Identification of a potent MAR element from the mouse genome and assessment of its activity in stable and transient transfections.

Matrix attachment regions are DNA sequences found throughout eukaryotic genomes that are believed to define boundaries interfacing heterochromatin and euchromatin domains, thereby acting as epigenetic regulators. When included in expression vectors, MARs can improve and sustain transgene expression, and a search for more potent novel elements is therefore actively pursued to further improve recombinant protein production. Here we describe the isolation of new MARs from the mouse genome using a modified in silico analysis. One of these MARs was found to be a powerful activator of transgene expression in stable transfections. Interestingly, this MAR also increased GFP and/or immunoglobulin expression from some but not all expression vectors in transient transfections. This effect was attributed to the presence or absence of elements on the vector backbone, providing an explanation for earlier discrepancies as to the ability of this class of elements to affect transgene expression under such conditions.

Niobium and rare earth minerals from the Virulundo carbonatite, Namibe, Angola

The Virulundo carbonatite in Angola, one of the biggest in the world, contains pyrochlore as an accessory mineral in all of the carbonatite units (calciocarbonatites, ferrocarbonatites, carbonatite breccias, trachytoids). The composition of the primary pyrochlore crystals is very close to fluornatrocalciopyrochlore in all these units. High-temperature hydrothermal processes caused the pseudomorphic replacement of the above crystals by a second generation of pyrochlore, characterized by lower F and Na contents. Low-temperature hydrothermal replacement of the above pyrochlores, associated with production of quartz-carbonates-fluorite veins, controled the development of a third generation of pyrochlore, characterized by high Sr contents. Finally, supergene processes produced the development of a secondary paragenesis in the carbonatite, consisting in late carbonates, goethite, hollandite and REE minerals (mainly synchysite-(Ce), britholite-(Ce), britholite-(La), cerite-(Ce)). Separation of Ce from the other REE was allowed by oxidizing conditions. Therefore, Ce4+ was also incorporated into a late generation of pyrochlore, which is also strongly enriched in Ba and strongly depleted in Ca and Na

A fucan from the brown seaweed Spatoglossum schröederi inhibits Chinese hamster ovary cell adhesion to several extracellular matrix proteins

Fucans, a family of sulfated polysaccharides present in brown seaweed, have several biological activities. Their use as drugs would offer the advantage of no potential risk of contamination with viruses or particles such as prions. A fucan prepared from Spatoglossum schröederi was tested as a possible inhibitor of cell-matrix interactions using wild-type Chinese hamster ovary cells (CHO-K1) and the mutant type deficient in xylosyltransferase (CHO-745). The effect of this polymer on adhesion properties with specific extracellular matrix components was studied using several matrix proteins as substrates for cell attachment. Treatment with the polymer inhibited the adhesion of fibronectin to both CHO-K1 (2 x 10(5))()and CHO-745 (2 x 10(5) and 5 x 10(5)) cells. No effect was detected with laminin, using the two cell types. On the other hand, adhesion to vitronectin was inhibited in CHO-K1 cells and adhesion to type I collagen was inhibited in CHO-745 cells. In spite of this inhibition, the fucan did not affect either cell proliferation or cell cycle. These results demonstrate that this polymer is a new anti-adhesive compound with potential pharmacological applications.

Brief daily postpartum separations from the litter alter dam response to psychostimulants and to stress

Neonatal handling induces several behavioral and neurochemical alterations in pups, including decreased responses to stress and reduced fear in new environments. However, there are few reports in the literature concerning the behavioral effects of this neonatal intervention on the dams during the postpartum period. Therefore, the aim of the current study was to determine if brief postpartum separation from pups has a persistent impact on the dam's stress response and behavior. Litters were divided into two neonatal groups: 1) non-handled and 2) handled [10 min/day, from postnatal day (PND) 1 to 10]. Weaning occurred at PND 21 when behavioral tasks started to be applied to the dams, including sweet food ingestion (PND 21), forced swimming test (PND 28), and locomotor response to a psychostimulant (PND 28). On postpartum day 40, plasma was collected at baseline for leptin assays and after 1 h of restraint for corticosterone assay. Regarding sweet food consumption, behavior during the forced swimming test or plasma leptin levels did not differ between dams briefly separated and non-separated from their pups during the postpartum period. On the other hand, both increased locomotion in response to diethylpropion and increased corticosterone secretion in response to acute stress were detected in dams briefly separated from their pups during the first 10 postnatal days. Taken together, these findings suggest that brief, repeated separations from the pups during the neonatal period persistently impact the behavior and induce signs of dopaminergic sensitization in the dam.

Intraoperative hemodynamic instability during and after separation from cardiopulmonary bypass : importance, mechanism and prevention

Chaque année, environ 1 à 1,25 million d’individus subiront une chirurgie cardiaque. [1] Environ 36 000 chirurgies cardiaques sont effectuées au Canada et 8000 procédures au Québec (http://www.ccs.ca). Le vieillissement de la population aura pour conséquence que la chirurgie cardiaque sera offerte à des patients de plus en plus à risque de complications, principalement en raison d’une co-morbidité plus importante, d’un risque de maladie coronarienne plus élevée, [2] d’une réserve physiologique réduite et par conséquent un risque plus élevé de mortalité à la suite d’une chirurgie cardiaque. L’une des complications significatives à la suite d’une chirurgie cardiaque est le sevrage difficile de la circulation extracorporelle. Ce dernier inclut la période au début du sevrage de la circulation extracorporelle et s’étend jusqu’au départ du patient de la salle d’opération. Lorsque le sevrage de la circulation extracorporelle est associé à une défaillance ventriculaire droite, la mortalité sera de 44 % à 86 %. [3-7] Par conséquent le diagnostic, l’identification des facteurs de risque, la compréhension du mécanisme, la prévention et le traitement du sevrage difficile de la circulation extracorporelle seront d’une importance majeure dans la sélection et la prise en charge des patients devant subir une chirurgie cardiaque. Les hypothèses de cette thèse sont les suivantes : 1) le sevrage difficile de la circulation extracorporelle est un facteur indépendant de mortalité et de morbidité, 2) le mécanisme du sevrage difficile de la circulation extracorporelle peut être approché d’une façon systématique, 3) la milrinone administrée par inhalation représente une alternative préventive et thérapeutique chez le patient à risque d’un sevrage difficile de la circulation extracorporelle après la chirurgie cardiaque.

INVESTIGATIONS ON THE RECOVERY OF TITANIUM VANADIUM AND IRON VALUES FROM THE WASTE CHILORIDE LIQUORS OF TITANIA INDUSTRY

The thesis entitled INVESTIDGATIONS ON THE RECOVERY OF TITANIUM VANADIUM AND IRON VALUES FROM THE WASTE CHILORIDE LIQUORS OF TITANIA INDUSTRY embodies the results of the investigations carried out on the solvent extraction separation of iron (III) vanadium(V) and titanium (IV) chlorides from the waste chloride liquors of titanium minerals processing industry by employing tributylphosphate (TBT) as an extractant. The objective of this study is to generate the knowledge base to achieve the recovery of iron, vanadium and titanium cvalues from multi- metal waste chloride liquors originating from ilmenite mineral beneficiation industries through selective separation and value added material development

Proteomic analysis of plasma membrane vesicles isolated from the rat renal cortex

Polarized epithelial cells are responsible for the vectorial transport of solutes and have a key role in maintaining body fluid and electrolyte homeostasis. Such cells contain structurally and functionally distinct plasma membrane domains. Brush border and basolateral membranes of renal and intestinal epithelial cells can be separated using a number of different separation techniques, which allow their different transport functions and receptor expressions to be studied. In this communication, we report a proteomic analysis of these two membrane segments, apical and basolateral, obtained from the rat renal cortex isolated by two different methods: differential centrifugation and free-flow electrophoresis. The study was aimed at assessing the nature of the major proteins isolated by these two separation techniques. Two analytical strategies were used: separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) at the protein level or by cation-exchange high-performance liquid chromatography (HPLC) after proteolysis (i.e., at the peptide level). Proteolytic peptides derived from the proteins present in gel pieces or from HPLC fractions after proteolysis were sequenced by on-line liquid chromatography-tandem mass spectrometry (LC-MS/MS). Several hundred proteins were identified in each membrane section. In addition to proteins known to be located at the apical and basolateral membranes, several novel proteins were also identified. In particular, a number of proteins with putative roles in signal transduction were identified in both membranes. To our knowledge, this is the first reported study to try and characterize the membrane proteome of polarized epithelial cells and to provide a data set of the most abundant proteins present in renal proximal tubule cell membranes.

Proteomic analysis of plasma membrane vesicles isolated from the rat renal cortex

Polarized epithelial cells are responsible for the vectorial transport of solutes and have a key role in maintaining body fluid and electrolyte homeostasis. Such cells contain structurally and functionally distinct plasma membrane domains. Brush border and basolateral membranes of renal and intestinal epithelial cells can be separated using a number of different separation techniques, which allow their different transport functions and receptor expressions to be studied. In this communication, we report a proteomic analysis of these two membrane segments, apical and basolateral, obtained from the rat renal cortex isolated by two different methods: differential centrifugation and free-flow electrophoresis. The study was aimed at assessing the nature of the major proteins isolated by these two separation techniques. Two analytical strategies were used: separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) at the protein level or by cation-exchange high-performance liquid chromatography (HPLC) after proteolysis (i.e., at the peptide level). Proteolytic peptides derived from the proteins present in gel pieces or from HPLC fractions after proteolysis were sequenced by on-line liquid chromatography-tandem mass spectrometry (LC-MS/MS). Several hundred proteins were identified in each membrane section. In addition to proteins known to be located at the apical and basolateral membranes, several novel proteins were also identified. In particular, a number of proteins with putative roles in signal transduction were identified in both membranes. To our knowledge, this is the first reported study to try and characterize the membrane proteome of polarized epithelial cells and to provide a data set of the most abundant proteins present in renal proximal tubule cell membranes.

Phase mixing and phase separation accompanying the catalytic oxidation of CO on Ir{100}

The co-adsorption of CO and O on the unreconstructed (1 x 1) phase of Ir {100} was examined by low energy electron diffraction (LEED) and temperature programmed desorption (TPD). When CO is adsorbed at 188 K onto the Ir{100} surface precovered with 0.5 ML O, a mixed c(4 x 2)-(2O + CO) overlayer is formed. All CO is oxidised upon heating and desorbs as CO2 in three distinct stages at 230 K, 330 K and 430 K in a 2:1:2 ratio. The excess oxygen left on the surface after all CO has reacted forms an overlayer with a LEED pattern with p(2 x 10) periodicity. This overlayer consists of stripes with a local p(2 x 1)-O arrangement of oxygen atoms separated by stripes of uncovered It. When CO is adsorbed at 300 K onto the surface precovered with 0.5 ML O an apparent (2 x 2) LEED pattern is observed. LEED IV analysis reveals that this pattern is a superposition of diffraction patterns from islands of c(2 x 2)-CO and p(2 x 1)-O structures on the surface. Heating this co-adsorbed overlayer leads to the desorption of CO, in two stages at 330 K and 430 K; the excess CO (0.1 ML) desorbs at 590 K. LEED IV structural analysis of the mixed c(4 x 2) O and CO overlayer shows that both the CO molecules and the O atoms occupy bridge sites. The O atoms show significant lateral displacements of 0.14 angstrom away from the CO molecules; the C-O bond is slightly expanded with respect to the gas phase (1.19 angstrom); the modifications of the Ir substrate with respect to the bulk-terminated surface are very small. (c) 2006 Elsevier B.V. All rights reserved.

Selective separation of the major whey proteins using ion exchange membranes

Synthetic microporous membranes with functional groups covalently attached were used to selectively separate beta-lactoglobulin, BSA, and alpha-lactalbumin from rennet whey. The selectivity and membrane performance of strong (quaternary ammonium) and weak (diethylamine) ion-exchange membranes were studied using breakthrough curves, measurement of binding capacity, and protein composition of the elution fraction to determine the binding behavior of each membrane. When the weak and strong anion exchange membranes were saturated with whey, they were both selective primarily for beta-lactoglobulin with less than 1% of the eluate consisting of alpha-lactalbumin or BSA. The binding capacity of a pure alpha-lactoglobulin solution was in excess of 1.5 mg/cm(2) of membrane. This binding capacity was reduced to approximately 1.2 mg/cm(2) when using a rennet whey solution (pH 6.4). This reduction in protein binding capacity can be explained by both the competitive effects of other whey proteins and the effect of ions present in whey. Using binary solution breakthrough curves and rennet whey breakthrough curves, it was shown that alpha-lactalbumin and BSA were displaced from the strong and weak anion exchange membranes by beta-lactoglobulin. Finally, the effect of ionic strength on the binding capacity of individual proteins for each membrane was determined by comparing model protein solutions in milk permeate (pH 6.4) and a 10 mM sodium phosphate buffer (pH 6.4). Binding capacities of beta-lactoglobulin, alpha-lactalbumin, and BSA in milk permeate were reduced by as much as 50%. This reduction in capacity coupled with the low binding capacity of current ion exchange membranes are 2 serious considerations for selectively separating complex and concentrated protein solutions.

First-dimension separation with the MicroRotoforTM cell prior to SDS-PAGE and LC-MS/MS analysis

Free-flow isoelectric focusing (IEF) is a gel-free method for separating proteins based on their isoelectric point (pl) in a liquid environment and in the presence of carrier ampholytes. this method has been used with the RotoforTM cell at the preparative scale to fractionate proteins from samples containing several hundred milligrams of protein; see the refeences listed in Bio-Rad bulletin 3152. the MicroRotofor cell applies the same method to much sl=maller protein samples without dilution, separating and recoverng milligram quantities of protein in a total volume of about 2 ml.

Experimental separation of the coherent component of X-ray scattering prior to RDF analysis of non-crystalline polymers

An experimental method is described which enables the inelastically scattered X-ray component to be removed from diffractometer data prior to radial density function analysis. At each scattering angle an energy spectrum is generated from a Si(Li) detector combined with a multi-channel analyser from which the coherently scattered component is separated. The data obtained from organic polymers has an improved signal/noise ratio at high values of scattering angle, and a commensurate enhancement of resolution of the RDF at low r is demonstrated for the case of PMMA (ICI `Perspex'). The method obviates the need for the complicated correction for multiple scattering.

Folding of graphene slit like pore walls—a simple method of improving CO2 separation from mixtures with CH4 or N2

We report for the first time a detailed procedure for creating a simulation model of energetically stable, folded graphene-like pores and simulation results of CO2/CH4 and CO2/N2 separation using these structures. We show that folding of graphene structures is a very promising method to improve the separation of CO2 from mixtures with CH4 and N2. The separation properties of the analyzed materials are compared with carbon nanotubes having similar diameters or S/V ratio. The presented results have potential importance in the field of CO2 capture and sequestration.

Regional and monthly and clear-sky aerosol direct radiative effect (and forcing) derived from the GlobAEROSOL-AATSR satellite aerosol product

Using the GlobAEROSOL-AATSR dataset, estimates of the instantaneous, clear-sky, direct aerosol radiative effect and radiative forcing have been produced for the year 2006. Aerosol Robotic Network sun-photometer measurements have been used to characterise the random and systematic error in the GlobAEROSOL product for 22 regions covering the globe. Representative aerosol properties for each region were derived from the results of a wide range of literature sources and, along with the de-biased GlobAEROSOL AODs, were used to drive an offline version of the Met Office unified model radiation scheme. In addition to the mean AOD, best-estimate run of the radiation scheme, a range of additional calculations were done to propagate uncertainty estimates in the AOD, optical properties, surface albedo and errors due to the temporal and spatial averaging of the AOD fields. This analysis produced monthly, regional estimates of the clear-sky aerosol radiative effect and its uncertainty, which were combined to produce annual, global mean values of (−6.7±3.9)Wm−2 at the top of atmosphere (TOA) and (−12±6)Wm−2 at the surface. These results were then used to give estimates of regional, clear-sky aerosol direct radiative forcing, using modelled pre-industrial AOD fields for the year 1750 calculated for the AEROCOM PRE experiment. However, as it was not possible to quantify the uncertainty in the pre-industrial aerosol loading, these figures can only be taken as indicative and their uncertainties as lower bounds on the likely errors. Although the uncertainty on aerosol radiative effect presented here is considerably larger than most previous estimates, the explicit inclusion of the major sources of error in the calculations suggest that they are closer to the true constraint on this figure from similar methodologies, and point to the need for more, improved estimates of both global aerosol loading and aerosol optical properties.

Automated artifact removal from the electroencephalogram; a comparative study

Contamination of the electroencephalogram (EEG) by artifacts greatly reduces the quality of the recorded signals. There is a need for automated artifact removal methods. However, such methods are rarely evaluated against one another via rigorous criteria, with results often presented based upon visual inspection alone. This work presents a comparative study of automatic methods for removing blink, electrocardiographic, and electromyographic artifacts from the EEG. Three methods are considered; wavelet, blind source separation (BSS), and multivariate singular spectrum analysis (MSSA)-based correction. These are applied to data sets containing mixtures of artifacts. Metrics are devised to measure the performance of each method. The BSS method is seen to be the best approach for artifacts of high signal to noise ratio (SNR). By contrast, MSSA performs well at low SNRs but at the expense of a large number of false positive corrections.