No Brasil acidentes por peixes peçonhentos como Thalassophryne nattereri, Cathorops spixii, Scorpaena plumieri e Potamotrygon gr orbygnyi vêm sendo relatados. O quadro clínico é caracterizado por efeitos locais e sistêmicos. Para esses acidentes não existe tratamento adequado e eficiente. A soroterapia é um eficiente tratamento para reverter os efeitos causados por venenos. Diante disso nosso objetivo é a obtenção de um soro poliespecifico através da junção de soros monoespecificos produzidos em camundongos e avaliar a sua capacidade de neutralização das atividades tóxicas induzidas pelos principais peixes peçonhentos brasileiros. As alterações induzidas na microcirculação foram abolidas na pré-incubação e no tratamento imediato, somente dano em fibra muscular induzido pelo C. spixii exigiu soro na proporção 1:2. Obtivemos o controle parcial da nocicepção e edema. O efeito sistêmico foi eficientemente neutralizado. Nossos dados nos permitem sugerir o uso de soro poliespecífico nos envenenamentos causados pelos peixes T. nattereri, S. plumieri, C. spixii e P. gr orbygnyi. / Envenoming induced by venomous fish represents a great cost brazilian communities. Victims develop local and systemic symptoms. The most frequent founded are Thalassophryne nattereri, Cathorops spixii, Scorpaena plumieri and Potamotrygon gr orbygnyi. The anti-inflammatory drugs used are not efficient. After the accident, the passive transfer of heterologous specific antibodies allows that immediate toxic effect can be neutralized in the victims. The aim of this work was to produce in mice polyspecific antivenom and evaluation the neutralization of the main toxic effects induced by the brazilian venomous fish. The effects induced in microcirculation were abolished by the polyspecific antiserum, except for the effects caused by C. spixii in muscular fibers where was necessary a proportion of 1:2. The polyspecific antiserum controlled the edematogenic and nociceptive activities and the airway inflammation.These data allow us to suggest the use of polyspecific antiserum in the treatment of the pathological effects provoked by the most frequent brazilian venomous fish.

Background - The first 1000 days of life (from conception to two years of age) is a critical period of nutritional vulnerability, affecting lifelong health. Iron deficiency (ID) and iron deficiency anaemia (IDA) are considered major public health problems that adversely affect development and growth, impair immunity, and increase morbidity and mortality in infants. ID and IDA in sub-Saharan Africa can be attributed to poor dietary, socioeconomic and disease conditions. One of the major obstacles in determining the prevalence of ID, using serum ferritin (SF) as marker of iron status, is that it not only reflects the amount of iron that is stored in the body, but also functions as an acute phase reactant that is raised in the presence of infection or inflammation. Aim - We conducted a re-analysis of the International Research on Infant Supplementation (IRIS) study’s baseline data to determine a more accurate estimation of the ID prevalence in apparently healthy four to thirteen-month-old infants from rural KwaZulu-Natal while accounting for the effect of chronic and acute inflammation on SF. Study design and methods - A cross-sectional analysis was performed on the baseline data (192 infants) of the IRIS study that was conducted in 2000. Infants’ haemoglobin (Hb), SF, C-reactive protein (CRP) and alpha-1 glycoprotein (AGP) concentrations were interpreted to determine the prevalence of ID. Literature of the past four years served as a guide to compare the ID prevalence obtained from four methods that account for the influence of inflammation on SF concentrations, to a reference method that does not take inflammation into consideration, and to what was reported in the original IRIS study. Weight and recumbent length measurements were converted to z-scores to interpret subjects’ anthropometric nutritional status. Results - A high prevalence of inflammation (52.6%) was present, with 11.5% of the subjects being in the incubation, 17.2% in the early convalescent, and 24% in the late convalescent phase of inflammation. SF was significantly associated with both CRP (ß = 0.200; P = 0.005) and AGP (ß = 0.223; P = 0.002) when adjusting for gender and age. The IRIS study reported an ID prevalence of 18.3%, whereas the results of this study ranged from 17.2 to 52.1%. We derived an IDA prevalence that ranged from 12 to 24.5% according to the different methods. The prevalence of stunting [length-for-age Z-score <-2SD] was 12.5%; while 25.1% of infants were overweight/obese [weight-for-length z-score >2SD]. Conclusion - A double burden of malnutrition was evident from the high prevalence of both overweight and ID, together with inflammation. The disconcertingly large variance in ID prevalence observed between the different methods that were employed highlights that iron supplementation interventions to treat anaemia must be based upon accurate estimates of IDA prevalence, otherwise they pose an increased risk of adverse effects to susceptible, iron-replete, but anaemic infants. Given the detrimental consequences of ID, it is imperative that governments, health care providers and parents must act to prevent or treat ID and IDA among vulnerable infants. / MSc (Dietetics), North-West University, Potchefstroom Campus, 2014

Les Bone Morphogenetic Proteins (BMP) jouent un rôle important dans de nombreux processus physiopathologiques, en particulier dans les processus d'angiogenèse. La mise au point de méthodes de détection sensibles et spécifiques pourrait être utiles pour des applications cliniques potentielles. Ce travail s'intéresse à BMP9, molécule récemment identifiée par notre équipe comme le ligand physiologique et spécifique du récepteur orphelin Activin Receptor-Like Kinase 1 (ALK1). BMP9 est présent dans le sang et joue un rôle de facteur de quiescence de l'endothélium vasculaire. Ces observations nous ont permis de proposer que la concentration sanguine de BMP9 pourrait être un marqueur de l'état d'activation de l'endothélium vasculaire. Afin de valider cette hypothèse, trois objectifs ont été définis : 1/ mettre au point une méthode de dosage sensible et spécifique de BMP9 circulant, 2/ caractériser les sites d'expression et les formes circulantes de BMP9 et 3/ évaluer la pertinence de BMP9 comme biomarqueur de l'angiogenèse physiologique et pathologique, en particulier tumorale. La première partie de mon travail de thèse a consisté à développer deux méthodes de dosage. La première consiste en une méthode biologique, basé sur l'utilisation d'un gène rapporteur sous le contrôle du promoteur BRE (BMP Responsive Element) qui répond à l'activation de la voie des BMPs. La seconde méthode est basée sur un système « sandwich » de type immunoenzymatique. Les caractéristiques de ces méthodes ont été établies. Dans un second temps, grâce à ces outils, nous avons montré que BMP9 est une protéine produite principalement par les hépatocytes et qu'elle circule majoritairement sous une forme active, complexée à son pro-domaine. Une forme inactive homo-dimérique non clivée est aussi synthétisée. Ce travail confirme également le rôle de BMP9 dans la quiescence de l'endothélium vasculaire. Dans la dernière partie de ce travail, le taux de BMP9 a été mesuré dans des fluides biologiques de patient(e)s présentant diverses situations physiopathologiques. Une augmentation de la concentration sanguine de BMP9 est observée peu de temps avant l'accouchement ; chez les patientes pré-éclamptiques des taux plus élevés de BMP9 sont aussi observés, ouvrant des perspectives dans le domaine obstétrical. Dans les pathologies tumorales, nos observations montrent que BMP9 n'a pas d'intérêt en tant que marqueur diagnostique ou pronostique. Toutefois, les données recueillies dans le cancer médullaire de la thyroïde traité par vandétanib suggèrent que la mesure du taux de BMP9 circulant pourrait avoir une valeur prédictive sur l'efficacité du traitement. Ainsi, l'utilisation du dosage de BMP9 comme marqueur prédictif ou pharmacodynamique des thérapies anti-angiogéniques doit être envisagé. En effet, à l'heure actuelle, l'index thérapeutique des traitements anti-angiogéniques pourrait être amélioré s'il était possible d'identifier et de valider des biomarqueurs susceptibles de prédire précocement l'efficacité clinique ou le développement de résistances au traitement. Cette perspective apparait d'autant plus intéressante que deux molécules, ciblant la voie de signalisation Alk1 ont récemment été développées et présentent des résultats prometteurs, en particulier chez les patients présentant une résistance aux anti-VEGF. / The Bone Morphogenetic Proteins (BMPs) play an important role in many pathophysiological processes, particularly in angiogenesis. The development of sensitive and specific detection methods could be useful for potential clinical applications. This work focuses on BMP9, a molecule recently identified by our team as the physiological and specific ligand of the orphan receptor Activin Receptor-Like Kinase 1 (ALK1). BMP9 is a circulating vascular quiescence factor. These observations have allowed us to propose that BMP9 blood concentration could be a biomarker of the activation state of the vascular endothelium. To validate this hypothesis, three objectives were defined: 1 / Develop a sensitive and specific assay of circulating BMP9, 2 / characterize expression sites and circulating forms of BMP9 and 3 / evaluate the relevance of BMP9 as a biomarker for physiological and pathological angiogenesis, in particular tumor. The first part of my thesis was to develop two assays. The first is a biological method, based on the use of a reporter gene under the control of the promoter BMP Responsive Element (BRE) which responds to the activation of the BMPs. The second method is based on a "sandwich"-type immunoassay. The characteristics of these methods have been established. In a second step, using these tools, we have shown that BMP9 is a protein produced mainly by hepatocytes and circulates predominantly in an active form complexed to its pro-domain. An inactive form homo-dimeric uncleaved is also synthesized. This work also confirms the role of BMP9 in the quiescence of vascular endothelium. In the last part of this work, the rate of BMP9 was measured in biological fluids of the patient (s) with various pathophysiological situations. An increase in the blood concentration of BMP9 is observed shortly before delivery and in preeclamptic patients higher rates of BMP9 were also observed, opening perspectives in obstetrics. In tumoral pathology, our observations show that BMP9 has no value as diagnostic or prognostic marker. However, the data collected in medullary carcinoma of the thyroid treated by vandetanib suggest that the measurement of circulating BMP9 rates could have a predictive value for the efficacy of treatment. So, the use of the BMP9 assay as a predictive marker or pharmacodynamic marker of anti-angiogenic therapies should be considered. Indeed, at present, the therapeutic index of anti-angiogenic treatments could be improved if it were possible to identify and validate biomarkers that may predict early clinical efficacy or the development of resistance to treatment. This perspective appears particularly interesting since two molecules targeting the signaling pathway Alk1 have recently been developed and show promising results, especially in patients with resistance to anti-VEGF

No Brasil acidentes por peixes peçonhentos como Thalassophryne nattereri, Cathorops spixii, Scorpaena plumieri e Potamotrygon gr orbygnyi vêm sendo relatados. O quadro clínico é caracterizado por efeitos locais e sistêmicos. Para esses acidentes não existe tratamento adequado e eficiente. A soroterapia é um eficiente tratamento para reverter os efeitos causados por venenos. Diante disso nosso objetivo é a obtenção de um soro poliespecifico através da junção de soros monoespecificos produzidos em camundongos e avaliar a sua capacidade de neutralização das atividades tóxicas induzidas pelos principais peixes peçonhentos brasileiros. As alterações induzidas na microcirculação foram abolidas na pré-incubação e no tratamento imediato, somente dano em fibra muscular induzido pelo C. spixii exigiu soro na proporção 1:2. Obtivemos o controle parcial da nocicepção e edema. O efeito sistêmico foi eficientemente neutralizado. Nossos dados nos permitem sugerir o uso de soro poliespecífico nos envenenamentos causados pelos peixes T. nattereri, S. plumieri, C. spixii e P. gr orbygnyi. / Envenoming induced by venomous fish represents a great cost brazilian communities. Victims develop local and systemic symptoms. The most frequent founded are Thalassophryne nattereri, Cathorops spixii, Scorpaena plumieri and Potamotrygon gr orbygnyi. The anti-inflammatory drugs used are not efficient. After the accident, the passive transfer of heterologous specific antibodies allows that immediate toxic effect can be neutralized in the victims. The aim of this work was to produce in mice polyspecific antivenom and evaluation the neutralization of the main toxic effects induced by the brazilian venomous fish. The effects induced in microcirculation were abolished by the polyspecific antiserum, except for the effects caused by C. spixii in muscular fibers where was necessary a proportion of 1:2. The polyspecific antiserum controlled the edematogenic and nociceptive activities and the airway inflammation.These data allow us to suggest the use of polyspecific antiserum in the treatment of the pathological effects provoked by the most frequent brazilian venomous fish.

Vid utförande av immunokemiska analyser används serum och plasma. Advia Centaur XPT är det instrument som används på Centrallaboratoriet Universitetssjukhuset Örebro inom immunokemi. Metoden baseras på kemiluminescens med mono- och/eller polyklonala antikroppar och akridiumester som utgör basen för detektion med direkt kemiluminescens. Instrumentet använder två typer av metoder: sandwichmetod och kompetitiv metod. Studien gick ut på att utföra analys av thyroideastimulerande hormon, fritt tyroxin, fritt trijodtyronin, thyroid peroxidas antikroppar, ferritin, pro B-typ natriuretisk peptid och högkänslig Troponin I på serum och plasma från 25 individer, vid 0-tidpunkt och efter 24, 48 och 72 timmar. Analysresultaten och skillnaderna mellan de två matriserna och tidpunkterna utreddes för statistiskt signifikanta och medicinskt betydelsefulla skillnader. De flesta analyterna visade stabilt resultat över tid och att de kan sparas och analyseras fram till 72 timmar, förutom TPO-antikroppar som verkar vara en instabil och känslig analys. Resultatet av jämförelsen mellan serum och plasma visade inte signifikant skillnad varför det går att övergå till alternativ matris serum/plasma i fall originalmatrisen (serum/plasma beroende på analys) saknas. / Serum and plasma are used in performing immunochemical analyzes. Advia Centaur XPT instruments are used in immunochemistry at the Central Laboratory at Örebro University Hospital. The method is based on chemiluminescence with the use of mono- and/or polyclonal antibodies, and acridium ester for direct chemiluminescence detection. The instrument uses two types of methods: sandwich method and competitive method. The study focused on performing analysis for thyroid stimulating hormone, free thyroxine, free triiodothyronine, thyroid peroxidase antibodies, ferritin, pro B-type natriuretic peptide and highly sensitive Troponin I on serum and plasma from 25 individuals, at time 0 and after 24, 48 and 72 hours. The results between the different time points and the two matrices, were examined to determine whether there was any statistically significant or medically unacceptable bias. Most analytes showed stable results over time and that they can be used and analyzed up to 72 hours, except for TPO antibodies which appear to be an unstable and sensitive analysis. Results in comparison between serum and plasma did not show a significant difference and therefore it is possible to switch to an alternative matrix    serum/ plasma in cases where the original matrix (serum / plasma depending on analysis) is missing.

Le Syndrome de Détresse Respiratoire Aiguë (SDRA) est la forme la plus sévère d’agression alvéolaire aiguë. Il estcaractérisé par un dommage alvéolaire diffus, suivi d’une phase de réparation nécessaire à la guérison. Bien queles monocytes/macrophages soient des acteurs incontournables de la pathogénie et de la résolution du SDRA, leurpolarisation et leur rôle dans la réparation alvéolaire restent mal connus chez l’Homme. L’hypothèse défendue parcette thèse est que l’environnement alvéolaire module la différenciation et la polarisation desmonocytes/macrophages au cours du SDRA, et que les macrophages alvéolaires ainsi polarisés participentactivement à la réparation et à sa régulation. Les principaux résultats de nos travaux ont permis d’établir que : 1)l’environnement alvéolaire de SDRA inhibe la différenciation des monocytes en fibrocytes (précurseursmésenchymateux associés à la fibroprolifération et à pronostic péjoratif). L’inhibition est majoritairement due à laSerum Amyloid protein P (SAP), provenant en partie du relargage de ses stocks matriciels pulmonaires à la faveurde la lésion alvéolaire ; 2) l’environnement alvéolaire de SDRA induit une polarisation anti-inflammatoire desmacrophages se rapprochant de la polarisation induite in vitro par l’IL-10 ; 3) les macrophages anti-inflammatoirespolarisés par le lavage broncho-alvéolaire (LBA) de patients SDRA favorisent la réparation alvéolaire épithéliale viala production polarisation-dépendante d’Hepatocyte Growth Factor (HGF). Cette production macrophagique d’HGFest amplifiée via une boucle autocrine PTGS2/PGE2 chez l’Homme ; 4) ces résultats semblent étayés par lesdonnées obtenues sur une cohorte clinique qui montrent que les concentrations de sCD163 (marqueur depolarisation anti-inflammatoire) et d’HGF rapportées au nombre de macrophages alvéolaires sont plus élevéeschez les patients survivants que chez les patients décédés de SDRA. L’ensemble de nos travaux démontrent pour lapremière fois chez l’Homme le rôle bénéfique de l’environnement alvéolaire sur les monocytes/macrophages,d’une part en inhibant leur différenciation en fibrocytes contribuant ainsi à limiter la fibroprolifération pulmonaire,et d’autre part en induisant un phénotype macrophagique anti-inflammatoire, contribuant à limiter l’inflammationengendrée par la lésion alvéolaire initiale et favorisant la réparation épithéliale via la production d’HGF. Lesdonnées physiopathologiques obtenues pourraient permettre d’envisager la reprogrammation anti-inflammatoiredes macrophages comme une cible thérapeutique du SDRA en cas d’excès d’inflammation ou de fibro-proliférationavec réparation aberrante. / Acute Respiratory Distress Syndrome (ARDS) is the most severe form of acute lung injury. ARDS is characterized bydiffuse alveolar damage followed by a phase of alveolar repair necessary to recovery. Althoughmonocytes/macrophages are key actors of pathogenicity and resolution of ARDS, little is known about theirpolarization and role on alveolar repair during human ARDS. The hypothesis of our studies was that ARDS alveolarenvironment modulates differentiation and polarization of monocytes and macrophages, and that polarizedmacrophages are involved in alveolar repair and its regulation. The main results of our work have shown that: 1)ARDS alveolar environment inhibited monocytes differentiation into fibrocytes (mesenchymal progenitorsassociated with fibroprolifération and a poor prognosis), mainly through its Serum Amyloid P (SAP) content,originating, at least in part, from the release of SAP associated with lung connective tissue during ARDS; 2) ARDSalveolar environment drove an anti-inflammatory macrophage polarization, close to that induced by IL-10 in vitro;3) anti-inflammatory macrophages polarized by broncho-alveolar lavage (BAL) from ARDS patients favored alveolarepithelial repair through a polarization-dependent production of Hepatocyte Growth Factor (HGF). This HGFproduction is amplified by an autocrine PTGS2/PGE2 dependent loop in human macrophages; 4) these results mayhave clinical relevance, since sCD163 (a marker of anti-inflammatory polarization) and HGF concentrations,expressed relatively to BAL macrophage count, were higher in ARDS survivors than non-survivors. Taken together,our works demonstrate for the first time the beneficial role of the ARDS alveolar environment onmonocytes/macrophages, inhibiting their differentiation into fibrocytes, thus limiting excessive lungfibroproliferation, and inducing an anti-inflammatory macrophage polarization, thus limiting the inflammationgenerated by the initial alveolar damage and favoring epithelial repair through HGF production. The datapresented in this thesis may allow considering anti-inflammatory macrophage repolarization as a potential newtherapeutic target of ARDS with excessive inflammation or fibro-proliferation with aberrant repair.

Steady advancements in diagnostics over the past century have propelled the world of medicine into the more advanced era of preventative medicine, an era with a resoundingly clear message: early detection can save lives. For patients who suffer from either pancreatic cancer or malignant hyperthermia susceptibility, early or preoperative diagnosis, respectively can save lives and minimize morbidity and mortality, in addition to offering cost-savings to hospitals and healthcare systems. Fortunately, significant progress have been made in the fields of metabolomics and biomarker identification. Given the benefits carried by serum biomarkers as targets of screening and diagnostic tool development, we applied functional nucleic acid technology and in vitro selection directly in whole human serum to search for disease-specific biomarkers and associated detection probes without a priori knowledge of the biomarkers pursued. This endeavour simultaneously serves as a proof-of-concept study to establish whether in vitro selection can be successfully performed in human serum. We specifically focused on the derivation of RNA-cleaving DNAzymes (RCD) through in vitro selection, or SELEX (systemic evolution of ligands through exponential exposure). DNAzymes constructed with a fluorogenic signalling molecule were incubated with human serum with the goal of identification of a functional nucleic acid probe capable of detecting the presence of a disease-specific biomarker. Two independent protocols have been designed and executed for the identification of DNAzyme sequences capable of detecting pancreatic cancer and malignant hyperthermia susceptibility, respectively. The first exploration was performed in serum obtained from cancer patients, with the goal of identifying DNAzymes capable of distinguishing pancreatic cancer from other cancer types. To do so, we employed in vitro selection, Next-Generation Sequencing, and bioinformatic analysis. We successfully demonstrated the feasibility of performing in vitro selection with DNAzymes in human serum, evidenced by distinct round-to-round enrichment of a DNA library towards the identification of DNAzymes capable of detecting pancreatic cancer. Additionally, we isolated two DNAzymes capable of distinguishing pancreatic cancer serum from healthy patient serum in fresh collected serum samples. Based on the positive results gathered in the pancreatic cancer in vitro selection project, we subsequently endeavoured to replicate the demonstrated feasibility of performing in vitro selection in human serum. By selecting malignant hyperthermia as the pathology investigated, we simultaneously sought to diversify the scope of DNAzyme detection by establishing whether successful DNAzyme selection can be achieved in a non-acute disease state. Thus, the second exploration was performed in serum obtained from patients who underwent evaluation for malignant hyperthermia susceptibility using the gold-standard caffeine-halothane contracture test. The goal of this project rested on the identification of DNAzymes capable of distinguishing malignant hyperthermia susceptibility in serum and approximating the performance of the gold standard test. We successfully isolated four DNAzyme candidates which demonstrated clinically relevant thresholds of sensitivity and specificity following thorough sensitivity and specificity analysis. In doing so, we once again demonstrated the ability to perform in vitro selection in human serum. Given the complexity of molecular interactions observed over the course of two in vitro selection protocols in human serum, it became clear that distinguishing meaningful target-mediated interactions from non-specific interactions would require advanced bioinformatic analysis. Consequently, using principles of computational biology, we performed a deep exploration of Next-Generation Sequencing results obtained from sequencing our recovered DNA libraries to extract additional data that would inform on the next required steps required to identify a DNAzyme specific for the pathology pursued. In doing so, we identified a two-step method to evaluate the progress of the in vitro selection protocol undertaken, and offered a systematic approach for choosing candidate sequences to undergo further testing based on promising performance in silico. Using this approach, we successfully identified a DNAzyme sequence capable of acting as a general cancer detection probe, with promising potential for diagnostic application. Ultimately, this thesis serves as a feasibility study of a novel approach to both in vitro selection and biomarker identification technique by combining the latest nanotechnology techniques with clinical data and real patient serum samples, and advanced computational biology tools. Despite the inability to identify a highly sensitive and specific DNAzyme capable of advancing towards biosensor construction, several important strides and lessons have been acknowledged, establishing the feasibility of performing in vitro selection in human serum, and outlining strategies for addressing and anticipating challenges with this technique. The hope is for this work to inspire and inform future efforts to apply functional nucleic acid technology to solve current gaps in both the diagnostic and therapeutic branches of medicine, and with the help of computational biology continue to bridge the gap between basic science and clinical medicine. / Dissertation / Doctor of Philosophy (PhD)

Atherosclerotic cardiovascular diseases (CVD), particularly coronary heart disease (CHD) and cerebrovascular disease, are today major causes of death in the industrialised parts of the world. There are evidence to suggest that the atherosclerotic process starts in childhood, implying that preventive measures should be implemented already in children and adolescents. The aim of this study was to examine CVD risk indicators and their determinants in healthy Swedish adolescents. The study population comprised 14- and 17-year-old boys and girls (n=1032), in the dty and surroundings of Umeå in northern Sweden. Biochemical, anthropometric, and physiological parameters associated to CVD (s- lipoproteins and s-apolipoproteins, s-insulin, s-ferritin, anthropometric measurements, blood pressure, and physical fitness) were evaluated in relation to family history of CVD, weight and length at birth, infant feeding regimen, physical growth during infancy and childhood, current diet, physical activity, smoking, and educational level and occupation of the parents. The main findings of the study were that, on average, total serum cholesterol (TC) values in boys and girls were at the same level as reported from other European countries. A family history of CVD, short duration of breast feeding, low attained height during infancy and childhood, high body mass index (BMI), and low physical fitness were all associated with an unfavourable serum lipid profile. The findings also showed that features typical of the insulin resistance syndrome are present already in adolescents. In boys, iron stores, estimated by serum ferritin, were related to BMI and physical fitness, in a similar way as well established CVD risk indicators. Compared to previous dietary studies in Sweden, mean relative (energy %) fat intake had decreased substantially although the mean relative intake of saturated fat was still rather high. For both boys and girls, reported relative energy intake (energy intake/estimated energy expenditure) decreased with increasing level of BMI. Furthermore, daily smoking was more common among adolescents from families with low socio-economic status (SES) but was most strongly associated to smoking in peers. Tobacco use was considerably higher among adolescents attending vocational programs at secondary high school as compared to theoretical programs. Daily smokers had a more unfavourable serum lipid profile compared to non-smokers. Low socio-economic status of the parents was related to higher BMI and low educational level to higher dietary fat intake in both boys and girls. In conclusion, the findings of the study show that parameters linked to adult CVD when examined in adolescents, are related to family history, infant nutrition, previous physical growth, current body composition, physical fitness, physical activity, smoking, and social status and educational level of the parents. / digitalisering@umu

Call number: LD2668 .T4 1956 L66 / Master of Science

Submitted in fulfilment of the requirements of the Degree of Master of Technology: Chemistry, Durban University of Technology, 2012. / The present work involves the interaction studies of chiral compounds with the Human Serum Albumin (HSA) protein using computational and experimental methods. The HSA protein has multiple binding sites that forms the basis for its exceptional ability to interact with many organic and inorganic molecules, which makes this protein an important regulator of intercellular fluxes and the pharmacokinetic behaviour of many drugs. This study was undertaken to evaluate the related pharmacokinetic and enantioselective binding parameters of the racemic catechin enantiomers with the HSA. Accordingly, this work involved a method development for the chiral separation of a racemic compound, by capillary electrophoresis-electrokinetic chromatography (CE-EKC) with a highly sulphated beta-cyclodextrin (HS--CD) as a chiral selector. The experimental work was supported by two molecular docking studies. The first included the mimicking of the host-guest interactions between a chiral selector and an enantiomeric compound. The second study included the estimation of the pseudo enantioselective (ES) binding of catechin to HSA. Overall, it was found that CE-EKC is the preferred method for the(±)-catechin binding to HSA protein evaluation. Moreover, the technique used in this work is not restricted to HSA or polyphenols, but can also be applied to other proteins and ligands that possess chirality. Furthermore, the molecular docking approaches also proved to be very useful for the evaluation of chiral recognition systems and for elucidation of the ligand-protein interactions.