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Current Search:
'Biochemistry' in all fields;
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Displaying Results 276 - 300 of 301 on page 12 of 13
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Protein glycosylation in the gram-negative gamma proteobacterium photorhabdus luminescens
(2011)
Fox, Mary Elizabeth
Protein glycosylation in the gram-negative gamma proteobacterium photorhabdus luminescens
(2011)
Fox, Mary Elizabeth
Abstract:
The objective of this research was to investigate the possibility that Photorhabdus luminescens produces glycoproteins and thus contains a protein glycosylation system. P. luminescens is a pathogen of insects and a symbiont of soil nematodes. Adhesion and invasion are very important in the life cycle of the organism and it is speculated that the bacteria may produce glycoproteins to facilitate infection of the host. Proteins from P. luminescens were analysed using lectins for the presence of glycoproteins. Many potential glycoproteins were isolated using lectin affinity chromatography (LAC) and one such protein was identified conclusively using mass spectrometry to be an outer membrane porin, OmpN. No glycoproteins have previously been identified in P. luminescens. The ompN gene was cloned and expressed in both Escherichia coli and P. luminescens. It was found that OmpN purified from P. luminescens was capable of binding the lectins WGA and GSL I, while OmpN purified from E. coli ...
http://doras.dcu.ie/16616/
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The cloning , expression and characterisation of bacterial chitin-binding proteins from pseudomonas aeruginosa , serratia marcescens, photorhabdus luminescens and photorhabdus asymbiotica.
(2011)
Larragy, Ruth
The cloning , expression and characterisation of bacterial chitin-binding proteins from pseudomonas aeruginosa , serratia marcescens, photorhabdus luminescens and photorhabdus asymbiotica.
(2011)
Larragy, Ruth
Abstract:
It is well recognised that most proteins are subject to post translational modifications and that these modifications can have specific effects on the biological properties and functions of these proteins. The majority of proteins secreted by cells are modified by the attachment of oligosaccharide chains. This glycosylation event has been shown to impact correct protein folding, protein stability, solubility, to aid in cell recognition and to help regulate cell processes. In order to gain a deeper understanding into the impact of altered glycosylation patterns on cellular processes and cell recognition it is necessary to develop new technologies to profile the glycan species displayed on the surface of protein molecules. The present study was dedicated to the development of prokaryotic chitin-binding proteins as novel carbohydrate-binding molecules. Prokaryotic chitin-binding proteins from Serratia marcescens, Pseudomonas aeruginosa, Photorhabdus asymbiotica and Photorhabdus lumines...
http://doras.dcu.ie/16310/
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Lipid cubic phase as a membrane mimetic for integral membrane protein enzymes
(2011)
LI, DIANFAN; CAFFREY, MARTIN
Lipid cubic phase as a membrane mimetic for integral membrane protein enzymes
(2011)
LI, DIANFAN; CAFFREY, MARTIN
Abstract:
The lipidic cubic mesophase has been used to crystallize important membrane proteins for high-resolution structure determination. To date, however, no integral membrane enzymes have yielded to this method, the in meso. For a crystal structure to be meaningful the target protein must be functional. Using the in meso method with a membrane enzyme requires that the protein is active in the mesophase that grows crystals. Because the cubic phase is sticky and viscous and is bicontinuous topologically, quantitatively assessing enzyme activity in meso is a challenge. Here, we describe a procedure for characterizing the catalytic properties of the integral membrane enzyme, diacylglycerol kinase, reconstituted into the bilayer of the lipidic cubic phase. The kinase activity of this elusive crystallographic target was monitored spectrophotometrically using a coupled assay in a high-throughput, 96-well plate format. In meso, the enzyme exhibits classic Michaelis–Menten kinetics and works with ...
http://hdl.handle.net/2262/61017
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Substrate Profiling and Aldehyde Dismutase Activity of the Kvβ2 Subunit of the Mammalian Kv1 Potassium Channel
(2010)
Kumari, A.; Ryan, Barry; Dolly, J.; Henehan, Gary
Substrate Profiling and Aldehyde Dismutase Activity of the Kvβ2 Subunit of the Mammalian Kv1 Potassium Channel
(2010)
Kumari, A.; Ryan, Barry; Dolly, J.; Henehan, Gary
Abstract:
<p>Voltage-dependent potassium channels (Kv) are involved in various cellular signalling processes by governing the membrane potential of excitable cells. The cytosolic face of these α subunit-containing channels is associated with β subunits that can modulate channel responses. Surprisingly, the β subunit of the mammalian Kv1 channels, Kvβ2, has a high level of sequence homology with the aldo-keto reductase (AKR) superfamily of proteins. Recent studies have shown that Kvβ2 can catalyze the reduction of aldehydes and, most significantly, that channel function is modulated when Kvβ2-bound NADPH is concomitantly oxidized. As a result, the redox chemistry of this subunit is crucial to understanding its role in K+ channel modulation. The present study has extended knowledge of the substrate profile of this subunit using a single turnover fluorimetric assay. Kvβ2 was found to catalyse the reduction of aromatic aldehyde substrates such as 2, 3 and 4-nitrobenzaldehydes, 4-hydroxybenz...
http://arrow.dit.ie/schfsehart/87
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Investigation of the intrinsic mechanism of drug resistance in multiple myeloma
(2011)
Ooi, Melissa
Investigation of the intrinsic mechanism of drug resistance in multiple myeloma
(2011)
Ooi, Melissa
Abstract:
The focus of this thesis was to evaluate the mechanisms whereby myeloma cells develop intrinsic resistance with a focus on resistance in the context of bortezomib treatment. The aims of this thesis were to examine multidrug resistance pumps as a mechanism of resistance in MM, to investigate the contribution of p53 signalling perturbations in resistance mechanism in MM, to study the AMPK pathway as an alternative target to overcome MM resistance and finally to characterise myeloma resistance to bortezomib treatment using 2D-DIGE analysis. Focussing on bortezomib resistance models, we found that that overexpression of P-gp attenuates bortezomib activity. Bortezomib is a P-gp substrate and a combination of P-gp inhibitor and bortezomib is able to overcome resistance. Bortezomib is also able to downregulate the expression and function of P-gp. Our findings therefore suggest that combination of a P-gp inhibitor and bortezomib in P-gp positive myeloma would be a reasonable treatment comb...
http://doras.dcu.ie/16601/
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Synergizing metabolic flux analysis and nucleotide sugar metabolism to understand the control of glycosylation of recombinant protein in CHO cells.
(2011)
DAVEY, GAVIN
Synergizing metabolic flux analysis and nucleotide sugar metabolism to understand the control of glycosylation of recombinant protein in CHO cells.
(2011)
DAVEY, GAVIN
Abstract:
BACKGROUND: The glycosylation of recombinant proteins can be altered by a range of parameters including cellular metabolism, metabolic flux and the efficiency of the glycosylation process. We present an experimental set-up that allows determination of these key processes associated with the control of N-linked glycosylation of recombinant proteins. RESULTS: Chinese hamster ovary cells (CHO) were cultivated in shake flasks at 0 mM glutamine and displayed a reduced growth rate, glucose metabolism and a slower decrease in pH, when compared to other glutamine-supplemented cultures. The N-linked glycosylation of recombinant human chorionic gonadotrophin (HCG) was also altered under these conditions; the sialylation, fucosylation and antennarity decreased, while the proportion of neutral structures increased. A continuous culture set-up was subsequently used to understand the control of HCG glycosylation in the presence of varied glutamine concentrations; when glycolytic flux was reduced ...
http://hdl.handle.net/2262/61047
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Understanding the Molecular Information Contained in Principal Component Analysis of Vibrational Spectra of Biological Systems
(2011)
Bonnier, Franck; Byrne, Hugh
Understanding the Molecular Information Contained in Principal Component Analysis of Vibrational Spectra of Biological Systems
(2011)
Bonnier, Franck; Byrne, Hugh
Abstract:
<p>K-means clustering followed by Principal Component Analysis (PCA) is employed to analyse Raman spectroscopic maps of single biological cells. K-means clustering successfully identifies regions of cellular cytoplasm, nucleus and nucleoli, but the mean spectra do not differentiate their biochemical composition. The loadings of the principal components identified by PCA shed further light on the spectral basis for differentiation but they are complex and, as the number of spectra per cluster is imbalanced, particularly in the case of the nucleoli, the loadings under-represent the basis for differentiation of some cellular regions. Analysis of pure bio-molecules, both structurally and spectrally distinct, in the case of histone, ceramide and RNA, and similar in the case of the proteins albumin, collagen and histone, show the relative strong representation of spectrally sharp features in the spectral loadings, and the systematic variation of the loadings as one cluster becomes r...
http://arrow.dit.ie/radart/34
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Cell Proliferation Tracking Using Graphene Sensor Arrays
(2012)
HEGNER, MARTIN; DUESBERG, GEORG STEFAN
Cell Proliferation Tracking Using Graphene Sensor Arrays
(2012)
HEGNER, MARTIN; DUESBERG, GEORG STEFAN
Abstract:
The development of a novel label-free graphene sensor array is presented. Detection is based on modification of graphene FET devices and specifically monitoring the change in composition of the nutritive components in culturing medium. Micro-dispensing of Escherichia coli in medium shows feasibility of accurate positioning over each sensor while still allowing cell proliferation. Graphene FET device fabrication, sample dosing, and initial electrical characterisation have been completed and show a promising approach to reducing the sample size and lead time for diagnostic and drug development protocols through a label-free and reusable sensor array fabricated with standard and scalable microfabrication technologies.
http://hdl.handle.net/2262/61289
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Identification of the synthetic cannabinoid R(+)WIN55,212-2 as a novel regulator of IFN regulatory factor 3 (IRF3) activation and IFN-{beta} expression: relevance to therapeutic effects in models of multiple sclerosis.
(2011)
FALLON, PADRAIC GERARD
Identification of the synthetic cannabinoid R(+)WIN55,212-2 as a novel regulator of IFN regulatory factor 3 (IRF3) activation and IFN-{beta} expression: relevance to therapeutic effects in models of multiple sclerosis.
(2011)
FALLON, PADRAIC GERARD
Abstract:
β-Interferons (IFN-βs) represent one of the first line treatments for relapsing-remitting multiple sclerosis, slowing disease progression while reducing the frequency of relapses. Despite this, more effective, well tolerated therapeutic strategies are needed. Cannabinoids palliate experimental autoimmune encephalomyelitis (EAE) symptoms and have therapeutic potential in MS patients although the precise molecular mechanism for these effects is not understood. Toll-like receptor (TLR) signaling controls innate immune responses and TLRs are implicated in MS. Here we demonstrate that the synthetic cannabinoid R(+)WIN55,212-2 is a novel regulator of TLR3 and TLR4 signaling by inhibiting the pro-inflammatory signaling axis triggered by TLR3 and TLR4, whereas selectively augmenting TLR3-induced activation of IFN regulatory factor 3 (IRF3) and expression of IFN-β. We present evidence that R(+)WIN55,212-2 strongly promotes the nuclear localization of IRF3. The potentiation of IFN-β expressio...
http://hdl.handle.net/2262/61589
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Caspase-dependent inactivation of proteasome function during programmed cell death in Drosophila and man
(2004)
MARTIN, SEAMUS JOSEPH; CREAGH, EMMA
Caspase-dependent inactivation of proteasome function during programmed cell death in Drosophila and man
(2004)
MARTIN, SEAMUS JOSEPH; CREAGH, EMMA
Abstract:
The caspase family of cysteine proteases plays a conserved role in the coordinate demolition of cellular structures during programmed cell death from nematodes to man. Because cells undergoing programmed cell death in nematodes, flies, and mammals all share common features, this suggests that caspases target a common set of cellular structures in each of these organisms. However, although many substrates for mammalian caspases have been identified, few substrates for these proteases have been identified in invertebrates. To search for similarities between the repertoires of proteins targeted for proteolysis by caspases in flies and mammals, we have performed proteomics-based screens in Drosophila and human cell lines undergoing apoptosis. Here we show that several subunits of the proteasome undergo caspase-dependent proteolysis in both organisms and that this results in diminished activity of this multicatalytic protease complex. These data suggest that caspase-dependent proteolysis...
http://hdl.handle.net/2262/61750
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Interchain proteolysis, in the absence of a dimerization stimulus, can initiate apoptosis-associated caspase-8 activation
(2004)
MARTIN, SEAMUS JOSEPH; CREAGH, EMMA
Interchain proteolysis, in the absence of a dimerization stimulus, can initiate apoptosis-associated caspase-8 activation
(2004)
MARTIN, SEAMUS JOSEPH; CREAGH, EMMA
Abstract:
Caspases coordinate the internal demolition of the cell that is seen during apoptosis. Proteolytic processing of caspases is observed during apoptosis, and this correlates with conversion of inactive caspase proenzymes into their active two-chain forms. However, recent studies have suggested that caspase-8 is activated through dimerization and that interchain proteolysis is not sufficient for activation of this caspase. This proposal casts doubt upon whether caspase-8 is productively activated by granzyme B during granule-dependent cytotoxic T lymphocyte or natural killer cell-mediated killing, for example. Contrary to the dimerization model, we show that direct proteolysis of caspase-8 by the cytotoxic T lymphocyte protease granzyme B, or by caspase-6, produces an active enzyme that displays robust proteolytic activity toward synthetic as well as natural caspase-8 substrates. These data suggest that enforced dimerization of caspase-8 zymogens by scaffold proteins such as Fas-associ...
http://hdl.handle.net/2262/61751
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Smac/Diablo antagonizes ubiquitin ligase activity of inhibitor of apoptosis proteins
(2004)
CREAGH, EMMA; MARTIN, SEAMUS JOSEPH
Smac/Diablo antagonizes ubiquitin ligase activity of inhibitor of apoptosis proteins
(2004)
CREAGH, EMMA; MARTIN, SEAMUS JOSEPH
Abstract:
Inhibitor of apoptosis proteins (IAPs) can block apoptosis through binding to active caspases and antagonizing their function. IAP function can be neutralized by Smac/Diablo, an IAP-binding protein that is released from mitochondria during apoptosis. In addition to their ability to interact with caspases, certain IAPs also display ubiquitin-protein isopeptide ligase activity because of the presence of a RING domain. However, it is not known whether the ubiquitin-protein isopeptide ligase activities of human IAPs contribute to their apoptosis inhibitory activity or whether this IAP property can be modulated through association with Smac/Diablo. Here we demonstrate that the ubiquitin ligase activities of XIAP, and to a lesser extent c-IAP-1 and c-IAP2, are potently repressed through binding to Smac/Diablo. We also show that mutation of the XIAP RING domain rendered this IAP a less effective inhibitor of apoptosis, suggesting that the ubiquitin ligase activity of XIAP contributes to it...
http://hdl.handle.net/2262/61797
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Apoptosis-associated release of Smac/DIABLO from mitochondria requires active caspases and is blocked by Bcl-2
(2001)
CREAGH, EMMA; MARTIN, SEAMUS JOSEPH
Apoptosis-associated release of Smac/DIABLO from mitochondria requires active caspases and is blocked by Bcl-2
(2001)
CREAGH, EMMA; MARTIN, SEAMUS JOSEPH
Abstract:
Smac/DIABLO is a mitochondrial protein that potentiates some forms of apoptosis, possibly by neutralizing one or more members of the IAP family of apoptosis inhibitory proteins. Smac has been shown to exit mitochondria and enter the cytosol during apoptosis triggered by UV- or γ-irradiation. Here, we report that Smac/DIABLO export from mitochondria into the cytosol is provoked by cytotoxic drugs and DNA damage, as well as by ligation of the CD95 death receptor. Mitochondrial efflux of Smac/DIABLO, in response to a variety of pro-apoptotic agents, was profoundly inhibited in Bcl-2-overexpressing cells. Thus, in addition to modulating apoptosis-associated mitochondrial cytochrome c release, Bcl-2 also regulates Smac release, suggesting that both molecules may escape via the same route. However, whereas cell stress-associated mitochondrial cytochrome c release was largely caspase independent, release of Smac/DIABLO in response to the same stimuli was blocked by a broad-spectrum caspase...
http://hdl.handle.net/2262/61798
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CARDINAL, a novel caspase recruitment domain protein, is an inhibitor of multiple NF-kappa B activation pathways.
(2001)
CREAGH, EMMA; MARTIN, SEAMUS JOSEPH
CARDINAL, a novel caspase recruitment domain protein, is an inhibitor of multiple NF-kappa B activation pathways.
(2001)
CREAGH, EMMA; MARTIN, SEAMUS JOSEPH
Abstract:
Proteins possessing the caspase recruitment domain (CARD) motif have been implicated in pathways leading to activation of caspases or NF-kappaB in the context of apoptosis or inflammation, respectively. Here we report the identification of a novel protein, CARDINAL, that contains a CARD motif and also exhibits a high degree of homology to the C terminus of DEFCAP/NAC, a recently described member of the Apaf-1/Nod-1 family. In contrast with the majority of CARD proteins described to date, CARDINAL failed to promote apoptosis or NF-kappaB activation. Rather, CARDINAL potently suppressed NF-kappaB activation associated with overexpression of TRAIL-R1, TRAIL-R2, RIP, RICK, Bcl10, and TRADD, or through ligand-induced stimulation of the interleukin-1 or tumor necrosis factor receptors. Co-immunoprecipitation experiments revealed that CARDINAL interacts with the regulatory subunit of the IkappaB kinase (IKK) complex, IKKgamma (NEMO), providing a molecular basis for CARDINAL function. Thus,...
http://hdl.handle.net/2262/61799
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Fungal proteomics: from identitcation to function
(2011)
Doyle, Sean
Fungal proteomics: from identitcation to function
(2011)
Doyle, Sean
Abstract:
Some fungi cause disease in humans and plants, while others have demonstrable potential for the control of insect pests. In addition, fungi are also a rich reservoir of therapeutic metabolites and industrially useful enzymes. Detailed analysis of fungal biochemistry is now enabled by multiple technologies including protein mass spectrometry, genome and transcriptome sequencing and advances in bioinformatics. Yet, the assignment of function to fungal proteins, encoded either by in silico annotated, or unannotated genes, remains problematic. The purpose of this review is to describe the strategies used by many researchers to reveal protein function in fungi, and more importantly, to consolidate the nomenclature of ‘unknown function protein’ as opposed to ‘hypothetical protein’ – once any protein has been identified by protein mass spectrometry. A combination of approaches including comparative proteomics, pathogen-induced protein expression and immunoproteomics are outlined, which, wh...
http://eprints.nuim.ie/3034/
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Kinetics in signal transduction pathways involving promiscuous oligomerizing receptors can be determined by receptor specificity: Apoptosis induction by TRAIL
(2012)
Szegezdi, Eva; Mahalingam, Devalingam
Kinetics in signal transduction pathways involving promiscuous oligomerizing receptors can be determined by receptor specificity: Apoptosis induction by TRAIL
(2012)
Szegezdi, Eva; Mahalingam, Devalingam
Abstract:
Here we show by computer modeling that kinetics and outcome of signal transduction in case of hetero-oligomerizing receptors of a promiscuous ligand largely depend on the relative amounts of its receptors. Promiscuous ligands can trigger the formation of non-productive receptor complexes, which slows down the formation of active receptor complexes and thus can block signal transduction. Our model predicts that increasing the receptor-specificity of the ligand without changing its binding parameters should result in faster receptor activation and enhanced signaling. We experimentally validated this hypothesis using the cytokine Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL) and its four membrane-bound receptors as an example. Bypassing ligand-induced receptor hetero-oligomerization by receptor-selective TRAIL variants enhanced the kinetics of receptor activation and augmented apoptosis. Our results suggest that control of signaling pathways by promiscuous ligands cou...
http://hdl.handle.net/10379/2584
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Crystal Structure of NAD-dependent Peptoniphilus Asaccharolyticus Glutamate Dehydrogenase Reveals Determinants of Cofactor Specificity
(2012)
KHAN, AMIR RAFIQ
Crystal Structure of NAD-dependent Peptoniphilus Asaccharolyticus Glutamate Dehydrogenase Reveals Determinants of Cofactor Specificity
(2012)
KHAN, AMIR RAFIQ
Abstract:
Glutamate dehydrogenases (EC 1.4.1.2–4) catalyse the oxidative deamination of l-glutamate to α-ketoglutarate using NAD(P) as a cofactor. The bacterial enzymes are hexamers and each polypeptide consists of an N-terminal substrate-binding (Domain I) followed by a C-terminal cofactor-binding segment (Domain II). The reaction takes place at the junction of the two domains, which move as rigid bodies and are presumed to narrow the cleft during catalysis. Distinct signature sequences in the nucleotide-binding domain have been linked to NAD vs. NADP specificity, but they are not unambiguous predictors of cofactor preferences. Here, we have determined the crystal structure of NAD–specific Peptoniphilus asaccharolyticus glutamate dehydrogenase in the apo state. The poor quality of native crystals was resolved by derivatization with selenomethionine, and the structure was solved by single-wavelength anomalous diffraction methods. The structure reveals an open catalytic cleft in the absence of...
http://hdl.handle.net/2262/62419
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Synthesis, structural characterisation and luminescent anion sensing studies of a Ru(II)polypyridyl complex featuring an aryl urea derivatised 2,2'-bpy auxiliary ligand
(2012)
GUNNLAUGSSON, THORFINNUR; KITCHEN, JONATHAN; BOYLE, ELAINE
Synthesis, structural characterisation and luminescent anion sensing studies of a Ru(II)polypyridyl complex featuring an aryl urea derivatised 2,2'-bpy auxiliary ligand
(2012)
GUNNLAUGSSON, THORFINNUR; KITCHEN, JONATHAN; BOYLE, ELAINE
Abstract:
The inclusion of a urea functionality into the coordination sphere of a Ru(II)-polypyridyl complex (Ru·L1) resulted in a system that can function as an effective long wavelength emissive fluorescent anion sensor. The MLCT emission of Ru·L1 is sensitive to the binding of acetate, phosphate and pyrophosphate but not fluoride in organic solvent. In addition, Ru·L1 can distinguish between phosphate and pyrophosphate with an emission increase upon binding of H2PO4- (“turn on” sensor) and an emission decrease upon binding of HP2O73- (“turn off” sensor), which occurs via hydrogen bonding to the urea receptor moiety as demonstrated by carrying out NMR titrations as well as by employing [Ru(II)bipy3](PF6−)2 as a model compound that lacks the anion receptor moiety.
http://hdl.handle.net/2262/62417
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New fluorescent bile acids: Synthesis, chemical characterization, and disastereoselective uptake by Caco-2 cells of 3-deoxy 3-NBD-amino deoxycholic and ursodeoxycholic acid
(2012)
LONG, AIDEEN; EHRHARDT, CARSTEN; KEAVENY, RAYMOND DOMINIC; GILMER, JOHN FRANCIS; WANG, ...
New fluorescent bile acids: Synthesis, chemical characterization, and disastereoselective uptake by Caco-2 cells of 3-deoxy 3-NBD-amino deoxycholic and ursodeoxycholic acid
(2012)
LONG, AIDEEN; EHRHARDT, CARSTEN; KEAVENY, RAYMOND DOMINIC; GILMER, JOHN FRANCIS; WANG, JUN; MAJER, FERENC; SHARMA, RUCHIKA
Abstract:
Deoxycholic acid (DCA), a secondary bile acid (BA), and ursodeoxycholic acid (UDCA), a tertiary BA, cause opposing effects in vivo and in cell suspensions. Fluorescent analogues of DCA and UDCA could help investigate important questions about their cellular interactions and distribution. We have prepared a set of isomeric 3α- and 3β-amino analogues of UDCA and DCA and derivatised these with the discrete fluorophore, 4-nitrobenzo-2-oxa-1,3-diazol (NBD), forming the corresponding four fluorescent adducts. These absorb in the range 465–470 nm and fluoresce at approx. 535 nm. In order to determine the ability of the new fluorescent bile acids to mimic the parents, their uptake was studied using monolayers of Caco-2 cells, which are known to express multiple proteins of the organic anion-transporting peptide (OATP) subfamily of transporters. Cellular uptake was monitored over time at 4°C and 37°C to distinguish between passive and active transport. All four BA analogues were taken up ...
http://hdl.handle.net/2262/62427
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Development and validation of software solutions to eliminate bottlenecks in the application of high-throughput quantitative immunohistochemistry
(2008)
Conway, Catherine
Development and validation of software solutions to eliminate bottlenecks in the application of high-throughput quantitative immunohistochemistry
(2008)
Conway, Catherine
http://doras.dcu.ie/188/
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Regulation and assembly of the associated network in human cells
(2011)
Quinn, Nadine
Regulation and assembly of the associated network in human cells
(2011)
Quinn, Nadine
Abstract:
Centromeres are the chromosomal loci which are responsible for the equal segregation of sister chromatids to daughter cells during mitosis, by directing the assembly of kinetochores. Despite being involved in the faithful inheritance of the genome, centromeres are themselves epigenetically determined. Centromeres of higher organisms are determined by their centromeric chromatin which consists of unique nucleosomes, in which canonical histone H3 is replaced by the histone H3 variant CENP-A. A large group of additional proteins associate with the centromere throughout the cell cycle, collectively known as the constitutive centromere associated network (CCAN), and these are required for kinetochore function. In this thesis I have investigated the regulation and assembly of these CCAN proteins in human cells. Chapter 3 focuses on the regulation of the abundance of CCAN components during the cell cycle. Our analysis of the relative transcript abundance of CCAN members using real-time PCR...
http://hdl.handle.net/10379/2598
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Development and Validation of a Novel Reporter Assay for Human Papillomavirus Type 16 Late Gene Expression
(2012)
Orru, Beatrice; Cunniffe, Ciaran; Ryan, Fergus X.; Schwartz, Stefan
Development and Validation of a Novel Reporter Assay for Human Papillomavirus Type 16 Late Gene Expression
(2012)
Orru, Beatrice; Cunniffe, Ciaran; Ryan, Fergus X.; Schwartz, Stefan
Abstract:
<p>To facilitate the investigations of HPV-16 late gene expression HPV-16 reporter plasmids were generated using previously described sub-genomic HPV-16 plasmids, named pBEL and pBELM, that, similar to the full viral genome, produce primarily HPV-16 early mRNAs and very little, if any, late mRNAs in cervical cancer cells. The HPV-16 late L1 gene was replaced by the chloramphenicol acetyltransferase (CAT) reporter gene, or green fluorescent protein (GFP), preceded by the poliovirus internal ribosome entry site (IRES). Results show that the reporter genes mimic the expression of L1 from these plasmids. For example, overexpression of adenovirus E4orf4 protein (E4orf4), polypyrimidine tract binding protein (PTB), arginine/serinerich SRp30c protein (SRp30c) or alternative splicing factor/splicing factor 2 (ASF/SF2) induced an increased expression of CAT or GFP. Stable cell lines with reporter plasmids pBELCAT and pBELMCAT were also generated. An induction of CAT was observed in HPV...
http://arrow.dit.ie/scschbioart/79
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Development and application of novel approaches to quantitate therapeutic drugs in human blood
(2012)
Fox, Deirdre
Development and application of novel approaches to quantitate therapeutic drugs in human blood
(2012)
Fox, Deirdre
Abstract:
The research described in this thesis utilised novel extraction phases and analytical approaches in order to develop new methods for the separation and determination of important pharmaceuticals in complex biological matrices. The project employed bioanalytical chemistry to solve real-world biological and clinical problems. The first project involved the development and validation of an analytical protocol for the extraction, separation and determination of rifampicin and efavirenz in HIV-positive patients. The protocol was developed to support the hypothesis that ‘concomitant treatment with rifampicin-containing anti-tuberculosis (TB) therapy and the anti- HIV therapy, efavirenz, results in a decreased rifampicin half life’. The emphasis in this work was the determination of both drugs simultaneously in one sample in a simple, rapid and cost effective assay. This was achieved using solid phase extraction and LC-UV analysis. The main challenge was developing a low cost assay to dete...
http://doras.dcu.ie/16751/
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Spiropryan-terthiophene polymers as multi-stimuli-responsive platforms for biomechanical interactions
(2012)
Zanoni, Michele; Gelmi, Amy; Higgins, Michael; Fraser, Kevin J.; Byrne, Robert; Wagner,...
Spiropryan-terthiophene polymers as multi-stimuli-responsive platforms for biomechanical interactions
(2012)
Zanoni, Michele; Gelmi, Amy; Higgins, Michael; Fraser, Kevin J.; Byrne, Robert; Wagner, Klaudia; Gambhir, Sanjeev; Officer, David; Wallace, Gordon; Diamond, Dermot
http://doras.dcu.ie/16903/
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The design and characterisation of biomimetic artificial receptors based on molecular imprinting technology
(2007)
Farrington, Keith
The design and characterisation of biomimetic artificial receptors based on molecular imprinting technology
(2007)
Farrington, Keith
http://doras.dcu.ie/16982/
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