Thesis (M.Sc. (Dietetics))--North-West University, Potchefstroom Campus, 2005.

Background: The role of dietary calcium in weight management is gaining support in the nutrition research community. It has been hypothesized that high calcium diets protect against fat gain by creating a balance of lipolysis over lipogenesis in adipocytes (Zemel et al., 2000) and that a diet deficient in calcium is associated with higher body weight and that augmenting calcium intake may reduce weight and fat gain or enhance fat loss (Shapses et al., 2004). Objectives: A lack of baseline data on the physical, physiological and mental effects of obesity on urban African women was the motivation for the POWIRS (Profiles of Obese Women with Insulin Resistance Syndrome) study. The aim of the study was to assess the effects of obesity on health determinants of urban African and white women by comparing the lifestyle and risk factors for non-communicable diseases (NCDs) of lean, overweight and obese subjects. This led to a multi-disciplinary cross-sectional case-control study in which health determinants and health status, as well as the underlying mechanistic relationships between these factors were measured in a sample of African women volunteers. The study was repeated a year later, done in a sample of white women volunteers, POWIRS II. The effect of calcium intake on body composition was assessed during this study. Methods: One hundred and two apparently healthy urban African women, between the ages of 20 and 50 years participated in the first phase of this case-control cross-sectional survey. For a period of about three weeks, each afternoon ten subjects were to report at a Metabolic Unit Facility (consisting of 10 single bedrooms, 2 bathrooms, a living room and kitchen). Each subject received a "participant sheet" which guided them through the different research 'stations' where the various measurements were done. During the course of the evening demographic questionnaires were filled in and all anthropometric measurements were taken, except weight and height measurements. All participants received an identical light supper which excluded alcohol and caffeine at 20h00, went to sleep before 23h00 and fasted overnight. From 06h00 in the morning weight, height and blood pressure measurements were taken. After a fasting blood sample was taken, a two-hour glucose tolerance test commenced. Subjects received a breakfast and afterwards habitual dietary intake questionnaires were completed. Results: Mean total dietary calcium intake as significantly higher in white women (POWIRS II), with a mean intake 1053.8 mg per day, as opposed to a mean intake of 494.8 mg calcium per day in the blacks subjects (POWIRS I). Mean fat intake in the black subjects was 59.3 g per day, and in the white women 103.1 g per day. Thus the calcium:fat ratio in white women was higher than in black women (11.0 and 8.4 respectively). After adjustment for age and total dietary energy intake, significant negative correlations were found between dietary calcium intake and various variables, only in the white subjects. These were BMI (r=-0.255, p=0.01), percentage body fat (r=-0.252, p=0.01), fasting insulin (r=-0.205, p=0.05) and fasting glucose (r=-0.199, p=0.046). The calcium:fat ratio correlated negatively with BMI (r=-0.378, p<0.0001), percentage body fat (r=-0.401, p<0.0001), fasting glucose (r=-0.229, p=0.02), fasting insulin (r=-0.212, p=0.04) and plasma leptin (r=-0.284, p=0.004). Adjustment for smoking resulted in slightly different correlation coefficients, but similar significant correlations were still found. The only significant association that was found in the black population, was a negative correlation between dietary calcium intake and systolic blood pressure (p=0.03) as well as diastolic blood pressure (p=0.04). After adjustment for age, smoking and dietary energy intake no significant correlations were found in the black subjects. Conclusion: The results from the POWIRS study in white women are consistent with the hypothesis that there may be an inverse relationship between adiposity and calcium intake. In our study higher calcium intakes were associated with lower body fat, lower BMI, lower fasting glucose and insulin, as well as plasma leptin in white women. The association seems to be significant in subjects with high intakes of fat and calcium (as seen in the white women). / Thesis (M.Sc. (Dietetics))--North-West University, Potchefstroom Campus, 2005.

Les omégas trois ont un bénéfice prouvé dans la prévention de maladie cardiovasculaire et l'inflammation. Un apport optimal peut être réalisé avec des modifications diététiques simples permettant d'avoir un enrichissement des membranes cellulaires et un effet métabolique. Le tissu adipeux de part son rôle important dans la genèse du syndrome métabolique semble être une cible importante du traitement par oméga trois. Les patients avec une maladie rénale chronique (MRC) ont un risque cardiovasculaire accru et cumulent les perturbations métaboliques comme le syndrome métabolique et un état micro inflammatoire. Des doses supra physiologiques d'oméga trois ont été utilisés dans le passé dans des études de prévention rénale ou traitement de dyslipidémie. Or l'effet métabolique en fonction de la dose d'oméga 3 n'est pas connu. En accord, avec les études chez le sujet sain, en fonction de la dose administrée, les omégas 3 ont un impact différent métabolique et sur l'expression génique. Des études complémentaires sont nécessaires pour vérifier la faisabilité et l'impact métabolique d'une modification de régime afin de diminuer le rapport n-6/n-3, ainsi que l'effet à long terme des omégas trois chez ces patients. Par ailleurs, les mécanismes impliqués dans les différences de dose réponse devront être caractérisés sur un modèle animal

The accumulation of androstenone in the fat of mature boars results in boar taint; the conjugation of androstenone would decrease this important meat quality problem by decreasing the accumulation and increasing the excretion of androstenone. Leydig cells and testis microsomes from mature boars were incubated with radiolabeled pregnenolone, and the free and conjugated metabolites were examined by HPLC. Sulfated androstenone with a mass of 367 m/z was directly identified by MS, with a novel tentative structure of 3-keto-4- sulfoxy-androstenone. Addition of enolase to the microsomal incubations increased the formation of 3-keto-4-sulfoxy-androstenone. Overexpression of SULT2A1 in HEK cells resulted in the sulfoconjugation of dehydroepiandrosterone, but not androstenone, suggesting that SULT2A1 may not be involved in sulfoconjugation of androstenone. This thesis describes the novel direct characterization of androstenone sulfate and the importance of enolase in its formation. The relevance to boar taint metabolism is discussed.

Background: The role of dietary calcium in weight management is gaining support in the nutrition research community. It has been hypothesized that high calcium diets protect against fat gain by creating a balance of lipolysis over lipogenesis in adipocytes (Zemel et al., 2000) and that a diet deficient in calcium is associated with higher body weight and that augmenting calcium intake may reduce weight and fat gain or enhance fat loss (Shapses et al., 2004). Objectives: A lack of baseline data on the physical, physiological and mental effects of obesity on urban African women was the motivation for the POWIRS (Profiles of Obese Women with Insulin Resistance Syndrome) study. The aim of the study was to assess the effects of obesity on health determinants of urban African and white women by comparing the lifestyle and risk factors for non-communicable diseases (NCDs) of lean, overweight and obese subjects. This led to a multi-disciplinary cross-sectional case-control study in which health determinants and health status, as well as the underlying mechanistic relationships between these factors were measured in a sample of African women volunteers. The study was repeated a year later, done in a sample of white women volunteers, POWIRS II. The effect of calcium intake on body composition was assessed during this study. Methods: One hundred and two apparently healthy urban African women, between the ages of 20 and 50 years participated in the first phase of this case-control cross-sectional survey. For a period of about three weeks, each afternoon ten subjects were to report at a Metabolic Unit Facility (consisting of 10 single bedrooms, 2 bathrooms, a living room and kitchen). Each subject received a "participant sheet" which guided them through the different research 'stations' where the various measurements were done. During the course of the evening demographic questionnaires were filled in and all anthropometric measurements were taken, except weight and height measurements. All participants received an identical light supper which excluded alcohol and caffeine at 20h00, went to sleep before 23h00 and fasted overnight. From 06h00 in the morning weight, height and blood pressure measurements were taken. After a fasting blood sample was taken, a two-hour glucose tolerance test commenced. Subjects received a breakfast and afterwards habitual dietary intake questionnaires were completed. Results: Mean total dietary calcium intake as significantly higher in white women (POWIRS II), with a mean intake 1053.8 mg per day, as opposed to a mean intake of 494.8 mg calcium per day in the blacks subjects (POWIRS I). Mean fat intake in the black subjects was 59.3 g per day, and in the white women 103.1 g per day. Thus the calcium:fat ratio in white women was higher than in black women (11.0 and 8.4 respectively). After adjustment for age and total dietary energy intake, significant negative correlations were found between dietary calcium intake and various variables, only in the white subjects. These were BMI (r=-0.255, p=0.01), percentage body fat (r=-0.252, p=0.01), fasting insulin (r=-0.205, p=0.05) and fasting glucose (r=-0.199, p=0.046). The calcium:fat ratio correlated negatively with BMI (r=-0.378, p<0.0001), percentage body fat (r=-0.401, p<0.0001), fasting glucose (r=-0.229, p=0.02), fasting insulin (r=-0.212, p=0.04) and plasma leptin (r=-0.284, p=0.004). Adjustment for smoking resulted in slightly different correlation coefficients, but similar significant correlations were still found. The only significant association that was found in the black population, was a negative correlation between dietary calcium intake and systolic blood pressure (p=0.03) as well as diastolic blood pressure (p=0.04). After adjustment for age, smoking and dietary energy intake no significant correlations were found in the black subjects. Conclusion: The results from the POWIRS study in white women are consistent with the hypothesis that there may be an inverse relationship between adiposity and calcium intake. In our study higher calcium intakes were associated with lower body fat, lower BMI, lower fasting glucose and insulin, as well as plasma leptin in white women. The association seems to be significant in subjects with high intakes of fat and calcium (as seen in the white women). / Thesis (M.Sc. (Dietetics))--North-West University, Potchefstroom Campus, 2005.

Persistent organic pollutants (POPs) and adipose tissue hypoxia have been shown to independently affect adipocyte functions. The goals of this study were to (1) determine the effect of PCB-77, PCB-153, and DDE on the differentiation of human preadipocytes, and (2) investigate the cross-talk between PCB-77 and hypoxia in differentiated human adipocytes. First, human preadipocytes were exposed to PCB-77, PCB-153, or DDE during the entire 14-day differentiation period. We found no effect of low POP levels on lipid accumulation. Second, differentiated human adipocytes were exposed to a combination of PCB-77 and hypoxia. We demonstrated gene-specific cross-talk between PCB-77 and hypoxia, showing an additive effect of PCB-77 on VEGF, MCP-1, and adiponectin, as well as an inhibition of PCB-77-induced expression of CYP1A1 by hypoxia. This work has expanded our understanding of the role of POPs and hypoxia in differentiated human adipocytes.

Recent studies suggested the persistence of brown adipocytes in adult humans, as opposed to being exclusively present in infancy. In this study, we investigated the presence of brown-like adipocytes in adipose tissue (AT) samples of children and adolescents aged 0 to 18 years and evaluated the association with age, location, and obesity. For this, we analysed AT samples from 131 children and 23 adults by histological, immunohistochemical and expression analyses. We detected brown-like and UCP1 positive adipocytes in 10.3% of 87 lean children (aged 0.3 to 10.7 years) and in one overweight infant, whereas we did not find brown adipocytes in obese children or adults. In our samples, the brown-like adipocytes were interspersed within white AT of perirenal, visceral and also subcutaneous depots. Samples with brown-like adipocytes showed an increased expression of UCP1 (>200fold), PRDM16 (2.8fold), PGC1α and CIDEA while other brown/beige selective markers, such as PAT2, P2RX5, ZIC1, LHX8, TMEM26, HOXC9 and TBX1 were not significantly different between UCP1 positive and negative samples. We identified a positive correlation between UCP1 and PRDM16 within UCP1 positive samples, but not with any other brown/beige marker. In addition, we observed significantly increased PRDM16 and PAT2 expression in subcutaneous and visceral AT samples with high UCP1 expression in adults. Our data indicate that brown-like adipocytes are present well beyond infancy in subcutaneous depots of non-obese children. The presence was not restricted to typical perirenal locations, but they were also interspersed within WAT of visceral and subcutaneous depots.

Diss. (sammanfattning) Linköping : Linköpings universitet, 2007. / Härtill 4 uppsatser.

Il existe deux populations d’adipocytes thermogéniques, les adipocytes bruns du tissu adipeux bruns et les adipocytes ‘’brites’’ qui apparaissent au sein du tissu adipeux blanc. Récemment, la caractérisation d’adipocytes bruns et brites fonctionnels chez l’homme adulte a permis d’envisager de nouvelles approches nutritionnelles et thérapeutiques pour traiter l’obésité. Mon projet de thèse a porté sur l’étude des effets des acides gras polyinsaturés ω6 et ω3 d’origine alimentaire sur la conversion des adipocytes blancs en brites, d’abord chez l’homme in vitro puis chez les rongeurs in vivo. Nous avons pu ainsi démontrer que l'acide arachidonique ω6 à l’origine de nombreux métabolites oxygénés, exerce 1) un effet inhibiteur sur la formation des adipocytes brites grâce aux prostaglandines E2 et F2α, et 2) un effet inducteur via la prostacycline. En effet, celle-ci active la conversion des adipocytes blancs en brites par une voie impliquant le récepteur membranaire IP et les récepteurs nucléaires PPARs. En nous basant sur les recommandations nutritionnelles actuelles chez l’Homme, qui ont pris en considération l’insuffisance de l’apport en acides garsω3 par rapport à l’excès des ω6, nous avons pu montrer chez la souris qu’une supplémentation en acides gras ω3 dans le régime alimentaire était capable d’inhiber l’effet néfaste des acides gras ω6 et d’activer le tissu adipeux brun. Nos résultats démontrent l’importance de la biodisponibilité de l’acide arachidonique dans la biologie du tissu adipeux et permettent de renforcer l’idée que le rééquilibrage du ratio ω6/ω3 est un outil de choix dans la prévention du surpoids et de l’obésité et les maladies métaboliques associées / There are two types of thermogenic adipocytes able to use fatty acids and glucose to produce heat. We distinguish brown adipocytes from the brown adipose tissue and ‘’brite’’ adipocytes which occur into the white adipose tissue. Recently, the characterization of functional brown and brite adipocytes in adult humans has led to the consideration of their use to treat obesity by increasing energy expenditure. My thesis project was to study the effect of dietary polyunsaturated fatty acids, on the conversion of white into brite adipocytes, in vitro and in vivo, in humans and rodents respectively. We demonstrated that arachidonic acid ω6, precursor of prostaglandins, has 1) an inhibitory effect on the recruitment of brite adipocytes via prostaglandins E2 and F2α and 2) an activatory effect via prostacyclin. In fact, prostacyclin induces the conversion of white into brite adipocytes through the IP receptor and the PPARs signaling pathways. Based on human present nutritional recommendations, we demonstrated that a supplementation of ω3 fatty acids in mice diet was able to inhibit the negative effect of ω6 fatty acids and activate brown adipose tissue. Our data highlights the importance of arachidonic acid bioavailability on the biology of adipose tissue and reinforce the idea that an equilibrate ω6/ω3 ratio is a tool that can be used to prevent overweight obesity and associated metabolic disorders

Obesity is associated with insulin resistance, dyslipidemia, and cardiovascular disease. The current obesity epidemic is the result of surplus energy consumption. Excess energy is stored in expanding adipose tissue. Adipose tissue growth entails the enlargement of existing adipocytes, the formation of new fat cells from preexisting progenitors, and the coordinated development of supporting vasculature. Identifying adipocyte progenitors and the mechanism of adipose tissue expansion is crucial for the development of new strategies to combat obesity and its complications. Though important progress has been made towards understanding the developmental origin of adipocytes, the identities of adipocyte progenitors are still not completely known. The main objective of this study is to determine whether endothelial cells of the adipose tissue can give rise to new adipocytes. Our results indicate that murine endothelial cells of adipose tissue are pluripotent and can potentially give rise to preadipocytes. Lineage tracing experiments using the VE-Cadherin-Cre transgenic mouse reveal localization of reporter genes in endothelial cells, preadipocytes and white and brown adipocytes. Moreover, capillary sprouts from human adipose tissue, which have predominantly endothelial cell characteristics, are found to express Zfp423, a preadipocyte determination factor. In response to PPARγ activation, endothelial characteristics of sprouting cells are progressively lost, and cells form structurally and biochemically defined adipocytes. Taken together, our data support an endothelial origin of a population of adipocytes. The ability of the vascular endothelium to give rise to adipocytes may explain how angiogenesis and adipogenesis can be temporally and spatially coordinated. Analysis of BAT and WAT revealed that adipose depots have distinct compositions of adipocyte progenitors. Of the CD45-CD29+Sca1+CD24+ progenitor population, only 17% and 52% express VE-Cadherin in WAT and BAT, respectively. Our data show that the number of these specific progenitors in BAT and WAT are highly variable and suggest that a considerable number of adipocytes progenitors may have a non-endothelial cell origin. Differences in composition and types of adipocyte progenitors may explain the differences in the adipocytes phenotypes that we observe in discrete depots. In brief, we find that the vascular endothelium gives rise to a population of brown and white fat cells, and that the number of endothelial-derived adipocyte progenitors residing in BAT and WAT is highly variable. These results expand our current understanding of adipose tissue growth, and, we hope, will accelerate the development of treatments for obesity-related complications.