Erythromelalgia is a neurovascular disorder which causes pain, swelling, erythema, and warmth of the distal extremities. Primary disease is due to a genetic mutation in the gene, but secondary erythromelalgia can be the consequence of a variety of underlying etiologies, including drug and toxin exposures. The disease is rare, occurring in only 1.3 out of every 100,000 in the United States, and symptoms can vary significantly in severity and presentation. Therefore, it can be difficult to recognize the disorder, identify the source, and promptly treat the condition. We report a reversible cause of erythromelalgia induced by the use of oral cyclosporine. This correlation is poorly documented in literature, with limited accounts identifying an association between erythromelalgia and cyclosporine. As drug-induced erythromelalgia represents a reversible cause of disease, physicians should obtain a detailed medication history during the diagnostic workup, specifically inquiring about the use of cyclosporine.

Peripheral arterial occlusive disease (PAOD) often presents as intermittent claudication, which may be caused by impaired vasodilation. Impairment of resistance vessels may contribute to the pathogenesis of PAOD, and explain the poor correlation between resting blood flow and limb function. Collateral function following arterial occlusion is not well defined, however collaterals and arterialized collateral capillaries (ACCs) in male and female animal models exhibit impaired vasodilation following arterial occlusion, which can potentially be improved with exercise training. Furthermore, resistance vessels in the ischemic tree and stem are likely involved in the pathogenesis of PAOD, however the relative importance of each is unknown. Therefore, we measured functional vasodilation in pre-existing collaterals, ACCs, the ischemic tree, and the stem region, 7 and 21-days following spinotrapezius feed artery ligation in male and female BALB/c mice, and with exercise therapy. Vasodilation in ACCs was more impaired in female mice than in males. Generally, vasodilation was impaired at day-7, likely due to impaired endothelium-dependent and smooth muscle-dependent vasodilation in maturing collaterals, and recovered by day-21. Exercise training appears to enhance collateral reactivity, more in ACCs in males than in females, suggesting that its therapeutic benefits are linked not only to structural adaptation but also to vessel functionality. Therefore, future research is required to determine the cause of sex differences in exercise therapy to treat peripheral arterial occlusive disease.

In birds like the Zebra finch (Taeniopygia guttata) the female, but not the male develop a brood patch upon incubation of eggs. The brood patch functions to increase heat exchange between the bird and the eggs. Development of the brood patch includes de-feathering, increased vascularization and edema formation. The increased vascularization is due to the development of arteriovenous anastomoses, AVA. The AVA are thermoregulatory vessels involved in cold induced vasodilation, CIVD, demonstrated to occur in the brood patch. Nitric oxide, NO, which is a well known vasodilator is a candidate substance for involvement in CIVD. In this study a NO-generating gel was applied to the brood patch of male and female zebra finches. Vasodilation was found to be markedly larger in females than in males. The larger vasodilation in the female brood patch is probably because NO vasodilate AVA selectively more than any other vessels. The study also investigated whether vasodilation would cause an increase in brood patch temperature. No definite changes in brood patch temperature could be observed and no conclusions could be drawn in the matter.

Chan Yau Chi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 157-177). / Abstracts in English and Chinese. / Contents / Declaration --- p.i / Acknowledgement --- p.ii / Abbreviations --- p.iii-iv / Abstract in English --- p.v-viii / Abstract in Chinese --- p.ix-xi / Contents --- p.xii-xvi / Chapter CHAPTER I - --- Introduction / Chapter 1.1. --- Selective Estrogen Receptor Modulators (SERMs) --- p.1 / Chapter 1.1.1. --- Raloxifene --- p.6 / Chapter 1.2. --- Mechanisms of Action of SERMs in Vascular System --- p.7 / Chapter 1.2.1. --- Estrogen --- p.7 / Chapter 1.2.2. --- Estrogen Receptors (ERs) --- p.8 / Chapter 1.2.3. --- General Mechanisms of Action of SERMs --- p.13 / Chapter 1.2.4. --- Actions of Raloxifene --- p.14 / Chapter 1.3. --- Effects of SERMs in Cardiovascular System --- p.14 / Chapter 1.3.1. --- Effects of SERMs on Endothelial Function --- p.15 / Chapter 1.3.2. --- Effects of SERMs on Vascular Smooth Muscle --- p.17 / Chapter 1.4. --- Effects of Raloxifene on Vascular Circulations --- p.18 / Chapter 1.4.1. --- Effects of Raloxifene on Systemic Circulation --- p.18 / Chapter 1.4.1.1. --- Preclinical Data --- p.18 / Chapter 1.4.1.1.1. --- Effects on Serum Lipids --- p.18 / Chapter 1.4.1.1.2. --- Effects on Inflammation Markers and Blood Coagulation --- p.19 / Chapter 1.4.1.1.3. --- Antioxidative Effects --- p.19 / Chapter 1.4.1.1.4. --- Effects on Nitric Oxide and Endothelial Function --- p.19 / Chapter 1.4.1.1.5. --- Effects on Vascular Smooth Muscle --- p.20 / Chapter 1.4.1.1.6. --- "Vascular Injury, Atherosclerosis and Ischaemia-Reperfusion Injury" --- p.20 / Chapter 1.4.1.2. --- Clinical Studies - Effects in Post-Menopausal Women --- p.21 / Chapter 1.4.1.2.1. --- "Effects on Serum Lipids, Lipoproteins and Triglycerides" --- p.21 / Chapter 1.4.1.2.2. --- Effects on Inflammation Markers and Homocysteine --- p.22 / Chapter 1.4.1.2.3. --- Effects on Coagulation Markers --- p.23 / Chapter 1.4.1.2.4. --- Effects on Endothelial Function --- p.23 / Chapter 1.4.1.2.5. --- Cardiovascular Events --- p.23 / Chapter 1.5. --- Myogenic Response and Vascular System --- p.24 / Chapter 1.5.1. --- Initiation and Development of Myogenic Response --- p.25 / Chapter 1.5.2. --- Regulation of Myogenic Response --- p.26 / Chapter 1.5.2.1. --- 20-hydroxyeicosatetraenoic acid (20-HETE) --- p.26 / Chapter 1.5.2.2. --- "Protein Kinase C, Rho/Rho-Kinase, and Tyrosine Kinase" --- p.27 / Chapter 1.5.3. --- Myogenic Response and Endothelium --- p.31 / Chapter 1.5.4. --- Estrogen and Myogenic Tone --- p.31 / Chapter 1.6. --- Objectives of the Present Study --- p.32 / Chapter CHAPTER II - --- Methods and Materials / Chapter 2.1. --- Tissue and Cell Preparation --- p.34 / Chapter 2.1.1. --- Vessel Preparation --- p.34 / Chapter 2.1.2. --- Removal of a Functional Endothelium --- p.36 / Chapter 2.2. --- Myograph and Pressure Myograph Setups --- p.36 / Chapter 2.2.1. --- Myograph 一 Isometric Tension Measurement --- p.36 / Chapter 2.2.2. --- Pressure Myograph - Isobaric Diameter Measurement --- p.37 / Chapter 2.3. --- Intracellular [Ca2+] Measurement in Vascular Smooth Muscle --- p.42 / Chapter 2.4. --- Chronic Raloxifene Therapyin Spontaneously Hypertensive Rats (SHRs) and Wistar-Kyoto Rats (WKYs) --- p.42 / Chapter 2.4.1. --- Surgical Procedure - Raloxifene Tubing Insertion --- p.42 / Chapter 2.4.2. --- "Body Weight, Mean Arterial Blood Pressure and Uterine Weight" --- p.42 / Chapter 2.4.3. --- Measurement of Raloxifene Tubing Consumption --- p.43 / Chapter 2.4.4. --- Effect of Chronic Raloxifene Treatment on Artery Reactivity --- p.43 / Chapter 2.5. --- Ovariectomy and Chronic Raloxifene Therapyin Syrian Golden Hamsters --- p.45 / Chapter 2.5.1. --- Surgical Procedure - Ovariectomy (OVX) --- p.45 / Chapter 2.5.2. --- Surgical Procedure - Raloxifene Tubing Insertion --- p.45 / Chapter 2.5.3. --- High-Cholesterol Food Preparation --- p.45 / Chapter 2.5.4. --- "Body Weight, Food Consumption and Uterine Weight" --- p.46 / Chapter 2.5.5. --- Measurement of Raloxifene Tubing Consumption --- p.46 / Chapter 2.5.6. --- Serum Lipid and Lipoprotein Determinations --- p.46 / Chapter 2.5.7. --- Effect of Chronic Raloxifene on Artery Reactivity --- p.46 / Chapter 2.6. --- Solutions and Drugs --- p.49 / Chapter 2.6.1. --- "Drugs, Chemicals and Enzymes" --- p.49 / Chapter 2.6.2. --- Solutions --- p.51 / Chapter 2.6.3. --- Diet Composition for Syrian Golden Hamsters --- p.51 / Chapter 2.7. --- Statistical Analysis --- p.52 / Chapter CHAPTER III - --- "Raloxifene Relaxes Rat Pulmonary Arteries and Veins: Roles of Gender, Endothelium, and Antagonism of Ca Influx" / Chapter 3.1. --- Abstract --- p.53 / Chapter 3.2. --- Introduction --- p.54 / Chapter 3.3. --- Methods and Materials --- p.55 / Chapter 3.3.1. --- Blood Vessel Preparation --- p.55 / Chapter 3.3.2. --- Protocols --- p.55 / Chapter 3.3.3. --- Measurement of Vascular Smooth Muscle [Ca2+]i --- p.56 / Chapter 3.3.4. --- Drugs --- p.57 / Chapter 3.3.5. --- Data Analysis --- p.53 / Chapter 3.4. --- Results --- p.58 / Chapter 3.4.1. --- Effects of Raloxifene on Pulmonary Arteries --- p.53 / Chapter 3.4.2. --- Effect of Raloxifene on CaCl2-induced Constrictionin Pulmonary Arteries --- p.59 / Chapter 3.4.3. --- Effects of Raloxifene on Pulmonary Veins --- p.59 / Chapter 3.4.4. --- Effect of Raloxifene on CaCl2-stimulated Increases in [Ca2+]i in Pulmonary Arteries --- p.60 / Chapter 3.5. --- Discussion --- p.67 / Chapter 3.6. --- Conclusion --- p.69 / Chapter CHAPTER IV - --- Raloxifene Modulates Pulmonary Vascular Reactivity in Spontaneously Hypertensive Rats / Chapter 4.1. --- Abstract --- p.70 / Chapter 4.2. --- Introduction --- p.71 / Chapter 4.3. --- Methods and Materials --- p.72 / Chapter 4.3.1. --- Raloxifene Treatment --- p.72 / Chapter 4.3.2. --- Blood Vessel Preparation --- p.72 / Chapter 4.3.3. --- Protocols --- p.73 / Chapter 4.3.4. --- Chemicals and Drugs --- p.73 / Chapter 4.3.5. --- Data Analysis --- p.74 / Chapter 4.4. --- Results --- p.74 / Chapter 4.4.1. --- Blood Pressure --- p.74 / Chapter 4.4.2. --- Vasocontraction in Spontaneously Hypertensive Rats --- p.75 / Chapter 4.4.3. --- Vasorelaxation in Spontaneously Hypertensive Rats --- p.75 / Chapter 4.4.4. --- Vasocontraction in Wistar-Kyoto rats --- p.76 / Chapter 4.4.5. --- Vasorelaxation in Wistar-Kyoto rats --- p.77 / Chapter 4.4.6. --- Comparison of contraction between WKY and SHR rats --- p.78 / Chapter 4.4.7. --- Comparison of relaxation between WKY and SHR rats --- p.78 / Chapter 4.5. --- Discussion --- p.93 / Chapter 4.6. --- Conclusion --- p.96 / Chapter CHAPTER V - --- Effects of Therapeutic Concentrations of Raloxifene in Pressurized Rat Small Mesenteric Artery / Chapter 5.1. --- Abstract --- p.98 / Chapter 5.2. --- Introduction --- p.99 / Chapter 5.3. --- Methods and Materials --- p.101 / Chapter 5.3.1. --- Blood Vessel Preparation --- p.101 / Chapter 5.3.2. --- Experimental Protocols --- p.102 / Chapter 5.3.2.1. --- Myogenic Tone Development --- p.102 / Chapter 5.3.2.2. --- Effects of Raloxifene and 17β-EstradioI on Myogenic Constriction --- p.102 / Chapter 5.3.2.3. --- Effects of Pharmacological Inhibitors on Raloxifene- or 17β-Estradiol-induced Myogenic Constriction --- p.103 / Chapter 5.3.3. --- Drugs and Solutions --- p.103 / Chapter 5.3.4. --- Expression of Results and Statistical Analysis --- p.104 / Chapter 5.4. --- Results --- p.104 / Chapter 5.4.1. --- Effects of Raloxifene and 17β-Estradiol on Rat Resistance Mesenteric Arteries1 --- p.104 / Chapter 5.4.2. --- Effects of Inhibitors of NOS --- p.105 / Chapter 5.4.3. --- Effect of CTX plus Apamin --- p.106 / Chapter 5.4.4. --- "Effect of ICI 182,780" --- p.106 / Chapter 5.4.5. --- "Effects of Wortmannin, LY 294002 and Cycloheximide" --- p.106 / Chapter 5.5. --- Discussion --- p.122 / Chapter 5.6. --- Conclusion --- p.125 / Chapter CHAPTER VI - --- Effects of Chronic Raloxifene Treatment on Vascular Reactivity in Pressurized Septal Coronary Arteries from Hamsters Fed with High-Cholesterol Diet / Chapter 6.1. --- Abstract --- p.127 / Chapter 6.2. --- Introduction --- p.128 / Chapter 6.3. --- Methods and Materials --- p.129 / Chapter 6.3.1. --- Preparatory Work --- p.129 / Chapter 6.3.1.1. --- Animals and Diets --- p.129 / Chapter 6.3.1.2. --- Preparation of High-Cholesterol (HC) Food --- p.129 / Chapter 6.3.1.3. --- Surgical Procedure - Ovariectomy (OVX) --- p.129 / Chapter 6.3.1.4. --- Surgical Procedure - Raloxifene Tubing Insertion --- p.130 / Chapter 6.3.1.5. --- Blood Vessel Preparation --- p.130 / Chapter 6.3.1.6. --- "Body Weight, Food Consumption and Uterine Weight" --- p.131 / Chapter 6.3.1.7. --- Measurement of Raloxifene Tubing Consumption --- p.131 / Chapter 6.3.1.8. --- Serum Lipid and Lipoprotein Determinations --- p.132 / Chapter 6.3.2. --- Experimental Protocols --- p.132 / Chapter 6.3.2.1. --- Development of Myogenic Tone --- p.132 / Chapter 6.3.2.2. --- Pressure-Diameter Relationships --- p.132 / Chapter 6.3.2.3. --- The Effect of Acetylcholine --- p.133 / Chapter 6.3.2.4. --- The Effect of U46619 --- p.133 / Chapter 6.3.2.5. --- The Effect of L-NAME --- p.133 / Chapter 6.3.3. --- Drugs and Solutions --- p.133 / Chapter 6.3.4. --- Expression of Results and Statistical Analysis --- p.134 / Chapter 6.4. --- Results --- p.135 / Chapter 6.4.1. --- Effects on Myogenic Response --- p.135 / Chapter 6.4.2. --- "Effects of Acetylcholine, U46619 and L-NAME" --- p.135 / Chapter 6.4.2.1. --- Comparison between OHHCD and OvxOHHCD --- p.135 / Chapter 6.4.2.2. --- Comparison between OvxOHHCD and OvxOHHCDRf --- p.135 / Chapter 6.4.2.3. --- Comparison between OHHCDRf and OvxOHHCDRf --- p.136 / Chapter 6.4.2.4. --- Comparison between OHHCD and OHHCDRf --- p.136 / Chapter 6.5. --- Discussion --- p.155 / Chapter 6.6. --- Conclusion --- p.156 / References --- p.157 / Publications --- p.176

Indivíduos portadores do glutamato na posição 27 do gene que codifica para o receptor beta2-adrenérgico têm resposta vasodilatadora muscular aumentada durante manobras fisiológicas. No entanto, o impacto do consumo agudo de gordura nessa resposta não é conhecido. Neste estudo, testou-se a hipótese de que o consumo gordura afetaria a resposta vasodilatadora aumentada destes indivíduos durante manobras fisiológicas. Vinte e cinco indivíduos saudáveis foram subdivididos em dois grupos: 11 homozigotos para o glutamato (Glu27Glu, 40+-3 anos; 65+-3kg) e 14 homozigotos para a glutamina (Gln27Gln, 40+-2 anos; 64+-2kg). O fluxo sangüíneo muscular foi medido por pletismografia de oclusão venosa. A resposta vasodilatadora muscular foi avaliada durante 3 minutos de exercício e estresse mental em jejum e 3 horas após consumo de 62 g de gordura. A condutância basal foi semelhante entre grupos (Glu27Glu=2,3+-0,1; Gln27Gln=2,2+-0,1; P=0,21). O aumento da condutância vascular durante exercício e durante o estresse mental foi maior no grupo Glu27Glu (0,73+-0,2 vs 0,22+-0,1; P=0,008 e 1,8?0,3 vs 1,2+-0,2; P=0,04, respectivamente). O consumo agudo de uma preparação rica em gordura eliminou esta diferença. A resposta de pressão arterial e freqüência cardíaca foi semelhante antes e após a ingestão de gordura. Os níveis de triglicérides, glicose e insulina foram semelhantes ao longo de todo período de estudo. O consumo agudo de gordura elimina a resposta aumentada do fluxo sangüíneo muscular durante manobras fisiológicas dos indivíduos portadores do genótipo Glu27Glu no receptorbeta2- adrenérgico. / Subjects who have glutamic acid at position 27 in gene encoding to beta2-adrenoceptor have increased muscle vasodilatory response during physiological maneuvers. However, the impact of a high-fat meal in this response is unknown. We tested the hypothesis that a high-fat meal would modify the increased muscle vascular reactivity during handgrip and mental stress in these subjects. Twenty-five healthy subjects were subdivided in two groups: 11 were homozygous to glutamic acid (Glu27Glu, 40?3 years; 65+-3kg) and 14 were homozygous to glutamine (Gln27Gln, 40+-2 years; 64+-2kg). Forearm blood flow was measured by venous occlusion pletysmography. Forearm blood flow was recorded for 3 minutes of handgrip and mental stress during fasting and three hours after 62g of fat consumption. Baseline forearm vascular conductance was similar between groups (Glu27Glu=2.3+-0.1; Gln27Gln=2.2+-0.1; P=0.21). Forearm vascular conductance during handgrip and mental stress was greater in the genotype Glu27Glu (0.73+-0.2 vs 0.22+-0.1; P=0.008 and 1.8+-0.3 vs 1.2+-0.2; P=0.04, respectively). Acute fat consumption eliminated the difference of vasodilatory response previously achieved. Blood pressure and heart rate response were similar before and after fat intake. Triglycerides, glucose and insulin levels were also similar between groups. We concluded that high-fat ingestion abolishes the augmented muscle blood flow responses during physiological maneuvers in individuals who are homozygous for the Glu27 allele of the beta2-adrenoceptor gene.

The relationship between the endocannabinoid system in the renal medulla and the long-term regulation of blood pressure is not well understood. To investigate the possible role of the endocannabinoid system in renomedullary interstitial cells, mouse medullary interstitial cells (MMICs) were obtained, cultured and characterized for their responses to treatment with a selective inhibitor of fatty acid amide hydrolase (FAAH), PF-3845. Treatment of MMICs with PF-3845 increased cytoplasmic lipid granules detected by Sudan Black B staining and multilamellar bodies identified by transmission electron microscopy. HPLC analyses of lipid extracts of MMIC culture medium revealed a 205nm-absorbing peak that showed responsiveness to PF-3845 treatment. The biologic activities of the PF-3845-induced product (PIP) isolated by HPLC were investigated in anesthetized, normotensive surgically-instrumented mice. Intramedullary and intravenous infusion of PIP at low dose rates (0.5-1 AU/10 min) stimulated diuresis and natriuresis, whereas at higher doses, these parameters returned toward baseline but mean arterial pressure (MAP) was lowered. Whereas intravenous bolus doses of PIP stimulated diuresis, GFR and medullary blood flow (MBF) and reduced or had no effect on MAP, an intraperitoneal bolus injection of PIP reduced MAP, increased MBF, and had no effect on urinary parameters. Genetic or pharmacological ablation of the cannabinoid type 1 receptors in mice completely abolished the diuretic and vasodepressor properties of intramedullary infused PIP, suggesting that the PF-3845-induced product requires the presence of CB1 receptors in order to elicit its renal effects. In a radioactive competition binding assay, using Chinese hamster ovary cells expressing CB1 receptors, PIP successfully displaced the CB1 selective inverse agonist [3H] SR141716A, revealing that the lipid extract was able to compete for binding to CB1 receptors. Finally, we investigated the tubular location of diuretic activity that the PF-3845-induced lipid fraction exhibits. In a renal function in vivo experiment, we pre-treated anesthetized mice with an intramedullary infusion of one of four well-known diuretics. This procedure was followed by an intramedullary infusion of PIP (1AU). Only inhibition of the proximal tubule sodium reabsorption diminished the diuretic activity of the PF-3845-induced product, suggesting that the lipid fraction requires a physiologically intact proximal tubular reabsorption mechanism for it to produce diuresis. These data support a model whereby PF-3845 treatment of MMICs results in increased secretion of a neutral lipid which acts directly to promote diuresis and natriuresis and indirectly through metabolites to produce vasodepression. Efforts to identify the structure of the PF-3845-induced lipid and its relationship to the previously proposed renomedullary antihypertensive lipids are ongoing.

The overall aim of this thesis was to combine optical methods into a system with the ability to simultaneously measure blood flow changes at different tissue depths. The goal of such a system was to reveal vascular mechanisms relevant to pressure ulcer etiology under clinically relevant conditions and in relation to the evaluation of pressure-redistribution support surfaces. This thesis consists of four quantitative, cross-sectional studies measuring blood flow responses before, during, and after pressure exposure of the sacral tissue. Two optical methods – photoplethysmography and laser Doppler flowmetry – were combined in a newly developed system that has the ability to discriminate blood flows at different tissue depths. Studies I and II explored blood flow responses at different depths in 17 individuals. In Study I the blood flow was related to tissue thickness and tissue compression during pressure exposure of ≥ 220 mmHg. In Study II, the sacral tissue was loaded with 37.5 mmHg and 50.0 mmHg, and the variation in blood flow was measured. Studies III and IV included 42 healthy individuals < 65 years, 38 healthy individuals ≥ 65 years, and 35 patients ≥ 65 years. Study III included between-subject comparisons of blood flow and pressure between individuals in the three study groups lying in supine positions on a standard hospital mattress. Study IV added within-subject comparisons while the individual was lying on four different types of mattress. The studies explored the vascular phenomena pressure-induced vasodilation (PIV) and reactive hyperemia (RH). The most common blood flow response to tissue exposure in this thesis was PIV, although a decrease in blood flow (a lack of PIV) was observed in some individuals. The patients tended to have higher interface pressure during pressure exposure than the healthy groups but no differences in blood flow responses were seen. Our results showed that pressure levels that are normally considered to be harmless could have a significant effect on the microcirculation in different tissue structures. Differences in individual blood flow responses in terms of PIV and RH were seen, and a larger proportion of individuals lacked these responses in the deeper tissue structures compared to more superficial tissue structures. This thesis identified PIV and RH that are important vascular mechanisms for pressure ulcer development and revealed for the first time that PIV and RH are present at different depths under clinically relevant conditions. The thesis also identified a population of individuals not previously identified who lack both PIV and RH and seem to be particularly vulnerable to pressure exposure. Further, this thesis has added a new perspective to the microcirculation in pressure ulcer etiology in terms of blood flow regulation and endothelial function that are anchored in clinically relevant studies. Finally, the evaluation of pressureredistribution support surfaces in terms of mean blood flow during and after tissue exposure was shown to be unfeasible, but the assessment of PIV and RH could provide a new possibility for measuring individual physiological responses that are known to be related to pressure ulcer development.

Thesis (Ph. D.)--University of Missouri--Columbia, 1999. / Typescript. Vita. Includes bibliographical references (leaves [174]-186). Also available on the Internet.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Nitric oxide (NO) is a reactive free radical, acting as a messenger molecule, mediating several functions, including vasodilation, platelet aggregation inhibition and neurotransmission. Because this radical to have a short half-life, its determination is difficult, therefore, measurement of metabolites nitrite/nitrate (NOx) is most often used to evaluate NO production. The Griess reaction is the most used method for NOx quantification due to its simplicity, speed and cost-effective. Due to the biological relevance of NO, it is extremely importance to its measurement. The objective of this study was to develop and validate an analytical method for the automated measurement of serum levels of NOx by the Griess method using the Cobas Mira clinical chemistry analyzer. This study provided the development of a protocol for the automated measurement of serum levels of NOx, which is linear method (r2= 0.993, P <0.001), precise, because it showed a coefficient of variation in intra-assay precision of 8.7% and inter-assay precision of 5.6%. Moreover, we observed a recovery of 114.6% which is considered accurate study. Thus, it was concluded that the automated method presented here is linear, precise, accurate, simple and low cost, likely to be adapted to Cobas Mira analyzer and other automated systems available in clinical routine laboratory. / O óxido nítrico (NO) é um radical livre reativo, agindo como uma molécula mensageira, mediando diversas funções, incluindo vasodilatação, inibição da agregação plaquetária e neurotransmissão. Pelo fato deste radical possuir uma meia-vida curta, a sua determinação torna-se difícil, consequentemente, a mensuração de seus metabólitos nitrito/nitrato (NOx) é mais frequentemente utilizado para avaliar a produção de NO. A reação de Griess é o metodo mais utilizado na quantificação do NOx, devido a sua simplicidade, rapidez e custo-benefício. Devido à relevância biológica do NO, é de suma importância a sua mensuração. Assim, o objetivo deste estudo foi desenvolver e validar um método analítico automatizado para a mensuração dos níveis séricos de NOx pelo método de Griess, utilizando o analizador Cobas Mira®. Este estudo propiciou o desenvolvimento de um protocolo automatizado para a quantificação dos níveis séricos de NOx, sendo este método linear (r2= 0,993, P<0,001), preciso, pois apresentou um coeficiente de variação na precisão intra-ensaio de 8,7% e na precisão inter-ensaio de 5,6%. Além disso, foi observada uma recuperação de 114,6%, sendo este estudo considerado exato. Dessa forma, foi possível concluir que o método automatizado apresentado é linear, preciso, exato, simples e de baixo custo, passível a ser adaptado ao analizador Cobas Mira® e outros sistemas automatizados disponíveis em laboratórios clínicos de rotina.

Submitted by Marlene Santos (marlene.bc.ufg@gmail.com) on 2018-08-14T17:49:15Z No. of bitstreams: 2 Tese- Jerônimo Raimundo de Oliveira Neto - 2017.pdf: 3024039 bytes, checksum: b381958f2c5d4a901e5c5cb791c482a7 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2018-08-16T11:36:48Z (GMT) No. of bitstreams: 2 Tese- Jerônimo Raimundo de Oliveira Neto - 2017.pdf: 3024039 bytes, checksum: b381958f2c5d4a901e5c5cb791c482a7 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-08-16T11:36:48Z (GMT). No. of bitstreams: 2 Tese- Jerônimo Raimundo de Oliveira Neto - 2017.pdf: 3024039 bytes, checksum: b381958f2c5d4a901e5c5cb791c482a7 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-05-05 / Beer is one of the oldest and most popular beverages consumed by mankind, the main classification of beers is the type of fermentation, divided into ale or lager, high and low fermentation, respectively. In this study, beer samples were divided into two portions, the first one for the spectrophotometric and electrochemical tests, which was used in natura form. And the second part was lyophilized, aiming at the concentration use standardization, alcohol withdrawal and guaranteeing an increase of durability, and then used in the pharmacological and chromatographic tests. The aim of this study was to investigate the antioxidant potential and vasodilator activity and correlate them with the phenolic profiles of twenty-two commercial beers. In addition, the antioxidant activity and the phenolic profile of hops and malts from different origins were also verified. From the raw materials, hops samples showed better antioxidant activity when compared to malt samples (p <0.05). The correlation between electrochemical index (EI) and total phenols (TPC), and radical scavenging methods, 1,1-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis (3-ethylbenzthiazoline -6-sulphonic acid) (ABTS) was 0.86, 0.77 and 0.85 respectively for the hop samples. From the beers, ale group showed better antioxidant activity and TPC values when compared to the lager group. However, according to the Pearson correlation matrix, the correlation between IE with TPC, DPPH and ABTS was 0.86, 0.89 and 0.96 respectively for the lager group. While for the ale group, the correlations were not statistically significant (p> 0.05), mainly due to the turbidity interference of the samples during spectrophotometric assays. This fact reiterates the use of electrochemical methods, which minimize this type of problem besides being more sensitive and quick. Following the proposed electrochemical methodology, using the IE calculation, LB10 and AB1 samples obtained the highest indices. In the results of the principal components analysis (PCA) three groups were observed, where group III confirms LB10 and AB1 as the best antioxidants potential. LB10, AB1 and AB6 showed a maximal vasodilator effect of 92 ± 4, 88 ± 3 and 79 ± 3%, respectively. Both the vasodilator effect and the lipid peroxidation inhibition capacity may be associated with the presence of phenolic compounds in the beer samples, identified by mass spectrometry. This study, therefore, verified that the proposed IE methodology is an important tool to evaluate antioxidant properties and showed a good correlation with radical scavenging assays. Moreover, the study showed that beers may have a beneficial effect on the cardiovascular system, showing a good correlation of the vasodilator effect and antioxidant potential, which may be useful for future research on health, sensorial properties and quality parameters. Still from the standpoint of the quality of the beers, hop extracts as well as malt exhibited statistically results, corroborating to the applicability of these tools in the choice of these major ingredients improve the quality of the final product. / A cerveja é uma das bebidas mais antigas e populares consumidas pela humanidade, a principal classificação das cervejas é quanto ao tipo de fermentação, dividindo-se em ale ou lager, alta e baixa fermentação, respectivamente. Neste estudo, as amostras de cerveja foram divididas em duas porções, a primeira para os ensaios espectrofotométricos e eletroquímicos, a qual foi utilizada de forma in natura. E a segunda parte foi liofilizada, para padronização da concentração de uso, retirada do álcool e garantir uma maior durabilidade, e então utilizada nos ensaios farmacológicos e cromatográficos. O objetivo deste estudo foi investigar o potencial antioxidante, a atividade vasodilatadora e correlacionar com os perfis fenólicos de vinte e duas cervejas comerciais. Adicionalmente foi também verificada a capacidade antioxidante e o perfil fenólico de lúpulos e maltes de diferentes origens. Das matérias-primas, as amostras de lúpulo mostraram uma melhor atividade antioxidante quando comparadas com as amostras de malte (p<0,05). A correlação entre o índice eletroquímico (IE) com os fenóis totais (TPC), e os ensaios de captura de radicais, 1,1- difenil-1-picril-hidrazil (DPPH) e 2,2’-azinobis (3-etilbenzotiazolina-6-ácido sulfônico) (ABTS) foi de 0,86, 0,77 e 0,85 respectivamente para as amostras de lúpulo. Enquanto que para as amostras de malte foi 0,49, 0,69 e 0,96, respectivamente. Análise de variância (ANOVA) foi utilizada para verificar diferenças dentro de cada grupo de matéria-prima, sendo observadas diferenças a nível de 95% de confiança (p<0,05). Das cervejas, o grupo ale mostrou melhores capacidade antioxidante e valores de fenóis totais quando comparado com o grupo lager. Porém, de acordo com a matriz de correlação de Pearson, a correlação entre o IE com TPC, DPPH e ABTS foi de 0,78, 0,82 e 0,89 respectivamente para o grupo lager. Enquanto que para o grupo ale as correlações não foram estatisticamente significativas (p>0,05), principalmente devido à interferência da turbidez das amostras durante ensaios espectrofotométricos. Este fato reitera o uso de métodos eletroquímicos, que minimizam este tipo de problema além de serem mais sensíveis e rápidos. Seguindo a metodologia eletroquímica proposta, através do cálculo do IE, as amostras LB10 e AB1 obtiveram os maiores índices. Nos resultados da análise de componentes principais foram observados três grupos, onde o grupo III confirma LB10 e AB1 como as de melhores potenciais antioxidantes. LB10, AB1 e AB6 mostraram efeito vasodilatador máximo de 92 ± 4, 88 ± 3 e 79 ± 3%, respectivamente. Tanto o efeito vasodilatador quanto a capacidade de inibição da peroxidação lipídica, podem estar associados à presença de compostos fenólicos nas cervejas, identificados via espectrometria de massa. Este estudo, portanto, verificou que a metodologia proposta do IE é uma importante ferramenta para avaliar as propriedades antioxidantes e mostrou uma boa correlação com ensaios de eliminação de radicais. Além disso, o estudo mostrou que as cervejas podem ter um efeito benéfico sobre o sistema cardiovascular, apresentando boa correlação do efeito vasodilatador e o potencial antioxidante, o que pode ser útil para pesquisas futuras sobre a saúde, propriedades sensoriais e parâmetros de qualidade. Ainda sobre o ponto de vista da qualidade das cervejas, os extratos de lúpulo, assim como de malte, apresentaram resultados estatisticamente diferentes entre si, corroborando para aplicabilidade destas ferramentas na escolha destes ingredientes majoritários em prol da qualidade do produto final.