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See detailIdentification of a Brucella spp. secreted effector specifically interacting with human small GTPase Rab2.
de Barsy, Marie; Jamet, Alexandre; Filopon, Didier et al

in Cellular microbiology (2011), 13(7), 1044-58

Bacteria of the Brucella genus are facultative intracellular class III pathogens. These bacteria are able to control the intracellular trafficking of their vacuole, presumably by the use of yet unknown ... [more ▼]

Bacteria of the Brucella genus are facultative intracellular class III pathogens. These bacteria are able to control the intracellular trafficking of their vacuole, presumably by the use of yet unknown translocated effectors. To identify such effectors, we used a high-throughput yeast two-hybrid screen to identify interactions between putative human phagosomal proteins and predicted Brucella spp. proteins. We identified a specific interaction between the human small GTPase Rab2 and a Brucella spp. protein named RicA. This interaction was confirmed by GST-pull-down with the GDP-bound form of Rab2. A TEM-beta-lactamase-RicA fusion was translocated from Brucella abortus to RAW264.7 macrophages during infection. This translocation was not detectable in a strain deleted for the virB operon, coding for the type IV secretion system. However, RicA secretion in a bacteriological culture was still observed in a DeltavirB mutant. In HeLa cells, a DeltaricA mutant recruits less GTP-locked myc-Rab2 on its Brucella-containing vacuoles, compared with the wild-type strain. We observed altered kinetics of intracellular trafficking and faster proliferation of the B. abortusDeltaricA mutant in HeLa cells, compared with the wild-type control. Altogether, the data reported here suggest RicA as the first reported effector with a proposed function for B. abortus. [less ▲]

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See detailIdentification of a continuous structure with a geometrical non-linearity. Part I: Conditioned reverse path method
Kerschen, Gaëtan ULg; Lenaerts, V.; Golinval, Jean-Claude ULg

in Journal of Sound & Vibration (2003), 262(4), 889-906

Particular effort has been spent in the field of identification of multi-degree-of-freedom non-linear systems. The newly developed methods permit the structural analyst to consider increasingly complex ... [more ▼]

Particular effort has been spent in the field of identification of multi-degree-of-freedom non-linear systems. The newly developed methods permit the structural analyst to consider increasingly complex systems. The aim of this paper and a companion paper is to study, by means of two methods, a continuous non-linear system consisting of an experimental cantilever beam with a geometrical non-linearity. In this paper (Part I), the ability of the conditioned reverse path method, which is a frequency domain technique, to identify the behaviour of this structure is assessed. The companion paper (Part II) is devoted to the application of proper orthogonal decomposition, which is an updating technique, to the test example. (C) 2002 Elsevier Science Ltd. All rights reserved. [less ▲]

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See detailIdentification of a continuous structure with a geometrical non-linearity. Part II: Proper orthogonal decomposition
Lenaerts, V.; Kerschen, Gaëtan ULg; Golinval, Jean-Claude ULg

in Journal of Sound & Vibration (2003), 262(4), 907-919

Particular effort has been spent in the field of identification of multi-degree-of-freedom non-linear systems. The newly developed methods permit the structural analyst to consider increasingly complex ... [more ▼]

Particular effort has been spent in the field of identification of multi-degree-of-freedom non-linear systems. The newly developed methods permit the structural analyst to consider increasingly complex systems. The aim of this paper and a companion paper is to study, by means of two methods, a continuous non-linear system consisting of an experimental cantilever beam with a geometrical non-linearity. In the companion paper (Part I) [1] the ability of the conditioned reverse path method, which is a frequency domain technique, to identify the behaviour of this structure-is assessed. This paper (Part II) is devoted to the application of proper orthogonal decomposition, which is an updating technique, to the test example. (C) 2002 Elsevier Science Ltd. All rights reserved. [less ▲]

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See detailIdentification of a family harboring a novel LHBéta subunit mutation associated with hypogonadism
Daly, Adrian ULg; Salvi, R.; Ménagé, J.-J. et al

in The Endocrine Society's - 88th Annual Meeting - Abstract book (2006, June)

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See detailIdentification of a fossil wood specimen in the Red Sandstone Group of Southwestern Tanzania: stratigraphical and tectonic implications.
Damblon, Freddy ULg; Gerrienne, Philippe ULg; Doutrelepont, Hughes et al

in Journal of African Earth Sciences (1998), 26

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See detailIdentification of a karyopherin alpha 2 recognition site in PLAG1, which functions as a nuclear localization signal.
Braem, Caroline V; Kas, Koen; Meyen, Eva et al

in Journal of Biological Chemistry (2002), 277(22), 19673-8

The activation of the pleomorphic adenoma gene 1 (PLAG1) is the most frequent gain-of-function mutation found in pleomorphic adenomas of the salivary glands. To gain more insight into the regulation of ... [more ▼]

The activation of the pleomorphic adenoma gene 1 (PLAG1) is the most frequent gain-of-function mutation found in pleomorphic adenomas of the salivary glands. To gain more insight into the regulation of PLAG1 function, we searched for PLAG1-interacting proteins. Using the yeast two-hybrid system, we identified karyopherin alpha2 as a PLAG1-interacting protein. Physical interaction between PLAG1 and karyopherin alpha2 was confirmed by an in vitro glutathione S-transferase pull-down assay. Karyopherin alpha2 escorts proteins into the nucleus via interaction with a nuclear localization sequence (NLS) composed of short stretches of basic amino acids. Two putative NLSs were identified in PLAG1. The predicted NLS1 (KRKR) was essential for physical interaction with karyopherin alpha2 in glutathione S-transferase pull-down assay, and its mutation resulted in decreased nuclear import of PLAG1. Moreover, NLS1 was able to drive the nuclear import of the cytoplasmic protein beta-galactosidase. In contrast, predicted NLS2 of PLAG1 (KPRK) was not involved in karyopherin alpha2 binding nor in its nuclear import. The residual nuclear import of PLAG1 after mutation of the NLS1 was assigned to the zinc finger domain of PLAG1. These observations indicate that the nuclear import of PLAG1 is governed by its zinc finger domain and by NLS1, a karyopherin alpha2 recognition site. [less ▲]

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See detailIdentification of a magnetic anomaly at Jupiter from satellite footprints
Grodent, Denis ULg

E-print/Working paper (2004)

Repeated imaging of Jupiter's aurora has shown that the northern main oval has a distorted 'kidney bean' shape in the general range of 90-140? System III longitude, which appears unchanged since 1994 ... [more ▼]

Repeated imaging of Jupiter's aurora has shown that the northern main oval has a distorted 'kidney bean' shape in the general range of 90-140? System III longitude, which appears unchanged since 1994. While it is more difficult to observe the conjugate regions in the southern aurora, no corresponding distortion appears in the south. Recent improved accuracy in locating the satellite footprint auroral emissions has provided new information about the geometry of Jupiter's magnetic field in this and other areas. The study of the magnetic field provides us with insight into the state of matter and the dynamics deep down Jupiter. There is currently no other way to do this from orbit. The persistent pattern of the main oval implies a disturbance of the local magnetic field, and the increased latitudinal separation of the locus of satellite footprints from each other and from the main oval implies a locally weaker field strength. It is possible that these phenomena result from a magnetic anomaly in Jupiter's intrinsic magnetic field, as was proposed by A. Dessler in the 1970's. There is presently only limited evidence from the scarcity of auroral footprints observed in this longitude range. We propose to obtain HST UV images with specific observing geometries of Jupiter to determine the locations of the auroral footprints of Io, Europa, and Ganymede in cycle 13 to accurately determine the magnetic field geometry in the suggested anomaly region, and to either confirm or refute the suggestion of a local magnetic anomaly. [less ▲]

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See detailIdentification of a microRNA landscape targeting the PI3K/Akt signaling pathway in inflammation-induced colorectal carcinogenesis
JOSSE, Claire ULg; Bouznad, Nassim ULg; Geurts, Pierre ULg et al

in American Journal of Physiology - Gastrointestinal and Liver Physiology (2014), 306

Inflammation can contribute to tumor formation; however, markers that predict progression are still lacking. In the present study, the well-established azoxymethane (AOM)/dextran sulfate sodium (DSS ... [more ▼]

Inflammation can contribute to tumor formation; however, markers that predict progression are still lacking. In the present study, the well-established azoxymethane (AOM)/dextran sulfate sodium (DSS)-induced mouse model of colitis-associated cancer was used to analyze microRNA (miRNA) modulation accompanying inflammation-induced tumor development and to determine whether inflammation-triggered miRNA alterations affect the expression of genes or pathways involved in cancer. A miRNA microarray experiment was performed to establish miRNA expression profiles in mouse colon at early and late time points during inflammation and/or tumor growth. Chronic inflammation and carcinogenesis were associated with distinct changes in miRNA expression. Nevertheless, prediction algorithms of miRNA-mRNA interactions and computational analyses based on ranked miRNA lists consistently identified putative target genes that play essential roles in tumor growth or that belong to key carcinogenesis-related signaling pathways. We identified PI3K/Akt and the insulin growth factor-1 (IGF-1) as major pathways being affected in the AOM/DSS model. DSS-induced chronic inflammation downregulates miR-133a and miR-143/145, which is reportedly associated with human colorectal cancer and PI3K/Akt activation. Accordingly, conditioned medium from inflammatory cells decreases the expression of these miRNA in colorectal adenocarcinoma Caco-2 cells. Overexpression of miR-223, one of the main miRNA showing strong upregulation during AOM/DSS tumor growth, inhibited Akt phosphorylation and IGF-1R expression in these cells. Cell sorting from mouse colons delineated distinct miRNA expression patterns in epithelial and myeloid cells during the periods preceding and spanning tumor growth. Hence, cell-type-specific miRNA dysregulation and subsequent PI3K/Akt activation may be involved in the transition from intestinal inflammation to cancer. [less ▲]

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See detailIdentification of a new aphid isoprenyl diphosphate synthase
Vandermoten, Sophie ULg; Beliveau, Catherine; Sen, Stéphanie et al

Conference (2006, November)

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See detailIdentification of a Nonlinear Wing Structure Using an Extended Modal Model
Platten, M. F.; Wright, J. R.; Cooper, J. E. et al

in Journal of Aircraft (2009), 46(5), 1614-1626

The nonlinear resonant decay method identifies a nonlinear dynamic system using a model based in linear modal space comprising the underlying linear system and a small number of additional terms that ... [more ▼]

The nonlinear resonant decay method identifies a nonlinear dynamic system using a model based in linear modal space comprising the underlying linear system and a small number of additional terms that represent the nonlinear behavior. In this work, the method is applied to an aircraftlike wing/store/pylon experimental structure that consists of a rectangular wing with two stores suspended beneath it by means of nonlinear pylons with a nominally hardening characteristic in the store rotation degree of freedom. The nonlinear resonant decay method is applied to the system using multishaker excitation. The resulting identified mathematical model features five modes, two of which are strongly nonlinear, one is mildly nonlinear, and two are completely linear. The restoring force surfaces obtained from the mathematical model are in close agreement with those measured from the system. This experimental application of the nonlinear resonant decay method indicates that the method could be suitable for the identification of nonlinear models of aircraft in ground vibration testing. [less ▲]

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See detailIdentification of a novel autoantigen in inflammatory bowel disease by protein microarray.
Vermeulen, Nathalie; de Beeck, Katrijn Op; Vermeire, Severine et al

in Inflammatory Bowel Diseases (2011), 17(6), 1291-300

BACKGROUND: Patients with inflammatory bowel disease (IBD) display immunoreactivity to self-antigens and microbial antigens. We used a protein microarray approach to identify novel autoantigens in IBD ... [more ▼]

BACKGROUND: Patients with inflammatory bowel disease (IBD) display immunoreactivity to self-antigens and microbial antigens. We used a protein microarray approach to identify novel autoantigens in IBD. METHODS: ProtoArray Human Protein Microarray v4.0 containing 8268 human proteins from Invitrogen (La Jolla, CA) was used. RESULTS: Twenty-five IBD patients and five healthy controls were screened for candidate autoantigens. For 256 antigens, IBD patients had a higher seroreactivity than controls. Twenty antigens were selected for further evaluation in a larger cohort (60 ulcerative colitis [UC] patients, 60 Crohn's disease [CD] patients, 60 healthy controls, and 60 gastrointestinal-diseased controls) by means of a customized protein microarray. Out of these 20 antigens, one antigen, family with sequence similarity 84 member A (FAM84A), was identified as a target antigen in IBD. Antibodies to FAM84A were significantly more prevalent in IBD patients (19%) than in gastrointestinal-diseased controls (1.7%) (P = 0.0008) and healthy controls (5%) (P = 0.01). Anti-FAM84A antibodies were found in 26.6% of UC patients and in 11.7% of CD patients. FAM84A was confirmed as target antigen in IBD by means of Western blotting in a large independent cohort (100 UC patients, 106 CD patients, 102 healthy controls, and 100 gastrointestinal-diseased controls). Antibodies to FAM84A were significantly more prevalent in IBD patients (20%) than in gastrointestinal-diseased controls (5%) (P = 0.0004) and healthy controls (0%) (P < 0.0001). Anti-FAM84A antibodies were found in 18% of UC patients and in 22% of CD patients. CONCLUSIONS: We identified FAM84A as a novel autoantigen in IBD. (Inflamm Bowel Dis 2011;). [less ▲]

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See detailIdentification of a novel snake peptide displaying high affinity and antagonist behaviour for the alpha2-adrenoreceptors
Rouget, Céline; Quinton, Loïc ULg; Maïga, Arhamatoulaye et al

in British Journal of Pharmacology (2010), 161

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See detailIdentification of a pharmacologically tractable Fra-1/ADORA2B axis promoting breast cancer metastasis
Desmet, Christophe ULg; Gallenne, Tristan; Prieur, Alexandre et al

in Proceedings of the National Academy of Sciences of the United States of America (2013)

Metastasis confronts clinicians with two major challenges: estimating the patient's risk of metastasis and identifying therapeutic targets. Because they are key signal integrators connecting cellular ... [more ▼]

Metastasis confronts clinicians with two major challenges: estimating the patient's risk of metastasis and identifying therapeutic targets. Because they are key signal integrators connecting cellular processes to clinical outcome, we aimed to identify transcriptional nodes regulating cancer cell metastasis. Using rodent xenograft models that we previously developed, we identified the transcription factor Fos-related antigen-1 (Fra-1) as a key coordinator of metastasis. Because Fra-1 often is overexpressed in human metastatic breast cancers and has been shown to control their invasive potential in vitro, we aimed to assess the implication and prognostic significance of the Fra-1-dependent genetic program in breast cancer metastasis and to identify potential Fra-1-dependent therapeutic targets. In several in vivo assays in mice, we demonstrate that stable RNAi depletion of Fra-1 from human breast cancer cells strongly suppresses their ability to metastasize. These results support a clinically important role for Fra-1 and the genetic program it controls. We show that a Fra-1-dependent gene-expression signature accurately predicts recurrence of breast cancer. Furthermore, a synthetic lethal drug screen revealed that antagonists of the adenosine receptor A2B (ADORA2B) are preferentially toxic to breast tumor cells expressing Fra-1. Both RNAi silencing and pharmacologic blockade of ADORA2B inhibited filopodia formation and invasive activity of breast cancer cells and correspondingly reduced tumor outgrowth in the lungs. These data show that Fra-1 activity is causally involved in and is a prognostic indicator of breast cancer metastasis. They suggest that Fra-1 activity predicts responsiveness to inhibition of pharmacologically tractable targets, such as ADORA2B, which may be used for clinical interference of metastatic breast cancer. [less ▲]

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See detailIdentification of a pharmacophore of SKCa channel blockers
Dilly, Sébastien ULg; Graulich, Amaury ULg; Farce, Amaury et al

in Journal of Enzyme Inhibition and Medicinal Chemistry (2005), 20(6), 517-523

Small conductance calcium-activated potassium channels ( SK) are widely expressed throughout the central nervous system ( CNS) and the periphery. Three subtypes of SK channels have so far been identified ... [more ▼]

Small conductance calcium-activated potassium channels ( SK) are widely expressed throughout the central nervous system ( CNS) and the periphery. Three subtypes of SK channels have so far been identified in different parts of the brain. Activation of the SK channels by a rise in intracellular calcium leads to the hyperpolarisation of the membrane, reducing cell excitability. Blocking the SK channels might be beneficial in the treatment of depression, Parkinson's disease and cognitive disorders. However, few blockers of SK channels have been characterized. In this study, a pharmacophoric model of SK channels blockers is presented. It is based on a series of nonpeptidic compounds and apamin, a peptidic blocker. To create the pharmacophore model, the conformational space of nonpeptidic blockers was investigated to generate a series of distance constraints applied to a simulated annealing study of apamin. The resulting conformation was superimposed with the nonpeptidic blockers to give a pharmacophore. [less ▲]

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See detailIdentification of a Time-varying Beam Using Hilbert Vibration Decomposition
Bertha, Mathieu ULg; Golinval, Jean-Claude ULg

in Proceedings of the International Modal Analysis Conference (IMAC) XXXII (2014, February)

The present work is concerned by modal identification of time-varying systems. For this purpose, a method based on instantaneous frequency identification and synchronous demodulation is used to extract ... [more ▼]

The present work is concerned by modal identification of time-varying systems. For this purpose, a method based on instantaneous frequency identification and synchronous demodulation is used to extract modal components from recorded signals. The proposed method of iterated sifting process is based on the Hilbert Vibration Decomposition (HVD) technique which is used to extract the instantaneous dominant vibrating component at each iteration. A source separation preprocessing step is introduced to treat multiple degree-of-freedom systems in an optimal way. Sources are used as reference signals to get a single instantaneous frequency of each mode for the demodulation on all the channels. The algorithm is presented and is applied to numerical simulation of a randomly excited time-varying structure for illustration purpose. The investigated structure is made up of a beam on which a non-negligible mass is traveling. The variable location of the mass results in changes in modal parameters. [less ▲]

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See detailIdentification of a universal VHH framework to graft non-canonical antigen-binding loops of camel single-domain antibodies
Saerens, Dirk; Pellis, Mireille; Loris, Remy et al

in Journal of Molecular Biology (2005), 352

Camel single-domain antibody fragments (VHHs) are promising tools in numerous biotechnological and medical applications. However, some conditions under which antibodies are used are so demanding that they ... [more ▼]

Camel single-domain antibody fragments (VHHs) are promising tools in numerous biotechnological and medical applications. However, some conditions under which antibodies are used are so demanding that they can be met by only the most robust VHHs. A universal framework offering the required properties for use in various applications (e.g. as intrabody, as probe in biosensors or on micro-arrays) is highly valuable and might be further implemented when employment of VHHs in human therapy is envisaged. We identified the VHH framework of cAbBCII10 as a potential candidate, useful for the exchange of antigen specificities by complementarity determining region (CDR) grafting. Due to the large number of CDRH loop structures present on VHHs, this grafting technique was expected to be rather unpredictable. Nonetheless, the plasticity of the cAbBCII10 framework allows successful transfer of antigen specificity from donor VHHs onto its scaffold. The cAbBCII10 was chosen essentially for its high level of stability (47 kJ/mol), good expression level (5 mg/l in E. coli) and its ability to be functional in the absence of the conserved disulfide bond. All five chimeras generated by grafting CDR-Hs, from donor VHHs belonging to subfamily 2 that encompass 75% of all antigen-specific VHHs, on the framework of cAbBCII10 were functional and generally had an increased thermodynamic stability. The grafting of CDR-H loops from VHHs belonging to other subfamilies resulted in chimeras of reduced antigen-binding capacity. [less ▲]

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See detailIdentification of accessible human cancer biomarkers using ex vivo chemical proteomic strategies
Kischel, Philippe ULg; Waltregny, David ULg; Castronovo, Vincenzo ULg

in Expert Review of Proteomics (2007), 4(6), 727-739

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

One promising avenue towards the development of more selective, better anticancer drugs lies 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 ideally use three criteria: accessibility from the bloodstream; expression at sufficient level, and 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. To address this limitation, our group recently developed two methodologies based on chemical proteomic modifications, enabling the discovery of proteins accessible from the bloodstream and the extracellular space in human pathological tissues. In this review, we will discuss the potential benefits of these methodologies for the fast discovery of therapeutically valuable biomarkers. [less ▲]

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