Dietetics, Nutrition and Biological Sciences
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Item The Inhibition of Free Radical Generation by Preparations of Harpagophytum procumbens in vitro(2009-01) Grant, L.; McBean, Douglas; Fyfe, Lorna; Warnock, MaryHarpagophytum procumbens (Hp), commonly known as Devil's Claw has been used as a traditional treatment for a variety of illnesses for centuries. Since the early twentieth century, it has become a popular antiinflammatory and analgesic preparation amongst European herbalists for supportive or adjuvant treatment of degenerative joint diseases. Extracts of Hp tubers have demonstrated antiinflammatory and analgesic effects in animal models of inflammation and in human trials. The mechanism(s) of action responsible for these attributes, however, remain to be elucidated. Reactive oxygen species generated in acute and chronic inflammatory diseases are known to be cytotoxic and can cause tissue damage. In this study, a root tuber extract (Hp extract) and commercially available tincture (Hp tincture) were investigated for antioxidant characteristics using in vitro test systems. Both preparations were found to effectively scavenge DPPH radical, inhibit nitrite levels in supernatants harvested from LPS-stimulated RAW 264.7 macrophages, and cause dose-dependent suppressions in the detection of fMLP-and AA-induced neutrophil MPO. The antioxidant effects demonstrated for both preparations of Hp may contribute to the antiinflammatory and analgesic properties observed for the plant. Copyright © 2008 John Wiley & Sons, LtdItem A review of the biological and potential therapeutic actions of harpagophytum procumbens(John Wiley & Sons, Ltd, 2007-11-24) McBean, Douglas; Grant, L.; Fyfe, Lorna; Warnock, MaryHarpagophytum procumbens (Hp), commonly known as Devil's Claw is a perennial plant which thrives in arid conditions. For centuries, it has been used as a traditional treatment for a variety of illnesses, including fevers, skin complaints, arthritis and diseases of the digestive tract as well as an appetite stimulant. Since its introduction to Europe in the early twentieth century, it has become a popular antiinflammatory and analgesic preparation amongst herbalists for supportive or adjuvant treatment of degenerative joint diseases, tendonitis, headache, backache and menstrual pain. The validity of Hp as an effective antiinflammatory and analgesic preparation, particularly in the relief of arthritic symptoms, has been investigated in numerous animal, clinical and in vitro studies. Although some contradictory evidence exists, the majority of animal studies appear to indicate Hp as an effective antiinflammatory and analgesic preparation in the treatment of acute and subacute inflammation. Clinical trials support Hp as a beneficial treatment for the alleviation of pain and improvement of mobility in a variety of musculoskeletal conditions. Analysis of the in vitro and ex vivo studies that currently exist, indicate that Hp has significant effects on numerous proinflammatory markers. However, the exact mechanism(s) by which Hp may reduce inflammation remain to be elucidated.Item The Roles of Opioid Receptors and Agonists in Health and Disease Conditions(Maxwell Scientific Organization,, 2011-04-30) Ibegbu, A. O.; Mullaney, I.; Fyfe, Lorna; McBean, DouglasOpioid receptors are found in the Central Nervous System (CNS) and are classified as mu (µ), kappa (κ), delta (δ) and sigma (σ) opioid receptors. Opioid receptors belong to the large family of G Protein Coupled Receptors (GPCRs), and have diverse and important physiological roles. The aim of the present review is to discuss the roles played by opioid receptors, their agonists and antagonists in health and disease conditions. Opioid receptors are not uniformly distributed in the CNS and are found in areas concerned with pain, with the highest concentration in the cerebral cortex, followed by the amygdala, septum, thalamus, hypothalamus, midbrain and spinal cord. Activated delta opioid receptors are coupled to Gi1 while activated mu opioid receptors are coupled to Gi3 in neuroblastoma cells. Mu opioid receptors are activated by mu receptor agonists and are coupled through the Gi1 and GoA. Both mu and kappa opioid receptors are coupled via both Gi and Gz and opioid receptors are important targets for thousands of pharmacological agents. GPCRs typically require activation by agonists for their signalling activity to be initiated but some of the GPCRs may display basal or spontaneous signalling activity in the absence of an agonist. The stimulation of these receptors triggers analgesic effects and affects the function of the nervous system, gastrointestinal tract and other body systems. Hundreds of analogs of opioid peptides have been synthesized in an effort to make the compounds more active, selective, and resistant to biodegradation than the endogenous ligands. All these modifications resulted in obtaining very selective agonists and antagonists with high affinity at mu-, delta-, and kappa-opioid receptors, which are useful in further studies on the pharmacology of opioid receptors in a mammalian organism.Item The Effect of Hypoxia on G Protein Coupled (Opioid) Receptor Gene Expression in Cortical B50 Neurons in Culture(Maxwell Scientific Organization,, 2011-04-30) Ibegbu, A. O.; Mullaney, I.; Fyfe, Lorna; McBean, DouglasHypoxia adversely affects cells and tissues, and neuronal cells in particular have been shown to be more susceptible to the injurious effects of hypoxia in which they may begin to die when oxygen supply is reduced or completely eliminated. Opioid receptor agonists have been shown to elicit several central nervous system effects, mediated via G protein-coupled receptors. The aim of this study was to study the effect of hypoxia on G protein coupled receptor gene expression using mu opioid receptor as a case study in cortical neuronal B50 cell lines in culture. The B50 cells were cultured in normoxia (21% O2; 5% CO2) and hypoxia (5% O2; 5% CO2), and were treated with opioid agonists to determine their effects on hypoxia-induced changes. Three opioid agonists {DAMGO(_), DSLET(*) and ICI--199,441(6)}, were administered to the cells as treatment for 48 hours after 48 hours of initial culture for a total of 96 hours of culture in hypoxic conditions at concentrations of 10, 50 and 100 :M. The levels of G-protein coupled receptor (mu opioid) mRNAs were assessed using RT-PCR. The results showed that hypoxia induced morphological changes in B50 cells in hypoxia while the mu opioid RT-PCR mRNA levels showed no appreciable changes in normal, hypoxic and treated cells. The results show that B50 neuronal cells are susceptible to damage and injurious effects of hypoxia, as are most brain cells and the opioid agonist treatments showed there were no changes in the level of mu opioid receptor gene expression due to hypoxia or agonist treatment in neuronal B50 cells in culture.Item The Effect of Hypoxia on G Protein Coupled (CB1) Receptor Gene Expression in Cortical B50 Neurons in Culture(Maxwell Scientific Organization,, 2011-02-10) Ibegbu, A. O.; Mullaney, I.; Fyfe, Lorna; McBean, DouglasHypoxia adversely affects cells and tissues, and neuronal cells in particular have been shown to be more susceptible to the injurious effects of hypoxia in which they may begin to die when oxygen supply is reduced or completely eliminated. Cannabinoid (CB1) receptor agonists have been shown to elicit several Central Nervous System (CNS) effects, mediated via G protein-coupled receptors. The aim of this study was to examine the effect of hypoxia on G protein coupled receptor (CB1) gene expression in cortical neuronal B50 cell lines in culture. The B50 cells were cultured in normoxia (21% O2; 5% CO2) and hypoxia (5% O2; 5% CO2), and were treated with cannabinoid agonists to determine their effects on hypoxia-induced changes. Three cannabinoid agonists [Win55,212-2 mesylate (Win), arachidonoylethanolamide (AEA) and 2- arachidonylglycerol (2-AG)], were administered to the cells as treatment for 48 hours after 48hours of initial culture for a total of 96hours of culture in hypoxic conditions at concentrations of 10, 50 and 100 nM. The levels of G-protein coupled receptor (CB1) mRNAs were assessed using RT-PCR. The results showed that hypoxia induced morphological changes in B50 cells in hypoxia while the CB1 RT-PCR mRNA levels showed no appreciable changes in normal, hypoxic and treated cells. The results show that B50 neuronal cells are susceptible to damage and injurious effects of hypoxia, as are most brain cells and the cannabinoid agonist treatments showed there were no changes in the level of CB1 receptor gene expression due to hypoxia or agonist treatment in neuronal B50 cells in culture.Item Oxidative stress does not predispose neuronal cells to changes in G protein coupled (opioid) receptor gene expression in cortical B50 neurons in culture(Kamla-Raj Enterprises, 2012-12) Ibegbu, A. O.; Mullaney, I.; Fyfe, Lorna; McBean, DouglasOxidative stress adversely affects neuronal cells in which they may die when oxygen supply is reduced or eliminated and opioid receptor agonists elicit several central nervous system effects. The aim of this study was to evaluate the effect of oxidative stress on opioid receptor gene expression in cortical B50 cells. The cells were cultured in normoxia, hypoxia and treated with opioid agonists; DAMGO (_), DSLET () and ICI-199,441 () for 48 hours after 48 hours of initial culture at dose of 10_M, 50_M and 100_M. The level of mu opioid receptor mRNA was assessed using RT-PCR. The results show that oxidative stress induced changes in B50 cells in hypoxia while mu opioid mRNA levels showed no change. The results show that B50 cells are susceptible to damage by oxidative stress and opioid agonist treatments showed no change in the level of mu opioid receptor gene expression in B50 cells. Kamla-Raj 2012.Item The Roles of Guanine Nucleotide Binding Proteins in Health and Disease(Maxwell Scientific Organization,, 2011-02-10) Ibegbu, A. O.; Mullaney, I.; Fyfe, Lorna; McBean, DouglasG-proteins are important mediators of cellular and tissue functions and are characterised by a recognition site for Guanine Triphosphate (GTP), Guanine Diphosphate (GDP) and possess intrinsic GTPase activity. They play important roles in signal transduction responsible for cytoskeletal remodelling, cellular differentiation and vesicular transport. They are made up of three types namely, the small G-proteins, the sensors and the heterotrimeric G-proteins. The G-protein heterotrimers consist of G-alpha (G), G-beta (G$) and G-gamma (G() subunits. Each heterotrimeric G-protein have different subunits and the combination of these subunits define the specific role of each G-protein. The activation of G subunits regulates the activity of effector enzymes and ion channels while G$( subunits function in the regulation of mitogen-activated protein kinase (MAP-kinase) pathway. The G-protein-mediated signal transduction is important in the regulation of a cells morphological and physiological response to external stimuli. MAPKs are involved in the phosphorylation of transcription factors that stimulate gene transcription. Gs stimulates adenylate cyclase, thereby increasing cyclic adenosine monophosphate (cAMP) leading to the phosphorylation and subsequent activation of Ca_+ channels. G proteins are involved in disease pathology through several mechanisms which interfere with the G protein activity. Other disease pathologies associated with abnormal mutations in G proteins can interfere with signal transduction pathways which may involve signal transmission that is either excessive, by augmentation of G protein function, or insufficient, via inactivation of G proteins.Item Oxidative stress-induced effects on pattern and pattern formation in cortical B50 neuronal cells in culture(2013-11) Ibegbu, A. O.; Fyfe, Lorna; McBean, Douglas; Mullaney, I.Oxidative stress adversely affects cells and tissues, and neuronal cells in particular have been shown to be more susceptible to the injurious effects of oxidative stress in which the cells may die when oxygen supply is reduced or completely eliminated. The aim of the present study was to study the effect of oxidative stress using hypoxia as a bench mark on the morphology of B50 neuronal cell lines cultured in hypoxia using neuronal pattern and pattern formation as case study. The B50 cells were cultured in normal incubator (21%O2; 5% CO2) as control group and hypoxic incubator (5%O2; 5% CO2) as the experimental group. Neuronal morphology, pattern and wellbeing were assessed using same field morphological assessment of cells and lactate dehydrogenase leakage (LDH). The result showed groups of dead and degenerating B50 neuronal cells, altered neuronal pattern and pattern formation and some significant changes (P<0.05) in cellular levels of LDH leakage in normal B50 cells and hypoxic cells. The changes in morphology, neuronal pattern and LDH release indicate that oxidative stress has induced morphological and cellular changes in cortical B50 cells in culture and that the B50 neuronal cells are susceptible to damage and injurious effects of oxidative stress represented by hypoxia as most brain cells.Item The Effects of Hypoxia and Opioid Receptor Agonists Treatment in Cortical B50 Neuronal Cells in Culture(Uludag University Applied Research Center for Agriculture (ARCA) and Applied Research Center for Environmental Problems (ARCEP), Uludag University Gorukle Campus, 16059 Bursa-Turkey, 2012-12) Ibegbu, A. O.; Fyfe, Lorna; McBean, Douglas; Mullaney, I.Hypoxia has been implicated in nerve cell deaths in many neurological disorders and opioid receptor agonists have some positive benefits on the nervous system. The aim of the present work was to investigate the effects of hypoxia and opioid receptor agonists' treatment on the morphology of B50 cells cultured in hypoxia using neuronal pattern and pattern formation as a case study. The B50 cells were cultured in normal incubator (21%O2; 5% CO2) as the control group and hypoxic incubator (5%O2; 5% CO2) as the experimental group and three opioid receptor agonists namely DAMGO (_), DSLET () and ICI-199,441 () were administered to the cells for 48 hours as treatment against hypoxia after 48 hours of culture at 10_M, 50_M and 100_M concentrations. Neuronal morphology and wellbeing was assessed using same field morphological assessment and lactate dehydrogenase leakage (LDH). The result showed groups of dead and degenerating B50 neuronal cells, altered neuronal pattern and pattern formation and some significant changes (P<0.05) in cellular levels of LDH leakage in normal, hypoxic cells and cells treated with different agonists. The changes in morphology, neuronal pattern and LDH release indicate that hypoxia induced morphological and cellular changes in B50 cells in hypoxia and opioid agonists have some potential benefits in the treatment of hypoxia-induced changes in B50 cells in culture.Item Effectiveness and safety of Devil's Claw tablets in patients with general rheumatic disorders(2007-12) Warnock, Mary; McBean, Douglas; Suter, Andreas; Tan, Jen; Whittaker, PatriciaArthritis and other rheumatic conditions (AORC) are the leading cause of disability, are associated with poor quality of life and incur considerable direct and indirect costs. It is considered that the instance of AORC will continue to increase. To assess the effectiveness, safety and tolerability of Harpagophytum (Bioforce) in the treatment of AORC, a single group open study of 8 weeks duration (259 patients) was performed in the United Kingdom. Effectiveness was assessed by numeric rating scales, the Western Ontario and McMasters Universities Osteoarthritis (WOMAC) Index and the Algofunctional Hand Osteoarthritis Index. Tolerance was measured by a numeric rating scale and safety by self-reporting, blood analysis and liver function tests. Quality of life was measured by SF-12 questionnaire. There were statistically significant (p < 0.0001) improvements in patient assessment of global pain, stiffness and function. There were also statistically significant reductions in mean pain scores for hand, wrist, elbow, shoulder, hip, knee and back pain. Quality of life measurements (SF-12) were significantly increased from baseline and 60% patients either reduced or stopped concomitant pain medication. Harpagophytum is an effective and well-tolerated serious treatment option for mild to moderate degenerative rheumatic disorders providing improved quality of life measure. Copyright 2007 John Wiley & Sons, Ltd.