1,720,971 research outputs found
Peroxynitrite formation from the simultaneous reduction of nitrite and oxygen by xanthine oxidase
No full textOne electron reductions of oxygen and nitrite by xanthine oxidase form peroxynitrite. The nitrite and oxygen reducing activities of xanthine oxidase are regulated by oxygen with K(oxygen) 26 and 100 microM and K(nitrite) 1.0 and 1.1 mM with xanthine and NADH as donor substrates. Optimal peroxynitrite formation occurs at 70 microM oxygen with purine substrates. Kinetic parameters: V(max) approximately 50 nmol/min/mg and K(m) of 22, 36 and 70 microM for hypoxanthine, pterin and nitrite respectively. Peroxynitrite generation is inhibited by allopurinol, superoxide dismutase and diphenylene iodonium. A role for this enzyme activity can be found in the antibacterial activity of milk and circulating xanthine oxidase activity
ROS generation in endothelial hypoxia and reoxygenation stimulates MAP kinase signaling and kinase-dependent neutrophil recruitment
No full textReactive oxygen species (ROS)-induced injury has been shown to occur during the reperfusion phase of ischemia–reperfusion and ROS are known to induce signaling events. We hypothesized that oxygen sensing in endothelial cells is also dependent on internal redox changes during hypoxia and that endothelial cells respond to changing oxygen environments via signaling, switching to an inflammatory phenotype. Endothelial cells exposed to relative hypoxia or the mitochondrial inhibitors rotenone, antimycin A, or FCCP show loss of mitochondrial membrane potential. During hypoxia, an increase in cytoplasmic ROS and glutathione S-transferase activity occurred, suggesting changes in intracellular redox state, mimicked with rotenone or FCCP but inhibited by antimycin A. Phosphorylation of stress-responsive mitogen-activated protein kinases occurred in hypoxia and was rapid and prolonged. Phosphorylation was inhibited by vitamin C, N-acetyl cysteine, or antimycin A. Chelation of intracellular calcium inhibits phosphorylation but the mitochondrial transition pore inhibitor cyclosporin A had no effect. Reoxygenation caused a further round of signaling, which was rapid but transient. Functionally, adhesion of neutrophils after hypoxia–reoxygenation under flow is ROS, P-selectin, and MAPK dependent. Therefore, changes in cellular signaling and phenotype are abrogated by ROS scavengers and suggest their use as therapeutic agents in ischemia–reperfusion
Xanthine oxidoreductase catalyses the reduction of nitrates and nitrite to nitric oxide under hypoxic conditions
No full textXanthine oxidoreductase (XOR) catalyses the reduction of the therapeutic organic nitrate, nitroglycerin (glyceryl trinitrate, GTN), as well as inorganic nitrate and nitrite, to nitric oxide (NO) under hypoxic conditions in the presence of NADH. Generation of nitric oxide is not detectable under normoxic conditions and is inhibited by the molybdenum site-specific inhibitors, oxypurinol and (-)BOF 4272. These enzymic reactions provide a mechanism for generation of NO under hypoxic conditions where nitric oxide synthase does not function, suggesting a vasodilatory role in ischaemia.<br/
Xanthine oxidase mediates cytokine-induced, but not hormone-induced bone resorption
No full textReactive oxygen species (ROS) such as hydrogen peroxide (H2O2) have been implicated as mediators of osteoclastic bone resorption. Xanthine oxidase (XO) a ubiquitous enzyme is widely known for its production of these ROS. We therefore evaluated the potential of XO as a source of ROS in cytokine-and hormone-induced bone resorption. XO activity in rat calvarial osteoblasts was found to be significantly elevated upon stimulation by the cytokines, TNFalpha and IL-1beta. These cytokines also caused a dose related increase in bone resorption of mouse calvariae, which was significantly inhibited by catalase (10 IU/ml). Allopurinol, the competitive inhibitor of XO, also caused a dose related (1-50 microM) inhibition of TNFalpha (20 ng/ml) and (0.01-10 microM) IL-1beta (50 IU/ml)-induced bone resorption, respectively. PTH- and 1,25-(OH)2 Vitamin D3-induced bone resorption could also be inhibited by catalase (100 IU/ml) but was unaffected by allopurinol, indicating that another mediator, other than XO, is required for hormone-induced bone resorption. These results demonstrate, that modulation of the redox balance in the bone microenvironment, which contains XO, can affect the bone resorbing process. Therefore, XO may play a pivotal role in cytokine-induced bone resorption and, if manipulated appropriately, could show a therapeutic benefit in inflammatory bone disorders such as RA.<br/
Manipulation of in vitro angiogenesis using peptide-coated gold nanoparticles
No full textWe demonstrate the deliberate activation or inhibition of in vitro angiogenesis using functional peptide coated gold nanoparticles. The peptides, anchored to oligo-ethylene glycol capped gold nanospheres, were designed to selectively interact with cell receptors responsible for activation or inhibition of angiogenesis. The functional particles are shown to influence significantly the extent and morphology of vascular structures, without causing toxicity. Mechanistic studies show that the nanoparticles have the ability to alter the balance between naturally secreted pro- and anti- angiogenic factors, under various biological conditions. Nanoparticle-induced control over angiogenesis opens up new directions in targeted drug delivery and therapy
Nanoparticles for inhibition of in vitro tumour angiogenesis: synergistic actions of ligand function and laser irradiation
No full textCareful design of nanoparticles plays a crucial role in their biomedical applications. It not only defines the stability of nanoparticles in a biological medium but also programs their biological functionality and specific interactions with cells. Here, an inorganic nanoparticulate system engineered to have a dual role as anti-angiogenic and hyperthermic agent is presented. The inorganic rod-shaped core is designed to strongly absorb near-infrared laser irradiation through the surface plasmon resonance and convert it into localized heat, while a peptide coating acts as an anti-angiogenic drug, altogether inhibiting vascular growth. The synergistic dual action provides an improved inhibition of the in vitro tumour angiogenesis, offering new possibilities for the development of nano-engineered anti-angiogenic drugs for therapies
Receptor-mediated interactions between colloidal gold nanoparticles and human umbilical vein endothelial cells
No full textA new strategy to manipulate cell operations is demonstrated, based on membrane-receptor-specific interactions between colloidal peptide-capped gold nanoparticles and human umbilical vein endothelial cells. It is shown that colloidal gold nanoparticles of similar charge and size but capped with different peptide sequences can deliberately trigger specific cell functions related to the important biological process of blood vessel growth known as angiogenesis. Specific binding of the peptide-capped particles to two endothelial-expressed receptors (VEGFR-1, NRP-1), which control angiogenesis, is achieved. The cellular fate of the functional nanoparticles is imaged and the influence of the different peptide-coated nanoparticles on the gene expression profile of hypoxia-related and angiogenic genes is monitored. The findings open up new avenues towards the deliberate biological control of cellular functions using strategically designed nanoparticle
Laser-induced damage and recovery of plasmonically targeted human endothelial cells
Full text linkLaser-induced techniques that employ the surface plasmon resonances of nanoparticles have recently been introduced as an effective therapeutic tool for destroying tumor cells. Here, we adopt a low-intensity laser-induced technique to manipulate the damage and repair of a vital category of noncancerous cells, human endothelial cells. Endothelial cells construct the interior of blood vessels and play a pivotal role in angiogenesis. The degree of damage and repair of the cells is shown to be influenced by laser illumination in the presence of gold nanoparticles of different morphologies, which either target the cellular membrane or are endocytosed. A pronounced influence of the plasmonic nanoparticle laser treatment on the expression of critical angiogenic genes is shown. Our results show that plasmon-mediated mild laser treatment, combined with specific targeting of cellular membranes, enables new routes for controlling cell permeability and gene regulation in endothelial cells.<br/
Dataset supporting the University of Southampton Doctoral Thesis "Development of poly(lactic-co-glycolic acid) electrospun membranes for incorporation into 3-D co-culture models of the airway-blood barrier"
No full textDataset supporting the University of Southampton Doctoral Thesis "Development of poly(lactic-co-glycolic acid) electrospun membranes for incorporation into 3-D co-culture models of the airway-blood barrier"
This dataset contains raw data and associated graphs and statistics in GraphPad Prism (version 10.2.0) files. Separate files are included for each results chapter.
Thesis_R1 - data for figures in the first results chapter, thesis chapter 3 - Development of an Electrospinning System and the Generation of PLGA Membranes
THesis_R2 - data for figures in the second results chapter, thesis chapter 4 - Biological characterisation of Epithelial Barrier Formation on Electrospun PLGA Membranes
Thesis_R3 - data for figures in the third results chapter, thesis chapter 5 - Development of a 3-D Co-culture Model using Electrospun Membrane
Data collection as described in associated thesis. Statistics are included with the data entry.
Software required to open filed: Graphpad Prism v10.2.0 (392)
The data is embargoed until 20/01/2025.
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