The aim of this thesis was to study the biomarker homocysteine and other novel potential vascular risk factors for dementia. In an out-patient based study of a cohort of 926 consecutive subjects referred to our Memory Unit during 1996―2000, serum-folate was lower and total plasma homocysteine (tHcy) and serum methyl malonate were higher in subjects being prescribed with B12. In the subgroup diagnosed with dementia and with a positive family history of dementia, tHcy was higher than in the subgroup diagnosed as non-demented. It is necessary to supplement subjects with vitamin B12 deficiency with B12, but our results indicate that it is not sufficient with B12 alone because this gives rise to intracellular folate deficiency. We also found indications of a genetic component in dementia because tHcy was higher in the group with a positive family history of dementia. These findings prompted further studies of homocysteine metabolism. The frequency of mutations in the gene for folate receptor-α (FOLR-1), and the fibrinolytic pattern in dementia and non-dementia were studied in the two cohorts DGM (n=300) and AS (n=389). The DGM cohort is a consecutive series of subjects attending our Memory Care Unit for investigation of suspected cognitive problems or dementia between 2003 - 2007. The AS (= active seniors) cohort comprises retired, apparently healthy subjects from central Sweden, actively participating in study circles. A rare haplotype in the FOLR-1, with mutations in two nearby loci, was discovered, possibly associated with lower serum-folate and higher tHcy concentrations and was more frequent in the DGM group. The transport of folate to the CSF was studied in the DGM-cohort. Dementia with a vascular component was associated with a lower CSF to serum folate ratio indicative of reduced transport of folate to the CSF and further to the brain. The vascular endothelial derived fibrinolytic markers tPA, tPA/PAI-1-complex, and vWF were not only higher in vascular dementia (VaD) but also in Alzheimer’s Disease (AD) when compared to the AS group. The impaired fibrinolytic activity in both vascular dementia and in AD is a novel finding, signifying a vascular component in the development of dementia. In conclusion we found that both hereditary and nutritional background factors were linked to dementia and furthermore that a dysregulated fibrinolysis was linked to both VaD and AD.

Abstract The aim of the present study was to examine associations between serum 25-hydroxyvitamin D [25(OH)D] and 1,25-dihydroxyvitamin D[(1,25(OH)2D]—the circulating and active forms of vitamin D—and periodontal infection. The data were gathered from a case-control study (63 periodontitis patients and 30 periodontally healthy controls) and an intervention study among individuals with type 1 diabetes mellitus (T1DM, 80 patients at the baseline and 65 after periodontal treatment). The periodontal data and the levels of serum 25(OH)D, 1,25(OH)2D and parathyroid hormone (PTH) were available. A third data set included periodontal data and the serum level of 25(OH)D of 1262 non-smoking and non-diabetic 30–49-year-old individuals (Health 2000 Survey). Serum 25(OH)D analyses were done using enzyme-linked immunoassay and radioimmunoassay, 1,25(O)2D analyses using enzyme-immunoassay after purification of 1,25(OH)2D by immunoextraction and PTH analyses using electrochemiluminescence immunoassay. In the case-control study individuals with a low serum 1,25(O)2D level were more likely to belong to the periodontitis group than to the periodontally healthy group and an inverse association was observed between serum 1,25(OH)2D and severity of periodontitis at the baseline of the intervention study. Serum 1,25(OH)2D increased significantly after periodontal treatment in the T1DM patients; a finding that was considered suggestive of a causal relationship between serum 1,25(OH)2D and periodontal infection. Also, serum PTH increased after periodontal treatment; this increase, which was statistically significant (p = 0.016) in patients with moderate or severe periodontitis, may partly account for the earlier observed post-treatment increase in serum 1,25(OH)2D level. Possible explanations for low serum 1,25(OH)2D in periodontal infection may be increased degradation of 1,25(OH)2D, increased use of 1,25(OH)2D, or decreased hydroxylation of 25(OH)D The association between serum 25(OH)D level and periodontal infection was weak, if existent. An inverse association between serum 25(OH)D and the severity of periodontal infection was observed only in the T1DM patients. Among individuals with low plaque level, those in higher 25(OH)D quintiles tended to have fewer teeth with deepened periodontal pockets than those in lower quintiles; a finding which was interpreted to mean a slight protective role of 25(OH)D against periodontal infection. / Tiivistelmä Tutkimuksen tarkoituksena oli selvittää seerumin 25-hydroksivitamiini D:n [25(OH)D, D-vitamiinin varastomuoto] ja 1,25-dihydroksivitamiini D:n [1,25(OH)2D, D-vitamiinin aktiivinen muoto] tasojen yhteyttä parodontiumin alueen infektiosairauksiin. Tulokset perustuvat kolmeen tutkimusasetelmaan: tapaus-verrokki-tutkimus (63 parodontiitti-potilasta, 30 verrokkia), interventio-tutkimus [80 tyypin 1 diabetes mellitus (T1DM) potilasta, joista 65 osallistui seurantaan parodontologisen hoidon jälkeen] ja poikittaistutkimus Terveys 2000 tutkimuksen osa-aineistossa (1262 30-49 vuotiasta tupakoimatonta ei-diabeetikkoa). Tapaus-verrokki- ja interventiotutkimuksissa tutkittiin myös seerumin parathormoonin (PTH) yhteyttä parodontaali-infektioon sekä PTH:n vaikutusta seerumin 1,25(OH)2D tasoon infektion hoidon jälkeen. D-vitamiinin ja PTH:n tasot määritettiin immunologisin menetelmin. Yhteyksiä tutkittiin käyttäen vakioituja monimuuttujamalleja. Tapaus-verrokki-tutkimuksessa yksilöt, joilla seerumin 1,25(OH)2D taso oli alhainen, kuuluivat todennäköisemmin parodontiitti- kuin verrokkiryhmään. Interventiotutkimuksen alkutilanteessa seerumin 1,25(OH)2D:n ja parodontaali-infektion vaikeusasteen välillä vallitsi tilastollisesti merkittävä käänteinen yhteys ja taso nousi merkittävästi infektion hoidon jälkeen. Myös seerumin PTH taso nousi parodontaali-infektion hoidon jälkeen; nousu oli tilastollisesti merkittävä (p = 0.016) pitkälle edennyttä parodontiittia sairastavilla. Interventiotutkimuksen tulokset viittaavat kausaaliseen yhteyteen 1,25(OH)2D:n ja parodontaali-infektion välillä. Alhainen seerumin 1,25(OH)2D pitoisuus infektion vallitessa voi selittyä sen suurella käytöllä immuunipuolustukseen infektion aikana tai lisääntyneellä hajoamisella. Tason nousu hoidon jälkeen tukee edellä mainittua. PTH on 25(OH)D:n hydroksylaation pääsäätelijä ja 1,25(OH)2D:n nousua hoidon jälkeen voi osittain selittää myös seerumin PTH tason kohoaminen. Seerumin 25(OH)D:n ja parodontaali-infektion välillä havaittu yhteys oli heikko, mutta ei täysin sulje pois 25(OH)D:n suojaavaa vaikutusta. Käänteinen yhteys löytyi vain interventiotutkimuksen alkutilanteessa T1DM potilailla. Infektion hoito ei vaikuttanut 25(OH)D tasoon. Terveys 2000 tutkimuksen osa-aineistossa havaittiin hyvän suuhygienian omaavilla jonkin verran alhaisempi määrä syventyneitä ientaskuja ylemmissä kuin alemmissa 25(OH)D kvintiileissä.

Lung cancer has been considered one of the leading causes of deaths while cancer re- mains the second most common cause of deaths in the USA. Understanding the behavior of a disease over time could yield important information to make decisions about the disease. Statistical models could provide crucial clues and help to make a decision about the dis- ease, budget allocation, evaluation, and implement prevention. Longitudinal trend analysis of the diseases helps to understand long term effects and nature. Cholesterol level is one of the most contributing risk factors for Coronary Heart Disease. Studying cholesterol statistically helps to know more about its nature and provides crucial information to mitigate its effectiveness in diagnosing its impact to public health. In our study, we have analyzed lung cancer mortality in the USA based on age at death, period at death, and birth cohort to investigate its nature in longitudinal effects. The attempt has been made to estimate mortality rate based on age for different age groups and to find the relative risk of mortality due to period effect and relative risk due to birth cohort for lung cancer in the United States. Our statistical analysis and modeling are based on the data obtained from Surveillance Epidemiology and End Results (SEER) program of the United States. We have also investigated the probabilistic behavior of average cholesterol level based on gender and ethnicity. The study reveals significant differences with respect to the distribution they follow and their basic inferences which could be beneficial to draw conclusions in various ways in addressing related issues. At the same time, the change of cholesterol level over time for an individual might be a good source to study the association of cholesterol level, coronary heart disease and their effects on age. The cholesterol data is obtained from inter-university Consortium for Political and Social Research and National Health and Nutrition Examination Survey (NHANS) of the United States. Understanding the average change in total serum cholesterol level over time as people get older could be vital to explore it. We have studied the longitudinal behavior of the association of sex and time with cholesterol level. It is observed that age, sex, and time have an individual effect and can impact differently upon collective considerations. Their adverse effect in increasing cholesterol level could promote to worsen the cholesterol re- lated issues and hence heart related diseases. We believe our study pivots knowing more about target population of cholesterol level and helps to have the useful inference about cholesterol levels for public health. Finally, we also analyzed the average cholesterol data using a functional data analysis approach to understand its nature and effect on age. Since functional data analysis approach presents more flexibility in modeling, it could provide more insight in studying cholesterol level.

Au laboratoire, une méthode originale d'encapsulation par transacylation entre l'alginate de propylène-glycol (PGA) et une protéine a été mise au point. Cette méthode est basée sur la création de liaisons amides entre les fonctions amines libres de la protéine et les groupes esters du PGA dans une phase aqueuse émulsionnée (E/H) après alcalinisation. Les microparticules obtenues, stables, biocompatibles et biodégradables, sont potentiellement intéressantes pour la délivrance de substances actives en thérapeutique ou en cosmétique.Le premier objectif de ce travail est d'étudier l'influence des propriétés physicochimiques des deux biopolymères (protéine et PGA) et de leurs solutions, ainsi que l'effet des paramètres de préparation sur la réaction de transacylation et sur les propriétés des microparticules obtenues. Pour cela, la sérumalbumine humaine (HSA) a servi de protéine modèle et les microparticules ont été préparées dans différentes conditions physicochimiques puis caractérisées. Différents liens ont été établis entre les propriétés physicochimiques des solutions initiales des deux polymères et les propriétés fonctionnelles des microparticules obtenues.Le deuxième objectif est de remplacer le PGA, seul polysaccharide utilisable jusqu'à présent pour la microencapsulation par transacylation, par d'autres polysaccharides naturels, dans la préparation de microparticules. Etant donné ses propriétés intrinsèques limitantes, le remplacement du PGA par d'autres esters polysaccharidiques parait avantageux dans le domaine d'application des microparticules.Dans ce travail, le PGA a été remplacé par une série d'esters semi-synthétiques d'alginate puis par d'autres polysaccharides estérifiés naturels (pectines) ou semi-synthétiques (esters polypectiques et esters de l'acide hyaluronique). Les conditions optimales pour l'utilisation de chaque ester ont été alors déterminées. / In our laboratory, an original method of microencapsulation was developed, based on the use of a transacylation reaction, creating covalent bonds between proteins and propylene glycol alginate (PGA). The covalent bonds are created after alkalization of the aqueous phase of a W/O emulsion, without using bifunctional crosslinking reagent.The resulting microparticles, which are stable, biocompatible and biodegradable, have potential applications for the delivery of active compounds for therapeutics or cosmetics.The first aim of this work is to study the influence of the physicochemical properties of the two polymers (protein and PGA) and of their solutions, as well as the effect of the preparation parameters on the transacylation reaction and on microparticle characteristics. For this purpose, human serum albumin (HSA) was picked as a model protein and microparticles were prepared using several physicochemical conditions then characterized. Several relationships were established between the physicochemical properties of the initial solutions of the two polymers and the functional properties of the resulting microparticles.The second purpose is to replace the PGA, only polysaccharide used for microencapsulation by transacylation so far, by other natural polysaccharides in the preparation of microparticles. Given its limiting intrinsic properties, the replacement of PGA by other polysaccharidic esters seems advantageous in the field of microparticle applications.In this work, the PGA was successfully replaced by a series of semisynthetic alginate esters, and then by other polysaccharidic esters, either natural esters (pectin) or semisynthetic esters (polypectate esters and hyaluronate esters). The optimal conditions for the use of each ester were then determined.

Introduction : Trauma is the leading cause of death in patients between the ages of 1-44 years in South Africa. Taking these statistics into consideration it is essential to better resuscitation strategies in order to improve outcome of trauma patients. Compensated and uncompensated haemorrhagic shock is frequently under diagnosed in trauma patients, which has a definitive effect on mortality. Concerns about inadequate monitoring of patients through the use of only physiological end points are raised. Methods : A comprehensive literature review was conducted on resuscitation strategies for patients presenting with haemorrhagic shock. Physiological and metabolic end points of resuscitation were identified to guide resuscitation strategies. A quantitative, retrospective, non-experimental, descriptive, correlational and crosssectional research design was chosen for this study. Data was collected by using biophysical measures, namely clinical audit checklists. Results : Lactate was identified as a good indicator to predicting mortality in patients presenting with shock caused by haemorrhage. Special consideration to patients’ age and physiological status should be made during resuscitation. The consequence of delayed resuscitation in haemorrhagic shock patients is associated with an increase in mortality that can be prevented. It is found that serum blood lactate levels taken over time are good predictors of patient survival rates. Patients presenting with a raised serum blood lactate level for more than 24-hours has an increase in mortality rate. Conclusion : Emergency nurse practitioners are responsible for the monitoring of patients admitted to the emergency unit with haemorrhagic shock. The use of serum lactate levels during the first 24-hours of the resuscitation of patients with haemorrhage can assist with the implementation of strategies to reverse the effect haemorrhagic shock on cellular level in these patients. / Dissertation (MCur)--University of Pretoria, 2011. / Nursing Science / unrestricted

Alpha 2 agonists are frequently used in equine medicine. This study focused primarily on α2 agonist-induced changes in PCV and TSP. A secondary aim of this study was to investigate the effects of α2 agonist on selected serum biochemical parameters and splenic size in order to identify potential causes for the changes seen in PCV and TSP. Four healthy adult mares were treated in a blinded, randomized, cross-over design with a single dose of xylazine (0.5 mg/kg), romifidine (0.04 mg/kg) or detomidine (0.01 mg/kg) intravenously, or detomidine (0.02 mg/kg) intramuscularly. A 1-week washout period was allowed between treatments. Haematology, TSP, COP, plasma osmolality, glucose, BUN, serum lactate, electrolytes, venous blood pH, ultrasonographic splenic size and degree of clinical sedation were evaluated at different time points post-injection and compared to baseline values. All treatments induced similar clinical sedation in the mares. A significant change over time in PCV and TSP following each treatment was identified, with overall median (range) maximal reductions compared to baseline of 20.9% (12.9 - 27.3%) and 5.8% (3.0 - 10.3%), respectively. Additionally, changes over time were significant for RBC count, BUN, COP and Ca2+, which decreased; and glucose, plasma osmolality, Na+ and splenic size, which increased, when compared to baseline. There was no significant main effect of treatment on PCV, TSP or any other parameters measured except for glucose. This study concluded that changes in PCV, TSP and other biochemical parameters induced by α2 agonists should be taken into consideration when assessing critically ill horses that received these drugs. There was evidence of splenic RBC sequestration as well as fluid shifts; therefore, the results suggest a multifactorial cause for the changes in PCV and TSP. / Dissertation (MSc)--University of Pretoria, 2011. / Companion Animal Clinical Studies / unrestricted

Infection is a leading cause of death in hemodialysis patients, partly due to dysfunctional immunity. Frequent dialysis therapy improves patient outcomes and quality of life. We hypothesize that extended home hemodialysis (EHHD) also improves immune function compared to conventional in-hospital hemodialysis (CHD); therefore, we designed a prospective matching-cohort clinical study to assess serum inflammatory markers and the functional capacity of monocyte-derived dendritic cells (MDDCs) and T-lymphocytes. Serum CRP was decreased in EHHD patients suggesting that extended dialysis may decrease inflammatory solute/cytokine levels. Compared to controls, MDDCs from hemodialysis patients had similar endocytic capacity, expression of co-stimulatory molecules, and T-cell activation capacity. However, CHD was associated with the highest expression of CD83 and CD40. Activated T-cells in CHD patients also produced significantly more immunosuppressive IL-10 compared to EHHD patients and controls. Therefore, EHHD may improve immune function by decreasing inflammation, MDDC pre-activation, and synthesis of immunosuppressive cytokines.

Le travail de la thèse présente une série de (bio)capteurs conductimétriques, à base de la clinoptilolite, pour la détermination de l’ammonium, de l’urée et de la L-arginine. La clinoptilolite, le matériau nanométrique, possédant des propriétés de la sorption intrinsèque et une capacité d’échange cationique vis-à-vis des espèces ammonium, a été d’abord utilisée pour la réalisation d’un microcapteur conductimétrique sélectif à NH4+. Ci-après, une application de ce nanomatériau dans les biocapteurs est favorable pour le fonctionnement dans les solutions tampons multicomposants. Parmi plusieurs variantes de biocapteurs à l’urée à base de la zéolite, la plus intéressante est le biocapteur, dans lequel la couche de la clinoptilolite, déposée sur le transducteur, a été recouverte par le dépôt de la couche de l’uréase et de la zéolite. Pour l’élaboration d’un biocapteur conductimétrique hautement sensible pour la détermination de la L-arginine, l’arginase et l’uréase ont été co-réticulées sur le transducteur. Une détermination quantitative de la L-arginine dans une solution buvable « Arginine Veyron » a montré un fort accord avec les données fournies par le producteur. Une procédure détaillée de l’optimisation du biocapteur conductimétrique pour la détection de la L-arginine dans le sérum bovin a été proposée. La clinoptilolite a été également appliquée comme un modificateur dans la co-immobilisation de l’arginase et l’uréase pour améliorer les caractéristiques analytiques de biocapteur conductimétriques pour la détermination de la L-arginine / Currentwork presents a serie of conductometric (bio)sensors based on clinoptilolite, for ammonium, urea and L-arginine determination. Clinoptilolite, a nanoscale material possessing exceptional sorption and cation-exchange properties toward ammonium species, was initially used for the development of NH4+-selective conductometric microsensor. The clinoptilolite-based microsensor was selective toward ammonium in the presence of interferences that are commonly found along with ammonium in natural waters. Hereafter, an application of this nanomaterial in biosensors is favorable for operation in multicomponent buffer solutions. Among the several variants of the urea biosensors based on zeolite, considerably better characteristics were obtained for the biosensor comprising a clinoptilolite adlayer and an upper layer of immobilized urease and zeolite. In the work, for first time was developed a highly sensitive conductometric biosensor for L-arginine determination based on arginase and urease co-immobilized in a single membrane. The results of a quantitative determination of L-arginine in a drinkable solution “Arginine Veyron”, obtained by the biosensor, were in high correlation with the data provided by the producer. The L-arginine conductometric biosensor was optimized for the serum analysis. Clinoptilolite was also applied as a modifier in co-immobilization of arginase and urease for the improvement of analytical characteristics of the conductometric biosensor for L-arginine determination

The one-humped or the dromedarian camel is a pseudo-ruminant mammal, well adapted to the hot and dry climates of the desert. Its ability to withstand torrid heat and extreme desiccation is of paramount importance to its survival. The studies presented in this thesis were designed to investigate and document the effect of dehydration in the presence or absence of angiotensin II (Ang II) AT1 receptor blocker (losartan) on blood constituents, electrolytes, hormones, neurotransmitters as well as liver and kidney enzymes in a subset of dehydrated camels and to compare them with hydrated camels. Additionally, we studied the response of atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP) and revealed for the first time the cardiac storage form of BNP in the camel heart. Dehydration induced significant increments in packed cell volume (PCV), white blood cells (WBC), gamma glutamyl-transferase (GGT), serum sodium, creatinine and urea levels, and a doubling in plasma cortisol and arginine vasopressin (AVP) levels. At the same time dehydration caused significant decrease in body weights, plasma insulin like growth factor-1 (IGF-1) and its binding protein-3 (IGFBP-3), and a 50% decrement in ANP and BNP levels. Moreover, dehydration with and without losartan resulted in significant changes in stress hormones and anti-oxidants in plasma, liver and kidney homogenates. Losartan on one hand enhanced the effect of dehydration resulting in significant increases in sodium, creatinine and urea levels. In addition losartan raised the  binding affinity of Ang II AT2 receptors in the small intestine with 8-fold and with 16-fold for liver AT1 receptors, indicating that Ang II AT1 and AT2 receptor binding sites were present in camel's small intestine while only AT1 receptor binding sites were found in the camel liver. One the other hand losartan resulted in significant decrease in body weights impaired the rise in anti-diuretic hormone and reduced aldosterone level. Finally, we showed that the proBNP is the storage form of BNP in the camel heart.

Interaction between bovine serum albumin (BSA) and thimerosal (TM), organic mercury compound, was investigated by spectroscopic methods. The results, by molecular fluorescence, show that the interaction takes place by static quenching with electrostatic forces spontaneously (ΔG = - 4.40 kJ mol-1 at 30°C). The binding constant (Kb) was 3.24 ± 0.01x103 L mol-1 (30°C) is considered a moderate interaction. Fluorescence in three dimensions revealed that TM causes structural involving the the polypeptide chain BSA changes in the polarity of the tryptophan and tyrosine residues confirmed by circular dichroism (CD) showed an increase in α-helix content after interaction with TM. In addition, the TM decreases the surface hydrophobicity of the protein. Bilirubin was used as a marker for the subdomain IB, confirming that TM interacts in this region of the protein. The study of the interaction mechanism proposed that TM is reacted with BSA through the free cysteine residue, forming the adduct BSA-HgEt release of thiosalicylic acid (ATS), which interacts with amino acids with side chain positive. Besides, it was seen that TM accelerates the protein fibrillation kinetics by 42%, with a possible indication of the toxicity of this compound in biological systems. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A interação entre albumina do soro bovino (BSA) e timerosal (TM), composto orgânico de mercúrio, foi investigada utilizando métodos espectroscópicos. Os resultados, por fluorescência molecular, evidenciam que a interação acontece por quenching estático através de forças eletrostáticas de forma espontânea (ΔG = - 4,40 kJ mol-1 a 30ºC). A constante de ligação (Kb) foi de 3,24 ± 0,01x103 L mol-1 (30ºC) sendo considerada uma interação moderada. A fluorescência em três dimensões revelou que TM causa mudanças estruturais envolvendo a cadeia polipeptídica da BSA assim como altera a polaridade dos resíduos de triptofano e tirosina, confirmada por dicroísmo circular (DC) que evidenciou aumento no conteúdo de α-hélice após interação com TM. Além disto, TM diminui a hidrofobicidade superficial da proteína. Bilirrubina foi utilizada como marcador para o subdomínio IB, confirmando que TM interage nesta região da proteína. O estudo do mecanismo de interação propôs que TM reage com BSA através do resíduo de cisteína livre, formando o aduto BSA-HgEt com liberação de ácido tiosalicílico (ATS), que interage com os aminoácidos com cadeia lateral positiva. Por fim, foi visto que TM acelera a cinética de fibrilação proteica em 42%, sendo um possível indício da toxicidade deste composto em sistemas biológicos.