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Combining Mbr And Nf/Ro Membrane Filtration For The Removal Of Trace Organics In Indirect Potable Water Reuse Applications, William Price, Nichanan Tadkaew, Long Nghiem, Stuart Khan, Abdulhakeem Alturki, James Mcdonald Dec 2012

Combining Mbr And Nf/Ro Membrane Filtration For The Removal Of Trace Organics In Indirect Potable Water Reuse Applications, William Price, Nichanan Tadkaew, Long Nghiem, Stuart Khan, Abdulhakeem Alturki, James Mcdonald

Long D Nghiem

The aim of this study was to demonstrate the complementarities of combining membrane bioreactor (MBR) treatment with nanofiltration (NF) or reverse osmosis (RO) membrane filtration for the removal of trace organic contaminants for potential indirect potable water recycling applications. Four commercially available NF/RO membranes, namely NF270, NF90, BW30 and ESPA2, were selected for this investigation. Challenge tests were conducted with 40 trace organic compounds at concentrations of approximately 2 ng/L in initial wastewater solutions using a laboratory scaleMBRsystem and a cross-flow NF/RO rig. The results suggest that the MBR system effectively removes hydrophobic and biodegradable trace organic compounds. The adsorption …


The Two-Faced Nature Of Small Heat Shock Proteins: Amyloid Assembly And The Inhibition Of Fibril Formation. Relevance To Disease States, Heath W. Ecroyd, S Meehan, John A. Carver Dec 2012

The Two-Faced Nature Of Small Heat Shock Proteins: Amyloid Assembly And The Inhibition Of Fibril Formation. Relevance To Disease States, Heath W. Ecroyd, S Meehan, John A. Carver

Heath Ecroyd

The ability of small heat-shock proteins (sHsps) such as alphaB-crystallin to inhibit the amorphous (disordered) aggregation of varied target proteins in a chaperone-like manner has been well described. The mechanistic details of this action are not understood. Amyloid fibril formation is an alternative off-folding pathway that leads to highly ordered beta-sheet-containing aggregates. Amyloid fibril formation is associated with a broad range of protein conformational diseases such as Alzhiemer's, Parkinson's and Huntington's and sHsp expression is elevated in the protein deposits that are characteristic of these disease states. The ability of sHsps to prevent fibril formation has been less well characterised. …


Molecular Dynamics Analysis Of Apolipoprotein-D - Lipid Hydroperoxide Interactions: Mechanism For Selective Oxidation Of Met-93, Aaron J. Oakley, Surabhi Bhatia, Heath Ecroyd, Brett Garner Dec 2012

Molecular Dynamics Analysis Of Apolipoprotein-D - Lipid Hydroperoxide Interactions: Mechanism For Selective Oxidation Of Met-93, Aaron J. Oakley, Surabhi Bhatia, Heath Ecroyd, Brett Garner

Heath Ecroyd

Background: Recent studies suggest reduction of radical-propagating fatty acid hydroperoxides to inert hydroxides by interaction with apolipoprotein-D (apoD) Met93 may represent an antioxidant function for apoD. The nature and structural consequences of this selective interaction are unknown. Methodology/Principal Findings: Herein we used molecular dynamics (MD) analysis to address these issues. Longtimescale simulations of apoD suggest lipid molecules are bound flexibly, with the molecules free to explore multiple conformations in a binding site at the entrance to the classical lipocalin ligand-binding pocket. Models of 5s- 12s- and 15s hydroperoxyeicosatetraenoic acids were created and the lipids found to wrap around Met93 thus …


Binding Of The Molecular Chaperone Alphab-Crystallin To Abeta Amyloid Fibrils Inhibits Fibril Elongation, Sarah L. Shammas, Christopher A. Waudby, Shuyu Wang, Alexander K. Buell, Tuomas P. Knowles, Heath W. Ecroyd, Mark E. Welland, John A. Carver, Christopher M. Dobson, Sarah Meehan Dec 2012

Binding Of The Molecular Chaperone Alphab-Crystallin To Abeta Amyloid Fibrils Inhibits Fibril Elongation, Sarah L. Shammas, Christopher A. Waudby, Shuyu Wang, Alexander K. Buell, Tuomas P. Knowles, Heath W. Ecroyd, Mark E. Welland, John A. Carver, Christopher M. Dobson, Sarah Meehan

Heath Ecroyd

The molecular chaperone αB-crystallin is a small heat-shock protein that is upregulated in response to a multitude of stress stimuli, and is found colocalized with Aβ amyloid fibrils in the extracellular plaques that are characteristic of Alzheimer's disease. We investigated whether this archetypical small heat-shock protein has the ability to interact with Aβ fibrils in vitro. We find that αB-crystallin binds to wild-type Aβ42 fibrils with micromolar affinity, and also binds to fibrils formed from the E22G Arctic mutation of Aβ42. Immunoelectron microscopy confirms that binding occurs along the entire length and ends of the fibrils. Investigations into the effect …


Nmr Spectroscopy Of 14-3-3zeta Reveals A Flexible C-Terminal Extension: Differentiation Of The Chaperone And Phosphoserine-Binding Activities Of 14-3-3zeta, H Fu, Danielle Williams, Heath Ecroyd, John Carver, Lixin Zhang, Huanqin Dai, Joanna Woodcock, K Goodwin Dec 2012

Nmr Spectroscopy Of 14-3-3zeta Reveals A Flexible C-Terminal Extension: Differentiation Of The Chaperone And Phosphoserine-Binding Activities Of 14-3-3zeta, H Fu, Danielle Williams, Heath Ecroyd, John Carver, Lixin Zhang, Huanqin Dai, Joanna Woodcock, K Goodwin

Heath Ecroyd

Intracellular 14-3-3 proteins bind to many proteins, via a specific phosphoserine motif, regulating diverse cellular tasks including cell signalling and disease progression. The 14-3-3 isoform is a molecular chaperone, preventing the stressinduced aggregation of target proteins in a manner comparable with that of the unrelated sHsps (small heat-shock proteins). 1H-NMR spectroscopy revealed the presence of a flexible and unstructured C-terminal extension, 12 amino acids in length, which protrudes from the domain core of 14-3-3 and is similar in structure and length to the C-terminal extension of mammalian sHsps. The extension stabilizes 14-3-3, but has no direct role in chaperone action. …


Enhanced Molecular Chaperone Activity Of The Small Heat-Shock Protein Alphab-Cystallin Following Covalent Immobilization Onto A Solid-Phase Support, V Bellotti, Heath Ecroyd, J Carver, H J Griesser, B Thierry, J G Shapter, S S Griesser, S Giorgetti, M R Nussio, J A Gerrard, J Garvey Dec 2012

Enhanced Molecular Chaperone Activity Of The Small Heat-Shock Protein Alphab-Cystallin Following Covalent Immobilization Onto A Solid-Phase Support, V Bellotti, Heath Ecroyd, J Carver, H J Griesser, B Thierry, J G Shapter, S S Griesser, S Giorgetti, M R Nussio, J A Gerrard, J Garvey

Heath Ecroyd

The well-characterized small heat-shock protein, alphaB-crystallin, acts as a molecular chaperone by interacting with unfolding proteins to prevent their aggregation and precipitation. Structural perturbation (e.g., partial unfolding) enhances the in vitro chaperone activity of alphaB-crystallin. Proteins often undergo structural perturbations at the surface of a synthetic material, which may alter their biological activity. This study investigated the activity of alphaB-crystallin when covalently bound to a support surface; alphaB-crystallin was immobilized onto a range of solid material surfaces, and its characteristics and chaperone activity were assessed. Immobilization was achieved via a plasma-deposited thin polymeric interlayer containing aldehyde surface groups and reductive …


Identification Of Phospholipids In Human Meibum By Nano-Electrospray Ionisation Tandem Mass Spectrometry, Jennifer T. Saville, Zhenjun Zhao, Mark Dp Willcox, Manjula A. Ariyavidana, Stephen J. Blanksby, Todd W. Mitchell Oct 2012

Identification Of Phospholipids In Human Meibum By Nano-Electrospray Ionisation Tandem Mass Spectrometry, Jennifer T. Saville, Zhenjun Zhao, Mark Dp Willcox, Manjula A. Ariyavidana, Stephen J. Blanksby, Todd W. Mitchell

Stephen Blanksby

Meibum is believed to be the major source of tear !lm lipids, which are vital in the prevention of excessevaporation of the aqueous phase. The complete lipid composition of meibum has yet to be established.While earlier studies reported the presence of phospholipids in human meibum, recent mass spectrometricstudies have not detected them. In this study we use electrospray ionisation tandem massspectrometry to investigate the presence of phospholipids in meibum and provide comparison to thephospholipid pro!le of tears.Lipids were extracted from human meibum and tear samples using standard biphasic methods andanalysed by nano-electrospray ionisation tandem mass spectrometry using targeted ion scans. …


Ozone Induced Dissociation: Elucidation Of Double Bond Position Within Mass-Selected Lipid Ions, Michael C. Thomas, Todd W. Mitchell, David G. Harman, Jane M. Deeley, Jessica R. Nealon, Stephen J. Blanksby Oct 2012

Ozone Induced Dissociation: Elucidation Of Double Bond Position Within Mass-Selected Lipid Ions, Michael C. Thomas, Todd W. Mitchell, David G. Harman, Jane M. Deeley, Jessica R. Nealon, Stephen J. Blanksby

Stephen Blanksby

Ions formed from lipids during electrospray ionization of crude lipid extracts have been mass-selected within a quadrupole linear ion trap mass spectrometer and allowed to react with ozone vapor. Gas-phase ion−molecule reactions between unsaturated lipid ions and ozone are found to yield two primary product ions for each carbon−carbon double bond within the molecule. The mass-to-charge ratios of these chemically induced fragments are diagnostic of the position of unsaturation within the precursor ion. This novel analytical technique, dubbed ozone-induced dissociation (OzID), can be applied both in series and in parallel with conventional collision-induced dissociation (CID) to provide near-complete structural assignment …


Understanding The Α-Crystallin Cell Membrane Conjunction, Shih-Ping Su, Jason D. Mcarthur, Michael G. Friedrich, Roger J. W Truscott, J. A. Aquilina Oct 2012

Understanding The Α-Crystallin Cell Membrane Conjunction, Shih-Ping Su, Jason D. Mcarthur, Michael G. Friedrich, Roger J. W Truscott, J. A. Aquilina

J. A. Aquilina

PURPOSE. It is well established that levels of soluble α-crystallin in the lens cytoplasm fall steadily with age, accompanied by a corresponding increase in the amount of membrane-bound α-crystallin. Less well understood, is the mechanism driving this age-dependent membrane association. The aim of this study was to investigate the role of the membrane and its associated proteins and peptides in the binding of α-crystallin. METHODS. Fibre cell membranes from human and bovine lenses were separated from soluble proteins by centrifugation. Membranes were stripped of associated proteins with successive aqueous, urea and alkaline solutions. Protein constituents of the respective membrane isolates …


Tandem Mass Spectrometry Reveals The Quaternary Organization Of Macromolecular Assemblies, J L Benesch, Andrew Aquilina, Brandon T. Ruotolo, Frank Sobott, C V Robinson Oct 2012

Tandem Mass Spectrometry Reveals The Quaternary Organization Of Macromolecular Assemblies, J L Benesch, Andrew Aquilina, Brandon T. Ruotolo, Frank Sobott, C V Robinson

J. A. Aquilina

The application of mass spectrometry (MS) to the study of progressively larger and more complex macromolecular assemblies is proving increasingly useful for structural biologists. The scope of this approach has recently been widened through the application of a tandem MS procedure. This two-step technique involves the selection of specific assemblies in the gas phase, and inducing their dissociation through collisions with argon atoms. Here we investigate the mechanism of this process and show that dissociation of subunits from a macromolecular assembly follows a sequential pathway, with the partitioning of charge between the dissociation products governed primarily by their relative surface …


3-Hydroxykynurenine Oxidizes Alpha-Crystallin: Potential Role In Cataractogenesis, Roger Truscott, Andrew Aquilina, Peter Hains, Anastasia Korlimbinis Oct 2012

3-Hydroxykynurenine Oxidizes Alpha-Crystallin: Potential Role In Cataractogenesis, Roger Truscott, Andrew Aquilina, Peter Hains, Anastasia Korlimbinis

J. A. Aquilina

No abstract provided.


Truncation, Cross-Linking And Interaction Of Crystallins And Intermediate Filament Proteins In The Aging Human Lens, Roger Truscott, Jason Mcarthur, Andrew Aquilina, Shi-Ping (Jim) Su Oct 2012

Truncation, Cross-Linking And Interaction Of Crystallins And Intermediate Filament Proteins In The Aging Human Lens, Roger Truscott, Jason Mcarthur, Andrew Aquilina, Shi-Ping (Jim) Su

J. A. Aquilina

The optical properties of the lens are dependent upon the integrity of proteins within the fiber cells. During aging, crystallins, the major intra-cellular structural proteins of the lens, aggregate and become water-insoluble. Modifications to crystallins and the lens intermediate filaments have been implicated in this phenomenon. In this study, we examined changes to, and interactions between, human lens crystallins and intermediate filament proteins in lenses from a variety of age groups (0-86 years). Among the lens-specific intermediate filament proteins, filensin was extensively cleaved in all postnatal lenses, with truncated products of various sizes being found in both the lens cortical …


Chemical Cross-Linking Of The Chloroplast Localized Small Heat-Shock Protein, Hsp21, And The Model Substrate Citrate Synthase, Emma Ahrman, W Lambert, Andrew Aquilina, C V Robinson, Cs Emanuelsson Oct 2012

Chemical Cross-Linking Of The Chloroplast Localized Small Heat-Shock Protein, Hsp21, And The Model Substrate Citrate Synthase, Emma Ahrman, W Lambert, Andrew Aquilina, C V Robinson, Cs Emanuelsson

J. A. Aquilina

The molecular mechanism whereby the small heat-shock protein (sHsp) chaperones interact with and prevent aggregation of other proteins is not fully understood. We have characterized the sHsp-substrate protein interaction at normal and increased temperatures utilizing a model substrate protein, citrate synthase (CS), widely used in chaperone assays, and a dodecameric plant sHsp, Hsp21, by chemical cross-linking with 3,3'-Dithiobis[sulfosuccinimidylpropionate] (DTSSP) and mass spectrometric peptide mapping. In the absence of CS, the cross-linker captured Hsp21 in dodecameric form, even at increased temperature (47 degrees C). In the presence of equimolar amounts of CS, no Hsp21 dodecamer was captured, indicating a substrate-induced Hsp21 …


Effects Of Glycosylation On The Structure And Function Of The Extracellular Chaperone Clusterin, Elise Stewart, Andrew Aquilina, Simon B Easterbrook-Smith, D Murphy-Durland, C Jacobsen, S Moestrup, Mark Wilson Oct 2012

Effects Of Glycosylation On The Structure And Function Of The Extracellular Chaperone Clusterin, Elise Stewart, Andrew Aquilina, Simon B Easterbrook-Smith, D Murphy-Durland, C Jacobsen, S Moestrup, Mark Wilson

J. A. Aquilina

Clusterin is the first well characterized, constitutively secreted extracellular chaperone that binds to exposed regions of hydrophobicity on non-native proteins. It may help control the folding state of extracellular proteins by targeting them for receptor-mediated endocytosis and intracellular lysosomal degradation. A notable feature of secreted clusterin is its heavy glycosylation. Although carbohydrate comprises approximately 20−25% of the total mass of the mature molecule, its function is unknown. Results from the current study demonstrate that deglycosylation of human serum clusterin had little effect on its overall secondary structure content but produced a small increase in solvent-exposed hydrophobicity and enhanced the propensity …


Protein-Bound And Free Uv Filters In Cataract Lenses. The Concentration Of Uv Filters Is Much Lower Than In Normal Lenses, Roger Truscott, Andrew Aquilina, Anastasia Korlimbinis Oct 2012

Protein-Bound And Free Uv Filters In Cataract Lenses. The Concentration Of Uv Filters Is Much Lower Than In Normal Lenses, Roger Truscott, Andrew Aquilina, Anastasia Korlimbinis

J. A. Aquilina

In human cataract lenses the UV filters, 3-hydroxykynurenine glucoside (3OHKG) and kynurenine (Kyn) were found to be covalently bound to proteins and the levels in the nucleus were much higher than in the cortex. The levels of the bound UV filters in cataract nuclei were much lower than those in age-matched normal lenses. 3-Hydroxykynurenine could not be detected in cataract lenses. As with normal lenses, protein-bound 3OHKG in cataract lenses was found at the highest levels followed by Kyn. Free UV filter concentrations were also markedly reduced in cataract lenses. This feature may well contribute to the lower protein-bound levels; …


The Major Toxin From The Australian Common Brown Snake Is A Hexamer With Unusual Gas-Phase Dissociation Properties, Andrew Aquilina Oct 2012

The Major Toxin From The Australian Common Brown Snake Is A Hexamer With Unusual Gas-Phase Dissociation Properties, Andrew Aquilina

J. A. Aquilina

Asymmetric dissociation of multiply charged proteins assemblies has been frequently reported. This phenomenon, which relies on the dissociation of one or more highly charged monomers, has been shown to provide insights into the structure and organization of large monodisperse and polydisperse assemblies. Here, the process of asymmetric dissociation is investigated using the multi-subunit protein, textilotoxin, which has unusually high structural constraints on its monomers due to multiple disulfide linkages. Initially, it is shown that, contrary to previous reports, textilotoxin is made up of 6, rather than 5 subunits. Furthermore, the hexamer exists as two isoforms, one of which is substantially …


Protein-Bound Uv Filters In Normal Human Lenses: The Concentration Of Bound Uv Filters Equals That Of Free Uv Filters In The Centre Of Older Lenses, Roger Truscott, Andrew Aquilina, Anastasia Korlimbinis Oct 2012

Protein-Bound Uv Filters In Normal Human Lenses: The Concentration Of Bound Uv Filters Equals That Of Free Uv Filters In The Centre Of Older Lenses, Roger Truscott, Andrew Aquilina, Anastasia Korlimbinis

J. A. Aquilina

No abstract provided.


Air-Drying Of Banana: Influence Of Experimental Parameters, Slab Thickness, Banana Maturity And Harvesting Season, Minh-Hue Nguyen, William E. Price Oct 2012

Air-Drying Of Banana: Influence Of Experimental Parameters, Slab Thickness, Banana Maturity And Harvesting Season, Minh-Hue Nguyen, William E. Price

William E. Price

Air-drying of banana slabs has been investigated and the influence of experimental parameters such as temperature, relative humidity and slab thickness has been studied. This was in part re-investigated because of inconsistencies in previous studies, particularly in relation to derived water diffusion coefficients. In addition, it is shown that harvest season and hence initial moisture content has a very marked influence on the drying kinetics. By contrast banana maturity (ripeness) has little influence on the kinetics despite there being significant differences in morphology and chemical composition between green and ripe bananas. The effect of these two variables on the drying …


Increased Probiotic Yogurt Or Resistant Starch Intake Does Not Affect Isoflavone Bioavailability In Subjects Consuming A High Soy Diet, Theresa A. Larkin, William E. Price, Lee B. Astheimer Oct 2012

Increased Probiotic Yogurt Or Resistant Starch Intake Does Not Affect Isoflavone Bioavailability In Subjects Consuming A High Soy Diet, Theresa A. Larkin, William E. Price, Lee B. Astheimer

William E. Price

Objective: Probiotics and prebiotics that affect gut microflora balance and its associated enzymeactivity may contribute to interindividual variation in isoflavone absorption after soy intake, possiblyenhancing isoflavone bioavailability. This study examined the effects of the consumption ofbioactive yogurt (a probiotic) or resistant starch (a known prebiotic) in combination with high soyintake on soy isoflavone bioavailability.Methods: Using a crossover design, chronic soy consumption was compared with soy plusprobiotic yogurt or resistant starch in older male and postmenopausal females (n 31). Isoflavonebioavailability was assessed at the beginning and end of each 5-wk dietary period by samplingplasma and urine after a standardized soy meal.Results: …


Fractionation Of Sedimentary Arsenic From Port Kembla Harbour, Nsw, Australia, Glennys A. O'Brien, William E. Price, Bryan E. Chenhall, Muhammad Damris Oct 2012

Fractionation Of Sedimentary Arsenic From Port Kembla Harbour, Nsw, Australia, Glennys A. O'Brien, William E. Price, Bryan E. Chenhall, Muhammad Damris

William E. Price

The binding of arsenic in sediments of the heavily industrialised Port Kembla Harbour, NSW, Australia, has been investigated. Both dredge and core samples have been used to develop a sieving/sequential extraction (SE) procedure. Dredge samples included oxic surficial and deeper anoxic sediment. The main core sample analysed was 18 cm deep, sliced at 2 cm intervals. Sediment was sieved to three size ranges (250 microm) and each of these was then subjected to a four step SE, sequentially solubilizing arsenic as ion exchangeable, 1 M HCl soluble, NH(2)OH.HCl soluble, and strong oxidising acid soluble. Concentrations of 50-500 mg As kg(-1) …


A Fragmentation Study Of Isoflavones In Negative Electrospray Ionization By Msn Ion Trap Mass Spectrometry And Triple Quadrupole Mass Spectrometry, Jinguo Kang, Larry A. Hick, William E. Price Oct 2012

A Fragmentation Study Of Isoflavones In Negative Electrospray Ionization By Msn Ion Trap Mass Spectrometry And Triple Quadrupole Mass Spectrometry, Jinguo Kang, Larry A. Hick, William E. Price

William E. Price

This study has elucidated the fragmentation pathway for deprotonated isoflavones in electrospray ionization using MSn ion trap mass spectrometry and triple quadrupole mass spectrometry. Genistein-d4 and daidzein-d3 were used as references for the clarification of fragment structures. To confirm the relationship between precursor and product ions, some fragments were traced from MS2 to MS5. The previous literature for the structurally related flavones and flavanones located the loss of ketene (C2H2O) to ring C, whereas the present fragmentation study for isoflavones has shown that the loss of ketene occurs elsewhere at ring A. In the further fragmentation of the [M-H-CH3]•- radical …


Effect Of Organic Solvents On The Separation Of Benzoic Acids By Capillary Electrophoresis, Young J. Lee, William E. Price, Margaret Sheil Oct 2012

Effect Of Organic Solvents On The Separation Of Benzoic Acids By Capillary Electrophoresis, Young J. Lee, William E. Price, Margaret Sheil

William E. Price

The effect of organic modifiers on the separation of a number of closely related isomeric benzoic acids by capillary electrophoresis is described. It is shown that while a single modifier concentration cannot help resolve the entire electropherogram, organic modifiers do significantly enhance the resolution of parts of the separation system by comparison with 40 mmol l-1 phosphate buffer. The effects on separation and retention times are discussed in terms of the effects on electroosmotic flow and the electrophoretic mobilities of the charged solutes. The effects were found to be modifier specific, although the trends were in the same direction (ie., …


Occurrence Of Phytoestrogens In Municipal Wastewater And Surface Waters, Jinguo Kang, William E. Price Oct 2012

Occurrence Of Phytoestrogens In Municipal Wastewater And Surface Waters, Jinguo Kang, William E. Price

William E. Price

Phytoestrogens (isoflavones, enterolignans and coumestrol) in wastewater samples and surface water samples have been analysed by LC-ESI-MSn. In wastewater samples, high levels of enterolactone (581 – 2111 ng/L), daidzein (341 – 1688 ng/L) and enterodiol (60 – 834 ng/L) were detected in raw sewage, but the vast majority of the analysed phytoestrogens were removed effectively in treatment process. The removal rates of the analysed phytoestrogens in the two advanced tertiary treatment plants were > 99%; a case study in one of the treatment plants showed that most of the residual phytoestrogens were removed by biological treatment using activated sludge. In surface …


Corrigendum To ‘‘The Chaperone Action Of Bovine Milk As1- And As2-Caseins And Their Associated Form As-Casein’’ [Arch. Biochem. Biophys. 510 (2011) 42–52], Teresa M. Treweek, David C. Thorn, William E. Price, John A. Carver Oct 2012

Corrigendum To ‘‘The Chaperone Action Of Bovine Milk As1- And As2-Caseins And Their Associated Form As-Casein’’ [Arch. Biochem. Biophys. 510 (2011) 42–52], Teresa M. Treweek, David C. Thorn, William E. Price, John A. Carver

William E. Price

No abstract provided.


The Key Importance Of Soy Isoflavone Bioavailability To Understanding Health Benefits, T. A. Larkin, William E. Price, L. Astheimer Oct 2012

The Key Importance Of Soy Isoflavone Bioavailability To Understanding Health Benefits, T. A. Larkin, William E. Price, L. Astheimer

William E. Price

Research over the past two decades has provided significant epidemiological and other evidence for the health benefits of the consumption of soy-based foods. A large number of dietary intervention studies have examined the effects of soy isoflavones on risk factors for cardiovascular disease and hormone-dependent cancers. However, these report large variability in outcome measures, very limited reproducibility between studies and in some cases, controversy between results of clinical trials using dietary soy or soy protein and isoflavone supplementation. This highlights a major gap in our understanding of soy isoflavone uptake, metabolism, distribution, and overall bioavailability. There are many potential factors …


Using Calibration Approaches To Compensate For Remaining Matrix Effects In Quantitative Liquid Chromatography Electrospray Ionization Multistage Mass Spectrometric Analysis Of Phytoestrogens In Aqueous Environmental Samples, Jinguo Kang, Larry A. Hick, William E. Price Oct 2012

Using Calibration Approaches To Compensate For Remaining Matrix Effects In Quantitative Liquid Chromatography Electrospray Ionization Multistage Mass Spectrometric Analysis Of Phytoestrogens In Aqueous Environmental Samples, Jinguo Kang, Larry A. Hick, William E. Price

William E. Price

Signal suppression is a common problem in quantitative LC-ESI-MSn analysis in environment samples, especially in highly loaded wastewater samples with highly complex matrix. Optimization of sample preparation and improvement of chromatographic separation are prerequisite to improve reproducibility and selectivity. Matrix components may be reduced if not eliminated by a series of sample preparation steps. However, extensive sample preparation can be time-consuming and risk the significant loss of some trace analytes. The best way to further compensate matrix effects is the use of internal standard for each analyte. However, in a multi-component analysis, finding appropriate internal standards for every analyte is …


Localization Of Low Molecular Weight Crystallin Peptides In The Aging Human Lens Using A Maldi Mass Spectrometry Imaging Approach, S. P. Su, Jason D. Mcarthur, J. A. Aquilina Jul 2012

Localization Of Low Molecular Weight Crystallin Peptides In The Aging Human Lens Using A Maldi Mass Spectrometry Imaging Approach, S. P. Su, Jason D. Mcarthur, J. A. Aquilina

J. A. Aquilina

Low molecular weight (LMW) peptides, derived from the breakdown of the major eye lens proteins, the crystallins, accumulate in the human lens with age. These LMW peptides are associated with age-related lens opacity and cataract, with some shown to inhibit the chaperone activity of α-crystallin. However, the mechanism(s) giving rise to the production of these peptides, as well as their distribution within the lens, are not well understood. In this study, we have mapped the distribution of these crystallin-derived peptides present in human lenses of different ages using matrix-assisted laser desorption/ionization-imaging mass spectrometry (MALDI-IMS). Our data showed that most of …


Defining The Structural Basis Of Human Plasminogen Binding By Streptococcal Surface Enolase, Amanda J. Cork, Slobodan Jergic, Sven Hammerschmidt, Bostjan Kobe, Vijay Pancholi, Justin L.P. Benesch, Carol V, Robinson, Nicholas E. Dixon, J Andrew Aquilina, Mark J. Walker Jul 2012

Defining The Structural Basis Of Human Plasminogen Binding By Streptococcal Surface Enolase, Amanda J. Cork, Slobodan Jergic, Sven Hammerschmidt, Bostjan Kobe, Vijay Pancholi, Justin L.P. Benesch, Carol V, Robinson, Nicholas E. Dixon, J Andrew Aquilina, Mark J. Walker

J. A. Aquilina

The flesh-eating bacterium group A Streptococcus (GAS) binds and activates human plasminogen, promoting invasive disease. Streptococcal surface enolase (SEN), a glycolytic pathway enzyme, is an identified plasminogen receptor of GAS. Here we used mass spectrometry (MS) to confirm that GAS SEN is octameric, thereby validating in silico modeling based on the crystal structure of S. pneumoniae -enolase. Site-directed mutagenesis of surface-located lysine residues (SENK252+255A, SENK304A, SENK334A, SENK344E, SENK435L and SEN434-435) was used to examine their roles in maintaining structural integrity, enzymatic function and plasminogen binding. Structural integrity of the GAS SEN octamer was retained for all mutants except SENK344E, as …


Mimicking Phosphorylation Of Alphab-Crystallin Affects Its Chaperone Activity, Heath W. Ecroyd, Sarah Meehan, J Horwitz, Andrew Aquilina, J L Benesch, C V Robinson, Cait Macphee, John Carver Jul 2012

Mimicking Phosphorylation Of Alphab-Crystallin Affects Its Chaperone Activity, Heath W. Ecroyd, Sarah Meehan, J Horwitz, Andrew Aquilina, J L Benesch, C V Robinson, Cait Macphee, John Carver

J. A. Aquilina

No abstract provided.


The Role Of Streptokinase As A Virulence Determinant Of Streptococcus Pyogenes - Potential For Therapeutic Targeting, Jason D. Mcarthur, Simon M. Cook, Carola Venturini, Mark J. Walker Mar 2012

The Role Of Streptokinase As A Virulence Determinant Of Streptococcus Pyogenes - Potential For Therapeutic Targeting, Jason D. Mcarthur, Simon M. Cook, Carola Venturini, Mark J. Walker

Faculty of Science - Papers (Archive)

Streptococcus pyogenes is a major human pathogen responsible for numerous diseases ranging from uncomplicated skin and throat infections to severe, life threatening invasive disease such as necrotising fasciitis and streptococcal toxic shock syndrome. These severe invasive infections progress rapidly and produce high rates of morbidity and mortality despite the implementation of aggressive treatment plans. The activation of plasminogen and the acquisition of plasmin activity at the bacterial cell surface is critical for the invasive pathogenesis of this organism. To facilitate this process, S. pyogenes secrete streptokinase, a potent plasminogen activating protein. Here, we describe the role of streptokinase in invasive …