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See detailCharacterization and dynamics of the auroral electron precipitation during substorms deduced from IMAGE-FUV
Meurant, M.; Gérard, Jean-Claude ULg; Hubert, Benoît ULg et al

in Journal of Geophysical Research. Space Physics (2003), 108(A6),

[1] The FUV imager on board the IMAGE satellite provides simultaneous images of the north polar aurora every 2 min in three spectral channels. The Wideband Imaging Camera (WIC) responds primarily to the N ... [more ▼]

[1] The FUV imager on board the IMAGE satellite provides simultaneous images of the north polar aurora every 2 min in three spectral channels. The Wideband Imaging Camera (WIC) responds primarily to the N-2 LBH bands while one of the Spectral Imagers (SI13) includes the OI 135.6 nm emission and nearby LBH bands. The third channel (SI12) is sensitive to Doppler-shifted Lyman-alpha emission at 121.8 nm generated by proton precipitation. The relative magnitude of the WIC and SI13 signals depends on the altitude distribution of the energy deposition, in response to the differential O-2 absorption and the altitude dependence of the neutral composition. The ratio of simultaneous images from WIC and SI13 is used to derive the spatial distribution of the characteristic energy of the precipitating auroral electrons and the energy flux they carry. The method is described and the uncertainties introduced by possible perturbations of the neutral composition known to occur in the auroral thermosphere are discussed. The first part of this study describes a validation of this method performed by comparing precipitation characteristics derived from FUV with in situ measurements from two coincident passes of the NOAA-16 satellite. They are shown to agree within about 45%. The second part applies this ratio method to analyze the time evolution of auroral activity which occured during two substorms on 28 October 2000. The time evolution is displayed in the form of magnetic local time and magnetic latitude keograms. It is shown that the pattern of the electron average energy distribution exhibits both spatial and temporal changes. Comparison with FAST in situ electron spectrograms confirms the ability of IMAGE to detect precipitation events with a similar to200 km spatial scale. However the characteristics of the physical process leading to electron acceleration cannot be identified with FUV. The highest values of the average energy are colocated in time and space with the largest electron precipitation fluxes. A dawnward motion of bright features is observed in the postmidnight at speeds on the order of 5 magnetic local time hours/UT hour. [less ▲]

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See detailCharacterization and impact of reduced copper plating overburden on 45 nm interconnect performances
Dubreuil, Olivier ULg; Caubet-Hilloutou, Véronique; Guillan, Julie et al

in Microelectronic Engineering (2010, March), 87(3),

During first metal level interconnects fabrication, a controlled modification of the electro-deposited copper over-deposition (overburden) is performed using a partial chemical–mechanical polishing (CMP ... [more ▼]

During first metal level interconnects fabrication, a controlled modification of the electro-deposited copper over-deposition (overburden) is performed using a partial chemical–mechanical polishing (CMP) step. Next, copper microstructure is stabilized with a short duration hot-plate anneal. Overburden is then removed during CMP end-of-step. Ionic microscopy and EBSD observations of overburden thickness reduction reveal that copper grain growth occurs differently, according to patterned geometries and with a strong (1 1 1) texture, as observed in modified films. Reduction of overburden thickness also reveals the capacity of anneal temperature to impact electrical performances. Reliability is impacted for thinnest wires. [less ▲]

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See detailCharacterization and modeling of a scroll expander with air and ammonia as working fluid
Mendoza, Luis Carlos; Navarro-Esbrí; Bruno, Joan Carles et al

in Applied Thermal Engineering (2014), 70

This paper presents the experimental characterization and modeling of a scroll expander. The expander used here is a scroll compressor modified to work as an expander. It is characterized in two ... [more ▼]

This paper presents the experimental characterization and modeling of a scroll expander. The expander used here is a scroll compressor modified to work as an expander. It is characterized in two experimental setups using air and ammonia as working fluids. The paper studies how the main operating variables (supply pressure and temperature, pressure ratio, rotational speed and lubrication) influence the performance of the scroll expander. A semi-empirical model is proposed to determine the scroll expander performance. This model uses some semi-empirical parameters (such as built-in volume ratio, leakage area and mechanical losses), obtained through experimentation, to calculate the mechanical power, supply mass flow rate and exhaust temperature. Using this semi-empirical model, the deviations in the calculated mechanical power, exhaust temperature and supply mass flow rate are ±9%, ±4 K and ±5% Hz. [less ▲]

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See detailCharacterization and Numerical Simulation of the Jovian UV Aurora Observed with the HST Cameras
Grodent, Denis ULg; Dols, V.; Gérard, Jean-Claude ULg et al

in Science with the Hubble Space Telescope - II (1996)

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See detailCharacterization and optimization of GMO-based gels with long term release for intraarticular administration
Réeff, J.; Gaignaux, A.; Goole, J. et al

in International Journal of Pharmaceutics (2013), 451(1-2), 95-103

Osteoarthritis is characterized by slow degenerative processes in the articular cartilage within synovial joints. It could be interesting to develop a sustained-release formulation that could be effective ... [more ▼]

Osteoarthritis is characterized by slow degenerative processes in the articular cartilage within synovial joints. It could be interesting to develop a sustained-release formulation that could be effective on both pain/inflammation and restoration of mechanical integrity of the joint. Recently, an injectable system based on glycerol monooleate (GMO), containing clonidine as a model hydrophilic analgesic/anti-inflammatory drug and hyaluronic acid as a viscoelastic scaffold, showed promising potential as a biodegradable and biocompatible preparation to sustain the drug activity. However, drug release from the system is relatively fast (complete within 1 week) and the underlying drug release mechanisms not fully understood. The aims of this study were: (i) to significantly improve this type of local controlled drug delivery system by further sustaining clonidine release, and (ii) to elucidate the underlying mass transport mechanisms. The addition of FDA-approved inactive ingredients such as sodium oleate or purified soybean oil was found to be highly effective. The release rate could be substantially reduced (e.g., 50% release after 10 days), due to the increased hydrophobicity of the systems, resulting in slower and reduced water uptake and reduced drug mobility. Interestingly, Fick's second law of diffusion could be used to quantitatively describe drug release. [less ▲]

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See detailCHARACTERIZATION AND OPTIMIZATION OF PEPTIDE ENCAPSULATION IN PEGYLATED LIPOSOMES
Ducat, Emilie ULg; Brion, Michael; Evrard, Brigitte ULg et al

Poster (2009)

Purpose: The purpose of this work is to study the peptide encapsulation into PEGylated liposomes. Two formulations (SPC:CHOL:mPEG-750-DSPE (47:47:6) or SPC:CHOL:mPEG-2000-DSPE (47:47:6)) were investigated ... [more ▼]

Purpose: The purpose of this work is to study the peptide encapsulation into PEGylated liposomes. Two formulations (SPC:CHOL:mPEG-750-DSPE (47:47:6) or SPC:CHOL:mPEG-2000-DSPE (47:47:6)) were investigated. Methods: Blank SUV liposomes were prepared by the lipid film hydration and the encapsulation was achieved by applying freeze-thawing cycles. Because many factors may influence peptide entrapment (number of freeze-thawing cycles (NC), lipid concentration (LC), peptide concentration (PC), mixing time (MT) and liposome composition (COMP)), a design of experiment (DOE) was performed. Results: The screening permitted to identify two factors having a positive and significant influence on the encapsulation efficiencies (NC and LC) while the liposome composition had a relatively weak effect. For the second part of the DOE, the positive factors were optimized for liposomes containing mPEG2000. The obtained results revealed a theoretical optimum at 64.75±3.55% when 11 cycles were applied and for the following LC: 36.1mM SPC, 36.1mM CHOL and 4mM mPEG-2000-DSPE. Experimental results showed an encapsulation efficiency of 62.68±2.93%. Conclusion: The DOE led to significant improvement of encapsulation for liposomes containing mPEG2000. Thereafter, an optimization design for liposomes containing mPEG750 will be started. Acknowledgements: This work was supported by the Ministry of the Walloon Region. [less ▲]

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See detailCHARACTERIZATION AND OPTIMIZATION OF PEPTIDE ENCAPSULATION IN PEGYLATED LIPOSOMES
Ducat, Emilie ULg; Brion, Michael; Evrard, Brigitte ULg et al

Poster (2009)

Purpose: Print 3G is a peptidic antagonist of oncoprotein involved in breast cancer, containing 25 amino acids. The purpose of this work is to study the peptide encapsulation into PEGylated liposomes. Two ... [more ▼]

Purpose: Print 3G is a peptidic antagonist of oncoprotein involved in breast cancer, containing 25 amino acids. The purpose of this work is to study the peptide encapsulation into PEGylated liposomes. Two formulations composed of SPC:CHOL:mPEG-750-DSPE (47:47:6) or SPC:CHOL:mPEG-2000-DSPE (47:47:6) were investigated. Methods: Unilamellar vesicles containing either mPEG750 or mPEG2000 were prepared by hydration of lipid films method. Unfortunately, a loss of Print 3G was observed during the different steps of this manufacturing technique giving rise to encapsulation efficiencies close to 0 %. Thus, the freeze-thawing method was used to enhance the amount of Print 3G encapsulated into blank liposomes prepared using the above procedure. Because many factors may influence the peptide entrapment into the vesicles (number of freeze-thawing cycles, lipid concentration, peptide concentration, mixing time and liposome composition), a design of experiment was performed (for the screening, a Plackett and Burman plan; for the optimization, a central composite design). Results: The encapsulation efficiencies obtained by the freeze-thawing method in standard conditions, varied between 17.26 ± 0.46 % (n=3) for liposomes containing mPEG750 and 26.20 ± 7.98 % (n=3) for those comprising mPEG2000. Among the different considered factors, the screening permitted to identify two factors having a positive and significant influence on the encapsulation efficiencies: the number of freeze-thawing cycles and the lipid concentration, while the presence of mPEG2000 or mPEG750 had a relatively weak effect on the encapsulation. Concerning the peptide concentration and the mixing time, no influence was revealed. For the second part of the DOE, the positive factors were optimized for the liposomes containing mPEG2000. The obtained results for liposomes containing mPEG2000 revealed a theoretical optimum at 64.75 ± 3.55 % when 11 freeze-thawing cycles were applied and for the following lipid concentrations: 36.1 mM SPC, 36.1 mM CHOL and 4mM mPEG-2000-DSPE. The experimental results showed an encapsulation efficiency of 62.68 ± 2.93 %. Conclusion: Changing the manufacturing technique permitted a significant encapsulation of Print 3G into liposomes. The DOE led to a significant improvement of encapsulation efficiencies for the liposomes containing mPEG2000. Thereafter, an optimization design for liposomes containing mPEG750 will be started. Acknowledgements: This work was supported by the Ministry of the Walloon Region. [less ▲]

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See detailCharacterization and partial amino acid sequencing of a 107-kDa procollagen I N-proteinase purified by affinity chromatography on immobilized type XIV collagen.
Colige, Alain ULg; Beschin, A.; Samyn, B. et al

in Journal of Biological Chemistry (1995), 270(28), 16724-30

Procollagen I N-proteinase (EC 3.4.24.14), the enzyme that specifically processes type I and type II procollagens to collagen, was isolated from extracts of fetal calf skin. After two chromatographic ... [more ▼]

Procollagen I N-proteinase (EC 3.4.24.14), the enzyme that specifically processes type I and type II procollagens to collagen, was isolated from extracts of fetal calf skin. After two chromatographic steps on concanavalin A-Sepharose and heparin-Sepharose, the semi-purified preparation was used to produce monoclonal antibodies. One reacting antibody was found to recognize not the enzyme itself but type XIV collagen on which the enzyme was bound. This binding, highly sensitive to ionic conditions (plH, salt concentrations) but not affected by non-ionic detergents, was used for affinity chromatography that strongly improved the purification procedure. The enzyme is extensively characterized: 1) it has a molecular mass of 107 kDa as determined by polyacrylamide gel electrophoresis in presence of SDS and of about 130 kDa when estimated by gel filtration on a Sephacryl-S300; 2) in standard assay (pH 7.5, 0.2 M NaCl, 35 degrees C), the activation energy for reaction with amino procollagen type I was 17,000 calories per mole. In the same conditions, Km and Vmax values were, respectively, 435 and 39 nM per hour but varied strongly with pH and salt concentration; 3) the enzyme cleaved the NH2-terminal propeptide of type I procollagen at the specific site, the Pro-Gln bond in the alpha 1 type I procollagen chain; 4) the enzyme contained a high proportion of Gly, Asx, and Glx residues but no Hyp or Hyl; 5) partial amino acid sequences obtained from internal peptides of the enzyme displayed no significant homology with known sequences. The association of procollagen I N-proteinase with a FACIT (fibril-associated collagens with interrupted triple helices) collagen as found here might be of physiological significance. [less ▲]

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See detailCharacterization and regulation of the angiotensin II type-1 receptor (binding and mRNA) in human adrenal fasciculata-reticularis cells.
Naville, D.; LEBRETHON, Marie-Christine ULg; Kermabon, A. Y. et al

in FEBS letters (1993), 321(2-3), 184-8

The classical concept of human adrenal physiology indicates that only glomerulosa cells are the target of A-II. Herein, we demonstrated that cultured human adrenal fasciculata-reticularis cells were also ... [more ▼]

The classical concept of human adrenal physiology indicates that only glomerulosa cells are the target of A-II. Herein, we demonstrated that cultured human adrenal fasciculata-reticularis cells were also responsive to this hormone. Indeed, these cells contained high affinity (Kd = 0.9-1.1 nM) and low capacity (8,000-13,000 sites/cell) A-II receptors, and more than 95% of them were of the type-1. These AT1 receptors are functional since A-II was able to increase cortisol production after 48 h of treatment. These effects were inhibited by losartan, an AT1 antagonist, but not by CGP42112A, an AT2 antagonist. The expression of the type-1 A-II receptor mRNA was detected in the whole adrenal in both adult and fetus, and in cultured human adrenal fasciculata-reticularis cells. In these cells A-II negatively regulated AT1 receptor mRNA, and this effect was also mediated through the AT1 receptor subtype. [less ▲]

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See detailCharacterization and regulation of the hb9/mnx1 beta-cell progenitor specific enhancer in zebrafish.
Arkhipova, Valeriya; Wendik, Bjorn; Devos, Nathalie et al

in Developmental Biology (2012), 365(1), 290-302

Differentiation of insulin producing beta-cells is a genetically well defined process that involves functions of various conserved transcription factors. Still, the transcriptional mechanisms underlying ... [more ▼]

Differentiation of insulin producing beta-cells is a genetically well defined process that involves functions of various conserved transcription factors. Still, the transcriptional mechanisms underlying specification and determination of beta-cell fate are poorly defined. Here we provide the description of a beta-cell progenitor specific enhancer as a model to study initial steps of beta-cell differentiation. We show that evolutionary non-conserved upstream sequences of the zebrafish hb9 gene are required and sufficient for regulating expression in beta-cells prior to the onset of insulin expression. This enhancer contains binding sites for paired-box transcription factors and two E-boxes that in EMSA studies show interaction with Pax6b and NeuroD, respectively. We show that Pax6b is a potent activator of endodermal hb9 expression and that this activation depends on the beta-cell enhancer. Using genetic approaches we show that pax6b is crucial for maintenance but not induction of pancreatic hb9 transcription. As loss of Pax6b or Hb9 independently results in the loss of insulin expression, the data reveal a novel cross-talk between the two essential regulators of early beta-cell differentiation. While we find that the known pancreatic E-box binding proteins NeuroD and Ngn3 are not required for hb9 expression we also show that removal of both E-boxes selectively eliminates pancreatic specific reporter expression. The data provide evidence for an Ngn3 independent pathway of beta-cell specification that requires function of currently not specified E-box binding factors. [less ▲]

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See detailCharacterization and tissue-specific expression of two lepidopteran farnesyl diphosphate synthase homologs: Implications for the biosynthesis of ethyl-substituted juvenile hormones
Cusson, M.; Beliveau, C.; Sen, Se. et al

in Proteins-Structure Function and Bioinformatics (2006), 65(3), 742758

The sesquiterpenoid juvenile hormone (JH) regulates insect development and reproduction. Most insects produce only one chemical form of JH, but the Lepidoptera produce four derivatives featuring ethyl ... [more ▼]

The sesquiterpenoid juvenile hormone (JH) regulates insect development and reproduction. Most insects produce only one chemical form of JH, but the Lepidoptera produce four derivatives featuring ethyl branches. The biogenesis of these JHs requires the synthesis of ethyl-substituted farnesyl diphosphate (FPP) by FPP synthase (FPPS). To determine if there exist more than one lepidopteran FPPS, and whether one FPPS homolog is better adapted for binding the builder ethyl-branched substrates/products, we cloned three lepidopteran FPPS cDNAs, two from Choristoneura fumiferana and one from Pseudaletia unipuncta. Amino acid sequence comparisons among these and other eukaryotic FPPSs led to the recognition of two lepidopteran FPPS types. Type-I FPPSs display unique active site substitutions, including several in and near the first aspartaterich motif, whereas type-II proteins have a more "conventional" catalytic cavity. In a yeast assay, a Drosophila FPPS clone provided full complementation of an FPPS mutation, but lepidopteran FPPS clones of either type yielded only partial complementation, suggesting unusual catalytic features and/or requirements of these enzymes. Although a structural analysis of lepidopteran FPPS active sites suggested that type-I enzymes are better suited than type-II for generating ethyl-substituted products, a quantitative real-time PCR assessment of their relative abundance in insect tissues indicated that type-I expression is ubiquitous whereas that of type-II is essentially confined to the JH-producing glands, where its transcripts are ∼20 times more abundant than those of type-I. These results suggest that type-II FPPS plays a leading role in lepidopteran JH biosynthesis in spite of its apparently more conventional catalytic cavity [less ▲]

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See detailCharacterization and transfection experiments of polyplexes targeting HDAC7
Frère, Antoine ULg; Kawalec, Michal; Collard, Laurence ULg et al

Poster (2012, October 22)

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See detailCharacterization and Tumorigenicity of Spheroids Composed of Pigmented or Non Pigmented B16 Melanoma Cells
Coucke, P. H.; De Pauw-Gillet, Marie-Claire ULg; Bonjean, K. A. et al

in In Vivo (Athens, Greece) (1994), 8(3, May-Jun), 333-8

A parental line of mouse B16 melanoma cells (B16) and two derived cloned lines, either pigmented (B16P) or non pigmented (B16NP), were cultured in vitro as spheroids. After 48 hrs, the pigmented cells ... [more ▼]

A parental line of mouse B16 melanoma cells (B16) and two derived cloned lines, either pigmented (B16P) or non pigmented (B16NP), were cultured in vitro as spheroids. After 48 hrs, the pigmented cells (B16, B16P) formed smaller and looser aggregates, with higher rates of cell proliferation and lower amounts of extracellular matrix as compared to B16NP spheroids. The three lines were more tumorigenic when inoculated subcutaneously as spheroids than as isolated cells. Furthermore, B16P or B16 spheroids developed richly vascularized subcutaneous tumors and metastases more rapidly than B16NP aggregates. After intravenous injection of spheroids, the measurement with an image analyzer of the area of sections in lung colonies indicated that B16P colonies were larger and more numerous than those induced by B16NP cells. [less ▲]

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See detailCharacterization at nanometric scale of the interaction between fluoroquinolones (ciprofloxacin) and membrane lipids
Bensikaddour, Helmet; Fa, Nathalie; Deleu, Magali ULg et al

Poster (2006)

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See detailCHARACTERIZATION OF 3 LABDANE DITERPENES FROM AFRAMOMUM-ALBOVIOLACEUM
Marlier, Michel; LEGUELLEC, G.; Lognay, Georges ULg et al

in Planta Medica (1993), 59(5), 455-457

Two labdane diterpenes were isolated from the seeds and the rhizomes of Aframomum alboviolaceum (Ridley) K. Schum (Zingiberaceae) and identified by GC-MS, H-1-, and C-13-NMR as (E)-labda-8(17),12-diene-15 ... [more ▼]

Two labdane diterpenes were isolated from the seeds and the rhizomes of Aframomum alboviolaceum (Ridley) K. Schum (Zingiberaceae) and identified by GC-MS, H-1-, and C-13-NMR as (E)-labda-8(17),12-diene-15,16-dial (1) and (E)-8beta,17-epoxylabd-12-ene-15,16-dial (2). A third minor compound could be the methyl (E)-14xi,15-epoxylabd-8(17), 12-dien-16-oate. The simultaneous occurrence of these three molecules has been mentioned only in one other species of the same genus, Aframomum daniellii (1). [less ▲]

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See detailCharacterization of 4-(2-hydroxyphenyl)-1-[2 '-[N-(2 ''-pyridinyl)-p-fluorobenzamido]ethyl]piperazine (p-DMPPF) as a new potent 5-HT1A antagonist
Defraiteur, Caroline ULg; Plenevaux, Alain ULg; Scuvée-Moreau, Jacqueline ULg et al

in British Journal of Pharmacology (2007), 152(6), 952-958

Background and purpose: The identification of potent and selective radioligands for the mapping of 5-HT receptors is interesting both for clinical and experimental research. The aim of this study was to ... [more ▼]

Background and purpose: The identification of potent and selective radioligands for the mapping of 5-HT receptors is interesting both for clinical and experimental research. The aim of this study was to compare the potency of a new putative 5-HT1A receptor antagonist, p-DMPPF, (4-(2-hydroxyphenyl)-1-[2'-[N-(2''-pyridinyl)-p-fluorobenzamido]-ethyl] piperazine) with that of the well-known 5-HT1A antagonists, WAY-100635(N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]-ethyl]-N-(2-pyridinyl) cyclohexanecarboxamide) and its fluorobenzoyl analogue, p-MPPF (4-(2-methoxyphenyl)-1-[2'-[N-(2''-pyridinyl)p-fluorobenzamido] ethyl] piperazine). Experimental approach: Single cell extracellular recordings of dorsal raphe (DR) neurones were performed in rat brain slices. The potency of each compound at antagonizing the effect of the 5-HT1A agonist, 8-OH-DPAT [8-hydroxy-2-(di-npropylamino)tetraline], was quantified using the Schild equation. The pharmacological profile of p-DMPPF was defined using competition binding assays. Key results: Consistently with a 5-HT1A receptor antagonist profile, incubation of slices with an equimolar (10 nM) concentration of each compound markedly reduced the inhibitory effect of 8-OH-DPAT on the firing rate of DR neurones, causing a significant rightward shift in its concentration-response curve. The rank order of potency of the antagonists was WAY-100635 > p-DMPPF >= p-MPPF. The sensitivity of DR neurones to the inhibitory effect of 8-OH-DPAT was found to be heterogeneous. The binding experiments demonstrated that p-DMPPF is highly selective for 5-HT1A receptors, with a K-i value of 7 nM on these receptors. Conclusions and implications: The potency of the new compound, p-DMPPF, as a 5-HT1A antagonist is similar to that of p-MPPF in our electrophysiological assay. Its selectivity towards 5-HT1A receptors makes it a good candidate for clinical development. [less ▲]

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See detailCharacterization of a cavern conduit system in Vietnam by time series correlation, cross-spectrum and wavelet analyses
Tam, V. T.; De Smedt, F.; Batelaan, Okke et al

in Hydrological Sciences Journal = Journal des Sciences Hydrologiques (2004), 49(5), 879-900

Time series analyses are applied to characterize the transient flow regimes of the Nam La cavern conduit, northwest Vietnam. The conduit transforms the input signal to an output signal, and the degree of ... [more ▼]

Time series analyses are applied to characterize the transient flow regimes of the Nam La cavern conduit, northwest Vietnam. The conduit transforms the input signal to an output signal, and the degree of transformation provides information on the nature of the flow system. The input for the analysis is net precipitation and the flow hydrograph at the cave entrance, while the output series is the flow hydrograph at the resurgence. Cross-correlation and cross-spectrum analysis are used to investigate the stationarity and linearity of the input-output transformation of the system, resulting in hydrodynamic properties such as system memory, response time, and mean delay between input and output. It is shown that during high flow periods, the flow in the conduit is pressurized. Consequently, the linear input-output assumption holds only for low flows. To highlight the hydrodynamics of the cavern conduit for the high flow periods, wavelet spectrum and wavelet cross-spectrum analyses are applied. [less ▲]

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See detailCharacterization of a cDNA encoding the mitochondrial alternative oxidase (AOX) in Chlamydomonas reinhardtii and assays of AOX inactivation by the antisense strategy
Dinant, Monique; Baurain, Denis ULg; Matagne, René-Fernand ULg

in Møller, I. M.; Gardeström, P.; Glimelius, K. (Eds.) et al Plant Mitochondria: From Gene to Function (1998)

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See detailCharacterization of a chimaeric plasminogen activator obtained by insertion of the second kringle structure of tissue-type plasminogen activator (amino acids 173 through 262) between residues Asp130 and Ser139 of urokinase-type plasminogen activator.
Lijnen, H. R.; Pierard, Luc ULg; Reff, M. E. et al

in Thrombosis research (1988), 52(5), 431-41

A chimaeric recombinant plasminogen activator (rscu-PA- K2) obtained by insertion of the second kringle (K2) of tissue-type plasminogen activator (t-PA) (amino acids 173-262) between residues Asp130 and ... [more ▼]

A chimaeric recombinant plasminogen activator (rscu-PA- K2) obtained by insertion of the second kringle (K2) of tissue-type plasminogen activator (t-PA) (amino acids 173-262) between residues Asp130 and Ser139 of single chain urokinase-type plasminogen activator (scu-PA) was purified from the conditioned medium of mouse myeloma cells transfected with the previously described plasmid pULB9137 (Pierard et al., J. Biol. Chem. 262, 11771-11778, 1987). Approximately 22 micrograms of purified protein was obtained per liter of conditioned medium with a yield of approximately 25 percent. On sodium dodecylsulfate gel electrophoresis under reducing conditions, rscu-PA- K2 migrated with an apparent Mr of 65,000. Plasmin caused a time- and concentration-dependent conversion to an amidolytically active two chain derivative (rtcu-PA- K2) with a specific activity of 45,000 IU/mg. Both rscu-PA- K2 and rtcu-PA- K2 activated plasminogen directly with Km = 2.0 microM and k2 = 0.00063 s-1 and Km = 100 microM and k2 = 4.1 s-1 respectively. rscu-PA- K2 did not bind extensively to fibrin. It caused concentration-dependent lysis of 125I-fibrin-labeled plasma clots immersed in human plasma with a comparable specific activity and fibrin-specificity as rscu-PA. It is concluded that insertion in scu-PA of the second kringle of t-PA, which is believed to be involved in its fibrin affinity, does not significantly alter the enzymatic properties of scu-PA, but does not confer marked fibrin-affinity to the molecule. [less ▲]

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See detailCharacterization of a family harboring a novel LHBéta subunit mutation associated with hypogonadism
Burlacu, M. C.; Daly, Adrian ULg; Salvi, R. et al

in 17th Meeting of the Belgian Endocrine Society : Bruxelles, 25 novembre 2007 (2007, November)

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