During atherosclerosis and restenosis, there is an extensive amount of collagen synthesis and degradation. Changes in the types of collagen present can have profound effects on vascular smooth muscle cell (SMC) proliferation and migration. Type VIII collagen, which is normally present at low levels within the mature vascular system, is greatly increased during atherogenesis. The central theme of this thesis is to determine the role of type VIII collagen in the pathogenesis of atherosclerosis and restenosis. In the first study, we demonstrated the importance of type VIII collagen in SMC migration and proliferation. SMCs from type VIII collagen-deficient mice display increased adhesion and decreased spreading, migration, and proliferation compared to SMCs from wild-type mice. Treatment of SMCs from type VIII collagen-deficient mice with exogenous type VIII collagen can rescue the defects. In the second study, we determined that type VIII collagen exerts its effects through regulation of MMP-2 expression. Type VIII collagen-deficient SMCs have decreased levels of MMP-2 and are impaired in chemotaxis toward PDGF-BB and in their ability to contract thick collagen gels. We found that decreasing endogenous MMP-2 levels in normal SMCs or adding exogenous collagen to type VIII collagen-deficient SMCs is sufficient to recapitulate the type VIII collagen-deficient or wild-type SMC phenotype, respectively. In the third study, we investigated the contribution of type VIII collagen to intimal hyperplasia after mechanical injury in the mouse. We found that type VIII collagen-deficient mice display a 35% reduction in intimal hyperplasia and attenuated vessel remodeling after femoral artery wire injury, establishing a role for type VIII collagen in restenosis. The results of the work presented in this thesis demonstrate that production of type VIII collagen confers an SMC phenotype with a greater propencity for proliferation and migration. These effects are in part mediated through regulation of MMP-2 expression and activation. We conclude that the increases in type VIII collagen production that occur during atherosclerosis and restenosis contribute to the capacity of SMCs to alter the existing extracellular matrix in a manner which permits enhanced migration.

Type II diabetes and hyperinsulinemia are associated with increased risks of developing colorectal cancer (CRC). Detailed mechanisms underlying this correlation, however, are yet to be explored. The present study demonstrates that insulin increases the expression of proto-oncogenes c-Myc and cyclin D1 via both translational and transcriptional mechanisms. We show here that insulin stimulates c-Myc gene translation via an Akt/PKB-dependent mechanism involving the mTOR signaling pathway. More importantly, we show for the first time that transcriptional stimulation of c-Myc and cyclin D1 expression by insulin involves a novel Akt/PKB-independent signal crosstalk between insulin and canonical Wnt signaling pathways. We then identified p21-activated protein kianse 1 (PAK-1) as a novel mediator for insulin and Wnt/beta-catenin (b-cat) molecular crosstalk, involving MEK/ERK signaling. Furthermore, we found that insulin treatment leads to increased b-cat phosphorylation at Ser675, and this is associated with increased b-cat nuclear content and increased b-cat interaction with Tcf/Lef-binding elements (TBEs) of the human c-Myc gene promoter. Lastly, we demonstrated that insulin signaling directly alters the expression levels of components of the Wnt signaling pathway, including fizzled homology 4 (Fdz-4) and TCF7L2 (=TCF-4). This study not only demonstrated the existence of signaling crosstalk between insulin and canonical Wnt signaling pathways at multiple levels, it reveals molecular mechanisms for observed correlation between CRC and hyperinsulinemia. The growing evidence implicating PAK-1 in various human tumorigenesis has emerged PAK-1 as a potential therapeutic target. Our discovery of PAK-1 functioning as a novel central mediator for insulin and Wnt signaling crosstalk in intestinal cells suggests that PAK-1 may potentially be a good target candidate for treating patients with CRC, especially those who have Type II diabetes or experience hyperinsulinemia.

Purpose: To examine the effectiveness of a community-based intensive multifactorial clinical intervention for patients with Type 2 diabetes, to evaluate the feasibility of achieving clinical targets for glycemic control in a community setting, and to identify factors that are predictive of glycemic control in this cohort (age, gender, disease duration, continuity of care, pharmacologic treatment, diabetes self-care and smoking status). Methods: Participants with Type 2 diabetes referred to the Diabetes Clinic following dissemination of the 2003 Clinical Practice Guidelines of Canadian Diabetes Association and who attended a minimum of two physician visits within a twelve month period were deemed eligible for participation. 70 patients were included in this retrospective study. Baseline and twelve month values for the following biomedical outcomes were collected via chart audit: BMI, hemoglobin A1c, blood pressure (systolic, diastolic) and lipid profile (HDL, LDL, triglycerides, total cholesterol, TC:HDL ratio). Data for identification of predictive factors for glycemic control were also retrieved by chart audit. Results: The results of the paired t-test yielded a significant improvement in hemoglobin A1c (p<0.05), systolic blood pressure (p<0.01), HDL-cholesterol (p<0.05), LDL-cholesterol (p<0.01), total cholesterol (p<0.05) and total cholesterol:HDL ratio (p<0.05) over twelve months. No significant difference in BMI, diastolic blood pressure or triglycerides was reported over twelve months. Over half the sample (52.9%) achieved clinical targets for glycemic control (hemoglobin A1c <7.0%) at twelve months. Logistic regression analysis identified disease duration (O.R. = 0.90, 95% CI Exp(B) = 0.079 - 0.773, p = 0.01) and continuity of care (O.R. = 0.25, 95% CI Exp(B) = 0.831 - 0.969, p = 0.02) as significant predictors of glycemic control at twelve months. Conclusions: These findings demonstrate the effectiveness of this community-based intensive multifactorial clinical intervention for patients with Type 2 diabetes and show that the implementation of CPGs related to glycemic control is feasible in a community-based setting. Additionally, patients in this cohort with increased disease duration and increased continuity of care were less likely to achieve clinical targets for glycemic control following a twelve month intensive multifactorial clinical intervention for Type 2 diabetes. In summary, health professionals should strive to implement similar intensive multifactorial interventions in community practice in order to decrease the likelihood of diabetes-related complications and improve the patients quality of life.

Dietary proteins from Atlantic salmon and cod have previously been shown to have antidiabetic effects. Since dietary proteins are digested into small peptides before being absorbed through the intestinal mucosa, it is reasonable to deduce that the antidiabetic effect is due to enzymatically-digested peptides rather than the proteins themselves. The aim of this study was to develop a protocol to recover peptides with antidiabetic effects from salmon and cod protein digests and then scale up and optimize the salmon protein hydrolysate production process for industrial-scale production. The peptide mixtures were screened using cell culture assays for insulin-modulating activities and were further fractionated and purified for the final identification. Total yields of salmon and cod protein hydrolysates (<1 kDa) as measured by Kjeldahl nitrogen were 16.9% and 40.1%, respectively. The production process used for the salmon protein hydrolysate (<1 kDa) showed good reproducibility and potential for the industrial-scale production.

ABSTRACT Objective: The objective of this review was to describe the effects of low carbohydrate diets in people with diabetes type 2. Methods: Review. An article search was made in the databases PubMed and Medline. The search resulted in 17 studies. After review and analysis of the studies main results five categories were highlighted; bloodsugar levels, HbA1c, weight, lipid levels and antidiabetic medications. Results: The results revealed that diets low in carbohydrates may reduce blood sugar levels, fasting blood glucose and HbA1c-levels in people with type 2 diabetes. A low carbohydrate diet may reduce body weight for people with type 2 diabetes, this was shown in 12 of 17 studies. Total cholesterol was reduced in six of 17 studies. A reduction of triglycerides was found in eight of 17 studies. LDL-cholesterol was reduced in four of the studies. HDL- cholesterol was increased in eight of 17 studies. Conclusion: Some of the studies showed positive effects of a low carbohydrate diet for people with type 2 diabetes, for instance improvements in blood sugar levels, bodyweight, total cholesterol, triglycerides, HDL-cholesterol and LDL-cholesterol. The study implies that a low carbohydrate diet may be an alternative for people with type 2 diabetes in short terms.Keywords: Diabetes type 2, diet, low-carbohydrate, ketogenic, HbA1c

Se estudiaron los efectos de la recurrencia del fuego en comunidades vegetales Mediterráneas dominadas por el árbol germinador P.halepensis Mill. (pino carrasco) a nivel regional en Cataluña. Entre 8 y 11 años tras la ocurrencia del fuego, se compararon la estructura y la composición florística de las comunidades vegetales, las poblaciones de P.halepensis, los horizontes orgánicos del suelo y el crecimiento de tres especies leñosas (Q.ilex, P.lentiscus, and R.officinalis) entre áreas quemadas una vez (en 1994) y dos veces (entre 1975 y 1993, y de nuevo en 1994) a lo largo de dos décadas. Se observó un patrón general de pérdida de resiliencia en las comunidades de P.halepensis quemadas dos veces a intervalos entre incendios menores de 17 años. / This work assessed the effects of fire recurrence on Mediterranean plant communities dominated by the seeder tree Pinus halepensis (Aleppo pine) at a regional level in Catalonia. It compared structure and floristic composition of plant communities, P.halepensis populations, soil organic horizons and plant growth of three woody species (Q.ilex, P.lentiscus, and R.officinalis) between areas burnt once (in 1994) and twice (between 1975 and 1993, and again in 1994) along 2 decades in the medium term after fire (between 8 and 11 years). A general trend of resilience loss was observed in P.halepensis communities burnt twice at fire intervals shorter than 17 years.

L'angiogenèse, soit la formation de nouveaux vaisseaux sanguins à partir de capillaires préexistants, est un processus essentiel au développement et à la croissance des tumeurs. Bon nombre de protéines ont été étudiées afin d'élucider les voies de signalisation impliquées dans l'angiogenèse, dont la métalloprotéase membranaire de type 1 (MT1-MMP). Cette protéine est reconnue pour jouer un rôle crucial dans la migration cellulaire, détruisant la matrice extracellulaire pour permettre aux cellules de migrer et ce, dans différents types cellulaires. Cependant, les mécanismes impliqués dans le contrôle de son activité demeurent incompris. Dans la présente étude, nous avons observé, en utilisant des procédures d'immunoprécipitation et de microscopie confocale, que la stimulation des cellules endothéliales de veines ombilicales humaines (HUVECs) avec la sphingosine-1-phosphate (S1P), un lipide qui induit la migration des cellules endothéliales (CEs), provoque le transfert de la MT1-MMP à la périphérie cellulaire et son association avec p130Cas (Crk-associated substrate). p130Cas est une protéine d'arrimage impliquée dans les voies de signalisation de la motilité cellulaire et est également reconnue pour se relocaliser dans des replis membranaires suite à une stimulation à la S1P. Ces résultats suggèrent fortement que l'identification du complexe MT1-MMP/p130Cas au front principal des CEs migrantes pourrait être un mécanisme efficace par lequel la protéolyse péricellulaire est reliée à l'activation des voies de signalisation, permettant la migration coordonnée des CEs au cours de l'angiogenèse. ______________________________________________________________________________ MOTS-CLÉS DE L’AUTEUR : Angiogenèse, MT1-MMP, p130Cas, Migration, Sphingosine 1-phosphate.

No description available.

Durant la progression du cancer, les cellules tumorales doivent envahir les tissus afin de se propager dans l'organisme. Pour ce faire, elles sécrètent ou expriment à leur surface des métalloprotéases matricielles, des enzymes capables de dégrader les composants de la matrice extracellulaire. Parmi ces enzymes, la métalloprotéase matricielle de type membranaire 1 (la MTI-MMP) joue un rôle primordial dans la progression tumorale. La MTI-MMP dégrade de nombreux composants de la matrice extracellulaire et des protéines de surface. Grâce à son activité protéolytique, la MTI-MMP joue un rôle important dans les différentes étapes de la progression tumorale. En effet, la MTl-MMP est importante dans l'angiogenèse tumorale, la formation de nouveaux capillaires à partir de vaisseaux pré-existants, car elle permet aux cellules endothéliales d'envahir la matrice extracellulaire. La MTI-MMP est également impliquée dans l'invasion tumorale et la formation de métastases. Bien que l'activité protéolytique de la MTI-MMP soit importante pour ses fonctions, plusieurs études ont démontré la participation de la séquence cytoplasmique de l'enzyme dans la migration et l'invasion tumorales. Toutefois, les mécanismes par lesquels le domaine intracellulaire de la MTI-MMP contribue aux fonctions de l'enzyme n'ont pas encore été élucidés. La présente thèse avait pour objectif principal d'étudier les mécanismes menant à l'implication du domaine cytoplasmique de la MTI-MMP dans la progression tumorale. Le premier objectif des travaux de recherche présentés dans cette thèse était d'identifier et de caractériser la phosphorylation de la MTI-MMP dans sa portion cytoplasmique. Pour ce faire, nous avons utilisé la mutagenèse dirigée afin d'identifier le site de phosphorylation de la MTI-MMP. Nos résultats démontrent que la MTI-MMP est phosphorylée sur son unique résidu tyrosine 573 situé dans la portion intracellulaire de l'enzyme. Cette phosphorylation a été confirmée par l'utilisation d'anticorps reconnaissant spécifiquement les formes phosphorylées de la MTI-MMP. De plus, la phosphorylation de cette enzyme requiert la présence de la kinase Src. La stimulation des cellules tumorales avec un facteur chimioattracteur induit la phosphorylation de la MTI-MMP endogène dans les cellules tumorales et endothéliales. Nous avons également montré que la phosphorylation de la MTI-MMP sur son résidu tyrosine intracellulaire est importante pour la migration des cellules tumorales et endothéliales. Le second objectif de cette thèse était d'étudier l'implication de la phosphorylation de la MTI-MMP dans la prolifération des cellules dans des matrices tridimensionnelles in vitro ainsi que dans la croissance tumorale in vivo. Nous avons constaté que la phosphorylation est importante pour la croissance des cellules tumorales dans des matrices tridimensionnelles alors qu'elle ne l'est pas dans des matrices en deux dimensions. Également, la phosphorylation de la MTI-MMP est nécessaire aux propriétés tumorigènes des cellules tumorales, à savoir la capacité de ces dernières à envahir de denses barrières de collagène fibrillaire et à former des colonies dans l'agar mou. Fait intéressant, nous avons constaté que l'inhibition de la phosphorylation de la MTI-MMP empêche la formation de xénogreffes chez la souris, suggérant que la MTl-MMP phosphorylée joue un rôle essentiel dans les mécanismes menant au développement tumoral. Le troisième objectif de cette thèse était de vérifier si la MTI-MMP phosphorylée était nécessaire pour la progression tumorale au niveau clinique. Nous avons donc étudié des spécimens provenant de biopsies de patients atteints de neuroblastome. Nos résultats ont démontré que la MTI-MMP phosphorylée était exprimée surtout dans les cas de neuroblastome avec un mauvais pronostic, alors que les cas qui avaient un bon pronostic exprimaient peu la MTI-MMP. Ces données indiquent que la phosphorylation de la MTI-MMP pourrait jouer un rôle important dans le développement du neuroblastome. Compte tenu du rôle majeur de la MTI-MMP phosphorylée dans la progression tumorale, le quatrième objectif de ces travaux de recherche était de concevoir une molécule pouvant inhiber la phosphorylation de la MTl-MMP afin de réduire le développement tumoral. Nous avons donc développé l'ACM-14, un peptide correspondant à la séquence cytoplasmique de la MTl-MMP dans laquelle la tyrosine a été remplacée par une phénylalanine et couplé à une séquence perméable aux cellules. Nos résultats démontrent que ACM-14 réduit la phosphorylation de la MTl-MMP et retarde par conséquent la croissance tumorale in vivo. En somme, les travaux présentés dans cette thèse apportent de nouvelles informations quant à l'implication du domaine intracellulaire de la MTI-MMP dans la progression tumorale. Les résultats présentés vont contribuer au développement et à l'élaboration de nouvelles stratégies afin d'inhiber le développement du cancer. ______________________________________________________________________________ MOTS-CLÉS DE L’AUTEUR : Cancer, Invasion tumorale, MTI-MMP, Phosphorylation, Traitement du cancer.

As awareness of the costs of workplace injury and illness continues to grow, there has been an increased demand for effective ergonomic interventions to reduce the prevalence of musculoskeletal disorders (MSDs). The goal of ergonomic interventions is to reduce exposures (mechanical and psychosocial); however there is conflicting evidence about the impact of these interventions as many studies produce inconclusive or conflicting results. In order to provide a clearer picture of the effectiveness of these interventions, we must find out if methodological issues, particularly statistical power, are limiting this research. The purpose of this study was to review and examine factors influencing statistical power in ergonomic intervention papers from five peer reviewed journals in 2008. A standardized review was performed by two reviewers. Twenty eight ergonomic intervention papers met the inclusion criteria and were fully reviewed. Data and trends from the reviewed papers were summarized specifically looking at the research designs used, the outcome measures used, if statistical power was mentioned, if a rationale for sample size was reported, if standardized and un-standardized effect sizes were reported, if confidence intervals were reported, the alpha levels used, if pair-wise correlation values were provided, if mean values and standard deviations were provided for all measures and the location of the studies. Also, the studies were rated based on the outcomes of their intervention into one of three categories (shown to be effective, inconclusive and not shown to be effective). Between these three groupings comparisons of post hoc power, standardized effect sizes, un-standardized effect sizes and coefficients of variation were made. The results indicate that in general, a lack of statistical power is indeed a concern and may be due to the sample sizes used, effect sizes produced, extremely high variability in some of the measures, the lack of attention paid to statistical power during research design and the lack of appropriate statistical reporting guidelines in journals where ergonomic intervention research may be published. A total of 69.6% of studies reviewed had a majority of measures with less than .50 power and 71.4% of all measures used had CVs of > .20.