by Priscilla Miu-kuen Poon. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1991. / Includes bibliographical references. / ACKNOWLEDGEMENT --- p.1 / ABSTRACT --- p.2 / CONTENTS / Chapter 1. --- INTRODUCTION --- p.4 / Chapter 1.1 --- Discovery of vitamin D / Chapter 1.2 --- Bioavailability of vitamin D and its metabolites / Chapter 1.3 --- Metabolism of vitamin D and its metabolites / Chapter 1.4 --- Mode of action of vitamin D / Chapter 1.5 --- Vitamin D-related diseases / Chapter 2. --- METHODS OF MEASURING VITAMIN D AND ITS METABOLITES --- p.32 / Chapter 2.1 --- Deproteinization / Chapter 2.2 --- Extraction / Chapter 2.3 --- Separation / Chapter 2.4 --- Quantitation / Chapter 3. --- OBJECTIVES --- p.51 / Chapter 4. --- MATERIALS & METHODS --- p.52 / Chapter 4.1 --- Materials / Chapter 4.2 --- General methods / Chapter 4.3 --- Blood collection / Chapter 4.4 --- Radioreceptor assay / Chapter 4.5 --- Serum treatment / Chapter 4.6 --- High Performance Liquid Chromatography (HPLC) / Chapter 4.7 --- Gas Chromatography-Mass Spectrometry (GC-MS) / Chapter 4.8 --- "Serum 1α,25-dihydroxyvitamin D3 analysis" / Chapter 4.9 --- Application of the established GC-MS method / Chapter 4.10 --- Study on hypercalcaemia of tuberculosis / Chapter 5. --- RESULTS --- p.66 / Chapter 5.1 --- Analysis of vitamin D3 standard / Chapter 5.2 --- "Analysis of 1α,25-dihydroxyvitamin D3 standard" / Chapter 5.3 --- Separation of vitamin D3 metabolites / Chapter 5.4 --- "Analysis of lα,25-dihydroxyvitamin D3 in serum samples" / Chapter 5.5 --- Study on hypercalcaemia of tuberculosis / Chapter 6. --- DISCUSSIONS --- p.118 / Chapter 6.1 --- Derivatization / Chapter 6.2 --- Optimization of GC-MS parameters / Chapter 6.3 --- Sample pre-treatment / Chapter 6.4 --- "GC-MS analysis of serum lα,25-dihydroxyvitamin D3" / Chapter 6.5 --- Study on hypercalcaemia of tuberculosis / Chapter 7. --- CONCLUSION --- p.129 / LIST OF ABBREVIATIONS --- p.131 / LIST OF FIGURES --- p.134 / LIST OF TABLES --- p.137 / REFERENCES --- p.139

Les leucémies aiguës myéloblastiques (LAM) sont un groupe hétérogène de pathologies malignes représentant environ 70% des leucémies aiguës. Il existe une prolifération, dans le cadre des LAM de cellules immatures appartenant à la lignée myéloïde appelées myéloblastes ou communément blastes. Les traitements actuels reposent essentiellement sur la chimiothérapie antimitotique. L’homéostasie du fer est une cible dans le traitement des LAM en induisant la différentiation des blastes. Le mécanisme implique la modulation des ROS. Leur action est synergique avec celle de la Vitamine D (VD) au travers de l’activation de la voie des MAPK. Cette association a été utilisée chez plusieurs patients avec succès permettant un doublement de leur espérance de vie. Nous avons ensuite montré que l’expression du récepteur expression vitamine D (VD) est altérée dans les états indifférenciés / immatures sous-types de LAM et que la diminution de l'expression du VDR et de ces gènes cibles est corrélée à un mauvais pronostic chez les patients. Le mécanisme moléculaire entraînant le blocage de l'expression VDR implique la méthylation de son promoteur. Les souris invalidées pour le VDR ont une expansion du compartiment des cellules souches hématopoïétiques demeurant à un état quiescent ainsi qu’une diminution des niveaux du stress oxydatif en leur sein. En outre, la transformation maligne des cellules déficientes en VDR a abouti à une différenciation myéloïde limitée, à l'augmentation du nombre de progéniteurs hématopoïétiques précoces et ces cellules présentaient un potentiel d'auto-renouvellement accru et étaient résistantes aux inhibiteurs de la méthyltransférase et à la chimiothérapie. Enfin, l'induction de l'expression du VDR dans les modèles de LAM par un traitement combinant des agents de déméthylation et les agonistes de VDR a permis de diminuer la séminalité, de promouvoir la différenciation cellulaire, de bloquer la croissance tumorale et de restaurer la sensibilité à la chimiothérapie. Par conséquent, nous proposons que le VDR est un gène maître contrôlant la séminalité et la prolifération / différenciation cellulaire des cellules souches hématopoïétiques normales et leucémiques. Ainsi, la combinaison d'agents déméthylants et d’agonistes de VDR pourrait à l’avenir être proposée en thérapeutique pour traiter les LAM. / Acute myeloid leukemia (AML) is a heterogeneous group of malignancies representing approximately 70% of acute leukemias. There is a proliferation of immature cells belonging to the myeloid lineage commonly called myeloblasts or blasts. Current treatments are mainly based on antimitotic chemotherapy. Iron homeostasis is a target for the treatment of AML blasts inducing cell differentiation. The mechanism involves the modulation of ROS. Their action is synergistic with that of Vitamin D (VD) through the activation of MAPK. This association has been used successfully in several patients for a doubling of life expectancy. Then, we show that Vitamin D receptor (VDR) expression was impaired in undifferentiated/immature AML subtypes and that decreased expression of VDR and VDR-targeted genes was correlated with a negative prognosis of patients. Molecular mechanism resulting in the blockade of VDR expression involved VDR promoter methylation. VDR-deficient mice showed an expansion of the hematopoietic stem cell compartment which presented an improved quiescent status and decreased ROS levels that have been shown to be involved in both AML differentiation and stem cells longevity. Moreover, malignant transformation of VDR-deficient cells resulted in limited myeloid differentiation, increased numbers of early hematopoietic progenitors and those cells presented an enhanced self-renewal potential and were resistant to DNA methyltransferase inhibitors and to chemotherapy. Finally, induction of VDR expression in AML models by combined treatment of demethylating agents and VDR agonists decreased stemness, promoted cell differentiation, blocked tumor propagation and restored sensitivity to chemotherapy. Therefore, we propose that VDR is a master gene controlling stemness and proliferation/cell differentiation of normal hematopoietic stem cells and leukemic cells. Thus, combination of demethylation agents and VDR agonists may be used therapeutically to treat AML.

Submitted by Rosivalda Pereira (mrs.pereira@ufma.br) on 2017-12-06T17:06:52Z No. of bitstreams: 1 JacquelineCantanhede.pdf: 6344249 bytes, checksum: c518cd1af256d3ecceca532df03539a7 (MD5) / Made available in DSpace on 2017-12-06T17:06:52Z (GMT). No. of bitstreams: 1 JacquelineCantanhede.pdf: 6344249 bytes, checksum: c518cd1af256d3ecceca532df03539a7 (MD5) Previous issue date: 2017-09-13 / The Prehypertension is characterized with systolic blood pressure levels between 120 - 139 mmHg and diastolic blood pressure 80-89mmHg, considered an intermediate state for the development of arterial hypertension. Detecting risk factors for prehypertension becomes important to prevent thousands of premature deaths. Vitamin D deficiency has been linked to high blood pressure and consequently to cardiovascular diseases that are responsible for high global morbidity and mortality. Thus, the analysis of the relationship between prehypertension and vitamin D is fundamental because it allows preventive intervention and avoids the progression to hypertension, thus reducing morbidity and mortality due to cardiovascular diseases. The present study aims to evaluate the association between serum 25 (OH) D levels and prehypertension. This is a cross-sectional study with a quantitative approach carried out at the Hospital Universitário da Universidade Federal do Maranhão in São Luís, Maranhão, Brazil. The 161 adults with prehypertensive and normotensive conditions participated in this study. Socio-demographic, anthropometric, behavioral and clinical data of the participants of both genders between 30 and 50 years old were used. Statistical analysis was performed using SPSS® software version 23. Data were treated using descriptive procedures. The Kolmogorov-Smirnov test was used to verify the normality of the variables. The results were considered statistically significant if p <0.05. In relation to the cardiometabolic risk factors, there was a statistically significant difference (p <0.05) between the control group and the study in the parameters evaluated (BMI, WC and WHtR). The prehypertensive group had a higher mean. Participants with excess weight have statistically higher odds of presenting prehypertension (OR = 3.62, 95% CI = 1.79-7.31 p <0.001). Regarding the Cardiometabolic Risk Factors stratified by gender, a statistically higher percentile was observed in females. Regarding systolic and diastolic blood pressure and vitamin D, there was a statistically significant difference (p <0.05) in all variables analyzed. For males, there was no statistically significant difference in the vitamin D variable. Mean SBP and DBP, and vitamin D (36.15 ± 12.31), were higher in the study group. Especially women (33.65 ± 10.41). In this study, the association of vitamin D and the presence of prehypertension was not observed. The serum vitamin D level of most participants was considered adequate. The female population had a higher prevalence of increased cardiometabolic levels and a higher prevalence of inadequate levels of vitamin D. There was no correlation between serum vitamin D levels with anthropometric data and blood pressure levels. / Pré - hipertensão é caracterizada com níveis de pressão arterial sistólica entre 120 -139 mmHg e pressão arterial diastólica 80-89mmHg, considerada um estado intermediário para o desenvolvimento da hipertensão arterial, representa grande fator de risco para as doenças cardiovasculares. Detectar fatores de risco para pré-hipertensão torna-se importante para evitar milhares de mortes prematuras. A deficiência de vitamina D têm sido relacionada com pressão arterial elevada e consequentemente com doenças cardiovasculares que são responsáveis por elevada morbimortalidade mundial. Desta forma, a análise da relação entre pré- hipertensão e vitamina D é fundamental, pois, pode permitir a intervenção preventiva e evita a progressão para hipertensão reduzindo assim a morbimortalidade por doenças cardiovasculares. O presente estudo tem por objetivo avaliar a associação entre os níveis séricos de 25 (OH)D e pré-hipertensão. Trata-se de um estudo transversal, realizado no Hospital Universitário da Universidade Federal do Maranhão no município de São Luís/ Maranhão. Participaram deste estudo 161 adultos em condições de pré-hipertensos e normotensos. Foram utilizados dados sóciodemográficos, antropométricos, comportamentais e clínicos dos participantes de ambos os sexos com idades entre 30 a 50 anos. A análise estatística foi realizada através do software SPSS versão 23. Os dados foram tratados por meio de procedimentos descritivos. O teste de Kolmogorov-Smirnov foi utilizado para verificar a normalidade das variáveis. Os resultados foram considerados estatisticamente significativos se p <0,05. Em relação, aos fatores de risco cardiometabólicos houve diferença estatisticamente significativa (p<0,05) entre o grupo controle e estudo nos parâmetros avaliados índice de massa corpórea, circunferência da cintura e relação cintura estatura. O grupo de pré-hipertensos apresentou maior média. Participantes com excesso de peso tem estatisticamente maior chance de apresentar pré - hipertensão (OR= 3,62; IC 95%=1,79-7,31 p<0,001). Em relação aos Fatores de Risco Cardiometabólicos estratificados por sexo. Observou-se um percentual estatisticamente maior no sexo feminino. Em relação, a pressão arterial sistólica e diastólica e vitamina D, houve diferença estatisticamente significativa (p<0,05) em todas as variáveis analisadas. Para sexo masculino não houve diferença estatisticamente significativa na análise da variável Vitamina D. A média PAS e PAD, e da vitamina D (36,15 ±12,31) foi maior no grupo estudo. Em especial as mulheres (33,65±10,41). Neste estudo não foi observado associação da vitamina D e a presença de pré- hipertensão. O nível sérico de vitamina D da maioria dos participantes foi considerado adequado. A população do sexo feminino apresentou maior prevalência dos níveis cardiometabólicos aumentados e maior prevalência dos níveis inadequado de vitamina D. Não houve correlação entre os níveis séricos de vitamina D com os dados antropométricos e níveis pressóricos.

Introdução: Nos últimos anos têm-se demonstrado que a vitamina D desempenha um papel crucial em muitas doenças agudas e crônicas, não só afetando as condições ósseas, mas também aumentando o risco de fraturas, doenças auto-imunes e câncer. A influência da vitamina D nas doenças do fígado tem sido amplamente discutida já que esta sofre metabolismo hepático. Indivíduos com doenças hepáticas, e em especial com hepatite C crônica, apresentam maior prevalência de deficiência de vitamina D. Objetivos: Avaliar, em duas amostras distintas, a influência dos níveis séricos de vitamina D e/ou dos polimorfismos envolvidos na sua rota metabólica na progressão da doença hepática crônica causada pelo vírus C. Métodos: A primeira amostra consistiu de um estudo transversal com 132 pacientes com hepatite C crônica genótipo 1 do Hospital de Clínicas de Porto Alegre (HCPA). Foi avaliada a influência de níveis séricos de vitamina D e dos polimorfismos rs7041 e rs4588 do gene GC no grau de fibrose hepática (escore METAVIR). As genotipagens foram feitas com ensaio TaqMan e as análises estatísticas foram realizadas no programa SPSS v.20.0. A segunda amostra consistiu da análise de dados extraídos da plataforma dbGaP. Foram investigados 681 pacientes com hepatite C crônica da coorte americana HALT-C, acompanhados pelo período quatro anos. Avaliou-se a relação de 40 polimorfismos dos genes DHCR7, GC, CYP2R1, CYP24A1, CYP27B1, VDR, SMAD3 e TGFB1 nos seguintes desfechos: piora da fibrose hepática; descompensação hepática (escore Child Pugh-Turcotte>7, ascite, encefalopatia hepática, peritonite bacteriana espontânea e/ou sangramento de varizes gastroesofágicas); desenvolvimento de carcinoma hepatocelular e morte do fígado. Os polimorfismos que não se encontravam disponíveis no banco de dados foram imputados com o programa Mach-Admix 2.0.203 e as análises foram no programa Plink 1.07 e foi realizada correção de Bonferroni. Nesta amostra, resultados com P<0,05 foram considerados 2 como tendência à associação. Resultados: Na amostra do HCPA, níveis diminuídos de vitamina D, bem como a deficiência grave de vitamina D, foram mais frequentes entre pacientes com fibrose intermediária/avançada (METAVIR 3 e 4). Embora os polimorfismos rs7041 e rs4588 e seus haplótipos tenham apresentado relação com os níveis séricos de vitamina D, estes não apresentaram associação com a gravidade da fibrose hepática. Na segunda amostra estudada, onze polimorfismos tiveram tendência à associação (P<0,05) com os desfechos analisados: quatro SNPs no gene DHCR7 com descompensação hepática (rs4944957, rs12800438, rs3829251 e rs4945008); dois no gene GC com piora da fibrose e morte do fígado (rs7041 e rs222020); dois no gene CYP2R1 com ascite ou carcinoma hepatocelular (rs7116978 e rs1562902); dois no gene VDR com sangramento de varizes gastresofágicas e carcinoma hepatocelular (rs4516035 e rs2239186); e um no gene SMAD3 com piora da fibrose e encefalopatia (rs2118610). Apenas um polimorfismo, rs1800469no gene TGFB1, apresentou associação com descompensação hepática após correção de Bonferroni (P<0,05/40). Conclusões: Nossos resultados demonstraram que níveis séricos menores de vitamina D estão associados à progressão da fibrose hepática na hepatite C crônica genótipo 1. Além disso, os onze polimorfismos da rota da vitamina D que apresentaram tendência à associação estatística, indicam que variantes genéticas da rota metabólica da vitamina D possuem fraca ou nenhuma relação com a progressão da hepatite C crônica, merecendo análises futuras. Já o polimorfismo rs1800469, do gene TGFB1, demonstrou potencial utilidade para auxiliar na identificação de pacientes com maior chance de descompensação hepática. / Introduction: In the past years it has been demonstrated that vitamin D plays a crucial role in many acute and chronic diseases, not only affecting bone conditions but increasing the chance of fractures, autoimmune diseases and cancer. Vitamin D influence in liver disease is also widely discussed since it undergoes trough hepatic metabolism. Subjects with liver diseases, especially chronic hepatitis C, present higher rates of vitamin D deficiency. Objectives: Evaluate, in two different samples, the influence of vitamin D serum levels and/or polymorphisms of vitamin D metabolic pathway in liver disease progression in patients with chronic hepatitis C. Methods: The first sample consisted of a transversal study with 132 patients from Hospital de Clínicas de Porto Alegre (HCPA) with chronic hepatitis C genotype 1. We evaluated the influence of serum levels of vitamin D and rs7041 and rs4588 GC polymorphisms over liver fibrosis (METAVIR scoring). Genotyping was performed with TaqMan probes and statistical analyses were conducted using SPSS v.20.0. The second sample was extracted from HALT-C cohort available in dbGap plataform. It included 681 patients with chronic hepatitis C, followed for 4 years. Forty polymorphisms in DHCR7, GC, CYP2R1, CYP24A1, CYP27B1, VDR, SMAD3 and TGFB1 genes were analyzed with the following outcomes: worsening of fibrosis; hepatic decompensation (gastric/esophageal bleeding, CTP > 7, ascites, spontaneous bacterial peritonitis, and/or encephalopathy); development of hepatocellular carcinoma; and liver death. Polymorphisms not available in dbGaP data were imputed using Mach-Admix 2.0.203 software and analyses were performed in Plink v.1.07 and a Bonferroni’s correction was performed. Results with P<0.05 were considered as having tendency towards association. Results: In HCPA sample low serum vitamin D, as well as severe vitamin D deficiency, were more frequent among patients with intermediate/severe fibrosis (METAVIR 3 and 4). Although rs7041 and rs4588 4 polymorphisms and its haplotypes presented association with serum levels of vitamin D, they were not associated with severity of liver fibrosis. In sample two, eleven polymorphisms presented tendency towards association (P<0.05) with the studied outcomes: four SNPs in DHCR7 with hepatic decompensation (rs4944957, rs12800438, rs3829251 and rs4945008); two in GC with worsening of fibrosis and liver death (rs7041 and rs222020); two in CYP2R1 with ascites or hepatocellular carcinoma (rs7116978 and rs1562902); two in VDR with gastric/esophageal bleeding and hepatocellular carcinoma (rs4516035 and rs2239186); and one in SMAD3 with worsening of fibrosis and encephalopathy (rs2118610). Only one polymorphism, rs1800469 in TGFB1, showed statistical significance with hepatic decompensation after Bonferroni’s correction (P<0.05/40). Conclusions: Our results demonstrated that low serum vitamin D has association with fibrosis progression in chronic hepatitis C genotype 1. Also, the eleven polymorphisms with tendency towards association indicate that genetic variants in vitamin D pathway possess weak or none relation with progression of chronic hepatitis C, deserving future analyses. Finally, polymorphism rs1800469 in TGFB1 demonstrated potential utility to help identify patients with higher odds of hepatic decompesantion.

The plasma vitamin D binding protein (DBP) is an a-glycoprotein, synthesised and secreted by the liver, which binds specifically vitamin D and its metabolites. The DBP molecule, has a single high affinity binding site for its ligands, and is present in blood in concentrations about 1000-fold greater than the sum of all its vitamin D ligands. Previous studies have not found any change in the concentration of DBP related to various derangements in mineral homeostasis. Therefore the general view is that DBP has a passive role in the physiology of vitamin D and its metabolites, and simply acts to solubilise and transport these hydrophobic ligands in the aqueous extracellular fluid. However, differences which have been described in its affinity for various vitamin D metabolites suggest that there have been evolutionary influences on the properties of this protein. Furthermore, plasma DBP concentration has been found to change in response to a number of physiological factors, such as changing sex steroid hormone secretion. The aim of the studies presented in this thesis was to investigate variation in the plasma concentration of the DBP in a range of vertebrate species, and in response to a variety of physiological factors. The results suggest that DBP may have an active role in regulating the bioavailability, and hence the utilisation and metabolism of its ligands. DBP concentration has traditionally been measured using immunological techniques. These techniques, although fast and simple, have a number of draw-backs which can be overcome by the use of assays which rely upon functional aspects of the DBP. A saturation binding assay was modified from those described previously. Using this technique, it was found that both the circulating concentration of the DBP and its affinity for 25-hydroxyvitamin D3 (25(OH)D3) varied significantly among a wide range of species of reptiles and birds. This variation did not reflect phylogenetic relationships among the study species, suggesting that the variation was more likely to be the result of selective pressure in response to individual ecological or physiological circumstance, rather than to random mutation. In support of this, both the plasma concentration of DBP, and its affinity for 25(OH)D3 were significantly associated with a number of ecological factors which might be considered to have some significance to vitamin D and calcium homeostasis. In addition, comparative binding data suggests that the ability of the DBP to bind 25-hydroxyvitamin D2 with equal affinity to 25(OH)D3 is an evolutionary innovation of mammalian vertebrates. In order to extend the idea of genetic variation in the concentration and affinity of plasma DBP, two strains of broiler (meat-type) chickens were studied. It was found that both the concentration and the affinity of plasma DBP for 25(OH)D3 was characteristic for each strain, emphasising the sensitivity of DBP to genetic variation. A number of factors have been found to modulate the genetically determined plasma concentration of DBP. Deficiencies of dietary protein and dietary energy, and variation in concentrations of sex steroids were found to affect the circulating concentration of DBP. However, species differences were still apparent, suggesting that the sensitivity of DBP to these physiological modifiers may have developed independently in different species, and may be secondary to genetic determinants of DBP properties. The plasma DBP concentration and specific binding affinity both determine the availability of its ligands for cellular uptake. It is likely that this process is complex, and involves a combination of protein mediated and non-mediated uptake events. This makes DBP a potentially important determinant of the biological actions of its ligands. The studies in this thesis have produced two main lines of argument supporting an active role for DBP in the regulation of vitamin D metabolism and utilisation. The first is that genetic variation in the properties of plasma DBP appears to be genetically determined, and is selected for, both at the between-species, and the within-species level, than it is to random mutation. Secondly, the ability of physiological and environmental factors to modify the circulating concentration of DBP suggests that this protein is responsive to homeostatic processes. It is proposed that DBP is an active regulator of the physiological economy of vitamin D and its metabolites by being itself regulated by a number of genetic and non-genetic factors.

Background: Determinants of vitamin D status are of interest when studying the epidemiology of disease in population groups because vitamin D is now recognized to decrease the risk of diseases such as osteoporosis, cancer, and cardiovascular disease. Understanding modifiable determinants of vitamin D status are important for managing vitamin D deficiency at the individual level and for addressing this issue at population level. Objective: The objective of this study was to evaluate the associations between serum vitamin D status (deficiency and insufficiency) and distinct demographic, dietary, and lifestyle characteristics of adults in the United States using a large, nationally representative sample survey, the National Health and Nutrition Examination Survey (NHANES) 2005-2006. Methods: The study sample consisted of 2340 adults aged 20-59 who had serum 25(OH)D measured and who had completed various questionnaires concerning dietary intake of vitamin D and other lifestyle factors. Multivariate logistic regression was used to estimate the odds ratio (OR) of vitamin D deficiency, insufficiency, and sufficiency in adults based on distinct demographic, dietary, and lifestyle characteristics. Statistical significance was set at α < 0.05. Results: The prevalence of vitamin D deficiency was higher in obese adults than in underweight to normal weight adults (50.9% ± 4.57 vs. 29.3% ± 3.57), higher in adults who reported no sunburns than in adults who reported ≥ 3 sunburns (49.9% ± 3.82 vs. 18.0% ± 3.07), and higher in adults who use sun protective measures regularly than in adults who do not (48.4% ± 3.93 vs. 27.0% ± 3.75). The prevalence of vitamin D deficiency increased as dietary intake of vitamin D decreased. Non-Hispanic black adults were significantly more likely to be vitamin D deficient (OR = 45.27, 95% CI = 17.27-118.64) and insufficient (OR = 9.37, 95% CI = 3.43-25.61) than non-Hispanic white adults. Significant positive associations were found between vitamin D deficiency and several characteristics, namely obesity (OR = 7.43, 95% CI = 4.33-12.77), physical inactivity (OR = 1.63, 95% CI = 1.03-2.58) poor dietary vitamin D intake (OR = 2.34, 95% CI = 1.44-3.81), non-supplement use or supplement use with a low amount of vitamin D (OR = 1.75, 95% CI = 1.05-2.89), and activities that decrease exposure to sunlight (from OR = 2.97, 95% CI = 2.14-4.13 to OR = 5.30, 95% CI = 3.17-8.85). Conclusion: The results of this nationally representative study demonstrate that obesity, physical inactivity, poor dietary intake of vitamin D, and low sunlight exposure increases the risk for vitamin D deficiency in U.S adults. Future studies are needed to investigate whether vitamin D supplementation, sunlight exposure, and vitamin D-fortified foods are efficient in correcting vitamin D deficiency and insufficiency among these groups.

Thesis--Music--Leeds--Univ., 1997. / Bibliogr. p. 260-265.

Th. Etat--Histoire de la musique et musicologie--Paris 4, 1997. / Nombreux exemples musicaux. Extraits de textes en arabe. Glossaire p. 523-530. Bibliogr. p. 532-556. Index.

Thèse d'université--histoire moderne et contemporaine--Paris 4, 2007. / Bibliogr. p. 863-873. Index p. 874-889.

Thesis (M.S.)--University of Wyoming, 2008. / Title from PDF title page (viewed on Dec. 4, 2009). Includes bibliographical references (p. 75-88).