Tesis sobre el tema "Blood proteins Physiology"

Ocular neovascularization (NV) associated with corneal NV, ischemic retinopathies and age-related macular degeneration is a leading cause of severe vision loss. While numerous contributing factors have been identified, the potential role of the CD36 scavenger receptor has been largely overlooked notwithstanding its crucial involvement in normal retinal function. Accordingly, the central aim of this work was to elucidate the contribution and regulation of CD36 during ocular NV using the cornea as a model.<br>Initial work investigating the role of CD36 10 maintaining corneal avascularity, an important feature of the normal cornea, revealed that genetic ablation of CD36 elicits age-related corneal NV. Subsequent studies using a pathophysiologically relevant model of inflammatory corneal NV showed constitutive expression of CD36 in the normal cornea with marked induction in the neovascularized cornea. Importantly, activation of CD36 suppressed and induced regression of corneal NV, effects that proceeded via concerted inhibition of VEGFA, JNK-1, and cJun.<br>Because hypoxia is a fundamental stimulus for angiogenesis, it was pertinent to explore the role and regulation of CD36 during hypoxia. We demonstrate that CD36 expression was significantly elevated in hypoxia-exposed corneal and retinal tissue and in hypoxic retinal pigment epithelial cells. Essential contributions of hypoxia-inducible factor (HIF)-1 and reactive oxygen species were also established. Functional consequences were depicted by augmentations in CD36 phagocytic and anti-angiogenic activities.<br>Collectively, data disclose CD36 as an important modulator of corneal avascularity and inflammatory corneal NV; this imparts several interesting avenues for future research on the involvement of CD36 in neovascular diseases of the eye. Novel data further identify CD36 as a hypoxia and HIF-1 regulated gene thus creating a framework for future elucidation of the regulatory aspects of this receptor.

Liver kinase B1 (LKB1), a serine/threonine kinase, is responsible for the activation of AMP-activated protein kinase (AMPK), the master regulator of energy metabolism. LKB1/AMPK signaling pathway possesses a wide range of biological functions in regulating cell cycle progression, cell polarity, senescence and inflammation. In cultured endothelial cells, the pro-senescence function of LKB1/AMPK signaling pathway has been observed. However, the mechanisms by which LKB1 is regulated in endothelial cells remain largely uncharacterized. Furthermore, little is known about the effects of activated endothelial LKB1/AMPK signaling pathway on vascular and energy homeostasis. The present study aimed to investigate the upstream molecular mechanisms regulating LKB1 protein stability during endothelial senescence and the downstream pathophysiological effects of hyperactivated AMPK signaling in endothelial cells. In cultured model of cellular senescence, the lysine (K) 64 residue of LKB1 was found to be crucial for mediating its pro-senescence activities. The protein stability and intracellular localization of LKB1 mutant with K64 replaced by arginine (R) was different from the wild type protein. K64R exhibited enhanced effects on promoting endothelial senescence. Moreover, mutation of this residue attenuated the binding to HERC2, a newly identified E3 ubiquitin ligase for LKB1, in turn preventing its ubiquitination and degradation. Using a transgenic mouse model that selectively over-expresses a constitutively active AMPK α1 subunit (EC-AMPK) in endothelial cells, the influence of hyperactivated AMPK signaling on metabolic and vascular functions was investigated. Under standard chow condition, the metabolic phenotypes were similar between wild type and EC-AMPK mice; under high fat diet condition, EC-AMPK mice showed more severe obesity-induced fatty liver injury. Selective activation of AMPK in endothelial cells caused vascular and hepatic inflammation. Cyclooxygenase-2 (COX-2) was found to be the mediator for the pro-inflammatory functions of AMPK in vascular endothelial cells and facilitated to the development of obesity-induced fatty liver injury in EC-AMPK mice. Evaluation using isolated arteries revealed an increased systolic blood pressure and abnormal endothelial function in EC-AMPK miceunder high fat diet. AMPK activation in endothelium of the blood vessel could not block vascular remodeling associated with dietary obesity. Taken in conjunction, the above findings suggest that continuous activation of LKB1/AMPK signaling elicits adverse effects on both energy and vascular homeostasis.<br>published_or_final_version<br>Pharmacology and Pharmacy<br>Doctoral<br>Doctor of Philosophy

Traumatic brain injury (TBI) is a leading cause of disability worldwide. Annually, 150 to 200/1,000,000 people become disabled as a result of brain trauma. Axonal degeneration is a critical, early event following TBI of all severities but whether axon degeneration is a driver of TBI remains unclear. Molecular pathways underlying the pathology of TBI have not been defined and there is no efficacious treatment for TBI. Despite this significant societal impact, surprisingly little is known about the molecular mechanisms that actively drive axon degeneration in any context and particularly following TBI. Although severe brain injury may cause immediate disruption of axons (primary axotomy), it is now recognized that the most frequent form of traumatic axonal injury (TAI) is mediated by a cascade of events that ultimately result in secondary axonal disconnection (secondary axotomy) within hours to days. Proposed mechanisms include immediate post-traumatic cytoskeletal destabilization as a direct result of mechanical breakage of microtubules, as well as catastrophic local calcium dysregulation resulting in microtubule depolymerization, impaired axonal transport, unmitigated accumulation of cargoes, local axonal swelling, and finally disconnection. The portion of the axon that is distal to the axotomy site remains initially morphologically intact. However, it undergoes sudden rapid fragmentation along its full distal length ~72 h after the original axotomy, a process termed Wallerian degeneration. Remarkably, mice mutant for the Wallerian degeneration slow (Wlds) protein exhibit ~tenfold (for 2–3 weeks) suppressed Wallerian degeneration. Yet, pharmacological replication of the Wlds mechanism has proven difficult. Further, no one has studied whether Wlds protects from TAI. Lastly, owing to Wlds presumed gain-of-function and its absence in wild-type animals, direct evidence in support of a putative endogenous axon death signaling pathway is lacking, which is critical to identify original treatment targets and the development of viable therapeutic approaches. Novel insight into the pathophysiology of Wallerian degeneration was gained by the discovery that mutant Drosophila flies lacking dSarm (sterile a/Armadillo/Toll-Interleukin receptor homology domain protein) cell-autonomously recapitulated the Wlds phenotype. The pro-degenerative function of the dSarm gene (and its mouse homolog Sarm1) is widespread in mammals as shown by in vitro protection of superior cervical ganglion, dorsal root ganglion, and cortical neuron axons, as well as remarkable in-vivo long-term survival (>2 weeks) of transected sciatic mouse Sarm1 null axons. Although the molecular mechanism of function remains to be clarified, its discovery provides direct evidence that Sarm1 is the first endogenous gene required for Wallerian degeneration, driving a highly conserved genetic axon death program. The central goals of this thesis were to determine (1) whether post-traumatic axonal integrity is preserved in mice lacking Sarm1, and (2) whether loss of Sarm1 is associated with improved functional outcome after TBI. I show that mice lacking the mouse Toll receptor adaptor Sarm1 gene demonstrate multiple improved TBI-associated phenotypes after injury in a closed-head mild TBI model. Sarm1-/- mice developed fewer beta amyloid precursor protein (βAPP) aggregates in axons of the corpus callosum after TBI as compared to Sarm1+/+ mice. Furthermore, mice lacking Sarm1 had reduced plasma concentrations of the phosphorylated axonal neurofilament subunit H, indicating that axonal integrity is maintained after TBI. Strikingly, whereas wild type mice exhibited a number of behavioral deficits after TBI, I observed a strong, early preservation of neurological function in Sarm1-/- animals. Finally, using in vivo proton magnetic resonance spectroscopy, I found tissue signatures consistent with substantially preserved neuronal energy metabolism in Sarm1-/- mice compared to controls immediately following TBI. My results indicate that the Sarm1-mediated prodegenerative pathway promotes pathogenesis in TBI and suggest that anti-Sarm1 therapeutics are a viable approach for preserving neurological function after TBI.

Chemerin is a chemoattractant adipokine that regulates adipogenesis and may induce insulin resistance. Chemerin serum concentrations are elevated in obese, insulin-resistant, and inflammatory states in vivo. Here we investigate the role of omental (OM) and subcutaneous (SC) adipose tissue chemerin and CMKLR1 messenger RNA (mRNA) expression in human obesity. In addition, we test the hypothesis that changes in chemerin serum concentrations are primarily associated with reduced body fat mass in the context of 3 weight loss intervention studies. Chemerin serum concentration was measured in 740 individuals in a cross-sectional (n = 629) study including a subgroup (n = 161) for which OM and SC chemerin mRNA expression has been analyzed as well as in 3 interventions including 12 weeks of exercise (n = 60), 6 months of calorie-restricted diet (n = 19) studies, and 12 months after bariatric surgery (n = 32). Chemerin mRNA is significantly higher expressed in adipose tissue of patients with type 2 diabetes mellitus and correlates with circulating chemerin, body mass index (BMI), percentage body fat, C-reactive protein, homeostasis model assessment of insulin resistance, and glucose infusion rate in euglycemic-hyperinsulinemic clamps. CMKLR1 mRNA expression was not significantly different between the 2 fat depots. Obesity surgery–induced weight loss causes a significant reduction on both OM and SC chemerin expression. All interventions led to significantly reduced chemerin serum concentrations. Decreased chemerin serum concentrations significantly correlate with improved glucose infusion rate and reduced C-reactive protein levels independently of changes in BMI. Insulin resistance and inflammation are BMI-independent predictors of elevated chemerin serum concentrations. Reduced chemerin expression and serum concentration may contribute to improved insulin sensitivity and subclinical inflammation beyond significant weight loss.

An injured vascular system has a substantial impact on an individuals overall health, and an understanding of the mechanisms that underlie blood vessel pathophysiology is required for the development of rational and effective treatment strategies. The phenotypic modulation of smooth muscle cells (SMC) during vascular injury, characterized by altered adhesion, migration and synthetic behavior, plays an important role in the eventual outcome. Specifically, the ability of SMCs to adhere to and remodel their extracellular environment via regulation of the syndecan class of cell adhesion molecules dictates the response of the vascular wall to local injury. The effect of in vitro syndecan-4 regulation on SMC adhesion was investigated through the use of a glass microsphere centrifugation assay, and an antisense-mediated reduction in gene expression was found to correlate with decreased adhesive strength. Regulation of syndecan-1, syndecan-2, and syndecan-4 gene expression was observed experimentally by mechanical strain of SMCs. Using real-time polymerase chain reaction (PCR), the kinetics of both static and cyclic mechanical strain were found to modify the gene expression in a time and strain magnitude-dependent manner unique to each syndecan. In particular, the responses of syndecan-4 were acute, but transient, while the evolution of syndecan-1 and syndecan-2 regulation was delayed by comparison. Mechanical strain also modulated syndecan-4 protein expression and ectodomain shedding, as measured by Western immunoblotting, and this effect was found, through selective inhibition, to be at least in part dependent on mitogen-activated protein (MAP) kinase signaling. In particular, intact extracellular signal-regulated MAP kinase (ERK) 1/2 and c-Jun NH2-terminal kinase / stress-activated protein kinase (JNK/SAPK) signaling pathways were found to be required for the observed strain-induced shedding. These findings offer a better understanding of syndecan function in response to mechanical strain and suggest potential new mechanisms by which physical forces may modulate vascular SMC behavior and regulation during normal physiology, pathologic conditions, and engineered arterial substitute development.

Caracterisation des lipoproteines plasmatiques et de leurs apolipoproteines chez salmo gairdneri. Determination de leur masse moleculaire et de leur densite. L'etude au long du cycle annuel de reproduction a permis de demontrer la presence de proteines vitellines ovulaires dans le plasma. De plus, le role des lipoproteines, plasmatiques dans la steroidogenese ovarienne a ete etudie

Bibliography: leaves 216-239.<br>xiv, 246 leaves : ill. ; 30 cm.<br>The aim of this thesis is to examine the relationship between vascular reactivity, contractile proteins and blood pressure development in the spontaneously hypertensive rat (SHR). In addition, the influence of angiotensin II on blood pressure and vascular structure and function is investigated.<br>Thesis (Ph.D.)--University of Adelaide, Dept. of Clinical and Experimental Pharmacology, 1997

Includes bibliographical references (leaves 188-217) Insulin-like growth factor-I circulates at high concentrations in blood, mainly complexed with IGF-binding proteins. The main objective of the thesis is to determine the general role played by plasma IGF-binding proteins in the regulation of IGF transfer from blood to tissues.

xxiii, 222 leaves : ill. ; 30 cm.<br>Insulin-like growth factor-I circulates at high concentrations in blood, mainly complexed with IGF-binding proteins. The main objective of the thesis is to determine the general role played by plasma IGF-binding proteins in the regulation of IGF transfer from blood to tissues.<br>Thesis (Ph.D.)--University of Adelaide, Dept. of Animal Science, 1994

The primary aim of this dissertation was to examine the role of short-term immobilization on muscle function recovery, excitability of the muscle, evoked contractile properties of the muscle, and muscle proteins in the blood after contraction-induced muscle injury. In Studies I and II, the effects of four days of immobilization on recovery of muscle function and serum creatine kinase (CK) activity after eccentric exercise was examined in 26 males, who were placed into one of three groups: immobilization, control, or light exercise. When the damaged elbow flexor muscles were immobilized or exercised for four consecutive days, force recovery over eight days was significantly enhanced compared to a control. In Study II, during the four-day treatment period after eccentric exercise, immobilization resulted in a significant blunting of the CK response compared to the light exercise or control groups. However, increasing activity with light exercise did not have any effect on the CK response compared to control. The data from Study II suggested that reduced lymphatic transport with decreased muscular activity may have contributed to the lower CK response in the immobilized muscle. In Study III, mechanisms to explain the observations in Studies I and II with immobilization were undertaken. Muscle excitability and evoked contractile properties of the muscle were examined to determine whether immobilization altered the mechanical properties of the muscle to favor an enhanced force response. After eccentric exercise, there were immediate and prolonged reductions in the evoked contractile properties of the muscle. Immobilization, however, had no effect on these measures. CK and myoglobin were assessed during the four-day treatment period as well as during the five-day recovery period. There was a significant difference in the CK response between groups, with the immobilization demonstrating significant blunting of the CK response during the treatment period. Upon remobilization of the arm, CK activity increased but not as high as was anticipated. The myoglobin response, however, was not different between groups. Because their routes of entry into the blood differ, taken together, the myoglobin and CK response suggest that lymph transport likely contributed to the blunting of the CK response observed with immobilization.

AIMS/HYPOTHESIS: We previously demonstrated that animals fed a high-fat (HF) diet for 10 weeks developed insulin resistance and behavioural inflexibility. We hypothesised that intervention with metformin would diminish the HF-feeding-evoked cognitive deficit by improving insulin sensitivity. METHODS: Rats were trained in an operant-based matching and non-matching to position task (MTP/NMTP). Animals received an HF (45% of kJ as lard; n = 24), standard chow (SC; n = 16), HF + metformin (144 mg/kg in diet; n = 20) or SC + metformin (144 mg/kg in diet; n = 16) diet for 10 weeks before retesting. Body weight and plasma glucose, insulin and leptin were measured. Protein lysates from various brain areas were analysed for alterations in intracellular signalling or production of synaptic proteins. RESULTS: HF-fed animals developed insulin resistance and an impairment in switching task contingency from matching to non-matching paradigm. Metformin attenuated the insulin resistance and weight gain associated with HF feeding, but had no effect on performance in either MTP or NMTP tasks. No major alteration in proteins associated with insulin signalling or synaptic function was detected in response to HF diet in the hypothalamus, hippocampus, striatum or cortex. CONCLUSIONS/INTERPRETATION: Metformin prevented the metabolic but not cognitive alterations associated with HF feeding. The HF diet protocol did not change basal insulin signalling in the brain, suggesting that the brain did not develop insulin resistance. These findings indicate that HF diet has deleterious effects on neuronal function over and above those related to insulin resistance and suggest that weight loss may not be sufficient to reverse some damaging effects of poor diet.

Collection of previously published articles<br>Includes Allozyme electrophoresis / B.J. Richardson, P.R. Baverstock and M. Adams (1986)<br>Includes bibliographies<br>2 v. :<br>Title page, contents and abstract only. The complete thesis in print form is available from the University Library.<br>Thesis (D. Sc.)--University of Adelaide, 1988

<p>Obesity has ascended to become the primary modifiable cause of death in the United States. New evidence has called into question the utility of BMI – the typical index of obesity – in predicting cardiometabolic disturbances. The distribution of body fatness may be just as important as the total quantity. Intermuscular adipose tissue (IMAT) has emerged as a distinct subset of adipose in skeletal muscle that may be particularly metabolically deleterious. Typically, sections of either the calf or thigh are used as proxy measurements for whole-body IMAT in investigations. However, IMAT dispersion may not be consistent across tissues, instead infiltrating specific muscle or muscle compartments, and these have may have different metabolic consequences. The study described in Chapter 2 was designed to address this possibility and investigate and compare associations among thigh and calf IMAT stores with indices of cardiometabolic health. The strength of the relationship between IMAT and glucose control-related indices of cardiometabolic health was dependent upon anatomic location. Specifically, thigh IMAT is a better predictor of cardiometabolic risk that calf IMAT. </p> <p>Skeletal muscle has gained increased recognition in recent years for its importance in promotion of health and wellness throughout the life course. While treatment models addressing issues of declining muscle mass and strength with age previously focused on older adults, the importance of utilizing a life course model to promote skeletal muscle health at all ages was more recently recognized. There is consistent evidence that higher-protein diets modestly improve body composition. However, women are at greater risk for not meeting protein requirements and seem to be less willing to adopt strategies to achieve greater protein intake, such as protein supplementation, for fear that it may cause ‘bulkiness’. Therefore, the study described in Chapter 3 was designed to critically evaluate the effect of whey protein supplementation on body composition changes in women via a systematic review & meta-analysis of published randomized controlled trials. It was hypothesized that whey protein supplementation would moderately improve body composition but would not cause excessive muscle hypertrophy. Consistent with our hypothesis, whey protein supplementation improved body composition by modestly (<1%) increasing lean mass, without influencing fat mass.</p> <p>Dietary protein and skeletal muscle are conceptually inseparable; protein is often only considered in terms of how it impacts skeletal muscle-related outcomes. However, it is of interest to determine if the proposed beneficial effects of increased dietary protein consumption extend beyond skeletal muscle. Consumption of higher protein diets result in lower resting blood pressure, but the potential for protein to attenuate acute exercise blood pressure responses is unclear. The study described in Chapter 4 was designed to investigate the effects of meals with different amounts of protein on blood pressure responses to exercise in a randomized, cross-over trial. We hypothesized that consuming the higher-protein meal would attenuate the blood pressure responses to exercise and result in a more robust post-exercise hypotensive response. Contrary to our hypothesis, a higher-protein meal does not attenuate exercise-induced blood pressure responses compared to a lower-protein meal. These findings build upon previous research suggesting that the beneficial effect of chronically elevated protein intake on blood pressure is typically not observed in an acute setting by extending these findings to encompass blood pressure responses to acute responses to exercise.</p> <p>The three studies packaged herein utilize different techniques and report on different outcomes, but conceptual threads unite these works which augment the collective findings. Future researchers investigating the effects of protein on skeletal muscle anabolism can: 1) learn of the importance of proper reflection on surrogate measures and potential for anatomic-specific effects from the IMAT findings (Chapter 2), 2) appreciate the relevance of energy and training states in modulating responses from the WP meta-analysis (Chapter 3), and 3) recognize the importance of holistic approaches and employing challenges to reveal heterogeneity from the protein and BP trial (Chapter 4). Taken together, the research presented in this dissertation forwards our understanding of the relations and effects of dietary protein with different components of body composition on cardiometabolic health. </p>

Indiana University-Purdue University Indianapolis (IUPUI)<br>Factors released from coronary perivascular adipose tissue (PVAT), which surrounds large coronary arteries, have been implicated in the development of coronary disease. However, the precise contribution of coronary PVAT-derived factors to the initiation and progression of coronary vascular dysfunction remains ill defined. Accordingly, this investigation was designed to delineate the mechanisms by which PVAT-derived factors influence obesity-induced coronary smooth muscle dysfunction. Isometric tension studies of coronary arteries from lean and obese swine demonstrated that both lean and obese coronary PVAT attenuate vasodilation via inhibitory effects on smooth muscle K+ channels. Specifically, lean coronary PVAT attenuated KCa and KV7 channel-mediated dilation, whereas obese coronary PVAT impaired KATP channel-mediated dilation. Importantly, these effects were independent of alterations in underlying smooth muscle function in obese arteries. The PVAT-derived factor calpastatin impaired adenosine dilation in lean but not obese arteries, suggesting that alterations in specific factors may contribute to the development of smooth muscle dysfunction. Further studies tested the hypothesis that leptin, which is expressed in coronary PVAT and is upregulated in obesity, acts as an upstream mediator of coronary smooth muscle dysfunction. Long-term administration (3 day culture) of obese concentrations of leptin markedly altered the coronary artery proteome, favoring pathways associated with calcium signaling and cellular proliferation. Isometric tension studies demonstrated that short-term (30 min) exposure to leptin potentiated depolarization-induced contraction of coronary arteries and that this effect was augmented following longer-term leptin administration (3 days). Inhibition of Rho kinase reduced leptin-mediated increases in coronary artery contractions. Acute treatment was associated with increased Rho kinase activity, whereas longer-term exposure was associated with increases in Rho kinase protein abundance. Alterations in Rho kinase signaling were also associated with leptin-mediated increases in coronary vascular smooth muscle proliferation. These findings provide novel mechanistic evidence linking coronary PVAT with vascular dysfunction and further support a role for coronary PVAT in the pathogenesis of coronary disease.

Chemerin is a chemoattractant adipokine that regulates adipogenesis and may induce insulin resistance. Chemerin serum concentrations are elevated in obese, insulin-resistant, and inflammatory states in vivo. Here we investigate the role of omental (OM) and subcutaneous (SC) adipose tissue chemerin and CMKLR1 messenger RNA (mRNA) expression in human obesity. In addition, we test the hypothesis that changes in chemerin serum concentrations are primarily associated with reduced body fat mass in the context of 3 weight loss intervention studies. Chemerin serum concentration was measured in 740 individuals in a cross-sectional (n = 629) study including a subgroup (n = 161) for which OM and SC chemerin mRNA expression has been analyzed as well as in 3 interventions including 12 weeks of exercise (n = 60), 6 months of calorie-restricted diet (n = 19) studies, and 12 months after bariatric surgery (n = 32). Chemerin mRNA is significantly higher expressed in adipose tissue of patients with type 2 diabetes mellitus and correlates with circulating chemerin, body mass index (BMI), percentage body fat, C-reactive protein, homeostasis model assessment of insulin resistance, and glucose infusion rate in euglycemic-hyperinsulinemic clamps. CMKLR1 mRNA expression was not significantly different between the 2 fat depots. Obesity surgery–induced weight loss causes a significant reduction on both OM and SC chemerin expression. All interventions led to significantly reduced chemerin serum concentrations. Decreased chemerin serum concentrations significantly correlate with improved glucose infusion rate and reduced C-reactive protein levels independently of changes in BMI. Insulin resistance and inflammation are BMI-independent predictors of elevated chemerin serum concentrations. Reduced chemerin expression and serum concentration may contribute to improved insulin sensitivity and subclinical inflammation beyond significant weight loss.