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Influence of haem availability on the viability of Porphyromonas gingivalis and Prevotella intermedia, following exposure to reactive oxygen speciesMackie, Tasha A, n/a January 2007 (has links)
Objectives: This investigation adapted the LIVE/DEAD� Baclight[TM] bacterial viability stain for the quantitative determination of bacterial cell viability of the aerotolerant anaerobes Porphyromonas gingivalis ATCC 33277 and Prevotella intermedia ATCC 25611. The Live/Dead stain was used to determine the influence of haem availability on the resistance of P. gingivalis and P. intermedia to the reactive oxygen species (ROS) superoxide anion and hydrogen peroxide and compare the sensitivities between the haem-requiring periodontal bacteria to ROS. Neutrophils use oxidative and non-oxidative killing mechanisms. During phagocytosis, neutrophils kill bacteria via a respiratory burst, producing ROS. P. gingivalis and P. intermedia are oxygen-tolerant gram-negative bacteria found in the gingival crevice. These bacteria express superoxide dismutase (SOD) activity, which extends some protection against superoxide radicals.
Methods: Initially, experiments were performed to validate the reliability and accuracy of the fluorogenic Live/Dead stain using Escherichia coli ATCC 10798 (K-12), followed by experiments using P. gingivalis. The Live/Dead stain distinguishes viable:non-viable proportions of bacteria using mixtures of green (SYTO 9) and red (propidium iodide) fluorescent nucleic acid stains respectively. Bacterial cell viability was assessed with fluorescence microscopy and subsequently quantitative measurement using a fluorescence microplate reader (BMG Fluorostar plus Optima). P. gingivalis and P. intermedia colonies were subcultured from frozen cultures, in Tryptic soy broth (TSB) (Difco) and incubated anaerobically for approximately five days. They were further subcultured in pre-reduced TSB, supplemented with menadione 0.5[mu]g/ml (TSB-M) and either 5 [mu]g/ml haemin (Haem 5), 50 [mu]g/ml haemin (Haem 50) or without supplemental haemin (Haem 0). Cultures were grown anaerobically at 37�C to early stationary phase (approximately 48 hours). For experimental purposes, bacteria were harvested, washed and resuspended in 10 mM Tris-buffered saline (pH 7.5) containing peptone (TBS-P) (0.1 mg/ml), with a final adjustment to OD₅₄₀ [approximately equals] 2.0 (which corresponds to 1 x 10⁹ bacteria/ml). Bacterial suspensions were diluted ([approximately equals] 10⁸/ml) into TBS-P containing the fluorogenic viability stain (BacLight, Molecular Probes). Either pyrogallol (0.02 - 2 mM) or hydrogen peroxide (0.01 - 100 mM) was added (except to control tubes); tubes were vortexed for ten seconds and incubated at 37�C. Viability was monitored fluorimetrically for three hours.
Results: For both P. gingivalis and P. intermedia, a pyrogallol concentration of 0.2 mM resulted in 80 to 90% cell death; and a hydrogen peroxide concentration of 10 mM killed approximately 80 to 90% of cells. Irrespective of the haem status, no significant difference was determined between the overall maximum rate of killing of P. gingivalis and P. intermedia, in their response to either superoxide or hydrogen peroxide; with the exception that the P. intermedia Haem 0 group was significantly less susceptible to hydrogen peroxide than the P. gingivalis Haem 0 group. For the majority of the experiments, there was no significant difference between final bacterial cell viability in the Haem 0 and Haem 5 cells for both species, after 3 hours exposure to various concentrations of ROS. However, the Haem 50 cells showed a significant increased susceptibility (albeit, a small difference) to both hydrogen peroxide and superoxide.
Conclusions: The Live/Dead bacterial viability stain provided a valuable method to monitor "real-time" killing, avoiding the difficulties associated with culture-based methods for assessing viability. Haem availability had no clear influence on the resistance to ROS of either P. gingivalis or P. intermedia Haem 0 and Haem 5 cells. The Haem 50 cells showed a very slight increase in susceptibility to hydrogen peroxide and superoxide. Although P. intermedia may be isolated in significant numbers from healthy gingivae, as well as from periodontally diseased sites, it was no more resistant to ROS than was P. gingivalis, which is associated with periodontal lesions and difficult to cultivate from relatively healthy (more oxygenated) sites. This suggests that resistance to ROS does not contribute to the ecological distinction between these two species. The finding that haem availability did not influence sensitivity implies that these bacteria do not accumulate haem for the purpose of protection from ROS.
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Quality characteristics of common carp (Cyprinus carpio) Surimi and Kamaboko and the role of Sarcaoplasmic ProteinsJafarpour Khozaghi, Seyed Ali, ali.jafarpour@rmit.edu.au January 2008 (has links)
This study was carried out to determine the characteristics of common carp surimi. In Australia, common carp (Cyprinus carpio) is an environmental pest, strongly coloured (dark-muscle fish), large (2-3 kg), low cost (AUD 2.5/kg) and not highly valued as it is every where else. Surimi could add value to carp, but the colour would have to be modified as surimi manufacturers prefer white coloured flesh. So, firstly the efficiency of Hydrogen peroxide (H2O2; 1-3% v/v) solution at alkaline side of pH (7.0-11.5) on whitening of light fillets of common carp was examined. The whiteness (L*-3b*) of surimi produced from treated (3% H2O2, pH 8.2) common carp light fillets was significantly (p less than 0.05) greater than that of threadfin bream surimi and was not significantly different to that of Alaska pollock. Based on a temperature sweep test, a similar pattern in G of tested surimi was observed which started at ca. 47?C and was completed at ca. 73-74?C. However, thread fin bream kamaboko showed better texture profile characteristics (hardness and gel strength) than that of the other kamaboko tested. To improve the quality of common carp surimi and kamaboko, alternative methods were applied such as modified conventional method (MCM), alkaline-aided method (AAM) and pH modified method (PMM) and the resultant surimi and kamaboko were compared with those produced by the traditional method (TM). In MCM each washing cycle was followed by a centrifugation step for a more effective dewatering and removal of sarcoplasmic proteins (Sp-P). Kamaboko prepared from MCM was whiter and had significantly (p less than 0.05) improved textural characteristics (hardness and gel strength) than that from TM, AAM and PMM. Furthermore, SEM of surimi and kamaboko showed higher number of polygonal structure/mm2 in the gel matrix of MCM kamaboko, as a result of more cross-linking of the myofibrillar proteins, than that recorded for TM, AAM and PMM samples tested. Finally, this study examined the effect of adding common carp sarcoplasmic proteins (Sp-P) on the gel characteristics of threadfin bream surimi and kamaboko. Based on the temperature sweep test, the depths of the valley in the G thermograph of the gels decreased as the concentration of added Sp-P increased from 5% to 35%. Storage modulus (G) of the gels showed greater elasticity in the samples with added Sp-P compared with the control samples without added Sp-P. Furthermore, the breaking force and breaking distance and consequently gel strength of the resultant kamaboko were improved, significantly (p less than 0.05) with added Sp-P. Thus, added Sp-P did not interfere with the gelling of myofibrillar proteins during sol-gel transition phase and was associated with textural quality enhancement for the resultant kamaboko. However, the addition of freeze-dried Sp-P from the dark muscle of the carp decreased the whiteness of the resultant surimi. Furthermore, the gel strength could not be associated with either the number of polygonal structures/mm2 or the area of the polygonal structures.
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Flavins as Biomimetic Catalysts for Sulfoxidation by H<sub>2</sub>O<sub>2</sub> : Catalyst Immobilization in Ionic Liquid for H<sub>2</sub>O<sub>2</sub> OxidationsLindén, Auri January 2005 (has links)
<p>This thesis deals with the development of catalytic oxidation reactions utilizing hydrogen peroxide as terminal oxidant. The main focus has been to find flavin catalysts that are easy to handle and stable to store but still able to perform the desired reaction. A variety of dihydroflavins were prepared and the electrochemical oxidation potentials were measured and compared with their catalytic activity.</p><p>A flavin catalyst was applied in the sulfoxidation of allylic and vinylic sulfides by H<sub>2</sub>O<sub>2</sub>. This transformation was highly chemoselective and the sulfoxides were obtained without formation of other oxidation products. The scope of the reaction was demonstrated by applying the method on substrates with a wide range of functional groups such as a tertiary amine. Another flavin catalyst was immobilized in the ionic liquid [BMIm]PF<sub>6</sub> and used for sulfoxidations by H<sub>2</sub>O<sub>2</sub>. The chemoselectivity was maintained in this system and the catalyst-ionic liquid system could be recycled several times.</p><p>Finally two bimetallic catalyst systems for the dihydroxylation of alkenes by H<sub>2</sub>O<sub>2</sub> were immobilized in the ionic liquid. These systems employed either vanadium acetylacetonate VO(acac)<sub>2 </sub>or methyl trioxorhenium (MTO) as co-catalysts together with the substrate-selective osmium catalyst. Good to excellent yields of the diols were obtained.</p>
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Determination of gp120 <em>&</em> Trx80 dependent production of hydrogen peroxide in cell free <em>&</em> cell-dependent systemsAlam, Sadaf Sakina January 2009 (has links)
<p>Hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>), a reactive oxygen specie (ROS), is most commonly associated with oxidative stress causing cytotoxic effects on living cells. Oxidative stress has been implicated in various conditions including neurodegenerative diseases, autoimmune diseases and cancer. In addition H<sub>2</sub>O<sub>2 </sub>is produced as a defense mechanism against pathogens, as being released by activated phagocytes.<em> </em>In recent years, H<sub>2</sub>O<sub>2</sub> has become established as an important regulator of signal transduction in eukaryotic cells. Hydrogen peroxide is generated both intracellularly and extracellularly in response to various stimuli including cytokines and growth factors. There are different mechanisms by which H<sub>2</sub>O<sub>2</sub> is generated, facilitating signal transduction in cells; through NOX-system in miyochondria, via singlet oxygen, receptor/ligand interaction or by redox active metal ions. The HIV glycoprotein 120 (gp120) is associated with HIV dementia and it is known as a neurotoxin that causes neuronal damage. It has been proposed that free radicals may be involved in the pathogenesis caused by gp120. In addition the truncated form of thioredoxin (Trx80) is known to stimulate HIV replication in HIV infected cells, however, the exact mechanism is not known. A possible way both proteins may mediate their activity is by inducing H<sub>2</sub>O<sub>2</sub> production. The aim of this study was to investigate H<sub>2</sub>O<sub>2</sub> production induced by the proteins gp120 and Trx80. In order to detect H<sub>2</sub>O<sub>2</sub> production an assay based on the fluorescent compound Amplex Red, was established. The assay was used to detect H<sub>2</sub>O<sub>2</sub> released by gp120 and Trx80 in a cell-free environment, in a cell-system and in the presence of metal ions (copper ions) with a physiological reductant (ascorbate). We did not detect H<sub>2</sub>O<sub>2</sub> production induced by gp120 and Trx80 respectively, using our assay, however, other ROS such as hydroxyl radicals may have been generated although they were not detectable with our method. Hence, further studies are needed in order to fully understand how gp120 and Trx80 mediate their activity.</p>
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An in vitro study of post-restorative bleaching : effect on microleakageManni, Hassan January 2010 (has links)
<p>Aim and Objectives: To assess the effect of bleaching on the marginal integrity of Class V composite resin restorations. To determine the effect of a 6% hydrogen peroxide over the counter and a 38% hydrogen peroxide in-office vital bleaching treatment products on the microleakage of Class V composite restorations.</p>
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On the bleachability of alkaline pulps. The influence of residual lignin structure.Wafa Al-Dajani, Waleed January 2001 (has links)
No description available.
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Flavins as Biomimetic Catalysts for Sulfoxidation by H2O2 : Catalyst Immobilization in Ionic Liquid for H2O2 OxidationsLindén, Auri January 2005 (has links)
This thesis deals with the development of catalytic oxidation reactions utilizing hydrogen peroxide as terminal oxidant. The main focus has been to find flavin catalysts that are easy to handle and stable to store but still able to perform the desired reaction. A variety of dihydroflavins were prepared and the electrochemical oxidation potentials were measured and compared with their catalytic activity. A flavin catalyst was applied in the sulfoxidation of allylic and vinylic sulfides by H2O2. This transformation was highly chemoselective and the sulfoxides were obtained without formation of other oxidation products. The scope of the reaction was demonstrated by applying the method on substrates with a wide range of functional groups such as a tertiary amine. Another flavin catalyst was immobilized in the ionic liquid [BMIm]PF6 and used for sulfoxidations by H2O2. The chemoselectivity was maintained in this system and the catalyst-ionic liquid system could be recycled several times. Finally two bimetallic catalyst systems for the dihydroxylation of alkenes by H2O2 were immobilized in the ionic liquid. These systems employed either vanadium acetylacetonate VO(acac)2 or methyl trioxorhenium (MTO) as co-catalysts together with the substrate-selective osmium catalyst. Good to excellent yields of the diols were obtained.
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Photochemical Degradation of ChlorobenzeneSycz, Mateusz 30 April 2013 (has links)
Persistent organic pollutants (POPs) are organic compounds of anthropogenic origin that have been linked to the development of cancer, neurobehavioural impairment, and immune system biochemical alterations. These chemicals have various industrial applications as well as acting as pesticides. Dioxins and furans are some of these compounds that are unintentionally produced in combustion and industrial processes. By definition these compounds have 4 common qualities: they are highly toxic, they are resistant to environmental degradation, they are introduced into the air and water where they travel long distances, and they accumulate in fatty tissues.
Photochemical degradation is a method that has been extensively researched in the last few decades. In the aqueous phase it has already been shown to be able to degrade a number of refractory organics, such as dioxins and furans. The ultimate products of this process tend to be carbon dioxide, water, and mineral anions. Air phase work has been also gaining attention in recent decades as a possible alternative to incineration methods in air pollution control. The advantages of photochemical degradation processes are that they can be initiated at low temperatures, are relatively low cost compared to incineration processes, environmentally benign, and have the potential for quick and complete degradation of organic compounds.
The main aim of the research is to investigate the photochemical degradation potential of PCDD/ PCDFs in gaseous air streams as a potential air pollution control technology. In order to do this, the photodegradation reaction kinetics were determined for chlorobenzene as a suitable surrogate for PCDD/PCDFs. Three different photodegradation schemes were employed: direct photolysis, UV/O3, and UV/H2O2. In addition, ozonolysis reaction rates were also determined to evaluate the effects of on the overall photodegradation rates for the UV/O3 process. Factors such as humidity levels and temperature were investigated to determine their effects on degradation rates.
Temperature and humidity were not greatly influential on the degradation rates of direct photolysis. The degradation rate of chlorobenzene at a temperature of 100°C and high humidity was noticeably reduced, but unchanged at the 10% RH and 60% RH levels for all temperatures.
Ozonolysis of chlorobenzene was negligible at 30°C for all humidity levels. Ozonolysis reactions at the 60°C and 100°C levels were higher than direct photolysis rates and in the 100°C case exceeded the UV/O3 degradation rates.
Ozone coupled with UV experiments proved to be the most destructive at the low temperature of 30°C and molar ratio of 10:1 ozone to chlorobenzene. There was a clear and positive relationship between the amount of ozone present in the reactor and the degradation rate. At lower ozone to chlorobenzene molar ratios the degradation rates were not much higher than those for direct photolysis of ozone. The 5:1 molar ratio saw a significant increase in degradation rates over the photolysis rates. The fastest degradation rate was achieved for the 10:1 molar ratio and high humidity, which was over 10 times the rate of direct photolysis. In addition, humidity had a noticeably significant positive effect in these reactions.
The effect of temperature on the UV/ozone reaction scheme was determined for the 5:1 ozone to chlorobenzene ratio. Temperature had an interesting effect on the degradation rates at higher temperatures. As the reactor temperature increased, the degradation rates from ozonolysis and UV/O3 began to converge at 60°C, ultimately leading to the ozonolysis reaction being faster than the UV/O3.
Exploratory experiments for the H2O2 scheme were performed. H2O2 had a positive influence on the degradation rate of chlorobenzene and was about 26% higher than the direct photolysis rates. However for similar conditions, the UV/O3 process had higher degradation rates as was expected from the difference in absorption values between ozone and hydrogen peroxide.
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Reactions of aqueous radiolysis products with oxide surfaces : An experimental and DFT studyLousada Patrício, Cláudio Miguel January 2013 (has links)
The reactions between aqueous radiolysis products and oxide surfaces are important in nuclear technology in many ways. In solid-liquid systems, they affect (and at the same time are dependent on) both the solution chemistry and the stability of materials under the influence of ionizing radiation. The stability of surface oxides is a factor that determines the longevity of the materials where such oxides are formed. Additionally, the aqueous radiolysis products are responsible for corrosion and erosion of the materials. In this study, the reactions between radiolysis products of water – mainly H2O2 and HO radicals – with metal, lanthanide and actinide oxides are investigated. For this, experimental and computational chemistry methods are employed. For the experimental study of these systems it was necessary to implement new methodologies especially for the study of the reactive species – the HO radicals. Similarly, the computational study also required the development of models and benchmarking of methods. The experiments combined with the computational chemistry studies produced valuable kinetic, energetic and mechanistic data. It is demonstrated here that the HO radicals are a primary product of the decomposition of H2O2. For all the materials, the catalytic decomposition of H2O2 consists first of molecular adsorption onto the surfaces of the oxides. This step is followed by the cleavage of the O-O bond in H2O2 to form HO radicals. The HO radicals are able to react further with the hydroxylated surfaces of the oxides to form water and a surface bound HO• center. The dynamics of formation of HO• vary widely for the different materials studied. These differences are also observed in the activation energies and kinetics for decomposition of H2O2. It is found further that the removal of HO• from the system where H2O2 undergoes decomposition, by means of a scavenger, leads to the spontaneous formation of H2. The combined theoretical-experimental methodology led to mechanistic understanding of the reactivity of the oxide materials towards H2O2 and HO radicals. This reactivity can be expressed in terms of fundamental properties of the cations present in the oxides. Correlations were found between several properties of the metal cations present in the oxides and adsorption energies of H2O, adsorption energies of HO radicals and energy barriers for H2O2 decomposition. This knowledge can aid in improving materials and processes important for nuclear technological systems, catalysis, and energy storage, and also help to better understand geochemical processes. / <p>QC 20130322</p>
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P38(MAPK) negatively regulates monoamine oxidase-A activity as well as its sensitivity to Ca2+Cao, Xia 04 January 2008
Monoamine oxidase (MAO) is a mitochondrial deaminating enzyme that exists as two isoforms, MAO-A and -B. The MAO-mediated reaction generates hydrogen peroxide (H2O2) as a normal by-product. Dysregulation of MAO has been implicated in a variety of neuropsychiatric and neurodegenerative disorders, as well as in the aging process. Endogenous regulators of MAO-A function include calcium (Ca2+) and the p38 mitogen-activated protein kinase (MAPK). Although the effect of p38(MAPK) is thought to rely on induction of mao-A gene expression, post-translational modification of the MAO-A protein is also possible. <p>Using standard biochemical approaches in combination with pharmacological interventions and recombinant DNA strategies, specific aspartic acid residues (within putative Ca2+-binding motifs) were demonstrated to contribute to MAO-A activity. Furthermore, MAO-A activity and its sensitivity to Ca2+ was negatively regulated by the p38(MAPK), which is usually activated during cell stress. The effect of p38(MAPK) on MAO-A function relies specifically on Serine209 in MAO-A, which resides in a p38(MAPK) consensus motif. The serine phosphorylation status of MAO-A determines its capacity for generating peroxy radicals and its toxicity in established cell lines (e.g. C6, N2a, HEK293A, HT-22) and in primary cortical neurons. p38(MAPK)-regulated MAO-A activity is also linked to neurotoxicity associated with the Alzheimer disease-related peptide, Ò-amyloid (AÒ). These data suggest a unique neuroprotective role for p38(MAPK) centered on a negative feedback regulation of the Ca2+-sensitive, H2O2-generating enzyme MAO-A.
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