During each cardiac cycle pulsatile arterial blood inflates the vascular bed of the brain, forcing cerebrospinal fluid (CSF) and venous blood out of the cranium. Excessive arterial pulsatility may be part of a harmful mechanism causing cognitive decline among elderly. Additionally, restricted venous flow from the brain is suggested as the cause of multiple sclerosis. Addressing hypotheses derived from these observations requires accurate and reliable investigational methods. This work focused on assessing the pulsatile waveform of cerebral arterial, venous and CSF flows. The overall aim of this dissertation was to explore cerebral blood flow and intracranial pulsatility using MRI, with respect to measurement, physiological and pathophysiological aspects. Two-dimensional phase contrast magnetic resonance imaging (2D PCMRI) was used to assess the pulsatile waveforms of cerebral arterial, venous and CSF flow. The repeatability was assessed in healthy young subjects. The 2D PCMRI measurements of cerebral arterial, venous and CSF pulsatility were generally repeatable but the pulsatility decreased systematically during the investigation. A method combining 2D PCMRI measurements with invasive CSF infusion tests to determine the magnitude and distribution of compliance within the craniospinal system was developed and applied in a group of healthy elderly. The intracranial space contained approximately two thirds of the total craniospinal compliance. The magnitude of craniospinal compliance was less than suggested in previous studies. The vascular hypothesis for multiple sclerosis was tested. Venous drainage in the internal jugular veins was compared between healthy controls and multiple sclerosis patients using 2D PCMRI. For both groups, a great variability in the internal jugular flow was observed but no pattern specific to multiple sclerosis could be found. Relationships between regional brain volumes and potential biomarkers of intracranial cardiac-related pulsatile stress were assessed in healthy elderly. The biomarkers were extracted from invasive CSF pressure measurements as well as 2D PCMRI acquisitions. The volumes of temporal cortex, frontal cortex and hippocampus were negatively related to the magnitude of cardiac-related intracranial pulsatility. Finally, a potentially improved workflow to assess the volume of arterial pulsatility using time resolved, four-dimensional phase contrast MRI measurements (4D PCMRI) was evaluated. The measurements showed good agreement with 2D PCMRI acquisitions. In conclusion, this work showed that 2D PCMRI is a feasible tool to study the pulsatile waveforms of cerebral blood and CSF flow. Conventional views regarding the magnitude and distribution of craniospinal compliance was challenged, with important implications regarding the understanding of how intracranial vascular pulsatility is absorbed. A first counterpoint to previous near-uniform observations of obstructions in the internal jugular veins in multiple sclerosis was provided. It was demonstrated that large cardiac- related intracranial pulsatility were related to smaller volumes of brain regions that are important in neurodegenerative diseases among elderly. This represents a strong rationale to further investigate the role of excessive intracranial pulsatility in cognitive impairment and dementia. For that work, 4D PCMRI will facilitate an effective analysis of cerebral blood flow and pulsatility.

With great appreciation, I would like to accentuate the substantial contributions of the following people who made this project possible: To Dr. CMT Fourie (my supervisor), Prof. JM van Rooyen (my co–supervisor) and Prof. AE Schutte (my co–supervisor) whose gracious advise, patient guidance, commitment and support have enabled me to plan, analyse, interpret and write this project in a scientific manner. It has been an educational experience for me, thank you. To Mr. LS Wyldbore for the language editing of this dissertation. I thank all the participants, researchers, field workers and supporting staff of the PURE study. The financial assistance of the National Research Foundation (DAAD–NRF) towards this research is hereby acknowledged. A special thanks to my parents, sister, Albert, family and friends, thank you for the never–ending love, support, patience and understanding that you gave me throughout this project. Last, but not the least, a special thank to God for giving me the opportunity, talent, determination and endurance to complete this project. / Thesis (M.Sc. (Physiology))--North-West University, Potchefstroom Campus, 2012.

With great appreciation, I would like to accentuate the substantial contributions of the following people who made this project possible: To Dr. CMT Fourie (my supervisor), Prof. JM van Rooyen (my co–supervisor) and Prof. AE Schutte (my co–supervisor) whose gracious advise, patient guidance, commitment and support have enabled me to plan, analyse, interpret and write this project in a scientific manner. It has been an educational experience for me, thank you. To Mr. LS Wyldbore for the language editing of this dissertation. I thank all the participants, researchers, field workers and supporting staff of the PURE study. The financial assistance of the National Research Foundation (DAAD–NRF) towards this research is hereby acknowledged. A special thanks to my parents, sister, Albert, family and friends, thank you for the never–ending love, support, patience and understanding that you gave me throughout this project. Last, but not the least, a special thank to God for giving me the opportunity, talent, determination and endurance to complete this project. / Thesis (M.Sc. (Physiology))--North-West University, Potchefstroom Campus, 2012.

Ao contrário dos indicadores estáticos, como a pressão venosa central e a pressão de oclusão da artéria pulmonar, a variação da pressão de pulso arterial tem se mostrado um indicador hemodinâmico preciso para a determinação da responsividade à administração de fluido. Objetivo: Este estudo objetivou mensurar a variação da pressão de pulso em cadelas mecanicamente ventiladas submetidas a expansão volêmica durante procedimento cirúrgico eletivo (ovário-salpingo-histerectomia) e determinar a relação deste indicador com os dados hemodinâmicos obtidos por meio do exame ecodopplercardiográfico transesofágico. Material e método: Quinze cadelas foram distribuídas aleatoriamente em dois grupos: sem pressão positiva ao final da expiração (GI - ZEEP, n = 7) e com pressão positiva ao final da expiração de 5cmH2O (GII - PEEP, n = 8). As avaliações hemodinâmicas incluindo freqüência cardíaca (FC), pressão arterial média invasiva (PaM), variação da pressão de pulso (VPP), pressão venosa central (PVC), volume sistólico (VE), índice cardíaco (IC), velocidade do fluxo aórtico (VFA) e gradiente de pressão do fluxo aórtico (Grad); análise de gases sangüíneos e parâmetros do sistema respiratório incluindo freqüência respiratória (FR) e pressão de pico inspiratória (PIP), foram mensuradas antes da aplicação da PEEP em cadelas do GII (M0); imediatamente antes (M1) e cinco minutos após (M2) a expansão volêmica com 10ml/Kg de solução de Ringer Lactato. Os grupos e tempos foram comparados por meio da análise de variância (ANOVA) seguida do teste de Tukey, com nível de significância de 5%. Resultados: A utilização da PEEP ocasionou um significante aumento dos seguintes parâmetros: VPP (M0 - 9.5 ± 2.92 vs M1 - 12.1 ± 2.19, P < 0.05); PVC ( 4.9 ± 2.47 vs 6.5 ± 2.73, P < 0.05); PIP (9.9 ± 1.36 vs 13.0 ± 2.2, P < 0.05) e PaO2 (336.5 ± 64.04 vs 373.6 ± 97.82, P < 0.05). Os animais do GI apresentaram um significante aumento dos seguintes parâmetros depois da expansão volêmica: VE (25.4 ± 3.92 vs 19.1 ± 3.22, P < 0.05), IC (3.8 ± 0.99 vs 2.4 ± 0.47, P < 0.05), VFA (93.8 ± 17.60 vs 74.5 ± 14.66, P < 0.05) e Grad (3.6 ± 1.30 vs 2.3 ± 0.92, P < 0.05). Alterações significativas também foram observadas nos seguintes parâmetros do GII depois da expansão volêmica: PaM (108.6 ± 20.6 vs 96.9 ± 19.97, P < 0.05), VPP (5.2 ± 1.8 vs 12.1 ± 2.19, P < 0.05), VE (26.6 ± 8.18 vs 20 ± 5.15, P < 0.05), IC (3.6 ± 0.90 vs 2.4 ± 0.56, P < 0.05), VFA (97.6 ± 25.49 vs 83.6 ± 24.12, P < 0.05) e Grad (4.0 ± 2.01 vs 3.0 ± 1.64, P < 0.05). Quando comparado M1 do GI e do GII, diferença significativa foi observada com a VPP (7.1 ± 1.64 vs 12.1 ± 2.19, P < 0.05). A FR foi maior no GII quando comparada ao GI (FR GII M1 e GII M2 - 16.9 ± 1.36 vs GI M1 - 14.9 ± 1.35 e GI M2 - 15.3 ± 1.5, P < 0.05). PIP foi também maior no GII quando comparada ao GI (GII M1 - 13.0 ± 2.20 vs GI M1 - 7.3 ± 2.21; GII M2 - 13.4 ± 2.50 vs GI M2 - 7.7 ± 2.14, P < 0.05). Conclusão: Este estudo mostrou que a variação da pressão de pulso é um índice adequado para avaliar o estado hemodinâmico durante a instituição de PEEP, sendo a avaliação ecodopplercardiográfica transesofágica uma excelente técnica de monitoração do estado hemodinâmico nos pacientes submetidos a procedimento cirúrgico. / Contrary to static parameters such as central venous pressure and pulmonary capillary wedge pressure, pulse pressure variation has proven itself as an accurate hemodynamic tool for the determination of fluid responsiveness. Objective: To measure pulse pressure variation in mechanically-ventilated bitches submitted to volume load during elective surgery (ovariohysterectomy) and to determine its relationship with hemodynamic parameters obtained with transesophageal Doppler echocardiography. Material and Methods: Fifteen bitches were randomly distributed between two groups: with zero end-expiratory pressure (GI - ZEEP, n=7) and with positive end-expiratory pressure of 5cmH2O (GII - PEEP, n=8). Hemodynamic evaluations including heart rate (HR), invasive mean arterial pressure (MAP), pulse pressure variation (PPV), central venous pressure (CVP), stroke volume (SV), cardiac index (CI), aortic blood flow velocity and aortic flow pressure gradient; blood-gas analysis and ventilatory parameters including respiratory rate (RR) and peak inspiratory pressure (PIP), were measured before employment of PEEP in bitches from GII (M0); immediately before (M1) and five minutes after (M2) volume expansion using 10ml/Kg lacted Ringers solution. Statistical analysis was based on ANOVA for repeated measures followed by Turkeys t-test with significance level of 0.05. Results: The use of PEEP induced a significant increase in the following parameters: PPV (M0 - 9.5 ± 2.92 vs M1 - 12.1 ± 2.19, P < 0.05); CVP ( 4.9 ± 2.47 vs 6.5 ± 2.73, P < 0.05); PIP (9.9 ± 1.36 vs 13.0 ± 2.2, P < 0.05) and PaO2 (336.5 ± 64.04 vs 373.6 ± 97.82, P < 0.05). Animals in GI presented a significant increase in the following parameters after fluid load: SV (25.4 ± 3.92 vs 19.1 ± 3.22, P < 0.05), CI (3.8 ± 0.99 vs 2.4 ± 0.47, P < 0.05), aortic blood flow velocity (93.8 ± 17.60 vs 74.5 ± 14.66, P < 0.05) and aortic flow pressure gradient (3.6 ± 1.30 vs 2.3 ± 0.92, P < 0.05). Statistical significance of the following parameters were observed in GII after fluid load: MAP (108.6 ± 20.6 vs 96.9 ± 19.97, P < 0.05), PPV (5.2 ± 1.8 vs 12.1 ± 2.19, P < 0.05), SV (26.6 ± 8.18 vs 20 ± 5.15, P < 0.05), CI (3.6 ± 0.90 vs 2.4 ± 0.56, P < 0.05), aortic blood flow velocity (97.6 ± 25.49 vs 83.6 ± 24.12, P < 0.05) and aortic flow pressure gradient (4.0 ± 2.01 vs 3.0 ± 1.64, P < 0.05). When comparing M1 of GI and GII, a statistical significant difference was observed with PPV (7.1 ± 1.64 vs 12.1 ± 2.19, P < 0.05). Respiratory rate was greater in GII than in GI (RR GII M1 and GII M2 - 16.9 ± 1.36 vs GI M1 - 14.9 ± 1.35 and GI M2 - 15.3 ± 1.5, P < 0.05). Peak inspiratory pressure was also greater in GII than in GI (GII M1 - 13.0 ± 2.20 vs GI M1 - 7.3 ± 2.21; GII M2 - 13.4 ± 2.50 vs GI M2 - 7.7 ± 2.14, P < 0.05). Conclusion: This study showed that the pulse pressure variation is an adequate indicator to evaluate the hemodynamic status during PEEP application, being the transesophageal Doppler echocardiography evaluation a great tool in monitoring the hemodynamic status in patients undergoing surgery.

Alterações das propriedades funcionais e estruturais de grandes artérias são correlacionadas à maior risco cardiovascular em diferentes populações. Recentes diretrizes europeias e brasileiras de hipertensão arterial incluem avaliações da espessura íntima medial (EIM) de carótida e da velocidade de onda de pulso (VOP) aórtica como métodos para avaliação de doença subclínica do paciente hipertenso. Estabelecer valores de referência dos métodos de avaliação da estrutura e da função arteriais e os principais determinantes clínicos e demográficos dos parâmetros obtidos com estes métodos, além de suas correlações, em indivíduos adultos saudáveis, é importante para melhor estratificação dos pacientes. Os objetivos do estudo foram avaliar em indivíduos adultos saudáveis: 1. a distribuição dos valores de VOP aórtica e do diâmetro, da espessura íntima medial e da distensão relativa da artéria carótida, para definir parâmetros de referência. 2. os principais determinantes demográficos e clínicos destes parâmetros e suas correlações.Foram avaliados 210 indivíduos (54% mulheres, idade média 44±13anos) sem história ou evidência de doença cardiovascular atual. A pressão arterial (PA) foi aferida por aparelho automático Omron (HEM 705 CP) com o indivíduo sentado. A medida automática da VOP carótida-femural foi feita pelo aparelho Complior®. As propriedades funcionais e estruturais da carótida (distensão, diâmetro e EIM) foram avaliadas por ultrassom de radiofrequência (WTS®). Todos os indivíduos realizaram avaliação bioquímica.As médias dos valores foram: VOP aórtica (m/s) = 8,7±1,5, diâmetro da carótida (m) = 6617±985, EIM da carótida (m) = 601±131 e distensão relativa da carótida (%) = 5,3±2,1. Não houve diferenças significativas nos parâmetros entre homens e mulheres, assim como entre as raças. Na análise multivariada, os fatores independentemente relacionados aos parâmetros vasculares foram: VOP aórtica = idade (r2 0,22/ p < 0,01), EIM da carótida = idade (r2 0,17 / p < 0,01), diâmetro da carótida = creatinina (r2 0,16 / p = 0,02), distensão relativa da carótida = idade (r2 0,37 / p < 0,01) e pressão arterial diastólica PAD - (r2 0,09 / p < 0,01). Observamos que a VOP teve correlação positiva com EIM (r² - 0,06 / p < 0,01), e correlação inversa com a distensão relativa da carótida (r² - 0,08 / p < 0,01). Em conclusão, em indivíduos saudáveis o principal fator relacionado à medida da rigidez aórtica é a idade, enquanto que a idade e a PAD foram relacionados com a medida funcional da carótida, e o nível de creatinina relacionado ao diâmetro. A estrutura da artéria carótida é diretamente relacionada à medida da rigidez aórtica, que por sua vez é inversamente relacionada à capacidade funcional da artéria carótida / Changes of structural and functional properties of large arteries are correlated with increased cardiovascular risk in different populations. Recent European and Brazilian guidelines for hypertension include assessment of carotid intima-media thickness (IMT) and aortic pulse wave velocity (PWV) for evaluation of subclinical disease in hypertensive patients. Thus, to establish reference values for methods that evaluate arterial properties and the main clinical and demographic determinants of the parameters obtained with these methods, as well as their correlation in healthy adult subjects, it is important for better stratification of patients. The objectives of the study were to evaluate in healthy adult subjects: 1. the distribution of aortic PWV values and of the diameter, intima-media thickness and distension of the carotid artery to obtain reference parameters. 2. the main demographic and clinical determinants of these parameters and their correlations. We evaluated 210 subjects (54% women, mean age 44 ± 13anos) with no history or current evidence of cardiovascular disease. Blood pressure (BP) was measured by automatic device Omron (HEM 705 CP) with the subject in a sitting position. The automatic measurement of carotid-femoral PWV was performed by Complior® device. The functional and structural properties of the carotid artery (distension, diameter and IMT) were assessed by ultrasound radiofrequency (WTS®). All subjects underwent biochemical evaluation. The mean values were: aortic PWV (m / s) = 8.7±1.5, carotid diameter (m) = 6617±985, carotid IMT (m) = 601±131 and distension on carotid (% ) = 5.3±2.1. There were no significant differences in parameters according gender or race. In multivariate analysis, the independent factors related to arterial parameters were: aortic PWV = age (r2 - 0.22 / p -< 0.01), carotid IMT = age (r2 - 0.17 / p <0.01 ), carotid diameter = creatinine (r2 - 0.16 / p = 0.02), carotid distension = age (r2 - 0.37 / p < 0.01) and diastolic blood pressure - DBP - (r2 - 0.09 / p< 0.01). We also found that aortic PWV was positively correlated with IMT (r² - 0.06 / p < 0.01) and inversely correlated with the relative distension of the carotid (r² - 0.08 / p < 0.01). In conclusion, in healthy subjects the main factor related to aortic stiffness is age, whereas age and DBP were related to functional measurement of the carotid artery, and creatinine levels related to the diameter. The structure of the carotid artery is directly related to the aortic stiffness, which in turn is inversely related to the functional capacity of the carotid artery.

The volume defined by the rigid cranium is shared by the brain, blood and cerebrospinal fluid (CSF). With every heartbeat the arterial blood volume briefly increases and venous blood and CSF are forced out of the cranium, leading to pulsatility in CSF flow and intracranial pressure (ICP). Altered CSF pulsatility has been linked to idiopathic normal pressure hydrocephalus (INPH), which involves enlarged cerebral ventricles and symptoms of gait/balance disturbance, cognitive decline and urinary incontinence that may be improved by implantation of a shunt. The overall aim of this thesis was to investigate the fluid dynamics of the CSF system, with a focus on pulsatility, and how they relate to INPH pathophysiology and treatment. Mathematical modelling was applied to data from infusion tests, where the ICP response to CSF volume manipulation is measured, to analyse the relationship between mean ICP and ICP pulse amplitude (AMP) before and after shunt surgery in INPH (paper I-II). The observed relationship, designated the pulsatility curve, was found to be constant at low ICP and linear at high ICP, corresponding to a shift from constant to ICP dependent compliance (paper I). Shunt surgery did not affect the pulsatility curve, but shifted baseline ICP and AMP along the curve towards lower values. Patients who improved in gait after surgery had significantly larger AMP reduction than those who did not, while ICP reduction was similar, suggesting that improving patients had baseline ICP in the linear zone of the curve before surgery. Use of this phenomenon for outcome prediction was promising (paper II). The fluid dynamics of an empirically derived pulsatility-based predictive infusion test for INPH was also investigated, with results showing strong influence from compliance (paper III). Clinical ICP data at different body postures was used to evaluate three models describing postural effects on ICP. ICP decreased in upright positions, whereas AMP increased. The model describing the postural effects based on hydrostatic changes in the venous system, including effects of collapse of the jugular veins in the upright position, accurately predicted the measured ICP (paper IV). Cerebral blood flow and CSF flow in the aqueduct and at the cervical level was measured with phase contrast magnetic resonance imaging, and compared between healthy elderly and INPH (paper V). Cerebral blood flow and CSF flow at the cervical level were similar in INPH patients and healthy elderly, whereas aqueductal CSF flow differed significantly. The pulsatility in the aqueduct flow was increased, and there was more variation in the net flow in INPH, but the mean net flow was normal, i.e. directed from the ventricles to the subarachnoid space (paper V). In conclusion, this thesis introduced the concept of pulsatility curve analysis, and provided evidence that pulsatility and compliance are important aspects for successful shunt treatment and outcome prediction in INPH. It was further confirmed that enhanced pulsatility of aqueduct CSF flow was the most distinct effect of INPH pathophysiology on cerebral blood flow and CSF flow. A new model describing postural and hydrostatic effects on ICP was presented, and the feasibility and potential importance of measuring ICP in the upright position in INPH was demonstrated. / <p>Forskningsfinansiär: </p><p>European Union, ERDF: Objective 2, Northern Sweden (grant no. 158715-CMTF). </p>

Idiopathic Normal Pressure Hydrocephalus (INPH) is a condition affecting gait, cognition and continence. Radiological examination reveals enlarged ventricles of the brain. A shunt that drains CSF from the ventricles to the abdomen often improves the symptoms. Much research on INPH has been focused on identifying tests that predict the outcome after shunt surgery. As part of this quest, there are attempts to find measurement methods of intracranial parameters that are valid, reliable, tolerable and safe for patients. Today's technologies for intracranial pressure (ICP) measurement are invasive, often requiring a burr-hole in the skull. Recently, a method for non-invasive ICP measurements was suggested: the Pulsatile Index (PI) calculated from transcranial Doppler data assessed from the middle cerebral artery. In this thesis the relation between PI and ICP was explored in INPH patients during controlled ICP regulation by lumbar infusion. The confidence interval for predicted ICP, based on measured PI was too large for the method to be of clinical utility. In the quest for better predictive tests for shunt success in INPH, recent studies have shown promising results with criteria based on cardiac related ICP wave amplitudes. The brain ventricular system, and the fluid surrounding the spinal cord are in contact. In this thesis it was shown that ICP waves could be measured via lumbar subarachnoid space, with a slight underestimation. One of the cardinal symptoms of hydrocephalus is cognitive impairment. Neuropsychological studies have demonstrated cognitive tests that are impaired and improve after shunt surgery in INPH patients. However, there is currently no standardized test battery and different studies use different tests. In response, in this thesis a fully automated computerized neuropsychological test battery was developed. The validity, reliability, responsiveness to improvement after shunt surgery and feasibility for testing INPH patients was demonstrated. It was also demonstrated that INPH patients were impaired in all subtests, compared to healthy elderly.

Alterações das propriedades funcionais e estruturais de grandes artérias são correlacionadas à maior risco cardiovascular em diferentes populações. Recentes diretrizes europeias e brasileiras de hipertensão arterial incluem avaliações da espessura íntima medial (EIM) de carótida e da velocidade de onda de pulso (VOP) aórtica como métodos para avaliação de doença subclínica do paciente hipertenso. Estabelecer valores de referência dos métodos de avaliação da estrutura e da função arteriais e os principais determinantes clínicos e demográficos dos parâmetros obtidos com estes métodos, além de suas correlações, em indivíduos adultos saudáveis, é importante para melhor estratificação dos pacientes. Os objetivos do estudo foram avaliar em indivíduos adultos saudáveis: 1. a distribuição dos valores de VOP aórtica e do diâmetro, da espessura íntima medial e da distensão relativa da artéria carótida, para definir parâmetros de referência. 2. os principais determinantes demográficos e clínicos destes parâmetros e suas correlações.Foram avaliados 210 indivíduos (54% mulheres, idade média 44±13anos) sem história ou evidência de doença cardiovascular atual. A pressão arterial (PA) foi aferida por aparelho automático Omron (HEM 705 CP) com o indivíduo sentado. A medida automática da VOP carótida-femural foi feita pelo aparelho Complior®. As propriedades funcionais e estruturais da carótida (distensão, diâmetro e EIM) foram avaliadas por ultrassom de radiofrequência (WTS®). Todos os indivíduos realizaram avaliação bioquímica.As médias dos valores foram: VOP aórtica (m/s) = 8,7±1,5, diâmetro da carótida (m) = 6617±985, EIM da carótida (m) = 601±131 e distensão relativa da carótida (%) = 5,3±2,1. Não houve diferenças significativas nos parâmetros entre homens e mulheres, assim como entre as raças. Na análise multivariada, os fatores independentemente relacionados aos parâmetros vasculares foram: VOP aórtica = idade (r2 0,22/ p < 0,01), EIM da carótida = idade (r2 0,17 / p < 0,01), diâmetro da carótida = creatinina (r2 0,16 / p = 0,02), distensão relativa da carótida = idade (r2 0,37 / p < 0,01) e pressão arterial diastólica PAD - (r2 0,09 / p < 0,01). Observamos que a VOP teve correlação positiva com EIM (r² - 0,06 / p < 0,01), e correlação inversa com a distensão relativa da carótida (r² - 0,08 / p < 0,01). Em conclusão, em indivíduos saudáveis o principal fator relacionado à medida da rigidez aórtica é a idade, enquanto que a idade e a PAD foram relacionados com a medida funcional da carótida, e o nível de creatinina relacionado ao diâmetro. A estrutura da artéria carótida é diretamente relacionada à medida da rigidez aórtica, que por sua vez é inversamente relacionada à capacidade funcional da artéria carótida / Changes of structural and functional properties of large arteries are correlated with increased cardiovascular risk in different populations. Recent European and Brazilian guidelines for hypertension include assessment of carotid intima-media thickness (IMT) and aortic pulse wave velocity (PWV) for evaluation of subclinical disease in hypertensive patients. Thus, to establish reference values for methods that evaluate arterial properties and the main clinical and demographic determinants of the parameters obtained with these methods, as well as their correlation in healthy adult subjects, it is important for better stratification of patients. The objectives of the study were to evaluate in healthy adult subjects: 1. the distribution of aortic PWV values and of the diameter, intima-media thickness and distension of the carotid artery to obtain reference parameters. 2. the main demographic and clinical determinants of these parameters and their correlations. We evaluated 210 subjects (54% women, mean age 44 ± 13anos) with no history or current evidence of cardiovascular disease. Blood pressure (BP) was measured by automatic device Omron (HEM 705 CP) with the subject in a sitting position. The automatic measurement of carotid-femoral PWV was performed by Complior® device. The functional and structural properties of the carotid artery (distension, diameter and IMT) were assessed by ultrasound radiofrequency (WTS®). All subjects underwent biochemical evaluation. The mean values were: aortic PWV (m / s) = 8.7±1.5, carotid diameter (m) = 6617±985, carotid IMT (m) = 601±131 and distension on carotid (% ) = 5.3±2.1. There were no significant differences in parameters according gender or race. In multivariate analysis, the independent factors related to arterial parameters were: aortic PWV = age (r2 - 0.22 / p -< 0.01), carotid IMT = age (r2 - 0.17 / p <0.01 ), carotid diameter = creatinine (r2 - 0.16 / p = 0.02), carotid distension = age (r2 - 0.37 / p < 0.01) and diastolic blood pressure - DBP - (r2 - 0.09 / p< 0.01). We also found that aortic PWV was positively correlated with IMT (r² - 0.06 / p < 0.01) and inversely correlated with the relative distension of the carotid (r² - 0.08 / p < 0.01). In conclusion, in healthy subjects the main factor related to aortic stiffness is age, whereas age and DBP were related to functional measurement of the carotid artery, and creatinine levels related to the diameter. The structure of the carotid artery is directly related to the aortic stiffness, which in turn is inversely related to the functional capacity of the carotid artery.

Motivation - The burden of cardiovascular diseases (CVD) is increasing in developing countries worldwide, but even more so in sub-Saharan Africa. Due to rapid urbanisation, black populations experience lifestyle changes (e.g. unhealthy diet, increased access to alcohol and tobacco) that predispose them to increased obesity and cardiovascular risk. In this study, attention will be given to cardiovascular alterations, specifically arterial calcification, in lean and overweight/obese women nearing or already experiencing menopause. These include elevated blood pressure, large artery stiffness (indicated by increased central pulse pressure (cPP)) and carotid intima-media thickness (CIMT). Other factors linked to arterial calcification include the level of obesity as well as low bone mineral density. Ectopic calcification plays a significant role in cardiovascular morbidity and mortality, especially in renal failure patients, osteoporotic and elderly women. Factors contributing to the development and progression of arterial calcification include calciotropic hormones and altered bone metabolism, particularly in older postmenopausal women. This is due to the lack of protective effects of oestrogen against vascular alterations and bone loss after menopause. Previous studies have shown that increased bone resorption indicated by elevated levels of c-telopeptide of type I collagen (CTX), parathyroid hormone (PTH), low 25- hydroxycholecalciferol (25(OH)D3) and parathyroid hormone to 25-hydroxycholecalciferol ratio (PTH:25(OH)D3) are independently linked to arterial stiffening, CIMT and vascular calcification. Knowledge on the contribution of altered bone metabolism and associated calciotropic hormones on cardiovascular health in Africans is limited. Previous studies on ectopic calcification in South Africans focused on men and renal failure patients. This study will explore the possible role of altered calcium regulation and bone metabolism in the development of arterial calcification and CVD in older African women. Aim - The aim of this study was to investigate the associations of brachial and central pressures and CIMT with PTH, PTH:25(OH)D3 and CTX, a marker of bone resorption, in lean and overweight/obese African women older than 46 years. Methodology - This sub-study forms part of the Prospective Urban Rural Epidemiology (PURE) study. A total of 434 urban and rural women older than 46 years were included in the study. Women infected with the human immunodeficiency virus (HIV) were excluded from the study. The study was reviewed and approved by the Ethics Committee of the North-West University (Potchefstroom campus) and all participants signed an informed consent form prior to enrolment into the project. Field workers administered demographic, general health and physical activity questionnaires in the participants’ home language. Anthropometric measurements included weight, height and waist circumference, while body mass index (BMI) was calculated in kg/m2. Cardiovascular measurements included brachial and central systolic blood pressure (SBP), brachial diastolic blood pressure (DBP), brachial and central pulse pressure (PP) as well as CIMT and carotid cross-sectional wall area (CSWA). Blood pressure measurements were performed on the right arm with the participant in the sitting position. Blood was drawn after an overnight fasting period. We performed biochemical analyses from serum and plasma samples for follicle stimulating hormone (FSH), PTH, 25(OH)D3, and CTX. HIV testing was performed according to standardised procedures. Since interactions existed for BMI with regards to associations of CIMT and cPP with PTH:25(OH)D3, the study population was divided into the lean (BMI <25 kg/m2) and overweight/obese (BMI ≥25 kg/m2) groups. We performed independent T-tests to compare means and used the chi-square test to compare proportions. Single and multiple regression analyses were performed to investigate the associations of markers of vascular structure and function with CTX and calciotropic hormones. Results - In this study, 90% of the women displayed an FSH concentration exceeding the cut-off value of 35 mIu/mL, indicating a postmenopausal state. When comparing lean and overweight/obese African women, we found that lean women had higher levels of CTX and 25(OH)D3 (both p<0.001), while the overweight/obese group was older (p=0.007) and presented with higher PTH and PTH:25(OH)D3 levels (both p<0.001). Brachial and central pressures did not differ between the groups (p≥0.23), except for DBP being higher in the overweight/obese group (p=0.017). Overweight/obese women had higher CIMT (p<0.001) and CSWA (p=0.001) as compared to their lean counterparts. A larger proportion of lean women smoked (63%) and self-reported on alcohol use (37%) than overweight/obese women (41% and 18%, respectively) (both p<0.001). Forty-one percent of overweight/obese women used antihypertensive medication, opposed to 25% in the lean group (p=0.001). In multivariate regression analyses, an independent positive association existed between CIMT and PTH:25(OH)D3 (R2=0.22; β=0.26; p=0.003) in lean women. In the overweight/obese group independent positive associations were confirmed between brachial SBP and PTH (p=0.013) and CTX (p=0.038), and between DBP and PTH (p=0.030). Brachial PP and central SBP remained positively associated with CTX (p=0.016 and p=0.024, respectively), while cPP was independently associated with PTH:25(OH)D3 (R2=0.20; β=0.17; p=0.017) and CTX (R2=0.20; β=0.17; p=0.025). Conclusion - Our results indicate that in older African women, large artery structure and function are associated with calciotropic hormones and bone resorption, suggesting that altered bone metabolism and associated calciotropic hormones play a role in the development of vascular calcification. The different associations in lean and overweight/obese women suggest different mechanisms at work regarding arterial calcification in states of low and high adiposity. These findings need confirmation in larger prospective and experimental studies. / MSc (Physiology), North-West University, Potchefstroom Campus, 2014

Motivation - The burden of cardiovascular diseases (CVD) is increasing in developing countries worldwide, but even more so in sub-Saharan Africa. Due to rapid urbanisation, black populations experience lifestyle changes (e.g. unhealthy diet, increased access to alcohol and tobacco) that predispose them to increased obesity and cardiovascular risk. In this study, attention will be given to cardiovascular alterations, specifically arterial calcification, in lean and overweight/obese women nearing or already experiencing menopause. These include elevated blood pressure, large artery stiffness (indicated by increased central pulse pressure (cPP)) and carotid intima-media thickness (CIMT). Other factors linked to arterial calcification include the level of obesity as well as low bone mineral density. Ectopic calcification plays a significant role in cardiovascular morbidity and mortality, especially in renal failure patients, osteoporotic and elderly women. Factors contributing to the development and progression of arterial calcification include calciotropic hormones and altered bone metabolism, particularly in older postmenopausal women. This is due to the lack of protective effects of oestrogen against vascular alterations and bone loss after menopause. Previous studies have shown that increased bone resorption indicated by elevated levels of c-telopeptide of type I collagen (CTX), parathyroid hormone (PTH), low 25- hydroxycholecalciferol (25(OH)D3) and parathyroid hormone to 25-hydroxycholecalciferol ratio (PTH:25(OH)D3) are independently linked to arterial stiffening, CIMT and vascular calcification. Knowledge on the contribution of altered bone metabolism and associated calciotropic hormones on cardiovascular health in Africans is limited. Previous studies on ectopic calcification in South Africans focused on men and renal failure patients. This study will explore the possible role of altered calcium regulation and bone metabolism in the development of arterial calcification and CVD in older African women. Aim - The aim of this study was to investigate the associations of brachial and central pressures and CIMT with PTH, PTH:25(OH)D3 and CTX, a marker of bone resorption, in lean and overweight/obese African women older than 46 years. Methodology - This sub-study forms part of the Prospective Urban Rural Epidemiology (PURE) study. A total of 434 urban and rural women older than 46 years were included in the study. Women infected with the human immunodeficiency virus (HIV) were excluded from the study. The study was reviewed and approved by the Ethics Committee of the North-West University (Potchefstroom campus) and all participants signed an informed consent form prior to enrolment into the project. Field workers administered demographic, general health and physical activity questionnaires in the participants’ home language. Anthropometric measurements included weight, height and waist circumference, while body mass index (BMI) was calculated in kg/m2. Cardiovascular measurements included brachial and central systolic blood pressure (SBP), brachial diastolic blood pressure (DBP), brachial and central pulse pressure (PP) as well as CIMT and carotid cross-sectional wall area (CSWA). Blood pressure measurements were performed on the right arm with the participant in the sitting position. Blood was drawn after an overnight fasting period. We performed biochemical analyses from serum and plasma samples for follicle stimulating hormone (FSH), PTH, 25(OH)D3, and CTX. HIV testing was performed according to standardised procedures. Since interactions existed for BMI with regards to associations of CIMT and cPP with PTH:25(OH)D3, the study population was divided into the lean (BMI <25 kg/m2) and overweight/obese (BMI ≥25 kg/m2) groups. We performed independent T-tests to compare means and used the chi-square test to compare proportions. Single and multiple regression analyses were performed to investigate the associations of markers of vascular structure and function with CTX and calciotropic hormones. Results - In this study, 90% of the women displayed an FSH concentration exceeding the cut-off value of 35 mIu/mL, indicating a postmenopausal state. When comparing lean and overweight/obese African women, we found that lean women had higher levels of CTX and 25(OH)D3 (both p<0.001), while the overweight/obese group was older (p=0.007) and presented with higher PTH and PTH:25(OH)D3 levels (both p<0.001). Brachial and central pressures did not differ between the groups (p≥0.23), except for DBP being higher in the overweight/obese group (p=0.017). Overweight/obese women had higher CIMT (p<0.001) and CSWA (p=0.001) as compared to their lean counterparts. A larger proportion of lean women smoked (63%) and self-reported on alcohol use (37%) than overweight/obese women (41% and 18%, respectively) (both p<0.001). Forty-one percent of overweight/obese women used antihypertensive medication, opposed to 25% in the lean group (p=0.001). In multivariate regression analyses, an independent positive association existed between CIMT and PTH:25(OH)D3 (R2=0.22; β=0.26; p=0.003) in lean women. In the overweight/obese group independent positive associations were confirmed between brachial SBP and PTH (p=0.013) and CTX (p=0.038), and between DBP and PTH (p=0.030). Brachial PP and central SBP remained positively associated with CTX (p=0.016 and p=0.024, respectively), while cPP was independently associated with PTH:25(OH)D3 (R2=0.20; β=0.17; p=0.017) and CTX (R2=0.20; β=0.17; p=0.025). Conclusion - Our results indicate that in older African women, large artery structure and function are associated with calciotropic hormones and bone resorption, suggesting that altered bone metabolism and associated calciotropic hormones play a role in the development of vascular calcification. The different associations in lean and overweight/obese women suggest different mechanisms at work regarding arterial calcification in states of low and high adiposity. These findings need confirmation in larger prospective and experimental studies. / MSc (Physiology), North-West University, Potchefstroom Campus, 2014