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Production dans Escherichia coli de vésicules enrichies en cavéoline-1(32-178) canine ou son fragment (76-178) / Production in Escherichia coli of Vesicles Enriched in Canin Caveolin-1(32-178) or its Fragment (76-178)Perrot, Nahuel 04 November 2015 (has links)
La cavéoline-1, une petite protéine membranaire de 21 kDa, est la protéinemembranaire majoritaire d'invaginations de la membrane cytoplasmique appeléescavéoles. Enrichis en cholestérol et sphingolipides, ces domaines ont un rôleimportant dans de nombreux aspects de la vie cellulaire et constituent une véritableplateforme d'interactions protéiques et lipidiques. Ces dernières années, malgré lenombre important de travaux concluant à l'implication de la cavéoline-1, ou descavéoles, dans de nombreux processus cellulaires ou pathologiques, les donnéesquant à l'organisation de cette protéine au sein de la membrane plasmique restent trèséparses. Aussi, l'objectif principal de ce travail est de contribuer à l'acquisition dedonnées structurales sur cette protéine. Ce travail se base sur les expressionshétérologues d'une isoforme de la cavéoline-1 canineou de l'un de ses fragments, dans un hôte bactérien, sous la forme de protéines defusion associées à la Maltose Binding Protein. Ces expressions induisent laformation de vésicules intracytosoliques composées majoritairement de la protéineexprimée. Aussi, la première partie du travail est consacré à la mise en place d'unprotocole de purification de ces vésicules dans des conditions natives, répondant aucritère de réaliser une étude structurale de cette protéine n'impliquant pas l'usage dedétergent. La deuxième partie porte sur une application potentielle de ces vésicules, eten particulier pour des essais de caractérisation d'une enzyme membranaire,la glutathion-S-transférase microsomale de rat. Enfin une troisième partie est dédiée àl'analyse de simulations de dynamique moléculaire dans le cadre d'étudesd'interactions au sein de systèmes membranaires. / Caveolin-1, a 21 kDa membrane protein, is the principal membrane protein ofcytoplasmic membrane domains named caveolae. Specifically enriched incholesterol and sphingolipids those domains are important in many aspect of the cell's life and make up a proteins and lipids interaction platform. In the past years, despite a large number of publications stating the implication of caveolin-1 or caveolae in many cell processes and pathologies, very few is known about theway this protein is organized at the cell membrane. Hence, the main purpose of thiswork is to contribute to the acquisition of structural data on this protein. At the base of this work is the heterologous expression within a bacterial host, and as a fusion protein, of the canin beta isoform of cavéolin-1 or one of its fragment. These expressions lead to the formation of cytoplasmic vesicles composed mainly ofthe expressed protein. Thence, the first part of this work focus on developping a method to purify those vesicles that does not rely on using any kind of detergent which could enable structural studies in a native environnment. The second part present a potentiel application of those vesicles and inparticular the use of those vesicles to characterize a membrane enzyme, namelythe murin microsomal glutathion-Stransferase. The last part will be a contribution to data analysis within the context of molecular dynamics simulationof membraneous systems.
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Removal of Heavy Metal Ions from Aqueous Solution by Alkaline FiltrationXu, Zitong 22 January 2020 (has links)
An innovative approach for the removal of heavy metal ions such as Pb2+ and Cd2+ from aqueous solution was evaluated. It was established that alkaline filtration, which is in essence the combination of alkaline precipitation and membrane filtration, could drastically increase both the efficiency and completeness of Pb2+ or Cd2+ ions removal, producing water whose metal concentration satisfying drinking water standard from a simulation wastewater containing 5 ppm or more Pb2+ or Cd2+ ions. Filtration with three different membranes, including microfiltration (MF), ultrafiltration (UF), and nanofiltration (NF) membranes, were studied at three different pH levels, i.e., 7.0, 8.5, and 10, in terms of metal ion rejection, flux, and permeate pH and at varied dissolved inorganic carbon (DIC) concentration. Increasing the pH of the feed in the tested range would lead to the decrease of metal ion concentration in permeate while flux was in general unaffected. When the feed pH was 10, the Pb2+ concentration in permeate was below 10 ppb regardless of the DIC concentration and membrane for filtration. The effects of DIC concentration were significant but complex. It was found that MF, UF, NF could all effectively reject Pb2+ ions at pH 8.5 and pH 10 although only NF was charged. A hypothesis was proposed to explain the mechanism of alkaline filtration based on experimental data.
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Poly-thiosemicarbazide Membrane for Gold Adsorption and In-situ Growth of Gold NanoparticlesParra, Luis F. 12 1900 (has links)
In this work the synergy between a polymer containing chelate sites and gold
ions was explored by the fabrication of a polymeric membrane with embedded gold
nanoparticles inside its matrix and by developing a process to recover gold from
acidic solutions. After realizing that the thiosemicarbazide groups present in the
monomeric unit of poly-thiosemicarbazide (PTSC) formed strong complexes with Au
ions, membrane technology was used to exploit this property to its maximum.
The incorporation of metal nanoparticles into polymeric matrices with current
technologies involves either expensive and complicated procedures or leads to poor
results in terms of agglomeration, loading, dispersion, stability or efficient use of raw
materials. The fabrication procedure described in this thesis solves these problems
by fabricating a PTSC membrane containing 33.5 wt% in the form of 2.9 nm gold
nanoparticles (AuNPs) by a three step simple and scalable procedure. It showed
outstanding results in all of the areas mentioned above and demonstrated catalytic
activity for the reduction of 4-Nitrophenol (4−NP) to 4-Aminophenol (4−AP).
The current exponential demand of gold for electronics has encouraged the development
of efficient processes to recycle it. Several adsorbents used to recover gold from
acidic solutions can be found in the literature with outstanding maximum uptakes,yet, poor kinetics leading to an overall inefficient process. The method developed
in this dissertation consisted in permeating the gold-containing solution through a
PTSC membrane that will capture all the Au ions by forming a metal complex with
them. Forcing the ions through the pores of the membrane eliminates the diffusion
limitations and the adsorption will only depended on the fast complexation kinetics,
resulting in a very efficient process. A flux as high as 1868 L/h m2 was enough to
capture >90% of the precious metal present in a solution of 100 ppm Au. The maximum
uptake achieved without sacrificing the mechanical stability was 5.4 mmol/g.
The selectivity between gold and copper (the most common unwanted metal present
along with gold) was 6.7 for 100 ppm initial concentration of both metals and 14.6
for 500 ppm.
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Natural Gas Sweetening by Ultra-Microporous Polyimides MembranesAlghunaimi, Fahd 05 1900 (has links)
Most natural gas fields in Saudi Arabia contain around 10 mol.% carbon dioxide. The present technology to remove carbon dioxide is performed by chemical absorption, which has many drawbacks. Alternatively, membrane-based gas separation technology has attracted great interest in recent years due to: (i) simple modular design, (ii) potential cost effectiveness, (iii) ease of scale-up, and (iv) environmental friendliness. The state-of-the-art membrane materials for natural gas sweetening are glassy cellulose acetate and polyimide, which were introduced in the 1980s. In the near future, the kingdom is planning to boost its production of natural gas for power generation and increase the feedstock for new petrochemical plants. Therefore, the kingdom and worldwide market has an urgent need for better membrane materials to remove carbon dioxide from raw natural gas.
The focus of this dissertation was to design new polyimide membrane materials for CO2/CH4 separation exhibiting high permeability and high selectivity relative to the standard commercial materials tested under realistic mixed-gas feed conditions. Furthermore, this study provided a fundamental understanding of structure/gas transport property relationships of triptycene-based PIM-polyimides. Optimally designed intrinsically microporous polyimide (PIM-PIs) membranes in this work exhibited drastically increased CO2/CH4 selectivities of up to ~75. In addition, a novel triptycene-based hydroxyl-containing polyimide (TDA1-APAF) showed 5-fold higher permeabilities over benchmark commercial materials such as cellulose acetate. Furthermore, this polyimide had a N2/CH4 selectivity of 2.3, thereby making it possible to simultaneously treat CO2- and N2-contaminated natural gas. Also, TDA1-APAF showed a CO2 permeability of 21 Barrer under binary 1:1 CO2/CH4 mixed-gas feed with a selectivity of 72 at a partial CO2 pressure of 10 bar which are significantly better than cellulose triacetate. These results suggest that TDA1-APAF polyimide is an excellent candidate membrane material for removal of CO2 and N2 from natural gas. Moreover, based on the collected data for CO2/CH4 mixed-gas experiments from this work and previously published reports, a new mixed-gas 2017 CO2/CH4 permeability/selectivity upper bound curve was initiated to reflect the actual performance including plasticization phenomena at high feed pressure and 10 bar CO2 partial pressure to simulate the real conditions of the wellhead pressure.
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Hyaline membrane disease: a study of lung function and treatmentHarrison, V C 07 April 2020 (has links)
At present, both the aetiology of hyaline membrane disease and a means of preventing it remain unknown. Recent studies indicate that a significant number of infants die of respiratory failure but there is no general agreement concerning the changes of pulmonary function which lead to this stage. Two approaches have been used in the treatment of respiratory decompensation. First it has been proposed that blood gas and acid base abnormalities which result from respiratory failure can be prevented by oxygen and intravenous alkali and secondly an attempt has been made to correct abnormal lung function itself by means of artificial ventilation. These methods are directed at different aspects of the problem and their efficacy is as yet not established. The application of artificial ventilation in particular must depend on the nature of any ventilation, diffusion or perfusion defect.
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Design, fabrication, and reduction to practice of milliscale membrane-free organ chip systemsJanuary 2021 (has links)
archives@tulane.edu / The goal of this research was to establish a novel digital manufacturing-based workflow for the fabrication of membrane free organ chip (MFOC) systems. This workflow is based on the implementation of top-down design, starting with CAD design of molds for MFOC components and can be conducted on a benchtop removing the need for cleanroom use. In conducting this research, a commercially available SLA printer was characterized and optimized for manufacturing molds suitable for MFOC fabrication. To achieve this, extensive research was required to determine printer resolution limits and work within the limitations of the resins available for printing. Specifically, the molds need to be flat and smooth in order to produce perfectly horizontal and transparent PDMS devices. Post-processing workflows were engineered to satisfy these MFOC design constraints. After establishing a reliable and reproducible workflow for MFOC fabrication, the focus of the research was reduction to practice, i.e. achieving a design that enables loading MFOC with patterned aqueous solutions with 100% success and a high degree of forgiveness. Key MFOC dimensions were systematically varied in a manner only possible with the rapid prototyping capability of DM in a series of experiments with a standardized injection test and success rate of loading as the primary output. With a robust MFOC design in place, more complex designs for tissue patterning applications were created, and advanced configurations for engineering patterned vascularized stromal tissues were tested and validated. Seqeuntial and simultaneous loading scenerios were imvestigated to better understand cell migration impedence in multi-gel lane devices. / 1 / William Bralower
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Transcriptional Homeostatic Control of Membrane Lipid CompositionThewke, Douglas, Kramer, Marianne, Sinensky, Michael S. 24 June 2000 (has links)
Plasma membranes have a structural property, commonly referred to as membrane fluidity, that is compositionally regulated. The two main features of plasma membrane lipid composition that determine membrane fluidity are the ratio of cholesterol to phospholipids and the ratio of saturated to unsaturated fatty acids that are incorporated into the phospholipids. These ratios are determined, at least in part, by regulation of membrane lipid biosynthesis-particularly that of cholesterol and oleate. It now appears that cholesterol and oleate biosynthesis are feedback regulated by a common transcriptional mechanism which is governed by the maturation of the SREBP transcription factors. In this article, we briefly review our current understanding of transcriptional regulation of plasma membrane lipid biosynthesis by sterols and oleate. We also discuss studies related to the mechanism by which the physical state of membrane lipids signals the transcriptional regulatory machinery to control the rates of synthesis of these structural components of the lipid bilayer.
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Biosynthesis, Transport, and Modification of Lipid ATrent, M. Stephen 01 February 2004 (has links)
Lipopolysaccharide (LPS) is the major surface molecule of Gram-negative bacteria and consists of three distinct structural domains: O-antigen, core, and lipid A. The lipid A (endotoxin) domain of LPS is a unique, glucosamine-based phospholipid that serves as the hydrophobic anchor of LPS and is the bioactive component of the molecule that is associated with Gram-negative septic shock. The structural genes encoding the enzymes required for the biosynthesis of Escherchia coli lipid A have been identified and characterized. Lipid A is often viewed as a constitutively synthesized structural molecule. However, determination of the exact chemical structures of lipid A from diverse Gram-negative bacteria shows that the molecule can be further modified in response to environmental stimuli. These modifications have been implicated in virulence of pathogenic Gram-negative bacteria and represent one of the molecular mechanisms of microbial surface remodeling used by bacteria to help evade the innate immune response. The intent of this review is to discuss the enzymatic machinery involved in the biosynthesis of lipid A, transport of the molecule, and finally, those enzymes involved in the modification of its structure in response to environmental stimuli.
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Radiative Heat Transfer in Free-Standing Silicon Nitridemembranes in the Application of Thermal Radiation SensingZhang, Chang 05 November 2020 (has links)
Thin-film silicon nitride (SiN) membranes mechanical resonators have been widely used for many fundamental opto-mechanical studies and sensing technologies due to their extremely low mechanical dissipation (high mechanical Q-factor). In this work, we experimentally demonstrate an opto-mechanical approach to perform thermal radiation sensing, using a SiN
membrane resonator. An important aspect of this work is to develop a closed-form analytical heat transfer model for assessing the thermal coupling conditionbetween free-standing membranes and their environment. We also derive analytical expressions for other important intrinsic thermal quantities of the membrane, such as thethermal conductance, the heat capacity and the thermal time constant. Experimental results show good agreement with our theoretical prediction. Of central importance, we show that membranes of realistic dimensions can be coupled to their environment more strongly via radiation than by solid-state conduction. For example, membranes with 100nm thickness (frequently encountered size) are predicted to be radiation dominated when their side length exceeds 6 mm. Having radiation dominated thermal coupling is a key ingredient for reaching the fundamental detectivity limit of thermal detectors. Hence, our work proves that SiN membranes are attractive candidates for reaching the fundamental limit. We also experimentally exhibit the high temperature responsivity of the SiN membranes resonance, in which we shift a 88.7 KHz resonance by over 1 KHz when temperature increment on the membrane is approximately 2 K.
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Synthesis and characterisation of sulphonated polyethersulphone membrane materialsBoukili, Aishah January 2020 (has links)
>Magister Scientiae - MSc / With current climate change, growing population, and rapid industrialization of developing countries, water is increasingly becoming a scare resource. Within a power plant, processes that consume most water are demineralized water production (boiler make-up), heat rejection (cooling) and emission control (wet flue gas desulfurization). Eskom’s fleet of existing coal-fired power plants are not equipped with SO2 abatement technologies and therefore retrofitting of the plants will be required to meet the compliance levels for SO2 emissions.
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