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Hypoxia influences the cellular cross-talk of human dermal fibroblasts. A proteomic approach.
Naldini Antonella, Tiozzo Roberta, Sommer Pascal, Carraro Fabio, Annovi Giulia, Boraldi Federica, Quaglino Daniela, 2007, izvirni znanstveni članek

Opis: The ability of cells to respond to changes in oxygen availability is critical for many physiological and pathological processes (i.e. development, aging, wound healing, hypertension, cancer). Changes in the protein profile of normal human dermal fibroblasts were investigated in vitro after 96 h in 5% CO2 and 21% O2 (pO2=140 mm Hg) or 2% O2 (pO2=14 mm Hg), these parameters representing a mild chronic hypoxic exposure which fibroblasts may undergo in vivo. The proliferation rate and the protein content were not significantly modified by hypoxia, whereas proteome analysis demonstrated changes in the expression of 56 proteins. Protein identification was performed by mass spectrometry. Data demonstrate that human fibroblasts respond to mild hypoxia increasing the expression of hypoxia inducible factor (HIF1a) and of the 150-kDa oxygen-regulated protein. Other differentially expressed proteins appeared to be related to stress response, transcriptional control, metabolism, cytoskeleton, matrix remodelling and angiogenesis. Furthermore, some of them, like galectin 1, 40S ribosomal protein SA, N-myc-downstream regulated gene-1 protein, that have been described in the literature as possible cancer markers, significantly changed their expression also in normal hypoxic fibroblasts. Interestingly, a bovine fetuin was also identified that appeared significantly less internalised by hypoxic fibroblasts. In conclusion, results indicate that human dermal fibroblasts respond to an in vitro mild chronic hypoxic exposure by modifying a number of multifunctional proteins. Furthermore, data highlight the importance of stromal cells in modulating the intercellular cross-talk occurring in physiological and in pathologic conditions.
Ključne besede: Human fibroblast, Primary cell culture, Hypoxia, Connective tissue, Proteome, 2D gel electrophoresis, Mass-spectrometry
Objavljeno: 22.07.2019; Ogledov: 175; Prenosov: 0
.pdf Polno besedilo (919,07 KB)

Matrix Gla protein (MGP) is involved in elastic fiber calcification in the dermis of Pseudoxanthoma Elasticum (PXE) patients.
Federica Boraldi, Annovi Giulia, Paolinelli Devincenzi Chiara, Schurgers Leon J, Vermeer Cees, Quaglino Daniela, Pasquali Ronchetti Ivonne, Gheduzzi Dealba, 2007, izvirni znanstveni članek

Opis: Mature MGP (Matrix g-carboxyglutamic acid protein) is known to inhibit soft connective tissues calcification. We investigated its possible involvement in pseudoxanthoma elasticum (PXE), a genetic disorder whose clinical manifestations are due to mineralization of elastic fibers. PXE patients have lower serum concentration of total MGP compared to controls (Po0.001). Antibodies specific for the noncarboxylated (Glu-MGP) and for the g-carboxylated (Gla-MGP) forms of MGP were assayed on ultrathin sections of dermis from controls and PXE patients. Normal elastic fibers in controls and patients were slightly positive for both forms of MGP, whereas Gla-MGP was more abundant within control’s than within patient’s elastic fibers (Po0.001). In patients’ calcified elastic fibers, Glu-MGP intensively colocalized with mineral precipitates, whereas Gla-MGP precisely localized at the mineralization front. Data suggest that MGP is present within elastic fibers and is associated with calcification of dermal elastic fibers in PXE.
Ključne besede: calcification, dermal fibroblast, elastic fiber, human skin, MGP, pseudoxanthoma elasticum
Objavljeno: 22.07.2019; Ogledov: 142; Prenosov: 0
.pdf Polno besedilo (689,32 KB)

Exhaled breath metabolomics reveals a pathogen-specific response in a rat pneumonia model for two human pathogenic bacteria: a proof-of-concept study
Iain R White, Pouline M van Oort, 2019, izvirni znanstveni članek

Opis: Volatile organic compounds in breath can reflect host and pathogen metabolism and might be used to diagnose pneumonia. We hypothesized that rats with Streptococcus pneumoniae (SP) or Pseudomonas aeruginosa (PA) pneumonia can be discriminated from uninfected controls by thermal desorption-gas chromatography-mass-spectrometry (TD-GC-MS) and selected ion flow tube-mass spectrometry (SIFT-MS) of exhaled breath. Male adult rats (n = 50) received an intratracheal inoculation of 1) 200 µl saline, or 2) 1 × 107 colony-forming units of SP or 3) 1 × 107 CFU of PA. Twenty-four hours later the rats were anaesthetized, tracheotomized, and mechanically ventilated. Exhaled breath was analyzed via TD-GC-MS and SIFT-MS. Area under the receiver operating characteristic curves (AUROCCs) and correct classification rate (CCRs) were calculated after leave-one-out cross-validation of sparse partial least squares-discriminant analysis. Analysis of GC-MS data showed an AUROCC (95% confidence interval) of 0.85 (0.73-0.96) and CCR of 94.6% for infected versus noninfected animals, AUROCC of 0.98 (0.94-1) and CCR of 99.9% for SP versus PA, 0.92 (0.83-1.00), CCR of 98.1% for SP versus controls and 0.97 (0.92-1.00), and CCR of 99.9% for PA versus controls. For these comparisons the SIFT-MS data showed AUROCCs of 0.54, 0.89, 0.63, and 0.79, respectively. Exhaled breath analysis discriminated between respiratory infection and no infection but with even better accuracy between specific pathogens. Future clinical studies should not only focus on the presence of respiratory infection but also on the discrimination between specific pathogens.
Ključne besede: biomarkers, exhaled breath analysis, infection, pneumonia
Objavljeno: 22.07.2019; Ogledov: 141; Prenosov: 0
.pdf Polno besedilo (320,58 KB)

Corso di formazione sulle strategie didattiche interculturali nelľarea transfrontaliera
ana toroš, prispevek na konferenci brez natisa

Opis: Coordinamento e ralatrice agli incontri di formazione per i docenti (WP 3.2.1) il 21/2/2019 presso ľISIS J. Stefan di Trieste, il 7/3/109 presso il Ginnasio di Tolmino, ĺ 11/3/2019 presso ľIC bilingue di San Pietro al Natisone, il 13/3/2019 presso ľIC G. B. Tiepolo di Pagnacco, il 14/3/2019 presso a SE Lucijana Bratkoviča Bratuša Renče e ľ 11/42019 presso ľISIS Nautico T. di Savoia Duca di Genova di Trieste
Ključne besede: EDUKA2, Trst, Gorica, Videm, Istra, literatura
Objavljeno: 22.07.2019; Ogledov: 131; Prenosov: 0
.pdf Polno besedilo (991,88 KB)

Real-time multi-marker measurement of organic compounds in human breath: Towards fingerprinting breath
Iain R White, Kerry A Willis, Christopher Whyte, Rebecca Cordell, Robert S Blake, Andrew J Wardlaw, 2013, izvirni znanstveni članek

Opis: The prospects for exploiting proton transfer reaction-time of flight-mass spectrometry (PTR-ToF-MS) in medical diagnostics are illustrated through a series of case studies. Measurements of acetone levels in the breath of 68 healthy people are presented along with a longitudinal study of a single person over a period of 1 month. The median acetone concentration across the population was 484 ppbV with a geometric standard deviation (GSD) of 1.6, whilst the average GSD during the single subject longtitudinal study was 1.5. An additional case study is presented which highlights the potential of PTR-ToF-MS in pharmacokinetic studies, based upon the analysis of online breath samples of a person following the consumption of ethanol. PTR-ToF-MS comes into its own when information across a wide mass range is required, particularly when such information must be gathered in a short time during a breathing cycle. To illustrate this property, multicomponent breath analysis in a small study of cystic fibrosis patients is detailed, which provides tentative evidence that online PTR-ToF-MS analysis of tidal breath can distinguish between active infection and non-infected patients.
Ključne besede: Volatile Organic Compounds, breath, proton transfer reaction mass spectrometry, Cystic Fibrosis
Objavljeno: 22.07.2019; Ogledov: 160; Prenosov: 0
.pdf Polno besedilo (1,37 MB)

Metabolite profiling of the ripening of Mangoes Mangifera indica L. cv. ‘Tommy Atkins’ by real-time measurement of volatile organic compounds
Paul S Monks, Andrew J Taylor, Robert S Blake, Iain R White, 2016, izvirni znanstveni članek

Opis: Real-time profiling of mango ripening based on proton transfer reaction-time of flight-mass spectrometry (PTR–ToF–MS) of small molecular weight volatile organic compounds (VOCs), is demonstrated using headspace measurements of ‘Tommy Atkins’ mangoes. VOC metabolites produced during the ripening process were sampled directly, which enabled simultaneous and rapid detection of a wide range of compounds. Headspace measurements of ‘Keitt’ mangoes were also conducted for comparison. A principle component analysis of the results indicated that several mass channels were not only key to the ripening process but could also be used to distinguish between mango cultivars. The identities of 22 of these channels, tentatively speciated using contemporaneous GC–MS measurements of sorbent tubes, are rationalized through examination of the biochemical pathways that produce volatile flavour components. Results are discussed with relevance to the potential of headspace analysers and electronic noses in future fruit ripening and quality studies.
Ključne besede: Mangifera indica, Tommy Atkins, PTR–ToF–MS, VOCs, Ripening, Mango
Objavljeno: 18.07.2019; Ogledov: 162; Prenosov: 0
.pdf Polno besedilo (1,12 MB)

Observations of the release of non-methane hydrocarbons from fractured shale
Roberto Sommariva, Robert S Blake, Robert J Cuss, Rebecca L Cordell, Jon F Harrington, Iain R White, Paul S Monks, 2014, izvirni znanstveni članek

Opis: The organic content of shale has become of commercial interest as a source of hydrocarbons, owing to the development of hydraulic fracturing ("fracking"). While the main focus is on the extraction of methane, shale also contains significant amounts of non-methane hydrocarbons (NMHCs). We describe the first real-time observations of the release of NMHCs from a fractured shale. Samples from the Bowland-Hodder formation (England) were analyzed under different conditions using mass spectrometry, with the objective of understanding the dynamic process of gas release upon fracturing of the shale. A wide range of NMHCs (alkanes, cycloalkanes, aromatics, and bicyclic hydrocarbons) are released at parts per million or parts per billion level with temperature- and humidity-dependent release rates, which can be rationalized in terms of the physicochemical characteristics of different hydrocarbon classes. Our results indicate that higher energy inputs (i.e., temperatures) significantly increase the amount of NMHCs released from shale, while humidity tends to suppress it; additionally, a large fraction of the gas is released within the first hour after the shale has been fractured. These findings suggest that other hydrocarbons of commercial interest may be extracted from shale and open the possibility to optimize the "fracking" process, improving gas yields and reducing environmental impacts.
Ključne besede: Environmental impact, Hydraulic fracturing, Mass spectrometry
Objavljeno: 18.07.2019; Ogledov: 145; Prenosov: 0
.pdf Polno besedilo (2,47 MB)

Validation of an assay for the determination of levoglucosan and associated monosaccharide anhydrides for the quantification of wood smoke in atmospheric aerosol
Rebecca L Cordell, Iain R White, Paul S Monks, 2014, izvirni znanstveni članek

Opis: Biomass burning is becoming an increasing contributor to atmospheric particulate matter, and concern is increasing over the detrimental health effects of inhaling such particles. Levoglucosan and related monosaccharide anhydrides (MAs) can be used as tracers of the contribution of wood burning to total particulate matter. An improved gas chromatography-mass spectrometry method to quantify atmospheric levels of MAs has been developed and, for the first-time, fully validated. The method uses an optimised, low-volume methanol extraction, derivitisation by trimethylsilylation and analysis with high-throughput gas chromatography-mass spectrometry (GC-MS). Recovery of approximately 90 % for levoglucosan, and 70 % for the isomers galactosan and mannosan, was achieved using spiked blank filters estimates. The method was extensively validated to ensure that the precision of the method over five experimental replicates on five repeat experimental occasions was within 15 % for low, mid and high concentrations and accuracy between 85 and 115 %. The lower limit of quantification (LLOQ) was 0.21 and 1.05 ng m-3 for levoglucosan and galactosan/mannosan, respectively, where the assay satisfied precisions of ≤20 % and accuracies 80-120 %. The limit of detection (LOD) for all analytes was 0.105 ng m. The stability of the MAs, once deposited on aerosol filters, was high over the short term (4 weeks) at room temperature and over longer periods (3 months) when stored at -20 °C. The method was applied to determine atmospheric levels of MAs at an urban background site in Leicester (UK) for a month. Mean concentrations of levoglucosan over the month of May were 21.4±18.3 ng m-3, 7.5±6.1 ng m-3 mannosan and 1.8±1.3 ngm-3 galactosan.
Ključne besede: Levoglucosan, Monosaccharide anhydrides, GC–MS, Wood burning
Objavljeno: 18.07.2019; Ogledov: 149; Prenosov: 0
.pdf Polno besedilo (555,64 KB)

A feasibility study of the use of reactive tracers to determine outdoor daytime OH radical concentrations within the urban environment
Iain R White, Damien Martin, K Fredrik Petersson, Stephen J Henshaw, Graham Nickless, Guy C Lloyd-Jones, Kevin C Clemitshaw, Dudley E Shallcross, 2014, izvirni znanstveni članek

Opis: Using a specifically designed chemical tracer to indirectly measure local atmospheric hydroxyl radical (OH) concentrations is a very appealing concept. Such a tracer will provide information on the amount of OH a tracer encounters, as it moves through the urban environment and provide a stringent test of models. However, to date an outdoor experiment such as this has not been conducted. This article discusses the reasons why this is so and examines the feasibility of using tracers to measure integrated urban OH levels over short (≤1km) distances.
Ključne besede: reactive tracers, OH radicals, NO3 radicals, oxidising rate, dispersion, urban
Objavljeno: 18.07.2019; Ogledov: 154; Prenosov: 0
.pdf Polno besedilo (748,36 KB)

Atmospheric chemistry and physics in the atmosphere of a developed megacity (London): An overview of the REPARTEE experiment and its conclusions
Gavin J Phillips, Carole Helfter, Chiara F Di Marco, Eiko Nemitz, Fay Davies, Janet F Barlow, Tyrone Dunbar, Iain R White, Dudley E Shallcross, Stephen J Henshaw, K Fredrik Peterson, Brian Davison, Damien Martin, Ben Langford, C Nicholas Hewitt, Stephen M Ball, Justin M Langridge, A K Benton, Roderick L Jones, Paul I Williams, John Whitehead, Martin W Gallagher, Claire Martin, James R Dorsey, Hugh Coe, James D Allan, William J Bloss, Alistair J Thorpe, David C S Beddows, Manuel DallOsto, Roy M Harrison, Steven Smith, 2012, pregledni znanstveni članek

Opis: The Regents Park and Tower Environmental Experiment (REPARTEE) comprised two campaigns in London in October 2006 and October/November 2007. The experiment design involved measurements at a heavily trafficked roadside site, two urban background sites and an elevated site at 160-190 m above ground on the BT Tower, supplemented in the second campaign by Doppler lidar measurements of atmospheric vertical structure. A wide range of measurements of airborne particle physical metrics and chemical composition were made as well as measurements of a considerable range of gas phase species and the fluxes of both particulate and gas phase substances. Significant findings include (a) demonstration of the evaporation of traffic-generated nanoparticles during both horizontal and vertical atmospheric transport; (b) generation of a large base of information on the fluxes of nanoparticles, accumulation mode particles and specific chemical components of the aerosol and a range of gas phase species, as well as the elucidation of key processes and comparison with emissions inventories; (c) quantification of vertical gradients in selected aerosol and trace gas species which has demonstrated the important role of regional transport in influencing concentrations of sulphate, nitrate and secondary organic compounds within the atmosphere of London; (d) generation of new data on the atmospheric structure and turbulence above London, including the estimation of mixed layer depths; (e) provision of new data on trace gas dispersion in the urban atmosphere through the release of purposeful tracers; (f) the determination of spatial differences in aerosol particle size distributions and their interpretation in terms of sources and physico-chemical transformations; (g) studies of the nocturnal oxidation of nitrogen oxides and of the diurnal behaviour of nitrate aerosol in the urban atmosphere, and (h) new information on the chemical composition and source apportionment of particulate matter size fractions in the atmosphere of London derived both from bulk chemical analysis and aerosol mass spectrometry with two instrument types.
Ključne besede: megacity, trace gas, urban atmosphere, atmospheric transport, chemical composition, aerosol
Objavljeno: 18.07.2019; Ogledov: 153; Prenosov: 0
.pdf Polno besedilo (4,66 MB)

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