Vitamin D plays an important role in over-all health. Few data exist on vitamin D deficiency related with intake for a Canadian population. The purpose of this study is to assess vitamin D intake and status in healthy young adults of diverse ancestry during the wintertime.<p> One hundred and seven young healthy adults living in Southern Ontario were recruited during the late winter of 2007. Their serum 25-hydroxyvitamin D [25(OH)D], skin melanin and anthropometric measures were determined. They completed a food frequency questionnaire (FFQ) (twice) and a 7-day food diary. Correlation analyses and t-test were used to validate the FFQ against the 7-day diary and 25(OH)D; one way ANOVA was used to determine ethnic group differences in vitamin D intake and status.<p> The results indicated that the FFQ used in this study was valid. Vitamin D deficiency [25(OH)D<50 nmol/L] was widespread and more apparent in the East and South Asian groups than in the European group (P<0.05). The dairy products were the greatest food source of vitamin D for each of the three groups and the European group exhibited higher total vitamin D intake (P<0.05). There was a trend for the European group to have higher consumption of dairy products, especially cow¡¯s milk (0.05<P<0.10). Combining subjects in the three ethnic groups, vitamin D intake but not BMI was closely related with serum 25(OH)D concentrations (r= 0.520, P<0.001; r=-0.018, P>0.05, respectively). The 25(OH)D levels were inversely related with parathyroid hormone (PTH) levels (r= -0.273, P= 0.009). With adequate calcium intake (¡Ý1000 mg/d), PTH levels were significantly lower when vitamin D was not deficient (P<0.05).<p> This study provides evidence that vitamin D deficiency is prevalent in healthy young adults living in Canada during wintertime, and non-European groups have a higher prevalence of this deficiency. Vitamin D intake varies with ethnicity, and dietary intake plays an important role in maintenance of serum vitamin D in wintertime. Compared with calcium intake, serum vitamin D levels may be a more important factor suppressing PTH levels.

Development of the secondary palate (SP) is a complex event despite the small area it encompasses. Problems with SP development can lead to a cleft palate, which is one of the most common birth disorders. The matrix metalloproteinases (MMPs) are required for proper SP development, but a functional role for any one of them remains unknown. MMP-25 is a candidate MMP to have a functional role in SP formation as genetic scans of the DNA of human cleft palate patients indicate a common mutation at a region upstream of the Mmp-25 gene. The purpose of this thesis is to investigate gene expression of Mmp-25 in the developing mouse SP, whether it has a functional role in mouse SP development and begin to identify factors potentially upstream of Mmp-25 expression.<p> Mmp-25 mRNA and protein is found at all SP developmental stages in mice with highest expression at embryonic day (E) 13.5 when analyzed by quantitative real-time PCR and western blotting. Immunohistochemistry localizes MMP-25 protein primarily to the plasma membranes of palate shelf epithelial cells with secondary expression in apical mesenchymal cells. Mmp-25 knockdown with siRNA in palatal cultures resulted in a significant decrease in palate shelf fusion and persistence of the medial edge epithelium in vitro. Mmp-25 mRNA and protein levels are significantly decreased in vitro when cultured palate shelves are incubated in growth medium with 5 ìg/ml of a TGFâ3-neutralizing antibody. Mmp-25 gene expression is highest at E12.5 and E13.5, which corresponds to increasing palate shelf growth downward alongside the tongue. Immunohistochemistry localized MMP-25 protein expression predominantly in the epithelium of the palate shelves, but also in areas of the mesenchyme that were immediately adjacent to the epithelium and apical in location. Knockdown of Mmp-25 expression resulted in palate shelf fusion being impaired and significant medial edge epithelium remaining in contacted areas. Bioneutralization of TGFâ3 resulted in a significant decrease in Mmp-25 gene expression. These data suggest a functional role for MMP-25 in mouse SP development by removing extra-cellular matrix barriers to increased palate shelf growth and place its expression downstream of TGF-â3 signaling. This is the first research to present a role for a single MMP in mouse SP development.

Que reproche-t-on aux experts? Selon la littérature scientifique sur le sujet, la crise de l'expertise semble généralisée. Parmi ses symptômes, on note l'abus de pouvoir des experts dans la prise de décision et leur perte de légitimité. Cela s'explique, notamment par le recours accru aux experts dans le cadre de procédures afin d'éclairer les décisions publiques complexes et par l'émergence de controverses dont le fort degré d'incertitudes face aux risques encourus rend d'autant plus ardue la tâche aux experts. Cette impasse s'insère dans le contexte plus large de la remise en cause de la prise de décision rationnelle qui suppose que les bonnes décisions sont fondées sur des faits provenant de détenteurs de savoirs spécialisés. Face à ce problème, plusieurs reconfigurations des relations entre les experts, le politique et les citoyens sont proposées. Ces modèles sont toutefois constitués selon des visions parfois aprioriques de ce que devraient être le rôle des experts. Nous privilégions donc de retourner la question en nous intéressant au point de vue des participants dans le cadre de procédure de prise de décision. Plus spécifiquement, ce mémoire porte sur l'étude du cas de la controverse du prolongement de l'autoroute 25 entre les villes de Laval et de Montréal, présenté devant la population par le Ministère des Transports du Québec en 2005. Récemment dans la région de Montréal, plusieurs controverses concernant des projets de transport ont émergé, signe de l'érosion de la décision rationnelle. Les controverses constituent un indice de l'incapacité des modes usuels à répondre à de nouveaux enjeux, ce qui illustre la pertinence d'étudier le rôle des experts dans ces situations. Ce mémoire explore, selon une posture inductive, les perceptions des acteurs par rapport aux experts mandatés afin de mieux cerner la crise de l'expertise dans un contexte précis et par conséquent, participe à la réflexion sur le rôle de l'expert et de la science dans la prise de décision. Pour ce faire, une analyse documentaire a été réalisée pour ancrer le projet à l'étude dans son contexte et une analyse lexicométrique des mémoires déposés par les participants aux consultations tenues par le Bureau d'audiences publiques sur l'environnement a été accomplie. À la lumière de nos analyses, on constate que la controverse du prolongement de l'autoroute 25 révèle principalement une crise de la décision publique. La crise de l'expertise semble davantage un appel à l'ouverture à d'autres acteurs et à d'autres connaissances dans les processus de prise de décision. Plusieurs participants réclament une légitimité qui ne provient ni de l'ordre scientifique ni du pouvoir politique. En raison du mode de réalisation du projet, soit une entente de partenariat public-privé, plusieurs enjeux d'expertise ont été évacués des consultations publiques. Cette particularité nous a permis de constater que l'expert est un élément essentiel au débat public. Selon ce constat, nous croyons qu'un regard plus approfondi s'impose sur les procédures qui encadrent le recours aux experts en vue de leur changement. ______________________________________________________________________________ MOTS-CLÉS DE L’AUTEUR : experts, expertise, controverse, prise de décision, science, évaluation environnementale, transport, autoroute 25

Background: The role of nutrients in mental health has recently been recognized and investigated. Vitamin D has been known to play a role in a wide range of diseases, such as bone, cardiovascular, and autoimmune diseases, and cancers. Recently, its role in cognitive function and mental health has been reported. Vitamin D receptor and hydroxylases have been mapped throughout the brain, suggesting a role for vitamin D in brain tissue. An inverse association between vitamin D and depression was observed in European epidemiologic studies. There is a paucity of data on the association between vitamin D concentrations and depression in the U.S. population. Objective: The objective of this study was to investigate the association between serum vitamin D concentrations and depression in a large, nationally representative sample survey, the third National Health and Nutrition Examination Survey 1988-1994 (NHANES III). Methods: The study sample included 7970 adults, ages 15-39 years, who completed the Diagnostic Interview Schedule for depression and had vitamin D concentrations measured. SAS and SUDAAN statistical software packages were used in data analysis. Multivariate logistic regression was used to estimate the likelihood of having depression in vitamin D deficient persons in relation to vitamin D sufficient persons, after taking several confounding variables into consideration. Significance was set at α < 0.05. Results: The prevalence of vitamin D deficiency was higher in women than in men (24 % vs. 15%), higher in African-Americans than in whites (60% vs. 10%), higher in people living in metropolitan rather than in rural areas (25% vs. 14%), and higher in subjects below the poverty threshold than in higher income subjects (29% vs. 14%). The prevalence of vitamin D deficiency increased as BMI increased. The diagnostic variables for depression did not show an association with vitamin D deficiency after adjusting for several confounding factors. However, subjects having a depressive episode at the time of the interview, were significantly more likely to exhibit vitamin D deficiencies (OR = 1.85; P = 0.0210). Conclusions: This is the first large epidemiologic study on the association between vitamin D and depression in a US representative sample survey. A significant positive association was found between subjects having an episode of depression and vitamin D deficiency. However, a causal relationship could not be established due to the cross-sectional nature of the study. Further studies need to investigate the mechanistic and causal relation between vitamin D and depression.

Background: Contribution of dietary sources to vitamin D status is not clearly known. Some studies have shown that dietary intake of certain vitamin D rich foods had a significant positive influence on serum 25-hydroxyvitamin D [25(OH)D] concentrations, whereas other studies have shown no effect. Although sunlight exposure is a major source of circulating serum 25(OH)D, children and adolescents have been advised on the dangers of sun exposure. Diet may therefore be an important contributor of circulating serum 25(OH)D in absence of or reduced sunlight exposure. Objective: The aim of this study was to determine whether serum 25(OH)D concentrations were associated with any specific dietary patterns in US children and adolescents using assay-adjusted serum 25(OH)D data from National Health and Nutrition Examination Survey (NHANES) 2003-2004 and 2005-2006. Methods: Data from 2 cycles of the NHANES 2003-2004 and 2005-2006 for individuals aged 2 to ≤19 y, were used to study the association between dietary patterns and serum 25(OH)D. Dietary patterns were established using factor analysis based on food-frequency questionnaire data. Eigenvalues and Scree plot were used to derive 2 major principal factors. They were labeled as High Fat Low Vegetable (HFLV) and Prudent dietary patterns. Results: Serum 25(OH)D was significantly lower in HFLV dietary pattern group compared to Prudent dietary pattern group (25.1 vs 27.0 ng/mL; P=0.001). The highest serum 25(OH)D concentrations for all subjects were in the low-intake HFLV group or medium and high-intake Prudent groups (P=0.003 and P=0.012, respectively). In multivariate adjusted analysis, children with higher Prudent dietary contribution scores to overall diet showed a significant positive relation with serum 25(OH)D (β=62.01, P=0.016). When data were stratified by sex, a significant positive relation was observed in girls who consumed the Prudent diet (β=86.34, P=0.014) and a significant negative relation was observed in girls who consumed the HFLV diet (β=-84.32, P=0.022). Conclusion: Overall, serum 25(OH)D concentrations were associated with Prudent dietary pattern but not with HFLV dietary pattern in US children and adolescents. When stratified by sex, the relation between dietary patterns and serum 25(OH)D was confined to only girls. Children consuming HFLV pattern diet may benefit from vitamin D supplementation and sunlight exposure (outdoor activities), and should be encouraged to consume more vitamin D fortified foods.

Title: Dormant companies – Taxation of closely held companies Problem: Many companies are facing a business transfer and fall within a smaller company with a few owners and therefore that tax jurisdiction is current. They risk facing high taxes but it can be avoid by setting the company in an inactive status for five years. Purpose: The purpose is to describe the inactive company rules and the verdict from the Swedish Supreme Administrative Court. Also to investigate how this verdict will affect company sellers. Method: Based on a case study and by analyzing the Swedish Law conclusions has been determinate. Result: A dormant company must be inactive for five years, and then the company can benefit from a different taxation. The owner and it´s relatives can´t be active in the company during this five year period. The tax benefit, in percentage, is at its highest level when lower transaction cost takes place. The higher transaction cost the lower percentage difference occurs. Even if the percentage difference is at its lowest for high transaction costs the difference in currency is higher the higher the transaction cost is. It’s up to every individual to decide if the five year inactivity time is worth the beneficial taxation.

Vitamin D plays an important role in over-all health. Few data exist on vitamin D deficiency related with intake for a Canadian population. The purpose of this study is to assess vitamin D intake and status in healthy young adults of diverse ancestry during the wintertime.<p> One hundred and seven young healthy adults living in Southern Ontario were recruited during the late winter of 2007. Their serum 25-hydroxyvitamin D [25(OH)D], skin melanin and anthropometric measures were determined. They completed a food frequency questionnaire (FFQ) (twice) and a 7-day food diary. Correlation analyses and t-test were used to validate the FFQ against the 7-day diary and 25(OH)D; one way ANOVA was used to determine ethnic group differences in vitamin D intake and status.<p> The results indicated that the FFQ used in this study was valid. Vitamin D deficiency [25(OH)D<50 nmol/L] was widespread and more apparent in the East and South Asian groups than in the European group (P<0.05). The dairy products were the greatest food source of vitamin D for each of the three groups and the European group exhibited higher total vitamin D intake (P<0.05). There was a trend for the European group to have higher consumption of dairy products, especially cow¡¯s milk (0.05<P<0.10). Combining subjects in the three ethnic groups, vitamin D intake but not BMI was closely related with serum 25(OH)D concentrations (r= 0.520, P<0.001; r=-0.018, P>0.05, respectively). The 25(OH)D levels were inversely related with parathyroid hormone (PTH) levels (r= -0.273, P= 0.009). With adequate calcium intake (¡Ý1000 mg/d), PTH levels were significantly lower when vitamin D was not deficient (P<0.05).<p> This study provides evidence that vitamin D deficiency is prevalent in healthy young adults living in Canada during wintertime, and non-European groups have a higher prevalence of this deficiency. Vitamin D intake varies with ethnicity, and dietary intake plays an important role in maintenance of serum vitamin D in wintertime. Compared with calcium intake, serum vitamin D levels may be a more important factor suppressing PTH levels.

Development of the secondary palate (SP) is a complex event despite the small area it encompasses. Problems with SP development can lead to a cleft palate, which is one of the most common birth disorders. The matrix metalloproteinases (MMPs) are required for proper SP development, but a functional role for any one of them remains unknown. MMP-25 is a candidate MMP to have a functional role in SP formation as genetic scans of the DNA of human cleft palate patients indicate a common mutation at a region upstream of the Mmp-25 gene. The purpose of this thesis is to investigate gene expression of Mmp-25 in the developing mouse SP, whether it has a functional role in mouse SP development and begin to identify factors potentially upstream of Mmp-25 expression.<p> Mmp-25 mRNA and protein is found at all SP developmental stages in mice with highest expression at embryonic day (E) 13.5 when analyzed by quantitative real-time PCR and western blotting. Immunohistochemistry localizes MMP-25 protein primarily to the plasma membranes of palate shelf epithelial cells with secondary expression in apical mesenchymal cells. Mmp-25 knockdown with siRNA in palatal cultures resulted in a significant decrease in palate shelf fusion and persistence of the medial edge epithelium in vitro. Mmp-25 mRNA and protein levels are significantly decreased in vitro when cultured palate shelves are incubated in growth medium with 5 ìg/ml of a TGFâ3-neutralizing antibody. Mmp-25 gene expression is highest at E12.5 and E13.5, which corresponds to increasing palate shelf growth downward alongside the tongue. Immunohistochemistry localized MMP-25 protein expression predominantly in the epithelium of the palate shelves, but also in areas of the mesenchyme that were immediately adjacent to the epithelium and apical in location. Knockdown of Mmp-25 expression resulted in palate shelf fusion being impaired and significant medial edge epithelium remaining in contacted areas. Bioneutralization of TGFâ3 resulted in a significant decrease in Mmp-25 gene expression. These data suggest a functional role for MMP-25 in mouse SP development by removing extra-cellular matrix barriers to increased palate shelf growth and place its expression downstream of TGF-â3 signaling. This is the first research to present a role for a single MMP in mouse SP development.

The vitamin D metabolite 1,25(OH)2D3 has long been known to inhibit growth of prostate cancer cells and this mainly through a VDR-mediated pathway controlling target gene expression, resulting in cell cycle arrest, apoptosis and differentiation. Another major way in which 1,25(OH)2D3 inhibits cell growth in prostate cancer is via membrane-initiated steroid signalling, which triggers activation of signal cascades upon steroid binding to a receptor complex, leading to induction of genes regulating cell growth, proliferation and apoptosis. The main prostate cancer inhibiting membrane-initiated route is the TGFβ signalling pathway, elicited by the protein TGFβ. Two other important proteins downstream in this cascade are Smad2 and Smad3. In this study the early effects of 1,25(OH)2D3 on activated Smad2/3 levelsin PC-3 prostate cancer cells were examined. PC-3 cells were incubated for 3, 5, 10, 30 and 60 minutes as well as 38 hours both together with 1,25(OH)2D3 of the concentrations 10-10 and 10-7 M and without. Western Blots were then performed on supernatants from the cells treated followed by treatment of the membranes with primary antibodies against phosphorylated Smad2/3 C-terminal linker regions, alkaline phosphatase conjugated secondary antibodies and finally visualization with BCIP/ NBT tablets. As the downstream cascade protein JNK is a proposed activator of Smad2/3, this procedure was also repeated with a JNK inhibitor. This is a follow-up to an earlier study which examined the influence of 1,25(OH)2D3 on TGFβ levels using the same doses and time points and which found that 1,25(OH)2D3 initially lowered the level of active TGFβ, then increased it. The results of this study indicated a 1,25(OH)2D3 mediated induction of the same pattern in the levels of active Smad2 and 3, both with and without JNK inhibitor. The results did not indicate that 1,25(OH)2D3 activates the Smad2/3 C-terminal linker region via the JNK pathway.

The vitamin D metabolite 1,25(OH)2D3 has long been known to inhibit growth of prostate cancer cells and this mainly through a VDR-mediated pathway controlling target gene expression, resulting in cell cycle arrest, apoptosis and differentiation. Another major way inwhich 1,25(OH)2D3 inhibits cell growth in prostate cancer is via membrane-initiated steroid signalling, which triggers activation of signal cascades upon steroid binding to a receptor complex, leading to induction of genes regulating cell growth, proliferation and apoptosis. The main prostate cancer inhibiting membrane-initiated route is the TGFβ signalling pathway, elicited by the protein TGFβ. Another important protein downstream in this cascade is Smad2. In this study the early effects of 1,25(OH)2D3 on activated Smad2 levels in PC-3 prostate cancer cells were examined. PC-3 cells were incubated for 5, 10, 30 and 60 minutes as well as 24 and 40 hours both together with 1,25(OH)2D3 of the concentrations 10-10 and 107 M and without. An ELISA assay scanning for activated Smad2 was then performed on supernatants from both treated and untreated cells. This is a follow-up to an earlier study which examined the influence of 1,25(OH)2D3 on TGFβ levels using the same doses and similar time points and which found that 1,25(OH)2D3 initially lowered the level of active TGFβ, then increased it. The results of this study showed a statistically insignificant, time delayed 1,25(OH)2D3 mediated induction of the same pattern in the levels of active Smad2. / Project Work in Biomedicine, Advanced Level, 7.5 ECTS