Bullying Involvement and Adolescent Substance Use: A Study of Multilevel Risk and Protective Factors

Bullying, frequent drunkenness, and frequent cannabis use are significant health-risk behaviours among youth. While many studies have demonstrated that bullying involvement may initiate a developmental pathway to both types of frequent substance use, there is a limited understanding of the connection between these behaviours. The presence of risk and protective factors within youths’ relationships and within their neighbourhoods may alter the associations between bullying involvement and both types of frequent substance use. A systemic approach is needed to assess the complex, social environments in which youth are embedded. The current thesis consists of two studies that examined the associations between bullying and both types of frequent substance use within the context of youths’ social environments. In Study 1, multilevel modeling was used to examine the associations between bullying and frequent substance use within the context of individual and neighbourhood risk factors. Our results indicated that the risk factors associated with both frequent drunkenness and frequent cannabis use exist at both levels, with neighbourhoods altering the association of individual risk factors. Moreover, bullying was a unique risk factor associated with both types of frequent substance use, whereas indirect associations were observed for victimization. Study 2 used a similar methodology to examine the association between bullying and both types of frequent substance use within the context of individual and neighbourhood protective factors. Once again, our results indicated that the protective factors associated with both types of frequent substance use exist at multiple levels, and that neighbourhoods altered the association of individual protective factors. Additionally, positive relationship characteristics interacted with the link between bullying and both types of frequent substance use. Together, these findings clarify the nature of the bullying-substance use link and emphasize the need to study adolescent development in context.

IMMUNE-ANGIOGENESIS MECHANISMS ASSOCIATED WITH PORCINE PREGNANCY SUCCESS AND FAILURE

Spontaneous fetal loss (25-40%) leading to decrease in litter size is a significant concern to the pork industry. A deficit in the placental vasculature has emerged as one of the important factors associated with fetal loss. During early pig pregnancy, the endometrium becomes enriched with immune cells recruited by conceptus-derived signals including specific chemokine stimuli. These immune cells assist in various aspects of placental development and angiogenesis. Recent evidence suggests that microRNAs (miRNAs: small non-coding RNAs that regulate gene expression) regulate immune cell development and their functions. In addition, intercellular communication including exchange of biomolecules (e.g. miRNAs) between the conceptus and endometrium regulate key developmental processes during pregnancy. To understand the biological significance of immune cell enrichment, regulation of their functions by miRNAs and transfer of miRNAs across the maternal fetal-interface, we screened specific sets of chemokines and pro- and anti-angiogenic miRNAs in endometrial lymphocytes (ENDO LY), endometrium, and chorioallantoic membrane (CAM) isolated from conceptus attachment sites (CAS) during early, gestation day (gd)20 and mid-pregnancy (gd50). We report increased expression of selected chemokines including CXCR3 and CCR5 in ENDO LY and CXCL10, CXCR3, CCL5, CCR5 in endometrium associated with arresting CAS at gd20. Some of these differences were also noted at the protein level (CXCL10, CXCR3, CCL5, and CCR5) in endometrium and CAM. We report for the first time significant differences for miRNAs involved in immune cell-derived angiogenesis (miR-296-5P, miR-150, miR-17P-5P, miR-18a, and miR-19a) between ENDO LY associated with healthy and arresting CAS. Significant differences were also found in endometrium and CAM for some miRNAs (miR-17-5P, miR-18a, miR-15b-5P, and miR-222). Finally, we confirm that placenta specific-exosomes contain proteins and 14 select miRNAs including miR-126-5P, miR-296-5P, miR-16, and miR-17-5P that are of relevance to early implantation events. We further demonstrated the bidirectional exosome shuttling between porcine trophectoderm cells (PTr2) and porcine aortic endothelial cells (PAOEC). PTr2-derived exosomes were able to modulate the endothelial cell proliferation that is crucial for the establishment of pregnancy. Our data unravels the selected chemokines and miRNAs associated with immune cell-regulated angiogenesis and reconfirm that exosome mediated cell-cell communication opens-up new avenues to understand porcine pregnancy.