18 enzymes that deacetylate lysine residues of both histone and nonhistone proteins and to a large extent govern the process of angiogenesis. Previous studies ... [more ▼]

Histone deacetylases (HDACs) are a family of 18 enzymes that deacetylate lysine residues of both histone and nonhistone proteins and to a large extent govern the process of angiogenesis. Previous studies have shown that specific inhibition of HDAC7 blocks angiogenesis both in vitro and in vivo. However, the underlying molecular mechanisms are not fully understood and hence preclude any meaningful development of suitable therapeutic modalities. The goal of the present study was to further the understanding of HDAC7 epigenetic control of angiogenesis in human endothelial cells using the proteomic approach. The underlying problem was approached through siRNA-mediated gene-expression silencing of HDAC7 in human umbilical vein endothelial cells (HUVECs). To this end, HUVEC proteins were extracted and proteomically analyzed. The emphasis was placed on up-regulated proteins, as these may represent potential direct epigenetic targets of HDAC7. Among several proteins, A-kinase anchor protein 12 (AKAP12) was the most reproducibly up-regulated protein following HDAC7 depletion. This overexpression of AKAP12 was responsible for the inhibition of migration and tube formation in HDAC7-depleted HUVEC. Mechanistically, H3 histones associated with AKAP12 promoter were acetylated following the removal of HDAC7, leading to an increase in its mRNA and protein levels. AKAP12 is responsible for protein kinase C mediated phosphorylation of signal transducer and activator of transcription 3 (STAT3). Phosphorylated STAT3 increasingly binds to the chromatin and AKAP12 promoter and is necessary for maintaining the elevated levels of AKAP12 following HDAC7 knockdown. We demonstrated for the first time that AKAP12 tumor/angiogenesis suppressor gene is an epigenetic target of HDAC7, whose elevated levels lead to a negative regulation of HUVEC migration and inhibit formation of tube-like structures. [less ▲]

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See detailEstimation of furan contamination across the Belgian food chain.
Scholl, Georges ULg; Scippo, Marie-Louise ULg; De Pauw, Edwin ULg et al

in Food Additives & Contaminants. Part A. Chemistry, Analysis, Control, Exposure & Risk Assessment (2012), 29(2), 172-9

This paper provides an estimate of the furan content of Belgian foods. The objective of the study was to achieve the best food chain coverage with a restricted number of samples (n = 496). The geographic ... [more ▼]

This paper provides an estimate of the furan content of Belgian foods. The objective of the study was to achieve the best food chain coverage with a restricted number of samples (n = 496). The geographic distribution, different market chains and labels, and consumption frequencies were taken into account in the construction of the sampling plan. Weighting factors such as contamination levels, consumption frequency and the diversity of food items were applied to set up the model. The very low detection capabilities (CC(beta)) of the analytical methods used (sub-ppb) allowed reporting of 78.2% of the overall dataset above CC(beta) and, in particular, 96.7% for the baby food category. The highest furan levels were found in powdered roasted bean coffee (1912 microg kg(-1)) with a mean of 756 microg kg(-1) for this category. Prepared meat, pasta and rice, breakfast cereals, soups, and baby food also showed high mean furan contents ranging from 16 to 43 microg kg(-1). Comparisons with contamination surveys carried out in other countries pointed out differences for the same food group and therefore contamination levels are related to the geographical origin of food items. [less ▲]

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See detailFibulin 3 peptides Fib3-1 and Fib3-2 are potential biomarkers of osteoarthritis.
Henrotin, Yves ULg; Gharbi, Myriam; Mazzucchelli, Gabriel ULg et al

in Arthritis and Rheumatism (2012), 64(7), 2260-7

OBJECTIVE: This study was undertaken to identify new biomarkers of osteoarthritis (OA) by proteomics analysis and to develop specific immunoassays to detect and quantify them. METHODS: Proteomics analysis ... [more ▼]

OBJECTIVE: This study was undertaken to identify new biomarkers of osteoarthritis (OA) by proteomics analysis and to develop specific immunoassays to detect and quantify them. METHODS: Proteomics analysis was performed in urine samples from 10 women (mean+/-SD age 76.0+/-5.0 years) undergoing knee replacement surgery due to severe OA and 5 healthy women (mean+/-SD age 25.6+/-2.6 years). Protein content was analyzed by 2-dimensional differential gel electrophoresis. Protein spots that exhibited an OA:control abundance ratio of >/=1.5 were identified by mass spectrometry. Specific enzyme-linked immunosorbent assays were developed and validated in serum obtained from 236 healthy subjects ages 20-64 years and from 76 patients with severe radiologic knee OA (mean+/-SD age 68.8+/-11.9 years). Immunohistochemical analysis was performed on articular cartilage from tibial plateaus. RESULTS: Thirteen proteins within spots that were significantly modified between groups were identified. Two peptides of fibulin 3, named Fib3-1 and Fib3-2, were of particular interest. Two antisera directed against these peptides were used to develop immunoassays. Compared with age-matched healthy subjects, median levels of serum Fib3-1 and Fib3-2 were elevated in OA patients (54.6 pM versus 85.1 pM [P<0.0001] and 144.4 pM versus 191.4 pM [P<0.0001], respectively). Using area under the receiver operating characteristic curve analysis, we demonstrated that Fib3-1 and Fib3-2 levels discriminate between OA and normal populations. Immunostaining revealed the presence of Fib3-1 and Fib3-2 in chondrocytes and in the extracellular matrix of the superficial layer of the fibrillated cartilage. CONCLUSION: Our findings indicate that Fib3-1 and Fib3-2 are potential biochemical markers for the diagnosis of OA. [less ▲]

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See detailPurification and characterisation of a 31 kDa chitinase from the Myzus persicae aphid, a target for Hemiptera biocontrol
Francis, Frédéric ULg; Saguez, Julien; Cherqui, Anas et al

in Applied Biochemistry and Biotechnology (2012), 166

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See detailUsing sets of behavioral biomarkers to assess short-term effects of pesticide: a study case with endosulfan on frog tadpoles
Denoël, Mathieu ULg; D'hooghe, Bastien; Ficetola, G. Francesco et al

in Ecotoxicology (2012), 21(4), 1240-1250

Pesticides and other chemicals often have detrimental effects at environmental concentrations. Many amphibian species are particularly threatened because of their susceptibility but also because wetlands ... [more ▼]

Pesticides and other chemicals often have detrimental effects at environmental concentrations. Many amphibian species are particularly threatened because of their susceptibility but also because wetlands are often polluted. Behavioral assessments of toxicity have the advantage of showing sublethal effects but quantitative measures at varied scales of integrations are rarely considered together. In this study, we aimed at showing that these behavioral endpoints could be differently affected across time and concentrations, and be biomarkers of toxicity. To this end, we tested the effects of an organochlorine pesticide (endosulfan) on amphibians during a standard 96 h test. We evaluated possible lag effects in continuing the analyses after removal of the pesticide. The study was based on 240 tadpoles (4 pesticide treatments: 0.4, 3, 22, and 282 lg/l, 1 control and 1 solvent-control). Abnormal behaviors such as lying and swirling rapidly were exhibited only in the presence of the pesticide. Essential functions such as breathing and feeding were deeply affected by the pesticide: contaminated tadpoles breathed and fed less than control tadpoles. They also moved less and occupied a more central position in the aquariums in the presence of the pesticide. A higher mortality was only found at the highest concentration. These results suggest that endosulfan is toxic to amphibians at environmental concentrations. Behavioral markers showed potential as early warning systems. They should thus be used in complement to other markers to detect sublethal effects only a few days after application of the pesticide and at concentrations where mortality does not occur. [less ▲]

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See detailRecent advances in analytical chemistry and mass spectrometry for organic selenium and tellurium compounds
Far, Johann ULg; De Pauw, Edwin ULg; Eppe, Gauthier ULg

in Rappoport, Zvi (Ed.) The Chemistry of Organic Selenium and Tellurium compounds Volume 3 Part 1 and 2 (2012)

The objective of this chapter is to present the recent developments in mass spectrometry for selenium (Se) and tellurium (Te) analysis. The state-of-the-art of mass spectrometry based techniques available ... [more ▼]

The objective of this chapter is to present the recent developments in mass spectrometry for selenium (Se) and tellurium (Te) analysis. The state-of-the-art of mass spectrometry based techniques available is critically evaluated with particular emphasis in ion sources and mass analyzers. The complexity of identification and characterization of Se, Te species including their speciation in the environment or in biological materials requires methodologies not only limited to mass spectrometry. Particular attention is given to sample preparation and hyphenated techniques with mass spectrometry for a comprehensive characterization of these trace elements. All these aspects are illustrated with up-to-date applications from the literature. Finally, future trends in mass spectrometry give the readers new insights in Se and Te species detection. [less ▲]

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See detailSurface and bio-adhesion properties of new hydrophobic and current materials for artificial intraocular lens
Bertrand, Virginie ULg; Svaldo Lanero, Tiziana ULg; Duwez, Anne-Sophie ULg et al

Poster (2012)

A high bio-adhesion appears to be one of the key factor for posterior capsular opacification (PCO) prevention. Indeed, the proteins adsorption and the lens epithelial cells (LEC) adhesion both contribute ... [more ▼]

A high bio-adhesion appears to be one of the key factor for posterior capsular opacification (PCO) prevention. Indeed, the proteins adsorption and the lens epithelial cells (LEC) adhesion both contribute to PCO development. We present in this work the comparison of a new glistening free hydrophobic material (GF® from Physiol) with benchmark hydrophobic and hydrophilic materials regarding their chemicophysical properties and their respective ability to interact with lens epithelial cells and proteins. For this purpose, we determined the hydrophobicity by contact angle measurement (assessed by water drop and air bubble methods), the surface adhesiveness by atomic force microscopy (AFM), the proteins adsorption by fluorescent measurement and the LEC adhesion by the determination of cell density. The new hydrophobic material presents comparable hydrophobicity, proteins adsorption and LEC adhesion to current commercial hydrophobic material. Its adhesiveness, measured with the AFM, is intermediate between hydrophilic and hydrophobic materials. In conclusion, the bio-adhesion properties of this new glistening free hydrophobic IOL material are similar to generic hydrophobic acrylic materials and therefore should to the same extent prevent PCO. [less ▲]

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See detailAn Unusual Family of Glycosylated Peptides Isolated from Dendroaspis angusticeps Venom and Characterized by Combination of Collision Induced and Electron Transfer Dissociation
Quinton, Loïc ULg; Gilles, Nicolas; Smargiasso, Nicolas ULg et al

in Journal of the American Society for Mass Spectrometry (2011), 22(11), 1891-1897

This study describes the structural characterization of a totally new family of peptides from the venom of the snake green mamba (Dendroaspis angusticeps). Interestingly, these peptides differ in several ... [more ▼]

This study describes the structural characterization of a totally new family of peptides from the venom of the snake green mamba (Dendroaspis angusticeps). Interestingly, these peptides differ in several points from other already known mamba toxins. First of all, they exhibit very small molecular masses, ranging from 1.3 to 2.4 kDa. The molecular mass of classical mamba toxins is in the range of 7 to 25 kDa. Secondly, the new peptides do not contain disulfide bonds, a post-translational modification commonly encountered in animal toxins. The third difference is the very high proportion of proline residues in the sequence accounting for about one third of the sequence. Finally, these new peptides reveal a carbohydrate moiety, indicating a glycosylation in the sequence. The last two features have made the structural characterization of the new peptides by mass spectrometry a real analytical challenge. Peptides were characterized by a combined use of MALDI- TOF/TOF and nanoESI-IT-ETD experiments to determine not only the peptide sequence but also the composition and the position of the carbohydrate moiety. Anyway, such small glycosylated and proline-rich toxins are totally different from any other known snake peptide and form, as a consequence, a new family of peptides. [less ▲]

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See detailReproductive impact and proteomic analysis of androgenic and anti-androgenic disruptors on the hermaphroditic freshwater gastropod Lymnaea stagnalis
Giusti, Arnaud ULg; Ducrot, Virginie; Joaquim-Justo, Célia ULg et al

Poster (2011, November)

In this study, e ects of two androgens (tributyltin and testosterone), one antiandrogen (cyproterone acetate) and one estrogen (chlordecone) on growth and reproduction were investigated in the ... [more ▼]

In this study, e ects of two androgens (tributyltin and testosterone), one antiandrogen (cyproterone acetate) and one estrogen (chlordecone) on growth and reproduction were investigated in the hermaphrodite gastropod Lymnaea stagnalis. In this study, exposure to a range of concentrations (ng/l to μg/l) of each chemical was performed during 21 days. e number of clutches and the number of eggs per clutch were monitored. A decrease in clutches laid per snail was observed after exposure to tributyltin (540, 1180 and 2600 ng/l) and chlordecone (10, 22, 50 and 110 μg/l). A signi cant decrease in egg laid per snail was observed after exposure to tributyltin (540, 1180 and 2600 ng/l) and chlordecone (50 and 110 μg/l). An increase of egg abnormalities ratio in exposed snails (atrophied albumen, polyembryonicity,...) was observed in L. stagnalis following exposure to testosterone (2, 22, 50 and 110 ng/l), cyproterone acetate (4,5 and 50 μg/l), tributyltin (110 and 244 ng/l) and chlordecone (4,5 and 10 μg/l). Investigation of alteration in protein expression in exposed snails was performed using proteomic analysis such as 2D-DIGE. Mass spectrometry identi cation was performed on proteins with altered expression. We could establish correlation between reproductive endpoints and changes in proteins involved in egg formation and in egg laying were underlined. Egg yolk ferritin, the main protein of egg yolk, was shown to be reduced signi cantly in relationship with a decrease of egg yolk quality after exposure to tributyltin 540 ng/l and cyproterone acetate 4,5 μg/l. Ovipostatin, a protein proved to reduce egg masses, was signi cantly over expressed in snails exposed to 50 μg/l of chlordecone and were in relationship with a reduction of clutches laid by individuals. Further western blot analysis on those proteins involved in the reproduction are underway. ese analysis will enable us to con rm and re ne with more speci city the 2D-DIGE results for the selected proteins. e results of this study can help to establish new biomarkers of exposure of endocrine disruptors in freshwater environment and can provide new insight on mode of action of endocrine disruptors in L. stagnalis. [less ▲]

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See detailDisulfide bond assignement and folding characterization of peptide toxins by Ion Mobility Mass Spectrometry
Echterbille, Julien ULg; Quinton, Loïc ULg; Rosu, Frédéric ULg et al

Conference (2011, October 11)

Main component of animal venoms is peptide toxins, which are highly structured by several disulfide bridges. Disulfide bridges fill different roles as increasing the toxins efficiency by lowering their ... [more ▼]

Main component of animal venoms is peptide toxins, which are highly structured by several disulfide bridges. Disulfide bridges fill different roles as increasing the toxins efficiency by lowering their immunogenicity or providing the adequate conformation to efficiently bind to the biological receptor. The sequencing and the determination of the cysteine pairing is still challenging and therefore an important step in structural analysis. In this work, we present a new strategy to sequence structured toxins and assign S-S bridges using ion mobility resolved MS/MS. The method relies on the analysis of partially reduced multiple-disulfide peptide. The mixture of the different forms is resolved by ion mobility, followed by MS/MS acquisition on each mobility separated species. The proof of concept has been successfully conducted on α-CnI, a toxin purified from the venom of Conus consors marine snail. The toxin’s sequence contains four cysteines linked together with two disulfide bridges. α-CnI was partially reduced by a small excess of tris(carboxyethyl)phosphine (10:1). The resulting mixture was purified before analysis by infusion nanoESI-Synapt-G2. Fragmentation was performed after the mobility cell, to obtain specific fragments of each species. Partial reduction of α-CnI results in a mixture of oxidized (the two disulfides are formed), reduced (the two disulfides have been reduced) and partially reduced forms (one of the two disulfides has been reduced). The arrival time distribution of triply charged ions reveals the presence of 4 different species, characterized by different relative cross sections in the gas-phase. Mass matching allows identifying the species: the first mobility (the most compact structure) was identified to be the oxidized folded toxin (M). The latest peak, corresponding to the larger cross-section, was identified as the fully reduced toxin (M+4Da). The second and the third mobility peaks were attributed to the two partially reduced forms in which only one disulfide bridge was reduced (M+2Da). The change in ion mobility depends on which S-S bridge is reduced. Ion mobility separated species give characteristic fragment ions upon fragmentation in the transfer cell (i.e. after ion mobility separator). Interestingly, fragment ions coming from partially reduced species, especially the C-S or S-S bond cleavages, clearly indicates that the disulfide linkage of α-CnI is (Cys1-Cys3) and (Cys2-Cys4) as expected from literature. The method is now being applied with success to more complex systems containing 3 or 4 disulfide bridges. The influence of the charge state on the mobility separation is systematically analyzed in terms of structural implications. [less ▲]

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See detailChanges in termites feeding diets for gut micro-organisms selection and further cultivation
Bauwens, Julien ULg; Brasseur, Catherine ULg; Matteotti, Christel ULg et al

Poster (2011, October 02)

Termites gut may overcome important dietary perturbations, initial diversity acting as key point buffering effects on host, although termites possess their own enzymatic system. Some artificial diets ... [more ▼]

Termites gut may overcome important dietary perturbations, initial diversity acting as key point buffering effects on host, although termites possess their own enzymatic system. Some artificial diets permitted a simplification of the lower termites gut symbiotic system, which could be used as first step in symbionts isolation and cultivation. Preliminary assay of cultivation actually gave encouraging results. Proteomic proved to be suitable tool to investigate such a complex system. Nevertheless, for some symbionts very few genes are sequenced, which should lead to more targeted proteomic studies. Protein chromatography will allow to split up the proteome and more accurate analysis. [less ▲]

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See detailMass spectrometry as an evolutive tool for toxinology: from sequencing to toxin shapes
Quinton, Loïc ULg; Gilles, Nicolas; Echterbille, Julien ULg et al

Conference (2011, September 12)

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See detailIdentification of novel accessible proteins bearing diagnostic and therapeutic potential in human pancreatic ductal adenocarcinoma
Turtoi, Andrei ULg; Musmeci, Davide; Wang, Yinghong et al

in Journal of Proteome Research (2011), 10(9), 4302-13

Pancreas ductal adenocarcinoma (PDAC) remains a deadly malignancy with poor early diagnostic and no effective therapy. Although several proteomic studies have performed comparative analysis between normal ... [more ▼]

Pancreas ductal adenocarcinoma (PDAC) remains a deadly malignancy with poor early diagnostic and no effective therapy. Although several proteomic studies have performed comparative analysis between normal and malignant tissues, there is a lack of clear characterization of proteins that could be of clinical value. Systemically reachable ("potentially accessible") proteins, suitable for imaging technologies and targeted therapies represent a major group of interest. The current study explores potentially accessible proteins overexpressed in PDAC, employing innovative proteomics technologies. In the discovery phase, potentially accessible proteins from fresh human normal and PDAC tissues were ex vivo biotinylated, isolated and identified using 2D-nano-HPLC-MS/MS method. The analysis revealed 422 up-regulated proteins in the tumor, of which 83 (including protein isoforms) were evaluated as potentially accessible. Eleven selected candidates were further confirmed as up-regulated using Western blot and multiple reaction monitoring protein quantification. Of these, transforming growth factor beta-induced (TGFBI), latent transforming growth factor beta binding 2 (LTBP2), and asporin (ASPN) were further investigated by employing large scale immunohistochemistry-based validations. They were found to be significantly expressed in a large group of clinical PDAC samples compared to corresponding normal and inflammatory tissues. In conclusion, TGFBI, LTBP2, and ASPN are novel, overexpressed, and potentially accessible proteins in human PDAC. They bear the potential to be of clinical value for diagnostic and therapeutic applications and merit further studies using in vivo models. [less ▲]

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See detailIon Mobility - Mass Spectrometry as a new approach for the screening of pesticide residues in food
Joly, Laure; Goscinny, Séverine ULg; Touilloux, Romain et al

Conference (2011, September)

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