Neste estudo, foram determinadas as atividades de duas enzimas de membrana: alanina-aminopeptidase (AAP) e Y-glutamil-transpeptidase (YGT) e da enzima lisossomal N-acetil-B-D-glucosaminidase (NAG), bem como os níveis de proteína total (PT), albumina (ALB) e ácido delta-aminolevulínico (ALA) em urinas de indivíduos expostos ocupacionalmente ao chumbo (grupo exposto) e de indivíduos não expostos ao chumbo e nem a outras substâncias químicas (grupo controle). Todos os indivíduos apresentavam creatinina sérica inferior a 1,5 mg/dL. Ao mesmo tempo foram determinados os níveis sangfiíneos de chumbo (Pb-S) e aferidas pressão arterial diastólica (PD) e pressa-o arterial sistólica (PS). O objetivo foi investigar a toxicidade renal crônica do chumbo e sua possível correlação com a pressão arterial em indivíduos com função renal normal. A mediana de chumbo no sangue no grupo controle foi de 11,5 pg/dL contra 36,8 pg/dL no grupo exposto, mostrando-se dependente do tempo de exposição (p < 0,001). O valor mediano da NAG-U corrigido pela creatinina foi mais alto no grupo exposto (31,74 U/g creatinina) e significativamente diferente do grupo controle (25,28 U/g creatinina), p < 0,001. A atividade NAG-U se correlacionou com o Pb-S (p < 0,001), tempo de exposição (p < 0,001) e ALA-U (p < 0,001), mas não houve correlação com a pressão arterial. Os demais indicadores de função renal, Y GT-U, AAP-U, PT-U e ALB-U não se mostraram mais elevados no grupo controle e nem mostraram correlação com o Pb-S, tempo de exposição ou pressão arterial. A pressão arterial diastólica foi maior no grupo exposto (83 mmHg contra 78 mmHg no grupo controle) mas não se observaram diferenças quanto à pressão sistólica. Observaram-se correlações das pressoes arteriais diastólica e sistólica com tempo de exposição (p < 0,001 e p < 0,05 ) e com chumbo no sangue (p < 0,05 e p < 0,05 ) respectivamente, enquanto somente a pressão diastólica se correlacionou com o ALA-U (p < 0,05). / In the present study we determined the activity of two renal tubule membrane enzymes: alanine-aminopeptidase (AAP) and Y-glutamyl-transpeptidase (YGT), and of the renal tubule lysosomal enzyme N-acetyl-B-D-glucosaminidase (NAG), as well as the levels of total protein (TP), albumin (ALB) and delta-aminolevulinic acid (ALA) in the urine of individuais occupationally exposed to lead (exposed group) and in individuais not exposed to lead or to any other chemical substances (control group). All individuais presented serum creatinine levels of less than 1.5 mg%. All subjects were also submitted to measurement of blood lead levels (B-Pb) and of diastolic and systolic arterial pressure. The objective was to investigate the chronic renal toxicity of lead and its possible correlation with arterial pressure in individuais with normal renal function. Median B-Pb levels were 11.5 pg/dL for the controls versus 36.8 Pg/dL for the exposed group, with the level being proportional to time o f exposure (p < 0.001). Median NAG-U values corrected for creatinine were higher in the exposed group (31.74 U/g creatinine) and significantly different from the control (25.28 U/g creatinine), p < 0.001. NAG-U activity was correlated with B-Pb (p < 0.001), time of exposure (p < 0.001) and ALA-U (p < O. 001), but not with arterial pressure. The remaining indicators of renal function, YGT-U, AAP-U, TP-U and ALB-U were not more elevated in the control group and were not correlated with B-Pb, time of exposure or arterial pressure. Diastolic arterial pressure was higher in the exposed group (83 mmHg versus 78 mmHg for the controls), but the two groups did not differ in systolic pressure. Diastolic and systolic pressure values were correlated with time of exposure (p < 0.001 and p < 0.05) and blood lead (p < 0,05 e 0,05) respectively but only diastolic pressure was correlated with ALA-U ( p < 0.05).

Lack of the enzyme 11β-hydroxysteroid dehydrogenase type 2 (11βHSD2) causes the syndrome of apparent mineralocorticoid excess (AME): low-renin hypertension, renal sodium (Na +) retention, hypokalaemic alkalosis and polyuria. This rare autosomal recessive disorder is observed in human kindreds carrying mutations in the HSD11B2 gene. Genetically modified mice, in which the homologue Hsd11b2 is rendered non-functional, have been used to study the pathogenesis of AME. Hitherto, data obtained from humans and mice have suggested that the physiological phenotype is a consequence of enhanced reabsorption of Na + through the epithelial sodium channel (ENaC) in the renal connecting tubule (CNT) and collecting duct. However, Hsd11b2 null mice exhibit epithelial hypertrophy in a different nephron segment, namely the distal convoluted tubule (DCT). The studies described herein aimed to characterise this structural phenotype and to examine the consequences for renal Na + reabsorption in AME. Hsd11b2 null mice exhibited hypertrophy and hyperplasia in the DCT, with an elevated rate of epithelial cell proliferation in this nephron segment at 60 days of age. Hsd11b2 null kidneys contained greater quantities of the thiazide-sensitive NaCl co-transporter (NCC), the dominant Na + transporter protein in the DCT. They also contained greater quantities of the phosphorylated forms of NCC that are associated with NaCl transport activity. Despite this, there was no increase in the proportion of filtered Na + that was reabsorbed in the DCT. This was assessed in anaesthetised mice, using clearance methodology to measure the thiazide-induced increment in the fractional excretion of Na + (FENa) during continuous ENaC blockade. Wild-type DCTs did not express 11βHSD2; therefore the structural and molecular changes were not a direct consequence of the loss of 11βHSD2 in affected cells. The discussion examines the likely mechanisms causing structural remodelling in the distal renal tubule of Hsd11b2 null kidneys and potential explanations for the dissociation between structural and functional phenotypes in the DCT. There are implications for our understanding of the cellular and molecular mechanisms underlying various renal phenomena including structural remodelling in the distal tubule, resolution of the ‘aldosterone paradox’ and escape from chronic aldosterone excess.

Sushi, von Willebrand factor type A, epidermal growth factor and pentraxin domain containing 1 (SVEP1) is an extracellular matrix protein which may bind to cell surface molecules such as integrins. A non-synonymous single amino acid polymorphism in the Svep1 gene is associated with a 14% increased risk of coronary heart disease, a 13% higher risk of type 2 diabetes and a 1mmHg increase in systolic blood pressure. Expression of the SVEP1 gene is increased in the kidney in the Cyp1a1mRen2 rat model of diabetes and hypertension previously developed in our lab. SVEP1 is also known to be upregulated in human diabetic nephropathy and is upregulated in rodent models of renal fibrosis. I hypothesized that Svep1 played a role in renal fibrosis, diabetes and blood pressure. Hence, the primary goal of this thesis was to investigate the role of SVEP1 and in the pathogenesis of diabetes, hypertension and renal fibrosis. Svep1 gene expression is increased in the kidney in the DOCA-salt-angII-uninephrectomy model of hypertension and following UUO. SVEP1 hemizygous mice showed no differences in expression of pro-fibrotic genes after UUO compared to wildtype littermates. No overt metabolic phenotype was exhibited by the Svep1 hemizygous mice, however there was a significant decrease in fat depot weights after high fat diet (HFD) and a significant increase in blood glucose concentrations during the glucose tolerance test at the 12 week time point in hemizygous Svep1 mice compared with wild-type controls. After telemetry analysis of blood pressure no difference was seen in blood pressure but SVEP1+/-animals had an increased heart rate of 100 beats per minute compared to wildtype animals. Svep1 expression is increased in the kidney in models of hypertension and fibrosis, however loss of one Svep1 allele did not alter the severity of fibrosis in the UUO model or significantly alter glucose tolerance after high fat diet. However, the high fat diet experiment was a pilot study and should be repeated with a larger number of animals. In addition, generation of a mouse with the human point mutation could determine the mechanisms by which this extracellular matrix protein confers risk of diabetes and hypertension.

Tight control of blood glucose and blood pressure (BP) reduces cardiovascular risk in early Type 1 diabetes mellitus (T1DM). Increased BP normally increases renal medullary perfusion and sodium excretion. This is called acute pressure natriuresis. Inadequate acute pressure natriuresis disrupts circadian regulation of BP, which predicts hypertension. The peptide, endothelin-1 (ET-1), regulates BP via ETA and ETB receptors. ETA receptor antagonists reduce BP and restore its circadian rhythm. Two hypotheses were investigated. First, that acute pressure natriuresis is impaired in early T1DM, prior to established nephropathy, and this is associated with elevated BP. Second, that the mechanism is an ETA receptor-mediated blunting of medullary perfusion which can be reversed with insulin and ETA receptor antagonism. Experimental acute pressure natriuresis was induced in young, early T1DM (2-3 weeks post streptozotocin) Sprague Dawley rats and healthy controls. Despite maintaining glomerular filtration rate, early T1DM suppressed urinary flow (UV, 22.9±2.9 v. 93.7±11.1μl/min/gkw) and sodium excretion (UNaV, 3.2±0.7 v. 22.7±3.3μmol/min/gkw) rates by >80%, and reduced gradients of pressure diuresis (linear, 1.9 to 0.3) and natriuresis (non-linear k, 0.05 to 0.01) curves. Insulin treatment lowered blood glucose (16.8±1.8 to 9.3±0.6mmol/l) and restored gradients of the responses. Tissue and urine analyses did not suggest structural nephropathy. In early T1DM rats, changes in BP on radiotelemetry were consistent with impaired circadian regulation of BP and precursors of hypertension: 24-hour diastolic BP rose (92.3±0.4 to 97.1±0.5mmHg), and the circadian dip in diastolic BP fell (6±1 to 2±1%). Atrasentan (ETA receptor antagonist, 5mg/kg/day orally) reduced diastolic dipping in early T1DM (3±1 to 1±1%) while additional ETB receptor antagonism (A-192621, 10mg/kg/day orally) reversed this, suggesting that ETA, and not ETB receptors, mediate impairment of acute pressure natriuresis. To address this, renal blood flow was measured during experimental acute pressure natriuresis and ET receptor antagonism. Early T1DM suppressed the normal rise in medullary perfusion (flux, 227.2±26.7 v. 115.4±10.3%) by ~90%. Suppressed medullary flux was unaffected by insulin (112.2±6.8%), despite restoration of UV and UNaV. In controls, atrasentan reduced UV (15.7±4.9 v. 38.6±6.2μl/min/gkw), UNaV (1.7±0.5 v. 16.7±1.4μmol/min/gkw), FENa (3.4±1.4 v. 15.0±2.4%) and medullary flux (122.2±26.7%) by 60 to 90% of control values, while A-192621 increased UNaV (26.6±6.9μmol/min/gkw) and FENa (21.6±3.4%), but not medullary flux, by ~50%. ET receptor antagonism did not modify early T1DM+/-insulin effects. Diabetic status had no effect on renal ET-1 and ET receptor expression. These results support the first hypothesis but disprove the second. Early T1DM blunts medullary perfusion and acute pressure natriuresis, and increases diastolic BP. Insulin restores natriuresis but not medullary flow. Therefore, targeting medullary perfusion may reduce cardiovascular risk in early T1DM, but this is not achievable with selective ETA receptor antagonists. Novel natriuretic (ETA) and anti-natriuretic (ETB) roles for ET receptors, which are not apparent in early T1DM during severe, experimental rises in BP, appear to contribute to daily regulation of BP, and may preclude the use of selective ETA receptor antagonists in T1DM prior to nephropathy.

<p>Potassium is an essential nutrient, that has been labeled a shortfall nutrient by recent Dietary Guidelines for Americans Advisory Committees. Increases in potassium intake have been linked to improvements in cardiovascular and other metabolic health outcomes. Blood pressure (BP) has often been cited as the primary criterion for determining potassium requirements. Hypertension (HTN), or high BP, is a primary risk factor for cardiovascular disease and other circulatory diseases. Cardiovascular disease (CVD) is responsible for the 31% of deaths worldwide. Findings from the Agency for Healthcare Research and Quality report (which informed the recently released Dietary Reference Intakes for sodium (Na) and potassium) on potassium intake and chronic disease concluded, with a moderate strength of evidence, that increasing potassium intake decreases BP, particularly among those with HTN. Although, of the 18 randomized controlled trials assessed by the AHRQ, only 4 were dietary interventions, the rest involved potassium supplementation. Observational studies also show a consistent bone benefit with increased potassium rich fruit and vegetable intakes in cohorts spanning adolescents to the elderly. In clinical trials, higher potassium intakes through supplementation have been associated with reduced urinary calcium (Ca) excretion and improvement in Ca balance. Although, similar to BP, intervention trials assessing the impact of dietary potassium on bone are lacking. Controlled feeding studies looking specifically at increases in potassium from food are sparse, leaving a large knowledge gap in the field for a nutrient with an important potential health impact. In general, little is known about whole-body potassium retention, with the few studies conducted lacking consistency and rigor in methods and design. What potassium retention means in terms of adequacy, or how higher or lower retention may influence specific health outcomes is understudied and not well understood. </p> <p>Utilizing a randomized, cross-over, controlled feeding clinical study with complete metabolic balance measures, our research aims to begin filling these gaps, looking specifically at the effects of potassium intake via potato sources and a potassium supplement on BP and vascular outcomes, as well as how the source of potassium may influence potassium, Na and Ca whole-body balance. </p> <p>This dissertation will discuss the physiology of potassium intake, how this may affect potassium, Na, and Ca retention, and in turn what influence this has on vascular and bone related health outcomes. Overall the goal of this research is to address the question: What is the importance of dietary potassium, and how can it benefit cardiovascular and skeletal health?</p>

Cervical vagal nerve stimulation (VNS) has been studied in the context of several conditions including epilepsy and depression. However, its effects on glucose metabolism, and its potentially beneficial effects in type II diabetes, have not yet been evaluated in humans. Efferent parasympathetic activation reduces hepatic glucose release and increases pancreatic insulin secretion, while afferent parasympathetic activation may increase hepatic glucose release and inhibit insulin secretion potentially through sympathetic activation. Thus, the effect of combined afferent and efferent cervical VNS is difficult to predict. We hypothesized that selective efferent VNS would decrease blood glucose concentration [Glu] and that selective afferent VNS would increase [Glu]. To investigate these potentially contrasting effects of efferent vs. afferent parasympathetic signaling, we recorded [Glu] and serum insulin and glucagon levels before and during 120 min of VNS in anesthetized rats. The nerve was left intact for combined afferent and efferent VNS (n=9) or sectioned proximal or distal from the stimulation electrode for selective efferent (n=8) of afferent (n=7) VNS, respectively. We found that afferent VNS caused a strong and sustained increase in [Glu] (+108.9±20.9% or +77.6±15.4% after 120 min of combined afferent and efferent VNS or selective afferent VNS) that was not accompanied by an increase in serum insulin concentration. Combined afferent and efferent VNS significantly increased serum glucagon concentration (57.6±23.4% at 120 min of VNS), while selective afferent VNS did not increase glucagon levels. Conversely, selective efferent VNS increased [Glu] only temporarily (+28.8±11.7% at 30 min of VNS). This response coincided with a transient increase in serum glucagon concentration at 30 min of VNS (31.6±8.3%) and a strong and sustained increase in serum insulin concentration (+71.2±27.0% after 120 min of VNS). These findings demonstrate that afferent VNS may increase [Glu] by suppressing pancreatic insulin release, while efferent VNS transiently increases [Glu] by stimulating glucagon secretion before reducing levels to or below baseline values by stimulating the release of insulin. Thus, selective efferent VNS may be potentially effective in the treatment of type II diabetes.

The present study documents that baroreceptors located in the pulmocutaneous artery (PCA) are key components in control of lymph heart rate in amphibians. A negative feedback control loop exists between arterial pressure and lymphatic heart rate. The recurrent laryngeal nerve (rLN), which innervates the PCA baroreceptors, transmits information on arterial pressure to integration centers in the central nervous system. Lymphatic heart rate (LHR) is reduced as a result of increases in arterial pressure. This loop was determined using three experimental protocols. First, the correlation between LHR reduction and hormonally induced vasoconstriction was determined. Increases in arterial pressure due to pressor actions of angiotensin II and arginine vasotocin at high concentrations was negatively correlated to LHR. Second, lymphatic heart rate changes due to natural increases in arterial pressure were compared to rate changes due to increase in arterial pressure after bilateral denervation of the rLN. Post-denervation LHR was not affected by natural increase in arterial pressure prior to the establishment of a new resting arterial pressure. Increase in arterial pressure due to administration of vasoconstricting hormones was negatively correlated with LHR following denervation. Third, the effect on LHR due to direct stimulation of the rLN was studied. Stimulation of the rLN caused LHR to stop without increases in arterial pressure. Presumably, this negative feedback loop is present to limit fluid return to the cardiovascular system from the lymphatic system during periods of acute hypertension. Reduction in the return of lymph volume to the cardiovascular system could eliminate potential damage to pulmonary tissues due to high arterial pressures.

The bond between pet and owner has been shown to decrease stress-related physical, mental, and emotional disorders by lowering blood pressure and heart rate. The purpose of this study was to examine whether a nonpet canine would have the same effect in reducing physical symptoms of stress as a known dog. Human-animal bond theory, Human pressor test. Data were analyzed using repeated measures, analyses of variance, and t tests. Results indicated that interaction with a strange dog lowered heart rate and blood pressure significantly faster in the experimental group than in the control group. The positive social change implications of this research include policy and procedural accommodations for the use of canines as comfort animals for individuals in stressful conditions.

Renin is an enzyme that catalyzes the rate-limiting step in the production of angiotensin peptides, and is thus a key regulator of processes controlled by angiotensin such as blood pressure, hydromineral balance, and metabolism. Our laboratory and others have previously identified a novel isoform of renin (icRen) which, as a result of the utilization of an alternate first exon, lacks the signal peptide and first third of the pro-segment of classical secreted renin (sRen). This alternate icRen isoform thus remains within the cytoplasm of the cell, but is constitutively active. Here, we report that while sRen is the predominant form of renin expressed in most tissues during development, icRen is the predominant form of renin within the adult brain. Thus, we hypothesized that sRen and icRen play distinct physiological roles in adult mice. To examine this hypothesis, we have utilized the Cre-LoxP system to selectively delete either isoform globally or within selected cell types such as neurons and glia. We have successfully developed a "sRen-flox" model, in which endogenous mouse sRen isoform can be selectively deleted, while not affecting endogenous icRen production. Breeding these mice against the E2A-Cre, Nestin-Cre, and GFAP-Cre mouse lines resulted in global-, neuronal-, and glial-specific knockouts of sRen, respectively. Physiological characterization of resulting mice has uncovered postnatal lethality, hypotension, renal atrophy, vascular dysfunction and decreased body weight and white adipose in the global knockouts. Depletion of sRen from only neuronal or glial cells does not appear to alter any of these phenotypes at baseline. From these data, we conclude that while peripheral sRen is of primary importance to blood pressure regulation, hydromineral balance, and metabolism, central expression of this isoform is unimportant. Further, comparison of our results to published findings from global total renin knockout models indirectly supports a role for icRen in the brain. We are currently in the process of generating icRen-flox and subsequent knockout mice, which will be useful models to directly analyze the physiological role(s) of icRen.

Abstract not available