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Regulatory Effect of Elastin Based Biomaterial on Cellular Behavior and Its Application on Wound Repair and RegenerationYuan, Yuan 17 March 2016 (has links)
Elastin-like peptides (ELPs) are stimulus-responsive protein-based polymers which are attractive material for biomedical research due to their biocompatibility and unique properties. The physical properties of ELPs are dependent on the chain length and the chosen amino acid at the guest residue position. This imparts unlimited flexibility in designing ELP based biomaterials with the desired physical properties.
We have shown that in addition to their physical properties, ELPs have biological activities that are conducive to tissue regeneration. Specifically, we found that ELPs induce fibroblast proliferation via cell surface heparan sulfate proteoglycans (HSPG). Furthermore, our data suggests that ELP based materials with differential proliferative potential can be designed by controlling the interaction of ELPs with HSPGs by incorporating either hydrophobic or positively charged residues within the ELP sequence. Fibroblast proliferation is important for granulation tissue formation which is important in chronic wounds as well as in healing of other tissues. The customizable biological activity of ELPs coupled with their unique physical properties will enable us to design novel, sustainable and cost effective therapies for different tissue regeneration applications.
ELPs can be genetically fused to biologically active peptides or proteins. These fusions can be expressed and readily purified since they maintain the phase transitioning property of the fused ELP domain. Moreover, depending on the ELP sequence chosen the chimeric fusion sequences can self-assemble into unique structures such as nanoparticles. These structures can then be applied to the injury site where they not only provide unique topographical cues or structural support but also act as delivery vehicles for the fused bioactive protein. We developed a multifunctional nanoparticle that is comprised of PMP-D2-ELP fusion protein and different functional peptide ELP fusion proteins to preserve the bioactivity of the functional group with the existence of elastase. These heterogeneous particles will be beneficial for the delivery of combination therapies to solve multiple problems that often existed in chronic wound healing or other tissue regeneration process.
In summary, this study adds to our understanding of the biological activity of ELP and the interaction mechanism that allow the regulation of cellular behavior. Furthermore this work also investigated the potential therapeutic application of ELP as a delivery platform for chronic wound healing.
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Using sodium bisulphite treatment and PCR to construct mammalian anti-HIV-1 long hairpin RNA expression cassettesLugongolo, Masixole Yvonne 03 May 2012 (has links)
M.Tech. / RNA interference (RNAi) is a gene silencing mechanism that uses short RNA duplexes to block gene expression. This mechanism has been widely explored to determine functions of genes. Furthermore, this phenomenon has been used to silence unwanted genes such as viral genes. RNAi has been successfully employed in non-mammalian organisms such as plants, where long dsRNAs (more than 30 bp) have been used without inducing non-specific effects. However, in mammalian cells, cytoplasmic dsRNAs of more than 30 bp trigger non-specific induction of many genes, which may result from the activation of dsRNA-dependent protein kinase (PKR) and 2’,5’-oligoadenylate synthetase (2’,5’-OAS), via the interferon response pathway. In this study, we describe a novel and simple strategy to overcome nonspecific effects induced by longer RNA duplexes. This strategy uses sodium bisulphite which is a mutagen that deaminates cytosine residue to uracil residues in order to introduce mutations in the sense strand of the duplex. Introduction of these mutations results in the formation of G:U pairings between the sense and antisense strands of the long hairpin RNA. RNA duplexes with mismatches have been shown to be able to prevent interferon induction in mammalian cells. According to the obtained results, long hairpins RNA with and without mismatches were unable to inhibit the expression of the target region, which was the U5 region of the HIV-1 subtype C LTR. The U5 region of the LTR is actively involved in the reverse transcription of HIV-1. Therefore silencing of this region would have led to the inhibition or reverse transcription blockage. Furthermore, data showed that the interferon response was induced when using these long hairpin RNA duplexes. Due to the sensitivity of mammalian cells, the action of sodium bisulphite could have stimulated certain genes of the interferon pathway. Even though hairpins constructed in this study were unable to prevent the induction of the interferon response pathway and also could not silence the target, this strategy of using sodium bisulphite has a great potential as shown by its ability to induce changes in cytosine residues and leaving other nucleotides unchanged.
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Etude du rôle des héparans sulfates protéoglycanes dans la mobilisation post-lesionnelle des progéniteurs oligodendrocytaires chez la souris adulte / Role of heparan sulphate proteoglycans in post-lesional mobilization of oligodendrocyte prgenitor cells in adult miceMacchi, Magali 12 November 2015 (has links)
La production physiologique continue de cellules myélinisantes dans le système nerveux (SN) de mammifère offre de nouvelles perspectives thérapeutiques. Lors d’une atteinte de la myéline, une régénération endogène impliquant la génération d’oligodendrocytes s’engage. Ce processus repose sur la mobilisation de progéniteurs oligodendrocytaires parenchymateux et de progéniteurs de la zone sous-ventriculaire (SVZ). Cette réparation ne permet cependant pas une récupération fonctionnelle systématique. Nos travaux ont pour but d’identifier les facteurs qui contrôlent les différentes étapes de régénération. Ils révèlent une réexpression du CNTF et une surexpression des héparans sulfates protéoglycanes (HSPGs) suite à une démyélinisation du corps calleux. Ces changements de l’environnement péri-lésionnel régulent positivement le processus de remyélinisation. Nous avons montré un impact direct de l’expression post-lésionnelle du CNTF sur la mobilisation des deux sources cellulaires. Différents tests in vitro ont identifié le CNTF comme facteur chémoattractant pour ces cellules. Nos données montrent également que des modifications de sulfatation des héparans sulfates (HS) protéoglycanes contrôlées par la N-désacétylase-Sulfotransférase 1 des cellules du lignage oligodendrocytaire s’établissent en bordure de lésion et créent un microenvironnement favorable à la régénération. Divers test fonctionnels in vivo et in vitro révèlent le rôle clef des HSPGs dans la cinétique de démyélinisation et de remyélinisation en régulant la mobilisation des cellules du lignage oligodendrocytaire et l’activation microgliale. / In the mammal’s nervous system, the ongoing production of new myelinating cells on has open news therapeutic perspectives for demyelinating diseases. An endogenous regeneration process involving the generation of oligodendrocytes can occur following demyelination. This process relies on the mobilization of an endogenous reservoir of progenitor cells located in the adult brain: The parenchymal oligodendrocyte precursors and the subventricular zone derived neural progenitors. However, these endogenous repair attempts do not permit an efficient functional recovery. These failures are mainly due to mobilization, differentiation or to the generation of a hostile environment for the repair process. Our work is focusing on the identification of factors regulating those events. Our data show the reexpression of CNTF and overexpression of heparan sulphate proteoglycans (HSPGs) following a focal demyelination of the corpus callosum in adult mice. These environmental changes favor myelin repair. We show a direct impact of the post-lesional expression of CNTF on the mobilization of both cellular sources. Using various in vitro assays, we showed that CNTF is acting on the two cellular sources as a chemoattractant factor. Our data also show that sulfation modifications of HSPGs performed by the deacetylase-N-sulfotransferase 1 (Ndst1) on oligodendrocyte lineage cells occurred around the lesion and created a permissive microenvironment for the regenerative process. Various in vitro and in vivo functional assays demonstrated the key role of HSPGs in demyelination and remyelination dynamic by controlling mobilization of the oligodendrocyte lineage cells and microglial activation.
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An evaluation of recirculating artificial stream designs for acute toxicity testing using two South African Ephemeroptera species exposed to sodium sulphateBinder, Markus January 2000 (has links)
Three artificial stream designs, termed Large Artificial Stream Units (LASUs), Raceways, and Channels, at two major scales (1700 L, 12.5 L and 20 L recirculated volume) were developed at the Institute for Water Research, Rhodes University, Grahamstown, in order to explore the possibilities of using indigenous rheophilic macroinvertebrates in routine toxicity tests. This study compared these systems, using 96h-EC50 values from sodium sulphate toxicity tests as the experimental response. Two local Ephemeroptera (Leptophlebiidae: Adenophlebia auriculata Eaton, and Baetidae: Afroptilum sudafricanum Lestage) were evaluated for their suitability in routine toxicity tests; and the possible effects of elevated salinity levels in South African rivers on the test species were assessed. Two sets of experiments with each mayfly species were conducted, following an unreplicated regression design. Dechlorinated tap water was used as the water source. Experiments in the Channels were repeated to determine experimental variability. Results were compared statistically by testing for overlap of 95% confidence limits (95%Cls) of EC50 values. The differences between A. auriculata EC50 values in the different systems were statistically significant (no overlap of 95%CLs), but they were not more variable than has been considered normal for biological systems (Coefficient of variation 20.1 %; ratio of greatest EC50 / smallest EC50 1.63). The differences were not related to the scale or the average current velocity characteristic of each stream design (average current velocity LASUs - Raceways - Channels 0.090 - 0.083 - 0.038 m/s). The Channels proved to be most efficient with regard to practical performance as they are portable and easily transportable, user-friendly, reliable, splash-free, cost effective to construct, and can easily be adapted to specific requirements. These systems are therefore recommended for regular use. The suitability of the two mayfly species for routine toxicity testing was evaluated. A. auriculata EC50 values showed a significant negative correlation with the corresponding average body-size (range 1476 - 1610 μm, mean 1555 μm). The different average body-sizes probably reflected the abundance of a certain size range present in the Palmiet River at the time of collection. Both species reacted similarly to Na₂S0₄ (similar slopes of the toxicity curves), identifying this salt as a slow acting toxicant. A. sudafricanum populations were more sensitive to Na₂S0₄ (EC50 3.404 g/L) than A. auriculata (EC50 8.090 g/L), probably because of its smaller body-size (mean 709 μm) and a lack of extremely tolerant individuals. In comparison to other freshwater macro invertebrates, including the standard toxicity test organism Daphnia spp., both mayfly species seemed to be moderately tolerant of Na₂S0₄; therefore there was no particular advantage to using these indigenous taxa rather than Daphnia spp. An assessment of the effects of elevated salinity/TDS levels on the test taxa yielded preliminary insights. A NaCI-EC50 for A. sudafricanum could be extrapolated and suggested a higher sensitivity to Na₂S0₄ than to NaCl. When Na₂S0₄ EC50 values of both species were compared to selected TDS levels of South African rivers, 4. auriculata would mostly not be affected, but A. sudafricanum might occasionally suffer from sub-lethal effects, depending on the sulphate proportion of the TDS. The South African guideline for TDS seemed to protect both species sufficiently.
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The effect of different levels and intervals of application of ammonium sulphate on the growth, chemical composition and yield of cayenne and queen pineapple plants under field conditionsVan Lelyveld, Louis Johannes January 1965 (has links)
From the Introduction. The fertilisation of pineapples in the Eastern Cape Province is still a relatively new practice which started approximately twelve years ago. Even after this time many growers still insist that pineapples can be grown without fertilisation on virgin soils. As the available virgin soil decreased, however, so the interest in fertiliser application increased. Very little basic research on the fertiliser requirements of pineapples in the Eastern Cape was done to serve as a guide to growers. The available advisory literature by le Roux (1951), Malan (1954) and Lewcock (1956) were based on overseas experience. When pineapple research was started on a large scale, in 1955, it was realised that emphasis should be placed on the nutritional requirements on virgin as well as replanted soils. From initial experiments it was clear that the main response in plant growth and yield was obtained from nitrogenous fertilisers.
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Electrodes positives à base de cuivre pour accumulateurs Li-ion / Copper-based positive electrodes for Li-ion batteriesVan Staen, Guilherme 19 February 2016 (has links)
Les accumulateurs Li-ion sont des systèmes de stockage électrochimique de l’énergie composés de deux électrodes, dans lesquelles les ions Li+ vont venir s’insérer réversiblement lors des cycles de charge et de décharge. Afin d’intégrer le domaine des véhicules électriques, leur densité d’énergie doit être augmentée pour apporter l’autonomie demandée. Ceci peut être réalisé en augmentant la d.d.p. entre les deux électrodes. Nous visons ici la synthèse de nouveaux matériaux polyanioniques d’électrode positive dans lesquels le lithium pourrait venir s’insérer à haut potentiel en faisant intervenir le couple Cu3+/Cu2+ (5,3 V vs Li+/Li). Parmi les phosphates de cuivre synthétisés, Li2CuP2O7 présente une oxydation non réversible à haut potentiel (> 5 V). Sa synthèse à basse température permet d’exacerber les réactions, en raison de la faible taille des particules obtenues ainsi que de la présence de carbone conducteur à leur surface, mais la phase s’avère instable à haut potentiel.En ce qui concerne les composés de type sulfate, une nouvelle phase Li4Cu4O2(SO4)4 est isolée, montrant une insertion réversible du lithium à une valeur moyenne de 4,7 V. Cependant, la capacité de ce matériau est très faible (15 mAh.g-1) et plusieurs substitutions chimiques avec du fluor, du magnésium ou du sodium sont étudiées dans le but d’augmenter la mobilité du lithium. / Li-ion batteries (LIBs) are energy storing electrochemical devices composed of two electrodes, in which Li+ ions are reversibly inserted during charge and discharge cycles. Their use in electric vehicles relies on the increase of their energy density, to provide enough autonomy. This can be reached by increasing the cell d.d.p. We thus aim the synthesis of new positive electrode polyanionic materials, in which lithium could be inserted at high potential, using the Cu3+/Cu2+ couple’s activity (5,3 V vs Li+/Li). Among the synthesized copper phosphates, Li2CuP2O7 presents a non-reversible oxidation at high potential (>5 V). Its low temperature synthesis intensifies the reaction, due to the smaller particle size achieved as well as the presence of a conductive carbon coating, but the phase is instable at high potential. Concerning sulfate-type compounds, a new phase Li4Cu4O2(SO4)4 is isolated, showing a reversible lithium insertion at an average value of 4.7 V. Nevertheless, its capacity is very low (15 mAh.g-1) and various chemical substitutions with fluorine, magnesium or sodium are attempted to increase lithium’s mobility.
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On the Role of the Pore Water Chemistry of Deficient Post-Tension Grout to Promote Steel CorrosionPermeh, Samanbar 30 March 2016 (has links)
Corrosion failure of post-tension tendons with pre-packaged thixotropic grout was documented in Florida Bridges in less than 10 years of service. Analysis of deficient grout in Florida post-tensioned (PT) bridges where severe corrosion developed indicated enhanced sulfate ion concentration, high pore water pH, enhanced moisture and low chloride content. The corrosion wasn’t consistent with the conventional causes of steel corrosion such as bleed water accumulation in grout void spaces, chloride contamination and pore water carbonation. Corrosion development was associated with deficient grout with enhanced sulfate levels. However, limited information is available on the corrosion behavior of PT strand in grout materials with enhanced sulfate content. Review of the technical literature has not shown consistent evidence of the role of sulfate ions in the corrosion of steel in solution and in hydrated cementitious material. In this research, the role of sulfates in cementitous materials was examined including identifying practical limits for sulfates in grout materials.
Laboratory samples were created with 15% and 20% excess water to enhance deficient grout materials and included enhanced sodium and chloride content. Enhanced sulfate and chloride ion presence was thought to provide aggressive corrosion condition of steel in segregated grout. Testing was intended to differentiate the corrosion of steel in pore water condition of deficient grout.
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Studies On The Bioremoval Of Zinc And Cadmium Using Desulfotomaculum nigrificansRadhika, V 08 1900 (has links) (PDF)
No description available.
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Tracing Biogeochemical Processes Using Sulfur Stable Isotopes: Two Novel ApplicationsCousineau, Mélanie L. January 2013 (has links)
Abstract Dissimilatory microbial sulfate reduction (MSR) The specific objectives of the study were to provide the first measurements of sulfur isotope fractionation associated with acidophilic sulfate reducing-microorganisms, and to examine whether pH influences sulfur fractionation during MSR. The fractionation associated with the strains investigated was comparable to that of neutrophilic strains with similar metabolisms (4-12‰), but varied with pH. Two fractionation regimes were identified: one regime is consistent with fractionation during exponential growth, while the other – not identified previously - is not linked to active sulfate reduction and may result from internal sulfate accumulation. This would represent the first measurement of sulfur fractionation during sulfate uptake, the first step of MSR. Geological processes at the Cretaceous-Paleogene (KPg) boundary The KPg boundary is associated with one of the largest biological extinctions in the history of our planet. Two major geologic events - the Chicxulub bolide impact with evaporite terrane and the eruption of the Deccan continental flood basalts - coincide with the KPg boundary and have been identified as possible triggers for the extinctions, but their relative timing remains unresolved. The objectives of this study were to identify the contribution of these processes to the sulfur burden in the sedimentary environment of two freshwater KPg sections, and to determine their relative timing. The results demonstrate that the peak of Deccan volcanism post-dates the Chicxulub impact and the associated abrupt KPg mass extinction, thus precluding a direct volcanic causal mechanism, but shedding light on the underlying causes for the delayed recovery of ecosystems in the early Paleogene.
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Stormwater Retention Ponds: Hydrogen Sulfide Production, Water Quality and Sulfate-Reducing Bacterial KineticsD'Aoust, Patrick Marcel January 2016 (has links)
Stormwater retention basins are an integral component of municipal stormwater management strategies in North America. The province of Ontario’s Ministry of the Environment and Climate Change obligates land developers to implement stormwater management in their land use and development plans to mitigate the effects of urbanization (Bradford and Gharabaghi, 2004). When stormwater retention ponds are improperly designed or maintained, these basins can fail at improving effluent water quality and may exasperate water quality issues.
Intense H2S production events in stormwater infrastructure is a serious problem which is seldom encountered and documented in stormwater retention ponds. This study monitored two stormwater retention ponds situated in the Riverside South community, Ottawa, Ontario, Canada for a period of 15 consecutive months to thoroughly characterize intense hydrogen sulfide (H2S) production in a stormwater retention pond under ice covered conditions during winter operation and during periods of drought under non-ice covered conditions during the summer.
Field experiments showed a strong relationship (p < 0.006, R > 0.58, n = 20+) between hypoxic conditions (dissolved oxygen (DO) concentration < 2 mg/L) and the intense production of H2S gas. Ice-capping of the stormwater ponds during winter severely hindered reaeration of the pond and led to significant production of total sulfides in the Riverside South Pond #2 (RSP2), which subsequently resulted in the accumulation of total sulfides in the water column (20.7 mg/L) during winter in this pond. There was a perceived lag phase between the drop in DO and the increase in total sulfides near the surface, which was potentially indicative of slow movement of total sulfides from the benthic sediment into the water column. These high-sulfide conditions persisted in RSP2 from early January 2015 until the spring thaw, in mid-April, 2015. Riverside South Pond #1 (RSP1), the reference pond studied in this work, showed significantly less production of total sulfides across a significantly shorter period of time. Analysis of the microbial communities showed that there was little change in the dominant bacterial populations present in the benthic sediment of the pond demonstrating significant total sulfide production (RSP2) and the pond that did not demonstrate significant total sulfide production (RSP1). Additionally, it was found that locations with the most accumulated sediment had the highest propensity for the production of H2S gas. Furthermore, there was no perceivable community shift in the two ponds throughout the seasons, indicating that the sulfate-reducing bacteria (SRB) in stormwater benthic sediment are ubiquitous, exist in an acclimatized microbial population and are robust. Study of the microbial abundances revealed that SRB represented approximately 5.01 ± 0.79 % of the microbes present in the benthic sediment of RSP2. Likewise, in the stormwater pond which did not experience intense H2S gas production, RSP1, 6.22 ± 2.11 % of microbes were of the SRB type, demonstrating that H2S gas production does not correspond to higher concentrations of SRB or the proliferation of dominant species, but rather is a symptom of increased bacterial activity due to favourable environmental conditions.
In addition, this work also covers the kinetics of sediment oxygen demand (SOD), ammonification and sulfate-reduction, and attempts to understand the processes leading to H2S gas production events.
In doing so, it was observed that kinetics obtained full-scale field studies were greater than in laboratory kinetic experiments. Laboratory experiments at 4°C identified total SOD, ammonification and sulfate-reduction kinetics to be 0.023 g/m2/day, 0.027 g N/m2/day and 0.004 g S/m2/day, respectively. Meanwhile, kinetics calculated from the field study of stormwater retention ponds for total SOD, ammonification and sulfate-reduction were of 0.491 g/m2/day, 0.120 g N/m2/day and 0.147 g S/m2/day, respectively. It is expected that this difference is due to the depth of active sediment influencing the total rates of production/consumption, making area-normalized daily rates of production/consumption (g/m2/day) unsuitable for the comparison of field and laboratory studies, without some scaling factor. This study also measured supplementary kinetic parameters such as the Arrhenius coefficients and the half-saturation coefficient, to add to existing knowledge of sulfate-reduction.
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