Corsten M.F.,Universiteitssingel 50 |
Dennert R.,Universiteitssingel 50 |
Jochems S.,Universiteitssingel 50 |
Kuznetsova T.,Catholic University of Leuven |
And 7 more authors.
Circulation: Cardiovascular Genetics | Year: 2010
Background - Small RNA molecules, called microRNAs, freely circulate in human plasma and correlate with varying pathologies. In this study, we explored their diagnostic potential in a selection of prevalent cardiovascular disorders. Methods and Results - MicroRNAs were isolated from plasmas from well-characterized patients with varying degrees of cardiac damage: (1) acute myocardial infarction, (2) viral myocarditis, (3) diastolic dysfunction, and (4) acute heart failure. Plasma levels of selected microRNAs, including heart-associated (miR-1, -133a, -208b, and -499), fibrosisassociated (miR-21 and miR-29b), and leukocyte-associated (miR-146, -155, and -223) candidates, were subsequently assessed using real-time polymerase chain reaction. Strikingly, in plasma from acute myocardial infarction patients, cardiac myocyte-associated miR-208b and -499 were highly elevated, 1600-fold (P<0.005) and 100-fold (P<0.0005), respectively, as compared with control subjects. Receiver operating characteristic curve analysis revealed an area under the curve of 0.94 (P<10-10) for miR-208b and 0.92 (P<10-9) for miR-499. Both microRNAs correlated with plasma troponin T, indicating release of microRNAs from injured cardiomyocytes. In viral myocarditis, we observed a milder but significant elevation of these microRNAs, 30-fold and 6-fold, respectively. Plasma levels of leukocyte-expressed microRNAs were not significantly increased in acute myocardial infarction or viral myocarditis patients, despite elevated white blood cell counts. In patients with acute heart failure, only miR-499 was significantly elevated (2-fold), whereas no significant changes in microRNAs studied could be observed in diastolic dysfunction. Remarkably, plasma microRNA levels were not affected by a wide range of clinical confounders, including age, sex, body mass index, kidney function, systolic blood pressure, and white blood cell count. Conclusions - Cardiac damage initiates the detectable release of cardiomyocyte-specific microRNAs-208b and -499 into the circulation. © 2010 American Heart Association, Inc.
Moksnes H.,Norwegian School of Sport Sciences |
Engebretsen L.,Norwegian School of Sport Sciences |
Engebretsen L.,University of Oslo |
Seil R.,Center Hospitalier Luxembourg |
Seil R.,Luxembourg Institute of Health
Knee Surgery, Sports Traumatology, Arthroscopy | Year: 2016
Purpose: To survey and describe the treatment of paediatric anterior cruciate ligament (ACL) injuries performed by orthopaedic surgeons affiliated with the European Society for Sports Traumatology, Knee Surgery and Arthroscopy (ESSKA). Methods: A closed e-survey was submitted to all registered members and affiliates of ESSKA in July 2013. All recipients were invited to participate in the survey by answering 34 questions online. The list of potential respondents was extracted from the ESSKA office database. Results: Invitation was sent to 2236 ESSKA members and affiliates, and received 491 (22 %) unique responses. Among the respondents, 445 (91 %) were orthopaedic surgeons, with 354 (72 %) stating that they were involved in treatment of paediatric ACL injuries. The main findings were that there are substantial differences with regard to preferred treatment algorithms, surgical techniques and long-term follow-up procedures. The summed estimate of skeletally immature children with ACL injury seen by the responders in 2012 was minimum 1923 individuals, and a minimum of 102 clinically relevant post-operative growth disturbances were registered. Conclusion: The present survey documents that the incidences of paediatric ACL injuries and idiopathic growth disturbances may be higher than previously estimated. Treatment algorithms and surgical techniques are highly diverse, and consensus could not be identified. It is worrying that only half the surgeons reported to follow-up children until skeletal maturity after surgical treatment. The results of this survey highlight the importance of international multicentre studies on paediatric ACL treatment and the development of an outcome registry to enable prospective data collections. Level of evidence: IV. © 2015, European Society of Sports Traumatology, Knee Surgery, Arthroscopy (ESSKA).
Crouzy S.,CNRS Chemistry and Biology of Metals Laboratory |
Schmit J.-C.,Center Hospitalier Luxembourg
Journal of Biomolecular Structure and Dynamics | Year: 2014
This article describes the construction and validation of a three-dimensional model of the human CC chemokine receptor 5 (CCR5) receptor using multiple homology modeling. A new methodology is presented where we built each secondary structural model of the protein separately from distantly related homologs of known structure. The reliability of our approach for G-protein coupled receptors was assessed through the building of the human C-X-C chemokine receptor type 4 (CXCR4) receptor of known crystal structure. The models are refined using molecular dynamics simulations and energy minimizations using CHARMM, a classical force field for proteins. Finally, docking models of both the natural agonists and the antagonists of the receptors CCR5 and CXCR4 are proposed. This study explores the possible binding process of ligands to the receptor cavity of chemokine receptors at molecular and atomic levels. We proposed few crucial residues in receptors binding to agonist/antagonist for further validation through experimental analysis. In particular, our study provides better understanding of the blockage mechanism of the chemokine receptors CCR5 and CXCR4, and may help the identification of new lead compounds for drug development in HIV infection, inflammatory diseases, and cancer metastasis. © 2013 Taylor & Francis.
Schmit J.-C.,Center Hospitalier Luxembourg
Biochemical Pharmacology | Year: 2011
Chemokines are small chemoattractive proteins involved in many important physiological and pathological processes such as leukocyte mobilisation, inflammation, cancer and HIV-1 infection. The N-terminus of chemokines was shown to be crucial for interaction and activation with their cognate receptors. Therefore, multiple strategies including elongation, truncation, mutagenesis or chemical modifications of chemokine N-terminus were developed to identify analogues with modified selectivity displaying antagonist or enhanced agonist activities. Library approaches allowed fast screening of a large number of such chemokine variants and led to the identification of promising therapeutic candidates. Additional studies were able to reduce the chemokine to the size of a peptide while retaining its receptor affinity and selectivity. In analogy to full length chemokines, peptides derived from the chemokine N-terminal sequence were improved by mutagenesis, elongation and truncation approaches to develop potential therapeutic molecules used in various clinical trials. Altogether these studies demonstrated the pharmacophore potential of the chemokine N-terminus and its vast modulation properties to develop analogues with great therapeutic value for a large set of pathologies. © 2011 Elsevier Inc. All rights reserved.
Devaux Y.,Laboratory of Cardiovascular Research |
Vausort M.,Laboratory of Cardiovascular Research |
Goretti E.,Laboratory of Cardiovascular Research |
Nazarov P.V.,Microarray Center |
And 7 more authors.
Clinical Chemistry | Year: 2012
BACKGROUND: Rapid and correct diagnosis of acute myocardial infarction (MI) has an important impact on patient treatment and prognosis. We compared the diagnostic performance of high-sensitivity cardiac troponin T (hs-cTnT) and cardiac enriched microRNAs (miRNAs) in patients with MI. METHODS: Circulating concentrations of cardiac-enriched miR-208b and miR-499 were measured by quantitative PCR in a case-control study of 510 MI patients referred for primary mechanical reperfusion and 87 healthy controls. RESULTS: miRNA-208b and miR-499 were highly increased in MI patients (>10 5-fold, P < 0.001) and nearly undetectable in healthy controls. Patients with ST-elevation MI (n=397) had higher miRNA concentrations than patients with non-ST-elevation MI (n =113) (P < 0.001). Both miRNAs correlated with peak concentrations of creatine kinase and cTnT (P <10 -9). miRNAs and hs-cTnT were already detectable in the plasma 1 h after onset of chest pain. In patients who presented <3 h after onset of pain, miR-499 was positive in 93% of patients and hs-cTnT in 88% of patients (P = 0.78). Overall, miR-499 and hs-cTnT provided comparable diagnostic value with areas under the ROC curves of 0.97. The reclassification index of miR-499 to a clinical model including several risk factors and hs-cTnT was not significant (P = 0.15). CONCLUSIONS: Circulating miRNAs are powerful markers of acute MI. Their usefulness in the establishment of a rapid and accurate diagnosis of acute MI remains to be determined in unselected populations of patients with acute chest pain. © 2011 American Association for Clinical Chemistry.
Arendt J.,Center Hospitalier Luxembourg
Bulletin de la Société des sciences médicales du Grand-Duché de Luxembourg | Year: 2012
In the case of early ovary extinction, the only way to have a child is either adoption or egg/embryo reception by donation. To day, egg donation is prohibited in Luxembourg by ministerial decision in 2003. Germ cell donation is part of artificial reproductive therapy. Oocyte donation, in particular, needs to be done by IVF treatment, which makes it more complicated then sperm donation What makes it more difficult is the fact that there are no oocyte bank yet. Today, prohibition encourages procreative tourism what only wealthy people can afford. Although donation programs are well established many questions arise about egg donation such as refunds, divulging information, women's age limit, health insurance participation.
Szpakowska M.,CRP Sante |
Fievez V.,CRP Sante |
Arumugan K.,CRP Sante |
Van Nuland N.,Vrije Universiteit Brussel |
And 3 more authors.
Biochemical Pharmacology | Year: 2012
Chemokines and their receptors play fundamental roles in many physiological and pathological processes such as leukocyte trafficking, inflammation, cancer and HIV-1 infection. Chemokine-receptor interactions are particularly intricate and therefore require precise orchestration. The flexible N-terminal domain of human chemokine receptors has regularly been demonstrated to hold a crucial role in the initial recognition and selective binding of the receptor ligands. The length and the amino acid sequences of the N-termini vary considerably among different receptors but they all show a high content of negatively charged residues and are subject to post-translational modifications such as O-sulfation and N- or O-glycosylation. In addition, a conserved cysteine that is most likely engaged in a receptor-stabilizing disulfide bond delimits two functionally distinct parts in the N-terminus, characterized by specific molecular signatures. Structural analyses have shown that the N-terminus of chemokine receptors recognizes a groove on the chemokine surface and that this interaction is stabilized by high-affinity binding to a conserved sulfotyrosine-binding pocket. Altogether, these data provide new insights on the chemokine-receptor molecular interplay and identify the receptor N-terminus-binding site as a new target for the development of therapeutic molecules. This review presents and discusses the diversity and function of human chemokine receptor N-terminal domains and provides a comprehensive annotated inventory of their sequences, laying special emphasis on the presence of post-translational modifications and functional features. Finally, it identifies new molecular signatures and proposes a computational model for the positioning and the conformation of the CXCR4 N-terminus grafted on the first chemokine receptor X-ray structure. © 2012 Elsevier Inc.
Delhalle S.,CRP Sante |
Schmit J.-C.,CRP Sante |
Schmit J.-C.,Center Hospitalier Luxembourg |
Chevigne A.,CRP Sante
International Journal of Molecular Sciences | Year: 2012
The complex hide-and-seek game between HIV-1 and the host immune system has impaired the development of an efficient vaccine. In addition, the high variability of the virus impedes the long-term control of viral replication by small antiviral drugs. For more than 20 years, phage display technology has been intensively used in the field of HIV-1 to explore the epitope landscape recognized by monoclonal and polyclonal HIV-1-specific antibodies, thereby providing precious data about immunodominant and neutralizing epitopes. In parallel, biopanning experiments with various combinatorial or antibody fragment libraries were conducted on viral targets as well as host receptors to identify HIV-1 inhibitors. Besides these applications, phage display technology has been applied to characterize the enzymatic specificity of the HIV-1 protease. Phage particles also represent valuable alternative carriers displaying various HIV-1 antigens to the immune system and eliciting antiviral responses. This review presents and summarizes the different studies conducted with regard to the nature of phage libraries, target display mode and biopanning procedures. © 2012 by the authors.
Lesur A.,CRP Sante |
Ancheva L.,CRP Sante |
Kim Y.J.,CRP Sante |
Berchem G.,CRP Sante |
And 3 more authors.
Proteomics - Clinical Applications | Year: 2015
Purpose: We report an immunocapture strategy to extract proteins known to harbor driver mutations for a defined cancer type before the simultaneous assessment of their mutational status by MS. Such a method bypasses the sensitivity and selectivity issues encountered during the analysis of unfractionated complex biological samples. Experimental design: Fast LC separations using short nanobore columns hyphenated with a high-resolution quadrupole-orbitrap mass spectrometer have been devised to take advantage of fast MS cycle times in conjunction with sharp chromatographic peak widths to accelerate the sample analysis throughput. Such an analytical platform is well suited to analyze simple protein mixtures obtained after immunoaffinity enrichment. Results: After establishing the technical performance of the platform, the method was applied to the quantitative profiling of cellular Ras and EGFR protein isoforms, as well as serum amyloid A isoforms in plasma. Conclusions and clinical relevance: Immunoaffinity purification combined with fast LC-MS detection for the detection of driver mutations in tissue and tumor biomarkers in plasma samples can assist clinicians to select an optimal therapeutic intervention for patients. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Ekosso L.,Center Hospitalier Luxembourg
Bulletin de la Société des sciences médicales du Grand-Duché de Luxembourg | Year: 2011
Intracoronary infusion of autologous bone marrow cells (CTX) has been shown to improve myocardial function in post infarct patients and in patients with chronic ischemic cardiomyopathy. Long term results of CTX are unknown. In this small pilot study, eleven patients with chronic ischemic cardiomyopathy and ejection fraction (EF) of 19 +/- 1% were treated with CTX and followed for 7 years. Four patients died during follow-up, all because of progressive heart failure. All patients received an implantable cardioverter defibrillator (ICD) during the course of the study but only 1 patients developed ventricular tachycardia after CTX. One patient received resynchronization therapy. The overall clinical benefit of CTX was modest (NYHA 3.0 +/- 0.1 pre and 2.5 +/- 0.2 post CTX, p= 0.06). CTX was not associated with reverse remodeling. However, left ventricular EF (19 +/- 1% pre and 18 +/- 6% post) and left ventricular end-diastolic volumes (289 +/- 71 ml pre and 294 +/- 123 ml post) remained remarkably stable over 7-year follow-up in the survivors of this very sick population. During 7-year follow-up, CTX was associated with stabilization of EF and ventricular volumes but without significant clinical benefit or evidence of reverse remodeling.