References of "De Pauw, Edwin"
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See detailThiaminylated adenine nucleotides — chemical synthesis, structural characterization and natural occurrence
Frederich, Michel ULiege; Delvaux, David ULiege; Gigliobianco, Tiziana ULiege et al

in FEBS Journal (2009), 276(12), 32563268

Thiamine and its three phosphorylated derivatives (mono-, di- and triphosphate) occur naturally in most cells. Recently, we reported the presence of a fourth thiamine derivative, adenosine thiamine ... [more ▼]

Thiamine and its three phosphorylated derivatives (mono-, di- and triphosphate) occur naturally in most cells. Recently, we reported the presence of a fourth thiamine derivative, adenosine thiamine triphosphate (AThTP), produced in E. coli in response to carbon starvation. Here, we show that the chemical synthesis of AThTP leads to another new compound, adenosine thiamine diphosphate (thiaminylated ADP, AThDP), as a side product. The structure of both compounds was confirmed by mass spectrometry and 1H-, 13C- and 31P-NMR and some of their chemical properties were determined. Our results show an upfield shifting of the C-2 proton of the thiazolium ring in adenosine thiamine derivatives compared to the conventional thiamine phosphate derivatives. This modification of the electronic environment of the C-2 proton might be explained by a through-space interaction with the adenosine moiety, suggesting an U-shaped folding of adenosine thiamine derivatives. Such a structure where the C-2 proton is embedded in a closed conformation can be located using molecular modeling as an energy minimum. In E. coli, AThTP may account for 15% of total thiamine under energy stress. It is less abundant in eukaryotic organisms, but is consistently found in mammalian tissues and in some cell lines. Using a HPLC method, we show for the first time that AThDP may also occur in small amounts in E. coli and in vertebrate liver. The discovery of two natural thiamine adenine compounds further highlights the complexity and diversity of thiamine biochemistry, which is not restricted to the cofactor role of thiamine diphosphate. [less ▲]

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See detailGCxGC coupled to fast scanning quadrupole MS for trace analysis of POPs
Kinet, Catherine ULiege; De Pauw, Edwin ULiege; Focant, Jean-François ULiege

in Organohalogen Compounds (2009), 70

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See detailLe ciblage thérapeutique : vers une guerre propre et efficace contre le cancer
Castronovo, Vincenzo ULiege; Waltregny, David ULiege; Detry, Olivier ULiege et al

in Revue Médicale de Liège (2009), 64

One promising avenue towards the development of more selective, better anticancer drugs consists in the targeted delivery of bioactive compounds to the tumor environment by means of binding molecules ... [more ▼]

One promising avenue towards the development of more selective, better anticancer drugs consists in the targeted delivery of bioactive compounds to the tumor environment by means of binding molecules specific for tumor-associated biomarkers. Eligibility of such markers for therapeutic use implies ideally three criteria : (i) accessibility from the bloodstream, (ii) expression at sufficient level and (iii) no (or much lower) expression in normal tissues. Most current discovery strategies (such as biomarker searching into body fluids) provide no clue as to whether proteins of interest are accessible, in human tissues, to suitable high-affinity ligands, such as systemically delivered monoclonal antibodies. Innovative proteomic technologies are able to identify such accessible biomarkers and represent a key step in the clinical development of such target therapies. [less ▲]

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See detailPositively Cooperative Binding of Zinc Ions to Bacillus cereus 569/H/9 beta-Lactamase II Suggests that the Binuclear Enzyme Is the Only Relevant Form for Catalysis
Jacquin, Olivier ULiege; Balbeur, Dorothée ULiege; Damblon, Christian ULiege et al

in Journal of Molecular Biology (2009), 392(5), 1278-1291

Metallo-beta-lactamases catalyze the hydrolysis of most beta-lactam antibiotics and hence represent a major clinical concern. While enzymes belonging to subclass B1 have been shown to display maximum ... [more ▼]

Metallo-beta-lactamases catalyze the hydrolysis of most beta-lactam antibiotics and hence represent a major clinical concern. While enzymes belonging to subclass B1 have been shown to display maximum activity as dizinc species, the actual metal-to-protein stoichiometry and the affinity for zinc are not clear. We have further investigated the process of metal binding to the beta-lactamase H from Bacillus cereus 569/H/9 (known as BcII). Zinc binding was monitored using complementary biophysical techniques, including circular dichroism in the far-UV, enzymatic activity measurements, competition with a chromophoric chelator, mass spectrometry, and nuclear magnetic resonance. Most noticeably, mass spectrometry and nuclear magnetic resonance experiments, together with catalytic activity measurements, demonstrate that two zinc ions bind cooperatively to the enzyme active site (with K-1/K-2 >= 5) and, hence, that catalysis is associated with the dizinc enzyme species only. Furthermore, competitive experiments with the chromophoric chelator Mag-Fura-2 indicates K-2 < 80 nM. This contrasts with cadmium binding, which is clearly a noncooperative process with the mono form being the only species significantly populated in the presence of 1 molar equivalent of Cd(II). Interestingly, optical measurements reveal that although the apo and dizinc species exhibit undistinguishable tertiary structural organizations, the metal-depleted enzyme shows a significant decrease in its alpha-helical content, presumably associated with enhanced flexibility. (C) 2009 Elsevier Ltd. All rights reserved. [less ▲]

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See detailElectron detachment dissociation (EDD) pathways in oligonucleotides
Kinet, Catherine ULiege; Gabelica, Valérie ULiege; Balbeur, Dorothée ULiege et al

in International Journal of Mass Spectrometry (2009), 283

Electron detachment dissociation (EDD) and electron photodetachment dissociation (EPD) are two novel fragmentation methods yielding radicals from negatively charged ions. With the goal of comparing EDD ... [more ▼]

Electron detachment dissociation (EDD) and electron photodetachment dissociation (EPD) are two novel fragmentation methods yielding radicals from negatively charged ions. With the goal of comparing EDD, EPD and the more traditional Collision-Induced Dissociation (CID) and Infrared Multiphoton Disscociation (IRMPD) fragmentation processes in oligonucleotides, we studied here the EDD fragmentation pathways of oligonucleotides of varying length. We chose polythymine oligonucleotides because these are the least prone to secondary structure formation, and found complete sequence coverage by EDD for up to dT20. We also found that the fragmentation pathways change with oligonucleotide length: electron detachment is a mandatory step in the fragmentation of larger sequences, while shorter oligonucleotides can also fragment via direct electronic or vibrational excitation by the electrons. This is supported by (1) the fact that continuous ejection of the charge reduced species does not totally prevent fragmentation of short oligonucleotides dT5 and dT6, (2) the fact that CID and EDD fragments are more similar for small oligonucleotides (although double resonance experiments show that they are not all issued from the same mechanisms), and (3) the fact that electron-induced dissociation (EID) of singly charged dT3 and dT4 gives similar fragments as EDD of doubly charged dT5 and dT6. Finally, the detachment efficiency as a function of the nature of the nucleobase was studied. The effect of base on electron detachment in EDD (G > T > A > C) is different than in EPD (G > A > C > T), indicating different electron loss mechanisms. [less ▲]

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See detailHybridization of short complementary PNAs to G-quadruplex forming oligonucleotides: An electrospray mass spectrometry study.
Amato, Jussara; Oliviero, Giorgia; De Pauw, Edwin ULiege et al

in Biopolymers (2009), 91(4), 244-255

We investigated the interaction of the short PNA strand [acccca]-PNA with oligodeoxynucleotides containing one, two, or four tracts of TGGGGT units. Electrospray ionization mass spectrometry allowed ... [more ▼]

We investigated the interaction of the short PNA strand [acccca]-PNA with oligodeoxynucleotides containing one, two, or four tracts of TGGGGT units. Electrospray ionization mass spectrometry allowed exploring the wide variety of complex stoichiometries that were found to coexist in solution. In water, the PNA strand forms short heteroduplexes with the complementary DNA sequences, but higher-order structures are also found, with PNA(2n)*DNA(n) triplex units, culminating in precipitation at very low ionic strength. In the presence of ammonium acetate, there is a competition between PNA*DNA heteroduplex formation and DNA G-quadruplex formation. Heteroduplex formation is favored when the PNA + DNA mixture in ammonium acetate is heated and cooled at room temperature, but not if the PNA is added at room temperature to the pre-formed G-quadruplex. We also found that the short [acccca]-PNA strand binds to G-quadruplexes. (c) 2008 Wiley Periodicals, Inc. Biopolymers, 2008. [less ▲]

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See detailA new method for the determination of the relative affinity of a ligand against various DNA sequences by electrospray ionization mass spectrometry. Application to a polyamide minor groove binder
Buchmann, W.; Boutorine, A.; Halby, L. et al

in Journal of Mass Spectrometry [=JMS] (2009), 44

A new method for the determination of the relative affinity of a ligand against various dsDNA sequences is presented by using electrospray ionization time-of-flight mass spectrometry (ESI-QTOF) mass ... [more ▼]

A new method for the determination of the relative affinity of a ligand against various dsDNA sequences is presented by using electrospray ionization time-of-flight mass spectrometry (ESI-QTOF) mass spectrometry. Thismethod does not require knowing the ligand concentration accurately. It allows determination of the relative affinity of a ligand against various dsDNA sequences for 1 : 1 complex stoichiometries in a quick manner without labeling. [less ▲]

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See detailPutative DNA G-quadruplex formation within the promoters of Plasmodium falciparum var genes
Smargiasso, Nicolas ULiege; Gabelica, Valérie ULiege; Damblon, Christian ULiege et al

in BMC Genomics (2009), 10

Background. Guanine-rich nucleic acid sequences are capable of folding into an intramolecular four-stranded structure called a G-quadruplex. When found in gene promoter regions, G-quadruplexes can ... [more ▼]

Background. Guanine-rich nucleic acid sequences are capable of folding into an intramolecular four-stranded structure called a G-quadruplex. When found in gene promoter regions, G-quadruplexes can downregulate gene expression, possibly by blocking the transcriptional machinery. Here we have used a genome-wide bioinformatic approach to identify Putative G-Quadruplex Sequences (PQS) in the Plasmodium falciparum genome, along with biophysical techniques to examine the physiological stability of P. falciparum PQS in vitro. Results. We identified 63 PQS in the non-telomeric regions of the P. falciparum clone 3D7. Interestingly, 16 of these PQS occurred in the upstream region of a subset of the P. falciparum var genes (group B var genes). The var gene family encodes PfEMP1, the parasite’s major variant antigen and adhesin expressed at the surface of infected erythrocytes, that plays a key role in malaria pathogenesis and immune evasion. The ability of the PQS found in the upstream regions of group B var genes (UpsB-Q) to form stable Gquadruplex structures in vitro was confirmed using 1H NMR, circular dichroism, UV spectroscopy, and thermal denaturation experiments. Moreover, the synthetic compound BOQ1 that shows a higher affinity for DNA forming quadruplex rather than duplex structures was found to bind with high affinity to the UpsB-Q. Conclusions. This is the first demonstration of non-telomeric PQS in the genome of P. falciparum that form stable G-quadruplexes under physiological conditions in vitro. These results allow the generation of a novel hypothesis that the G-quadruplex sequences in the upstream regions of var genes have the potential to play a role in the transcriptional control of this major virulence-associated multi-gene family. [less ▲]

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See detailA Simple Method to Determine Electrospray Response Factors of Noncovalent Complexes
Gabelica, Valérie ULiege; Rosu, Frédéric ULiege; De Pauw, Edwin ULiege

in Analytical Chemistry (2009), 81(16), 6708-6715

The quantitative study of noncovalent complexes by electrospray mass spectrometry requires the determination of the relative response of each species. The method proposed here to determine the ... [more ▼]

The quantitative study of noncovalent complexes by electrospray mass spectrometry requires the determination of the relative response of each species. The method proposed here to determine the electrospray response factors is based on the use of (1) an internal standard and (2) the mass balance equation applied to one binding partner M, for which different complexes MxLy are detected in the electrospray mass spectra. A set of experiments providing various ratios between the complexes (e.g. different ligand concentrations in a titration experiment or different time points in a kinetics experiment) is used to generate a set of independent linear equations that can be solved using simple matrix algebra to find the response factors of each MxLy complex relative to that of the internal standard. The response factors can then be used to determine equilibrium dissociation constants or for the quantitative monitoring of reaction kinetics. The first is illustrated with a study of DNA-ligand complexes, where we show that neither minor groove binding nor intercalation dramatically affects the DNA response factor. The second is illustrated with a study of the association kinetics of the telomeric G-quadruplex dGGG(TTAGGG)3 with its complementary strand, where the response factors allow correcting for the relative response of the quadruplex and the long duplex and obtaining reproducible association rate constants independently of the source tuning potentials. [less ▲]

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See detailAdvances in quality control for dioxins monitoring and evaluation of measurement uncertainty from quality control data.
Eppe, Gauthier ULiege; De Pauw, Edwin ULiege

in Journal of Chromatography. B : Analytical Technologies in the Biomedical & Life Sciences (2009), 877

This paper describes an application of multivariate and multilevel quality control charts with the aim of improving the internal quality control (IQC) procedures for the monitoring of dioxins and dioxin ... [more ▼]

This paper describes an application of multivariate and multilevel quality control charts with the aim of improving the internal quality control (IQC) procedures for the monitoring of dioxins and dioxin-like PCBs analysis in food. Dioxin analysts have to use the toxic equivalent concept (TEQ) to assess the toxicity potential of a mixture of dioxin-like compounds. The TEQ approach requires quantifying individually 29 dioxin-like compounds. Monitoring the congeners separately on univariate QC charts is misleading owing to the increase of false alarm rate. We propose to subdivide the TEQ value into 3 sub-groups and to control simultaneously the 3 variables in a T(2) chart. When a T(2) exceeds the upper control limit, it acts as a warning to trigger additional investigations on individual congeners. We discuss the minimum number of runs required to reliably estimate the QC chart parameters and we suggest using data from multilevel QC charts to properly characterize the standard deviations and the correlation coefficients. Moreover, the univariate QC chart can be sensitised to detect systematic errors by using exponentially weighted moving average (EWMA) technique. The EWMA chart provides an additional guidance on setting appropriate criteria to control the method bias and to support trend analysis. Finally, we present an estimate of measurement uncertainty by computing the accuracy profile in a retrospective way with the QC data generated and we discuss assessment of compliance with regulatory maximum levels. [less ▲]

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See detailTranscriptomic and proteomic analyses of seasonal photoperiodism in the pea aphid
LE TRIONNAIRE, G.; Francis, Frédéric ULiege; JAUBERT-POSSAMAI, S. et al

in BMC Genomics (2009), 29

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See detailMCF-7/BOS cells membrane proteome: comparison of two isolation methods using mass spectrometry
Bertrand, Virginie ULiege; Massart, Anne-Cécile ULiege; De Pauw-Gillet, Marie-Claire ULiege et al

Poster (2009)

Membrane proteins play an important role in biological processes but their isolation and quantification using classical techniques is often limited due to their poor solubility and relatively low ... [more ▼]

Membrane proteins play an important role in biological processes but their isolation and quantification using classical techniques is often limited due to their poor solubility and relatively low abundance. These membrane markers have to be accessible to antibodies and should be potential therapeutic targets. The tests were conducted on MCF-7 / BOS cell line, immortal and easier to cultivate. The goal of this work is to obtain a pure membrane fraction to facilitate the analysis of the sample. To isolate transmembrane proteins, we compared two methods. The first one used different extraction cycles characterized by different buffers to isolate membrane proteins. The second method labelled accessible extracellular domains at the surface of MCF-7 cells with biotin prior to differential centrifugation. [less ▲]

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See detailMCF-7/BOS cells membrane proteome: comparison of two isolation methods using mass spectrometry
Bertrand, Virginie ULiege; Massart, Anne-Cécile ULiege; De Pauw-Gillet, Marie-Claire ULiege et al

Poster (2009)

Membrane proteins (MP) play an important role in biological processes. Isolation and quantification of these MP using classical techniques is often limited due to their poor solubility and relatively low ... [more ▼]

Membrane proteins (MP) play an important role in biological processes. Isolation and quantification of these MP using classical techniques is often limited due to their poor solubility and relatively low abundance. These membrane proteins enclosed markers which could be potential therapeutic targets. These potential therapeutic targets have to be accessible to antibodies and need to be presented in the plasmic membrane. Assays were conducted on MCF-7 / BOS cell line, immortal and easier to cultivate. The goal of this work is to obtain an enriched membrane fraction to facilitate the analysis of the sample and to simplify the complex proteins mixture. To isolate transmembrane proteins, we compared two methods. The first one used different extraction cycles characterized by different buffers to isolate membrane proteins. The second method labelled accessible extracellular domains at the surface of MCF-7 cells with biotin prior to differential centrifugation. The obtained enriched membrane proteome was digested with trypsin and/or Lysyl Endopeptidase. Obtained peptides were separated by 2D-HPLC chromatography and on-line analysed ion trap mass spectrometer, the Esquire HCT. [less ▲]

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See detailMembrane and cytoplasmic proteome biology of anterior mice hippocampus
Jackers, Pascale ULiege; Massart, Anne-Cécile; Depaulis, Antoine et al

Poster (2008, October 10)

The prevalence of neurodegenerative diseases increases steadily and proteomics on disease-related brain areas become more and more complex, whereas suitable samples are scarce. Due to this difficulty ... [more ▼]

The prevalence of neurodegenerative diseases increases steadily and proteomics on disease-related brain areas become more and more complex, whereas suitable samples are scarce. Due to this difficulty, animal models are indispensable in understanding human biology and disease, and the most commonly used model is the mouse (Mus musculus). Mice share many genetic and physiological characteristics with humans, breed rapidly, and can be genetically modified leading to the functional characterisation of many proteins. As for human, the mouse hippocampus is one of the most important areas of the brain and has therefore been an object of intensive investigation for many years. Mouse hippocampus has served as models for degenerating brain diseases associated with Alzheimer's disease, Parkinson's disease, ischemia, epilepsy, synaptic plasticity and underlying mechanisms of learning and memory. Using 2 anterior parts of normal mouse hippocampi, we intend to characterize membrane and cytoplasmic proteome using a modified procedure previously established (WISNIEWSKI et al., 2008). This method permit the preparation and analysis of 2 individual fractions enriched in membrane and cytoplasmic proteins. The method for separation of membrane and cytoplasm fractions comprises a stepwise depletion of non-integral membrane proteins from entire tissue homogenate by high-salt, carbonate, and urea washes. The cytoplasmic proteins obtained from high-salt depletion are considered as the soluble fraction. The membrane proteins obtained from the stepwise depletion are considered as the insoluble fraction. Enzymatic digestion of both membrane and cytoplasmic fractions are carried out without use of detergents by double digestion with endoproteinase Lys-C and trypsin. Digested peptide fractions are loaded on StageTips for rapid desalting and are separated by online reversed-phase (RP) nanoscale capillary liquid chromatography (nanoLC). Separated peptides are analyzed by electrospray tandem mass spectrometry (HPLC-ESI-MS/MS). The entire procedure allows rapid processing and preparation of samples from minute amounts as 30-40 mg frozen tissue leading to about 70 g of membrane proteins and 500 g of cytoplasmic proteins. This can be extremely helpful for proteomic profiling of small pieces of tissue and clinical material. Analyses of membrane fraction identified about 45% total membrane proteins. This fraction includes glutamate receptor 1 and 2, proteins involved into the trafficking of synaptic vesicles, lipid-anchor proteins, G proteins involved in various transmembrane signalling systems, NMDAR signalling complex proteins, voltage channel proteins… Analyses of the cytoplasmic fraction identify various proteins belonging to different compartments and/or pathways. A large amount of these proteins are specifically expressed into the hippocampus and/or into the nervous system. [less ▲]

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See detailFunctionalized plasmonic gold nanoparticles for optoacoustic cancer detection
Schol, Daureen ULiege; Fleron, Maximilien ULiege; Greisch, Jean-François et al

Poster (2008, September 12)

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See detailStudies of sequence specific recognition and interaction of bis-hairpin polyamide minor groove binders with target DNA duplexes
Boutorine, A. S.; Buchmann, W.; Halby, L. et al

in Nucleic Acids Symposium Series (2008), 52

Study of the binding of bis-MGB using DNA footprinting, native gel shift, thermal denaturation, mass spectrometry and circular dichroism. Elaboration of a method to determin

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See detailVALAPODYN: A new systems biology approach to develop predictive dynamic models of complex intracellular networks for neurological disease
Sanoudou, Despina; Depaulis, Antoine; Vafiadaki, Elizabeth et al

Poster (2008, August 22)

Objective: VALAPODYN, a European Commission funded research network, is using an original systems biology approach for the development of an innovative dynamic model of molecular interaction networks (MIN ... [more ▼]

Objective: VALAPODYN, a European Commission funded research network, is using an original systems biology approach for the development of an innovative dynamic model of molecular interaction networks (MIN) in relation to cell death and survival for the detection of new therapeutic targets for human neurological diseases. To this end, a comprehensive multidisciplinary strategy has been established combining functional genomics, proteomics and bioinformatics. Results: Using a mouse model of induced hippocampal sclerosis associated with focal epilepsy, dynamic expression analyses are conducted at different time points. Proteomic databases are being used along with advanced microarray and proteomics platform systems to investigate protein-protein interactions and regulation networks, identify and validate biological targets in complex intracellular pathways. The first phase involves whole genome and proteome analysis, integrating biological and statistical data in order to functionally annotate genes and proteins. Using Affymetrix microarrays, 2D-DIGE and MALDI/TOF-TOF, we are evaluating whole genome and proteome expression profiles bringing to light critical new pathways and molecular targets implicated in neurodegeneration. Conclusion: VALAPODYN develops a dynamic and quantitative analysis method for new therapeutic targets through MIN dynamic models and specifically addresses the systems biology of complex cellular pathways and transcriptional networks. Novel predictive dynamic models will be validated by testing the selected drug targets on innovative in vivo and in vitro models of CNS pathologies. VALAPODYN will provide a cutting-edge highly accurate in silico tool for identifying novel and effective therapeutic targets in a faster, more efficient and more economical way than it is possible today. [less ▲]

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See detailDeveloping predictive dynamic models of complex intracellular networks for neurological disease
Vafiadaki, Elizabeth; Depaulis, Antoine; Jackers, Pascale ULiege et al

in FEBS Journal (2008, June 28), 275(Issue s1), 206

Introduction: VALAPODYN, a European Commission funded research network, is an original systems biology approach for the development of an innovative model on the dynamics of molecular interaction networks ... [more ▼]

Introduction: VALAPODYN, a European Commission funded research network, is an original systems biology approach for the development of an innovative model on the dynamics of molecular interaction networks (MIN) in relation to cell death and survival for the detection of new therapeutic targets for human neurological diseases. To this end, a comprehensive multidisciplinary strategy has been established combining functional genomics, proteomics and bioinformatics. Methods and Results: Using a mouse model of induced hippocampal sclerosis associated with focal epilepsy, dynamic expression analyses are conducted at different time points. Proteomic databases are being used along with advanced microarray and proteomics platform systems to investigate protein-protein interactions and regulation networks, identify and validate biological targets in complex intracellular pathways. The first phase involves whole genome and proteome analysis, integrating biological and statistical data in order to functionally annotate genes and proteins. Using Affymetrix microarrays, 2D-DIGE and MALDI/TOF-TOF, we are evaluating whole genome and proteome expression profiles bringing to light critical new pathways and molecular targets implicated in neurodegeneration. Discussion: VALAPODYN develops a dynamic and quantitative analysis method for new therapeutic targets through MIN dynamic models and specifically addresses the systems biology of complex cellular pathways and transcriptional networks. Novel predictive dynamic models will be validated by testing the selected drug targets on innovative in vivo and in vitro models of CNS pathologies. VALAPODYN will provide a cutting-edge highly accurate in silico tool for identifying novel and effective therapeutic targets in a faster, more efficient and more economical way than it is possible today. [less ▲]

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