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See detailHigh-throughput sequencing of toxins with pharmacological interest: proof of concept and first applications
Echterbille, Julien ULg; Degueldre, Michel ULg; Boulanger, Madeleine ULg et al

Conference (2015, September 28)

Animal venoms are complex chemical cocktails, comprising wide ranges of biologically active reticulated peptides that target with high selectivity and efficacy varieties of membrane receptors. Assuming ... [more ▼]

Animal venoms are complex chemical cocktails, comprising wide ranges of biologically active reticulated peptides that target with high selectivity and efficacy varieties of membrane receptors. Assuming the fact that each of the 170,000 venomous species reported can produce more than 250 bioactive toxins, at least 40,000,000 bioactive peptides and proteins may be discovered. Among the four described species of mambas, Eastern Jameson’s mamba (Dendroaspis jamesonii kaimosae) venom is the less characterized since only 9 peptides are referenced in database. This work aims at developing a new strategy devoted to the deep analysis of animal venoms. Our approach consists in a first separation of the venom using cation exchange chromatography. Each primary fraction is then purified a second time by classical RP-HPLC. A total of 328 fractions, containing amongst 1 and 4 toxins, are finally collected. MALDI-MS analysis of each fraction is done in order (1) to obtain information about masses and (2) to obtain sequences of toxins thanks to MALDI-In Source Decay (ISD) dissociation coupled with on MALDI target plate reduction of the peptides. ISD has already been demonstrated efficient for toxin sequencing1, and especially when using 1,5-DAN as reducing matrix2. ISD yields to sequences that cover more than 50% of peptide sequences by series of singly charged c-type ions. Thanks to this methodology, we were able to obtain 85% of satisfactory results i.e. spectra giving quite long tags of amino acids (up to 20 residues). As a way to validate our method, a tag coming from ISD spectrum interpretation has found a match in database for an Eastern Jameson’s mamba toxin. The global sequence has then been obtained by extrapolation on the ISD spectrum. Since ISD spectra are simpler than classical MS/MS spectra, automation of spectra interpretation, difficult with other fragmentation techniques (CID, ETD…), is implementable. In the near future, sequences obtained with this approach will be used to direct tests of biological activity through sequence homologies with already known ligands for different kinds of membrane receptors. [less ▲]

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See detailHigh-throughput synthesis of functional oxide films
Dhanapal, Pravarthana ULg

Doctoral thesis (2014)

Epitaxy of metal oxides is of great interest since it provides a way to obtain desired novel properties for the applications such as electronics and energy. However, earlier epitaxy research's have been ... [more ▼]

Epitaxy of metal oxides is of great interest since it provides a way to obtain desired novel properties for the applications such as electronics and energy. However, earlier epitaxy research's have been restricted because of the limited range of compositions and low-index of commercially available single crystal substrates. Consequently, novel epitaxy synthesis methods need to be developed in order to go beyond the present demands of of single crystal substrates in terms of phase, composition, size, orientation and symmetry. In this research work, we have developed a high-throughput synthesis process, called combinatorial substrate epitaxy (CSE), where an oxide film is grown epitaxially on a polycrystalline substrate. As a proof-of-concept, we firstly fabricated Ca3Co4O9 films on Al2O3 ceramics. Films have a good local epitaxial registry, and the Seebeck coefficient is about 170 µV/K at 300 K. High quality BiFeO3/La0.7Sr0.3MnO3 thin film heterostructures were secondly deposited on dense LaAlO3 ceramics prepared by spark plasma sintering. Piezoforce microscopy was used to confirm the local ferroelectric properties. Thirdly, we investigate the growth of of metastable monoclinic Dy2Ti2O7 epitaxial films on polycrystalline La2Ti2O7 substrates. We conclude that CSE approach opens the way towards unexpected electronic properties in oxide films. [less ▲]

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See detailHigh-throughput synthesis of thermoelectric Ca3Co 4O9 films
Dhanapal, Pravarthana ULg; Lebedev, O. I.; Hebert, S. et al

in Applied Physics Letters (2013), 103

Properties of complex oxide thin films can be tuned over a range of values as a function of mismatch, composition, orientation, and structure. Here, we report a strategy for growing structured epitaxial ... [more ▼]

Properties of complex oxide thin films can be tuned over a range of values as a function of mismatch, composition, orientation, and structure. Here, we report a strategy for growing structured epitaxial thermoelectric thin films leading to improved Seebeck coefficient. Instead of using single-crystal sapphire substrates to support epitaxial growth, Ca3Co 4O9 films are deposited, using the Pulsed Laser Deposition technique, onto Al2O3 polycrystalline substrates textured by spark plasma sintering. The structural quality of the 2000 Å thin film was investigated by transmission electron microscopy, while the crystallographic orientation of the grains and the epitaxial relationships were determined by electron backscatter diffraction. The use of a polycrystalline ceramic template leads to structured films that are in good local epitaxial registry. The Seebeck coefficient is about 170 μV/K at 300 K, a typical value of misfit material with low carrier density. This high-throughput process, called combinatorial substrate epitaxy, appears to facilitate the rational tuning of functional oxide films, opening a route to the epitaxial synthesis of high quality complex oxides. © 2013 AIP Publishing LLC. [less ▲]

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See detailHigh-volume versus standard-volume haemofiltration for septic shock patients with acute kidney injury (IVOIRE study): a multicentre randomized controlled trial
JOANNES-BOYAU, Olivier; HONORE, Patrick; PEREZ, Paul et al

in Intensive Care Medicine (2013), 39(9), 1535-1546

PURPOSE : Septic shock is a leading cause of death among critically ill patients, in particular when complicated by acute kidney injury (AKI). Small experimental and human clinical studies have suggested ... [more ▼]

PURPOSE : Septic shock is a leading cause of death among critically ill patients, in particular when complicated by acute kidney injury (AKI). Small experimental and human clinical studies have suggested that high-volume haemofiltration (HVHF) may improve haemodynamic profile and mortality. We sought to determine the impact of HVHF on 28-day mortality in critically ill patients with septic shock and AKI. METHODS: This was a prospective, randomized, open, multicentre clinical trial conducted at 18 intensive care units in France, Belgium and the Netherlands. A total of 140 critically ill patients with septic shock and AKI for less than 24 h were enrolled from October 2005 through March 2010. Patients were randomized to either HVHF at 70 mL/kg/h or standard-volume haemofiltration (SVHF) at 35 mL/kg/h, for a 96-h period. RESULTS: Primary endpoint was 28-day mortality. The trial was stopped prematurely after enrolment of 140 patients because of slow patient accrual and resources no longer being available. A total of 137 patients were analysed (two withdrew consent, one was excluded); 66 patients in the HVHF group and 71 in the SVHF group. Mortality at 28 days was lower than expected but not different between groups (HVHF 37.9 % vs. SVHF 40.8 %, log-rank test p = 0.94). There were no statistically significant differences in any of the secondary endpoints between treatment groups. CONCLUSIONS: In the IVOIRE trial, there was no evidence that HVHF at 70 mL/kg/h, when compared with contemporary SVHF at 35 mL/kg/h, leads to a reduction of 28-day mortality or contributes to early improvements in haemodynamic profile or organ function. HVHF, as applied in this trial, cannot be recommended for treatment of septic shock complicated by AKI. [less ▲]

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See detailHigh-yield radiosynthesis and preliminary in vivo evaluation of p-[18F]MPPF, a fluoro analog of WAY-100635.
Le Bars, Didier; Lemaire, Christian ULg; Ginovart, N. et al

in Nuclear Medicine & Biology (1998), 25(4), 343-50

No-carrier-added 4-[18F]fluoro-N-[2-[1-(2-methoxyphenyl)-1 piperazinyl]ethyl-N-2-pyridinyl-benzamide (p-[18F]MPPF) was synthesized by nucleophilic substitution of the corresponding nitro compound in the ... [more ▼]

No-carrier-added 4-[18F]fluoro-N-[2-[1-(2-methoxyphenyl)-1 piperazinyl]ethyl-N-2-pyridinyl-benzamide (p-[18F]MPPF) was synthesized by nucleophilic substitution of the corresponding nitro compound in the presence of Kryptofix 222 and K2CO3 by microwave heating (3 min, 500 W) using a remotely controlled radiosynthesis. Baseline separation of p-[18F]MPPF from the nitro derivative was performed on a semipreparative HPLC C18 column. After Sep-Pak formulation, the radiopharmaceutical was obtained with a radiochemical yield of 25% (EOS) in about 70 min. Specific radioactivity averaged between 1-5 Ci/micromol EOS. Labelling of the ortho and meta derivatives was also attempted. Brain uptake of p-[18F]MPPF was studied with PET on fluothane-anesthetized cats. Following intravenous injection of p-[18F]MPPF, high accumulation of radioactivity was observed in the hippocampus and cerebral cortex. Low levels of radioactivity were observed in cerebellum. At 30 min, the mean hippocampus/cerebellum and cortex/cerebellum ratios were 5 and 3.8, respectively. The accumulation of the tracer was blocked by prior administration of reference WAY-100635, demonstrating the specificity of the ligand. [less ▲]

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See detailHigh-Yield Synthesis of a (3H)Ethylenediamine Ditetrodotoxin Derivative
Bontemps, José; Cantineau, Robert; Grandfils, Christian ULg et al

in Analytical Biochemistry (1984), 139

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See detailHigh-Yield Synthesis Of The Bovine Leukemia-Virus (Blv) P24 Major Internal Protein In Saccharomyces-Cerevisiae
Dumont, Jacques; Legrain, M.; Portetelle, Daniel ULg et al

in Gene (1989), 79(2),

Bovine leukemia virus (BLV) p24 gene was expressed in Saccharomyces cerevisiae under the control of the PHO5 (encoding repressible acid phosphatase, rAPase) promoter. Yeast cells were transformed by a ... [more ▼]

Bovine leukemia virus (BLV) p24 gene was expressed in Saccharomyces cerevisiae under the control of the PHO5 (encoding repressible acid phosphatase, rAPase) promoter. Yeast cells were transformed by a yeast-E. coli shuttle vector carrying the PHO5 promoter, the p24 gene and the CYC1 transcription terminator. After low inorganic phosphate (Pi) induction of the PHO5 promoter, p24 accumulated in the producing cells up to a concentration representing 10% of total soluble proteins. The expression level of p24 gene was not increased by insertion of the positive regulatory gene PHO4 on the p24 expression vector. The p24 produced in this system and incubated in crude yeast extract showed a remarkably high resistance to proteolytic degradation, a feature that presumably correlates with the compact globular conformation of the protein combined to the stabilizing effect of the N-terminal residue. [less ▲]

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See detailHigher frontal EEG synchronization in young women with major depression: a marker for increased homeostatic sleep pressure?
Birchler-Pedross, Angelina; Frey, Sylvia; Chellappa, Sarah Laxhmi ULg et al

in Sleep (2011), 34(12), 1699-706

STUDY OBJECTIVES: Major depressive disorder (MDD) is often associated with disturbances in circadian and/or sleep-wake dependent processes, which both regulate daytime energy and sleepiness levels. DESIGN ... [more ▼]

STUDY OBJECTIVES: Major depressive disorder (MDD) is often associated with disturbances in circadian and/or sleep-wake dependent processes, which both regulate daytime energy and sleepiness levels. DESIGN: Analysis of continuous electroencephalographic (EEG) recordings during 40 h of extended wakefulness under constant routine conditions. Artifact-free EEG samples derived from 12 locations were subjected to spectral analysis. Additionally, half-hourly ratings of subjective tension and sleepiness levels and salivary melatonin measurements were collected. SETTING: Centre for Chronobiology, Psychiatric Hospitals of the University of Basel, Switzerland. PARTICIPANTS: Eight young healthy women and 8 young untreated women with MDD. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: MDD women exhibited higher frontal low-frequency (FLA) EEG activity (0.5-5.0 Hz) during extended wakefulness than controls, particularly during the night. Enhanced FLA was paralleled by higher levels of subjective sleepiness and tension. In MDD women, overall FLA levels correlated positively with depression scores. The timing of melatonin onset did not significantly differ between the two groups, but the nocturnal secretion of salivary melatonin was significantly attenuated in MDD women. CONCLUSIONS: Our data imply that young women with MDD live on a higher homeostatic sleep pressure level, as indexed by enhanced FLA during wakefulness. Its positive correlation with depression scores indicates a possible functional relationship. High FLA could reflect a use-dependent phenomenon in depression (enhanced cognitive rumination or tension) and/or an attenuated circadian arousal signal. [less ▲]

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See detailHigher long-lasting ethanol sensitization after adolescent ethanol exposure in mice
Quoilin, Caroline; Didone, Vincent ULg; Tirelli, Ezio ULg et al

in Psychopharmacology (2014), 231

Rationale. Due to their maturing brain, adolescents are suggested to be more vulnerable to the long-term consequences of chronic alcohol use. Increased sensitization to the stimulant effects of ethanol is ... [more ▼]

Rationale. Due to their maturing brain, adolescents are suggested to be more vulnerable to the long-term consequences of chronic alcohol use. Increased sensitization to the stimulant effects of ethanol is a possible consequence of ethanol exposure during adolescence. Objectives. The aim of this study was to characterize the long-term alterations in the stimulant effects of ethanol and in the rate of ethanol sensitization in mice pre-exposed to ethanol during adolescence in comparison to mice pre-exposed to ethanol in adulthood. Methods. Adolescent and adult female SWISS mice were injected with saline or ethanol (2.5 or 4 g/kg) during 14 consecutive days. After a three weeks period of ethanol abstinence, mice were tested as adults before and after a second exposure to daily repeated ethanol injections. Results. All mice pre-exposed to ethanol as adults or adolescents showed higher stimulant effects when re-exposed to ethanol three weeks later. However, this enhanced sensitivity to the stimulant effects of ethanol was of significantly higher magnitude in mice repeatedly injected with high ethanol doses (4g/kg) during adolescence. Furthermore, the increased expression of ethanol stimulant effects in these mice was maintained even after a second procedure of ethanol sensitization. Conclusions. Adolescence is a critical period for the development of a sensitization to ethanol stimulant properties providing that high intermittent ethanol doses are administered. These results might contribute to explain the relationship between age at first alcohol use and risks of later alcohol problems and highlight the dangers of repeated consumption of high alcohol amounts in young adolescents. [less ▲]

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See detailHigher male than female recombination rate in cattle is controlled by genetic variants effective in both sexes
Kadri, Naveen Kumar ULg; Harland, Chad ULg; Coppieters, Wouter ULg et al

Poster (2015, May 05)

We herein study genetic recombination in three dairy cattle populations from France, New-Zealand and the Netherlands. We apply a new phasing algorithm extracting familial information suited for large half ... [more ▼]

We herein study genetic recombination in three dairy cattle populations from France, New-Zealand and the Netherlands. We apply a new phasing algorithm extracting familial information suited for large half-sib families to reconstruct haplotypes and detect cross-overs (CO). The software is robust to genotyping and map errors. We identify more than 2,000,000 CO events in sperm cells transmitted by 3008 sires to 94,603 offspring, and more than 500,000 CO events in oocytes transmitted by 11,497 cows to 25,390 offspring. When measured in identical family structures, the average number of CO in males (24.0) was found to be larger than in females (21.8). In males, recombination rates were higher closer to telomeres whereas in females, recombination rates dropped at both centromeres and telomeres (probably as a result of lower informativity). The heritability of the global recombination rate (GRR) was close to 0.20 in males and to 0.08 in females. Genetic correlation ranged from 0.38 to 0.69 depending on the population, indicating that shared variants are influencing GRR in both genders. Haplotype-based genome-wide association studies revealed four genome-wide significant QTL, including two previously identified ones (involving REC8 and RNF212). For all QTLs, there was a positive correlation between haplotype effects across sexes, ranging from 0.35 to 0.68. We selected two reference panels of respectively 122 and 215 bulls sequenced at cover > 15x to impute variants in the New-Zealand and French populations. All variants identified by next-generating sequencing in 5 Mb windows encompassing the QTL peaks were imputed with Beagle in order to perform a sequence-based association study. For three QTLs, we identified missense mutations in genes known to be involved in meiosis among the most significantly associated variants. These variants were perfectly associated with the haplotypes underlying the QTL effects. The variant identified in RNF212 had already been reported, whereas missense mutations in MLH3 (N408S) and HFM1 (S1189L) are new findings. Surprisingly, variants previously identified in REC8 did not capture the QTL effect whereas variants in RNF212B, PPP1R3E, BCL2L2, HOMEZ and PABPN1 had much stronger association with the phenotype. The three missense mutations were significant in both genders with two of them accounting for approximately 10% of the genetic variance in males (the allelic substitution effect being approximately equal to one additional CO per genome). Our results are very different from reports of recombination in other species. For instance, in human, recombination rate is higher in females, distinct variants affect recombination rate in males and females and the genetic correlation is close to 0 whereas in cattle, we observed a higher recombination rate in males controlled by shared variants effective in both sexes. [less ▲]

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See detailHigher Octonions
Kreusch, Marie ULg

Conference (2015, June 11)

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See detailHigher order noise-shaping filters for high-performance micromachined accelerometers
Dong, Yufeng; Kraft, Michaël ULg; Redman-White, William

in IEEE Transactions on Instrumentation and Measurement (2007), 56(5), 1666-1674

Micromachined inertial sensors that have been incorporated in sigma-delta force-feedback loops have been proven to improve linearity, dynamic range, and bandwidth, and also provide a direct digital output ... [more ▼]

Micromachined inertial sensors that have been incorporated in sigma-delta force-feedback loops have been proven to improve linearity, dynamic range, and bandwidth, and also provide a direct digital output. Previous work mainly focused on using the sensing element only to form a second-order single-loop sigma-delta modulator (SigmaDeltaM); however, the advantages of higher order single-loop electromechanical SigmaDeltaM have not been fully explored. High-performance inertial sensors require higher signal-to-quantization noise ratio (SQNR). This paper presents topologies for higher order single-loop electromechanical SigmaDeltaM with optimal stable coefficients that lead to a better SQNR. The topologies have good immunity to fabrication tolerances, which was verified by Monte Carlo analysis. The topologies are applicable not only to accelerometers but also to other inertial sensors such as gyroscopes. [less ▲]

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See detailHigher order sigma-delta modulators interfaces for capacitive sensors (Invited)
Kraft, Michael ULg

in The IEE Seminar and Exhibition on MEMS Sensor Technologies (2005)

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See detailHigher order sigma-delta modulators interfaces for capapcitive sensors
Kraft, Michaël ULg; Dong, Yufeng

in 2005 NSTI Nanotechnology Conference and Trade Show : NSTI Nanotech 2005 (2005, May)

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See detailHigher plant-like subunit composition of mitochondrial complex I from Chlamydomonas reinhardtii: 31 conserved components among eukaryotes
Cardol, Pierre ULg; Vanrobaeys, F.; Devreese, B. et al

in Biochimica et Biophysica Acta-Bioenergetics (2004), 1658(3), 212-224

The rotenone-sensitive NADH:ubiquinone oxidoreductase (complex I) is the most intricate membrane-bound enzyme of the mitochondrial respiratory chain. Notably the bovine enzyme comprises up to 46 subunits ... [more ▼]

The rotenone-sensitive NADH:ubiquinone oxidoreductase (complex I) is the most intricate membrane-bound enzyme of the mitochondrial respiratory chain. Notably the bovine enzyme comprises up to 46 subunits, while 27 subunits could be considered as widely conserved among eukaryotic complex I. By combining proteomic and genomic approaches, we characterized the complex I composition from the unicellular green alga Chlamydomonas reinhardtii. After purification by blue-native polyacrylamide gel electrophoresis (BN-PAGE), constitutive subunits were analyzed by SDS-PAGE coupled to tandem mass spectrometry (MS) that allowed the identification of 30 proteins. We compared the known complex I components from higher plants, mammals, nematodes and fungi with this MS data set and the translated sequences from the algal genome project. This revealed that the Chlamydomonas complex I is likely composed of 42 proteins, for a total molecular mass of about 970 kDa. In addition to the 27 typical components, we have identified four new complex I subunit families (bovine ESSS, PFFD, B16.6, B12 homologues), extending the number of widely conserved eukaryote complex I components to 31. In parallel, our analysis showed that a variable number of subunits appears to be specific to each eukaryotic kingdom (animals, fungi or plants). Protein sequence divergence in these kingdom-specific sets is significant and currently we cannot exclude the possibility that homology between them exists, but has not yet been detected. (C) 2004 Elsevier B.V. All rights reserved. [less ▲]

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See detailHigher prevalence of clinically relevant pituitary adenomas confirmed.
Beckers, Albert ULg

in Clinical Endocrinology (2010), 72(3), 290-291

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See detailHigher risk of death among MEN1 patients with mutations in the JunD interacting domain. A Groupe d'étude des Tumeurs Endocrines (GTE) cohort study
Thevenon, Julien; Bourredjem, Abderrahmane; Faivre, Laurence et al

in Human Molecular Genetics (2013)

BackgroundMultiple Endocrine Neoplasia syndrome type 1 (MEN1), which is secondary to mutation of the MEN1 gene, is a rare autosomal-dominant disease that predisposes mutation carriers to endocrine tumors ... [more ▼]

BackgroundMultiple Endocrine Neoplasia syndrome type 1 (MEN1), which is secondary to mutation of the MEN1 gene, is a rare autosomal-dominant disease that predisposes mutation carriers to endocrine tumors. Although genotype-phenotype studies have so far failed to identify any statistical correlations, some families harbor recurrent tumor patterns. The function of MENIN is unclear but has been described through the discovery of its interacting partners. Mutations in the interacting domains of MENIN functional partners have been shown to directly alter its regulation abilities.MethodsWe report on a cohort of MEN1 patients from the Groupe d'etude des Tumeurs Endocrines. Patients with a molecular diagnosis and a clinical follow-up, totalling 262 families and 806 patients were included. Associations between mutation type, location or interacting factors of the MENIN protein and death as well as the occurrence of MEN1-related tumors were tested using a frailty Cox model to adjust for potential heterogeneity across families.ResultsAccounting for the heterogeneity across families, the overall risk of death was significantly higher when mutations affected the JunD interacting domain (adjusted HR=1.88: 95%-CI=1.15- 3.07). Patients had a higher risk of death from cancers of the MEN1 spectrum (HR=2.34; 95%-CI=1.23- 4.43).ConclusionThis genotype-phenotype correlation study confirmed the lack of direct genotype-phenotype correlations. However, patients with mutations affecting the JunD interacting domain had a higher risk of death secondary to a MEN1 tumor and should thus be considered for surgical indications, genetic counseling and follow-up. [less ▲]

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See detailHigher sensitivity of Adamts12-deficient mice to tumor growth and angiogenesis.
El Hour, Mehdi ULg; Moncada-Pazos, A.; Blacher, Silvia ULg et al

in Oncogene (2010), 29(20), 3025-32

ADAMTS (a disintegrin and metalloproteinase domain with thrombospondin motifs) constitute a family of endopeptidases related to matrix metalloproteinases. These proteases have been largely implicated in ... [more ▼]

ADAMTS (a disintegrin and metalloproteinase domain with thrombospondin motifs) constitute a family of endopeptidases related to matrix metalloproteinases. These proteases have been largely implicated in tissue remodeling and angiogenesis associated with physiological and pathological processes. To elucidate the in vivo functions of ADAMTS-12, we have generated a knockout mouse strain (Adamts12−/−) in which Adamts12 gene was deleted. The mutant mice had normal gestations and no apparent defects in growth, life span and fertility. By applying three different in vivo models of angiogenesis (malignant keratinocyte transplantation, Matrigel plug and aortic ring assays) to Adamts12−/− mice, we provide evidence for a protective effect of this host enzyme toward angiogenesis and cancer progression. In the absence of Adamts-12, both the angiogenic response and tumor invasion into host tissue were increased. Complementing results were obtained by using medium conditioned by cells overexpressing human ADAMTS-12, which inhibited vessel outgrowth in the aortic ring assay. This angioinhibitory effect of ADAMTS-12 was independent of its enzymatic activity as a mutated inactive form of the enzyme was similarly efficient in inhibiting endothelial cell sprouting in the aortic ring assay than the wild-type form. Altogether, our results show that ADAMTS-12 displays antiangiogenic properties and protect the host toward tumor progression. [less ▲]

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See detailHigher Symmetries of the conformal Laplacian
Radoux, Fabian ULg

Conference (2014, July 14)

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