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Molecular detection and characterisation of biologically relevant free radicals during surgical ischaemia-reperfusionGutowski, Mariusz January 2011 (has links)
Oxygen is one of the most important molecules in human beings. Our research is focused on how the human body can respond and adapt to the physiological challenge posed by a lack of oxygen. Ascorbic acid (Vitamin C) is one of the most important and considered the most effective water-soluble, chain-breaking antioxidant in human plasma, with the capacity to prevents damage by free radicals. This thesis presents four studies investigating the phenomenon of Reactive Oxygen Species (ROS) generation in the many different surgical conditions in the animal and in the human. Study one investigated the geometry and thermodynamic properties of vitamin C. Calculations were carried out at the restricted and unrestricted B3LYP/6-31+G(d,p), B3LYP/6-311++G(d,p) and B3LYP/EPR-II levels for two conformers (1 and 2) of L-ascorbic acid and their respective oxidation products to monodehydroascorbates of ab-initio methods by Gaussian O3W package. Conformer 1, free radical properties are compared with previously published calculations in the gaseous and aqueous solution states and with experimental EPR values. Calculated molecular structures, EPR (electron paramagnetic resonance spectroscopy), the vibration spectral and energetic properties and all are reported including some proposed changes to previous EPR assignments. Conformer 2 of L-ascorbic acid is predicted to have lower energy than Conformer 1, under the method and basis sets used, by between 11 and 26 kJ mol-1 and is stabilised by internal hydrogen bonding. Relaxed potential energy surface (PES) scans were carried out for two proton transfer processes and relative energies of stable minima and barriers between them determined. Hydrogen transfer is predicted in two systems with favourable spatial arrangements of O–H and O groups for which relaxed potential energy surface scans are reported. Calculated vibrational wavenumber values are provided for selected C=C, C=O, C–H and O–H modes assigned to particular groups and significant calculated EPR hyperfine coupling constants (HCC) values for splitting by H(1) and C(13) for radical species are also reported. These calculations contribute to a better understanding of the complex role of L-ascorbic acid and its various oxidised, neutral, ionic and radical forms in biochemistry and medicine. Study two examined if vitamin C could ameliorate the damaging effects of I-R on myocardium and we postulated that the mechanism of vitamin C protection against iii I-R-induced cell death involved quenching of ROS. In the vitamin C group after 5 min of reperfusion a significant, sudden increase of diastolic pressure in the heart was noted and reached a maximum of 77 mmHg after 12 min of reperfusion and then gradually decreased to 51 mmHg after 60 min of reperfusion period but was quicker than in Control group reaching 37 mmHg by the end of the reperfusion period. The level of A·− (ascorbate free radicals) sudden and massive increased at the time of reperfusion in the Vitamin C group. This increase was associated with poor mechanical function in hearts as indicated by the significantly depressed recovery process. After 30 min of global, now-flow ischaemia and 60min of reperfusion infarct size averaged 33% ± 1 in Control group and 30 % ± 1 in Vitamin C group, respectively, (P<0.05). There is strong evidence that oxygen centered radicals contribute to postischaemic dysfunction after global ischaemia. Our data unquestionably suggest that the large production of A·− was associated with a greater depression in myocardial contractile function, therefore could represent a marker of oxidative stress during I-R and could be related to the functional impairment during reperfusion. In summary, we have used the animal models of isolated heart perfusion to provide evidence that vitamin C did not reduce the infarct size, however “tendency” towards a decrease (↓) in infarct size with ascorbate and it protects from oxidative damage during global I-R as manifested by decreased concentrations of A·− and enhance recovery of mechanical function such as diastolic pressure and LVDP in postischaemic working rat hearts. Study three was designed to test the hypothesis that the physiological trauma associated with venous cannulation may artefactually stimulate systemic free radical formation in the acute phase that if not accounted for may under-estimate the oxidative stress response to exercise. The relationship between the time of venepuncture and the level of free radical generation during normoxic conditions was further investigated. The venous cannulation in Phase I, increased plasma A·− by 347 ± 173 AU/√G, P <0.05 after 2min of venepuncture with further increases observed after 5min and 10min of venous cannulation, respectively (403 ± 178 AU/√G; 462 ± 93 AU/√G, P < 0.05) vs baseline point time. After this time the level of A·− slightly blunted as to achieve a similar level to baseline point control after 30 minutes. In phase II the exerciseinduced increase in A·− was subsequently shown to be 48% greater (30min as opposed to the 2min post-cannulation resting baseline)(1754 ± 361 vs. 1979 ± 375 AU, P <0.05). Our findings demonstrate and confirm that venous cannulation per se stimulates iv the systemic formation of free radicals as an acute phase response which peaks at 10min and require approximately 15min to normalise. This has important interpretive implications for future studies that employ catheterisation. The final Study examined if the combination of exercise and inspiratory hypoxia would further compound regional tissue de-oxygenation that is frequently encountered during the ischaemic phase of surgery and thus, by consequence increase oxidative stress. The aim of the study was to further understand a potential relationship between oxidative stress and alterations in muscle oxygenation. Clear significant increases in the plasma concentration of A·− were detected in the peripheral blood of patients (normoxia(baseline) vs 6 data points of reperfusion after 5min of global ischaemic condition, P<0.05),(baseline vs immediate after ischaemia; 2337±525 vs 2633±508, AU, respectively). During global ischaemia the regional muscle oxygenation significantly decreased (↓∆O2Hb-oxyhaemoglobin), ↑∆HHb- deoxyhaemoglobin ), although increased regional blood volume (↑∆tHb- total haemoglobin). From the end of global ischaemia to 10 min after the regional muscle oxygenation progressively back to the start data point (↓∆HHb, ↑∆O2Hb). This study demonstrates for the first time that the I-R has got a big influence on the muscle oxygenation to increased ROS and the return of values towards baseline period in reperfusion stage appears to coincide with increased oxidative stress. Moreover, the present study has also demonstrated increased A·− level as early as the ischaemic phase of experiment independent of perioperative changes in the partial pressure of oxygen (pO2), elucidate a potentially important role for oxidative stress in provoking an appropriate vasodilation (NO-bioavailability) during the I-R period. This work demonstrates that; - Ascorbate is an antioxidant that can scavenge tissue and blood borne free radical, is essential in controlled amounts and is capable of initiating protective adaptation in the face of oxidative stress for the maintenance of physiological homeostasis. - Reperfusion is always associated with a sudden and massive release of ascorbate free radicals, with a maximal liberation within the first minutes of reperfusion. Vitamin C tended to reduce infarct size and protects from oxidative damage during global ischaemia and reperfusion. - The venous cannulation alone is enough per se stimulates the systemic formation of free radicals as a acute phase response. If this baseline artefact is not taken into account, the true magnitude of the exercise-induced oxidative stress response will be under-estimated. / The I-R has got a major influence on the muscle oxygenation to increased ROS and the return of values towards baseline period in reperfusion stage appears to coincide with increased oxidative stress. Using the state-of-the-art molecular techniques that include Electron Paramagnetic Spectroscopy (EPR) for the direct detection of free radicals and Near Infrared Spectroscopy (NIRS) for the direct detection of muscle oxygenation these studies have attempted to translate the basic mechanisms associated with free radical formation during I-R and have provided unique insight into the basic mechanisms responsible for the oxidative stress with the ultimate objective of developing novel antioxidant interventions that can provide effective prophylaxis.
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Optogenetic investigation of the neural network underlying the oxygen modulation of C. elegans locomotionSoltesz, Zoltan January 2014 (has links)
No description available.
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Mitochondrial respiratory transportation is the key determinant of aging in Caenorhabditis elegansFeng, Jinliu, 1974- January 2001 (has links)
No description available.
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Oxidative stress and antioxidant intake in HIV-related wastingCallow, Lisa Jane. January 2000 (has links)
No description available.
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The influence of temperature, salinity, and dissolved oxygen on juvenile salmon distributions in a nearshore estuarine environmentMesa, Kathryn A. January 1985 (has links)
This study examines the effects of a low oxygen environment, in concert with fluctuating temperature and salinity conditions, on the nearshore depth distributions (0-1 m) and flood tide movements of juvenile chinook (Oncorhynchus tshawytscha) and chum (O. keta) salmon. Comparisons are made between an unpolluted and a sewage polluted estuarine intertidal flat in the Fraser River estuary, British Columbia, the polluted area being characterized by the regular occurrence of low dissolved oxygen levels. Results are based on 380 beach seine samples taken between April and June of 1984.
In general, chum and chinook salmon of increasing length were captured in increasing depths, though this pattern was modified by seasonal changes in water temperature. Low dissolved oxygen conditions in deeper waters may have been responsible for the presence of larger, and often sluggishly swimming fish in higher oxygenated surface water layers or in shallow waters near the shore. In both areas, the risk of aerial predation was high.
On a flood tide, the likelihood of capturing a chinook salmon was reduced as temperatures increased and oxygen levels decreased. A combination of avoidance behaviour and a
regularity in the movement patterns of chinook onto the study area in the later stages of the flood tide may account for their rare occurrence in low oxygen concentrations (<6 mg/1) and high temperatures (>20 °C). Fish mortalities were most likely to occur on the ebb tide when fish were forced into waters of low oxygen content by the drainage patterns characteristic of the polluted study area. Though wide ranges in salinity were recorded on both tidal flats, this factor was not strongly correlated to Chinook distributions. However, significantly higher salinity levels in the unpolluted area may account for the greater numbers of chum salmon captured there. An understanding of the influence of estuarine water quality conditions on the distribution of juvenile salmonids may assist in the identification of significant sources of mortality in their early marine life. This knowledge is particularly important in the evaluation of water quality changes as caused by human activity. / Science, Faculty of / Zoology, Department of / Graduate
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Responses of Selected Texas Fishes to Abiotic Factors, and an Evaluation of the Mechanisms Controlling Thermal Tolerance of the Sheepshead MinnowBennett, Wayne A. (Wayne Arden) 05 1900 (has links)
Low oxygen tolerances of ten fishes were estimated using an original nitrogen cascade design, and reciprocally transformed to express responses as ventilated volume necessary to satisfy minimal oxygen demand (L·mg O2^-1). Values ranged from 0.52 to 5.64 L·mg^-1 and were partitioned into three statistically distinct groups. Eight stream fishes showed moderately high tolerances reflecting metabolic adaptations associated with stream intermittency. Juvenile longear sunfish and two mollies comprised the second group. High tolerance of hypoxia may allow juvenile sunfish to avoid predation, and mollies to survive harsh environmental oxygen regimens. The sheepshead minnow was the most tolerant species of low oxygen, of those examined, explaining its presence in severely hypoxic environments.
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Effects of Reactive Oxygen Species on Life History Traits of Caenorhabditis elegansSmith, Samson William 01 January 2012 (has links)
Evolutionary life history theory predicts that tradeoffs among fitness-related phenotypes will occur as a result of resource limitations and/or physiological constraints. Such tradeoffs are defined as the cost(s) incurred on one component of fitness (e.g., reproduction) by the increased expression of another fitness-related trait (e.g., longevity). Only recently have researchers begun to investigate the mechanistic bases of life history tradeoffs. A recent proposal is that reactive oxygen species (ROS) have a central role in shaping life history traits and tradeoffs. Research on disparate animal taxa has highlighted strong correlations between oxidative stress resistance and fitness-related life history traits, for example. Here, I use the model organism Caenorhabditis elegans to test several hypotheses concerning the effects of ROS on life history traits and the manifestation of life history tradeoffs. Additionally, I use heat stress and an alternate food source to explore the responses of life history traits to other forms of physiological stress. Relative fitness and other traits related to reproduction were found to be affected in mostly negative ways by increasing oxidative insult. Lifespan was surprisingly unaffected by oxidative stress, but was modified by temperature. In vivo ROS levels as measured by fluorescent microscopy reveal a tradeoff between antioxidant production and reproduction in this species.
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Men and women in hypoxia : the influence of tissue oxygenation on repeated-sprint abilitySmith, Kurt, University of Lethbridge. Faculty of Arts and Science January 2010 (has links)
This thesis examined the impact of oxygen (O2) availability on prefrontal cortex and muscle tissue oxygenation during repeated-sprint exercise (RSE) in men and women. Men and women matched for initial-sprint mechanical work performed during ten, 10-s sprints (30s of rest) in normoxia (21% FIO2) and acute hypoxia (13% FIO2). Mechanical work and arterial O2-saturation (SPO2) were obtained for every sprint. Oxy- and deoxygenated haemoglobin concentrations (O2Hb, HHb) were obtained via near-infrared spectroscopy. Hypoxia elicited lower SPO2 and work (14.8% & 7.4%, P < 0.05), larger (45.1%, P < 0.05) and earlier reductions in cortical oxygenation, and no differences between sexes. Cortical de-oxygenation and work decrement were strongly correlated (R2=0.85, P < 0.05). Muscle de-oxygenation was greater in men than women (67.3%, P < 0.05). These results show that O2 availability influences cortical oxygenation and performance equally in men and women, and suggest a more efficient muscle O2 uptake in women. / ix, 108 leaves : ill. ; 29 cm
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Evaluation of oxygen uptake and heart rate responses during rest and exercise in Chinese adolescents =: 華裔靑少年於休息及運動狀態中的氧攝氧量及心率反應之評量. / 華裔靑少年於休息及運動狀態中的氧攝氧量及心率反應之評量 / VO₂ and HR responses / Evaluation of oxygen uptake and heart rate responses during rest and exercise in Chinese adolescents =: Hua yi qing shao nian yu xiu xi ji yun dong zhuang tai zhong de yang she yang liang ji xin lü fan ying zhi ping liang. / Hua yi qing shao nian yu xiu xi ji yun dong zhuang tai zhong de yang she yang liang ji xin lü fan ying zhi ping liangJanuary 2001 (has links)
Chan Wan-Sze. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 64-76). / Text in English; abstracts in English and Chinese. / Chan Wan-Sze. / Table of Content --- p.II / List of Table --- p.VI / List of Figure --- p.VII / Abstract --- p.1 / Chapter Chapter 1 --- Introduction --- p.4 / Chapter 1.1 --- Background of study --- p.4 / Chapter 1.1.1 --- Oxygen consumption and heart rate --- p.4 / Chapter 1.1.2 --- V02Rand HRR --- p.7 / Chapter 1.1.3 --- Application of the association between V02 and HR- --- p.8 / Chapter 1.2 --- Purpose and significance --- p.9 / Chapter 1.3 --- Operational Definitions --- p.10 / Chapter 1.4 --- Hypotheses --- p.11 / Chapter Chapter 2 --- Review of Literature --- p.12 / Chapter 2.1 --- Exercise prescription for Cardiorespiratory Fitness --- p.14 / Chapter 2.1.1 --- Mode of exercise --- p.14 / Chapter 2.1.2 --- Frequency --- p.15 / Chapter 2.1.3 --- Duration --- p.16 / Chapter 2.1.4 --- Intensity --- p.17 / Chapter 2.2 --- Measurement of exercise intensity --- p.19 / Chapter 2.2.1 --- Rate of Perceived Exertion (RPE) --- p.21 / Chapter 2.2.2 --- Heart Rate (HR) --- p.23 / Chapter 2.3 --- Children and exercise intensity --- p.29 / Chapter 2.4 --- Summary --- p.30 / Chapter Chapter 3 --- Methodology --- p.32 / Chapter 3.1 --- Participants --- p.32 / Chapter 3.2 --- Procedures --- p.32 / Chapter 3.2.1 --- Resting measures --- p.34 / Chapter 3.2.2 --- Peak oxygen uptake (V02peak) testing --- p.34 / Chapter 3.3 --- Equipment and instrumentation --- p.36 / Chapter 3.3.1 --- Body mass and height --- p.36 / Chapter 3.3.2 --- Expired gases collection --- p.37 / Chapter 3.3.3 --- Heart rate measurement --- p.38 / Chapter 3.3.4 --- Determination of oxygen uptake (V02peak) and heart rate (HR) values --- p.38 / Chapter 3.4 --- Statistical Analysis --- p.39 / Chapter Chapter 4 --- Results --- p.41 / Chapter 4.1 --- Descriptive statistics --- p.41 / Chapter 4.2 --- Resting and maximal measures --- p.43 / Chapter 4.3 --- Submaximal exercise --- p.45 / Chapter 4.3.1 --- Phase I: %HRmax versus %V02peak --- p.47 / Chapter 4.3.2 --- Phase II: %HRR versus %V02peak --- p.48 / Chapter 4.3.3 --- Phase III: %HRR versus %V02R --- p.50 / Chapter 4.4 --- Summary of results --- p.53 / Chapter Chapter 5 --- Discussion and Conclusion --- p.54 / Chapter 5.1 --- Phase I: Preliminary findings from basic measures --- p.54 / Chapter 5.2 --- Phase II: %HRmax versus %V02peak --- p.57 / Chapter 5.3 --- Phase III: %HRR versus %V02peak --- p.58 / Chapter 5.4 --- Phase IV: %HRR versus %V02R --- p.60 / Chapter 5.5 --- Limitations of the study --- p.62 / Chapter 5.6 --- Conclusion and implications --- p.62 / Chapter 5.7 --- Recommendations and further studies --- p.63 / References --- p.64 / Appendix A --- p.77 / Appendix B --- p.79 / Appendix C --- p.80 / Appendix D --- p.81 / Appendix E --- p.82 / Appendix F --- p.83
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Bone Metabolism: The Role of STAT3 and Reactive Oxygen SpeciesNewnum, America Bethanne 14 August 2013 (has links)
Indiana University-Purdue University Indianapolis (IUPUI) / Signal Transducers and Activators of Transcription 3 (STAT3), a transcription factor expressed in many cell types, including osteoblasts and osteoclasts, is emerging as a key regulator of bone mass and strength. STAT3 mutations cause a rare human immunodeficiency disease characterized by extremely elevated levels of IgE in serum that have associated craniofacial and skeletal features, such as reduced bone mineral density and recurrent pathological fractures. Our microarray data and immunohistochemical staining using a normal rat model have shown that STAT3 mRNA and protein levels markedly increase in response to mechanical loading. In addition, as indicated by STAT3 phosphorylation in MC3T3-E1 osteoblastic cells, STAT3 activity significantly increases in response to 30 to 90 minutes fluid shear stress. In order to further study the role that STAT3 plays in bone responsiveness to loading, tissue-selective STAT3 knockout (KO) mice, in which inactivation of STAT3 occurs in osteoblasts, were generated by breeding the transgenic mice in which Cre recombinase cDNA was cloned downstream of a 3.6 or 2.3 kb fragment of the rat Col1a1 promoter (Col3.6-Cre and Col2.3-Cre, respectively) with a strain of floxed mice in which the two loxP sites flank exons 18-20 of the STAT3 gene were used. Mice engineered with bone selective inactivation of STAT3 in osteoblasts exhibited significantly lower bone mineral density (7-12%, p<0.05) and reduced ultimate force (21-34%, p<0.01) compared to their age-matched littermate controls. The right ulnae of 16-week-old bone specific STAT3 KO mice and the age-matched control mice were loaded with peak forces of 2.5 N and 2.75 N for female and male mice, respectively, at 2 Hz, 120 cycles/day for 3 consecutive days. Mice with inactivation of STAT3 specific in bone were significantly less responsive to mechanical loading than the control mice as indicated by decreased relative mineralizing surface (rMS/BS, 47-59%, p<0.05) and relative bone formation rate (rBFR/BS, 64-75%, p<0.001). Bone responsiveness was equally decreased in mice in which STAT3 is inactivated either in early osteoblasts (Col3.6-Cre) or in mature osteoblasts (Col2.3-Cre).
Accumulating evidence indicates that bone metabolism is significantly affected by activities in mitochondria. For instance, although STAT3 is reported to be involved in bone formation and resorption through regulation of nuclear genes, inactivation of STAT3 is shown to disrupt mitochondrial activities and result in an increased level of reactive oxygen species (ROS). Inactivation of STAT3 suppressed load-driven mitochondrial activity, which led to an elevated level of ROS in cultured primary osteoblasts. Oxidative stress induced by administration of buthionine sulfoximine (BSO) significantly inhibits load-induced bone formation in wild type mice. Taken together, the results support the notion that the loss-of-function mutation of STAT3 in osteoblasts and osteocytes diminishes load-driven bone formation and impairs the regulation of oxidative stress in mitochondria.
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