The core intent of this study is to learn about the challenges faced by women survivors of sex trafficking in Nepal after their return. The study has attempted to answer questions about challenges survivors face, how they cope with the challenges, and resources available to these women to cope with the trauma and move ahead in life. In doing so, the study first considers demand and supply theory examining the influences of patriarchy, structural violence, the feminization of poverty and the social practices that support sex trafficking. The study also considers basic needs theory, increased opportunities to empower women, enhance participation and ensure their basic human rights. Despite all the hardships, survivors’ determination to rise from the ashes demonstrates courage and resilience. Throughout the study, empowerment is recognised as the driving force for these women to survive and thrive post-return. Economic independence, family’s love and acceptance, support and care from non-governmental organizations, and breaking silence regarding the ordeal these women survive foster empowerment. The study stresses the need to increase public awareness about sex trafficking in order to enable a respectful and dignified environment for the survivors. Survivors and NGO workers’ insight and experiences emphasize that for plans and policies to work effectively, the government bodies should work hand in hand with non-governmental organizations and increase the involvement of survivors throughout the reintegration process. / October 2015

Cellular pH homeostasis plays crucial roles in cellular functions, and it is now widely recognized that Na⁺/H⁺ exchangers are among the most prominent players in this process. Although recently described mammalian Na⁺/H⁺ exchanger NHE7 has attracted much attention, its biological functions remain largely unknown. Most proteins exist as protein complexes in the cell and elicit their unique functions in collaboration with their binding partners. Therefore, identification and characterization of binding proteins will often unveil unexpected functions of the protein of interest. To begin to elucidate biological roles of the novel class of Na⁺/H⁺ exchanger NHE7, yeast two-hybrid screening was conducted and several binding candidates were identified. Among these candidates, I show that Secretory Carrier Membrane Proteins (SCAMPs) are novel NHE7 binding proteins and that SCAMPs regulate endocytic trafficking of NHE7 from the recycling endosomes to the trans-Golgi network (TGN). In agreement with this finding, I found that NHE7 can also be targeted to the plasma membrane and then internalized. Caveolins, structural proteins for caveolae, were identified as NHE7-binding proteins and it was initially hypothesized that caveolins might regulate NHE7-internalization. Interestingly, caveolins bound to NHE7 through a novel binding domain and facilitated its association to caveolae/lipid rafts, but did not affect NHE7-internalization. I also show that SCAMP2 associates with the heterotrimeric G protein β subunit (Gβ) and regulates the ERK1/2 signaling. Moreover, NHE7 was found to associate with both SCAMP2 and Gβ in the cell, suggesting that ERK1/2 signaling mediated by the SCAMP2-Gβ complex might regulate NHE7.

The Saccharomyces cerevisiae oxysterol binding protein homologue Kes1/Osh4 is a member of an enigmatic class of proteins found throughout Eukarya. This family of proteins is united by a ?-barrel structure that binds sterols and oxysterols. An N-terminal lid is thought to both sequester sterols inside the core and promote localization of Kes1 to regions of high membrane curvature via a predicted ArfGAP lipid packing sensor motif. Additionally, a phosphoinositide-binding region on a discrete surface of Kes1 has also been identified. In this thesis, structure-function analysis of Kes1 determined that phosphoinositide binding is required for membrane association in vitro, and in vivo phosphoinositide binding is required for localization to the Golgi. Ergosterol, the major sterol in S. cerevisiae, and membrane curvature had minimal effects on membrane association. This study also revealed a role for Kes1 in the regulation of both phosphatidylinositol-4-phosphate and phosphatidylinositol-3-phosphate homeostasis. Phosphoinositide and sterol binding by Kes1 are necessary for it to alter phosphatidylinositol-4-phosphate, but not phosphatidylinositol-3-phosphate homeostasis. Misregulation of phosphatidylinositol-4-phosphate homeostasis by Kes1 manifested itself in an inability of the v-SNARE Snc1 to traffic properly and was consistent with a defect in trans-Golgi/endosome trafficking. I went on to demonstrate a role for Kes1 in regulating the conversion of phosphatidylinositol-4-phosphate to phosphatidylinositol for the synthesis of sphingolipids, and I present a model for the role of Kes1 at the Golgi. Kes1 acts as a sterol sensor that regulates phosphatidylinositol-4-phosphate to sphingolipids metabolism, which ultimately regulates the delivery of proteins that assemble into lipid rafts for their transport from the Golgi to the plasma membrane. I also uncovered a previously unknown role for Kes1 in the regulation of the cytoplasm-to-vacuole and autophagy trafficking pathways, which is mediated by the ability of Kes1 to regulate phosphatidylinositol-3-phosphate homeostasis.

The RET proto-oncogene encodes a receptor tyrosine kinase (RTK) that is widely expressed in neuroendocrine tissues and is essential for embryonic development of the kidney and enteric nervous system. Mutations leading to constitutive activation of the RET protein underlie various tumours of endocrine tissues. Conversely, loss-of-function mutations of RET lead to Hirschsprung disease, a congenital disorder characterized by a loss of enteric neurons throughout the colon and small intestine. Intracellular trafficking of RTKs through multiple cellular compartments has been shown to impact on downstream signalling. To date, the intracellular trafficking of RET has not been investigated. Here, we show that RET is rapidly internalized after activation and that trafficking to cytoplasmic endosomes plays an important role in downstream signalling. RET is alternatively spliced into multiple isoforms that are co-expressed in cells; therefore, we further investigated RET internalization in an isoform-specific context. This study revealed a number of differences between RET isoforms including differences in sub-cellular localization pre-activation, rate of internalization, and ability to recycle to the plasma membrane. Differential trafficking of RET isoforms alter their downstream signalling properties, providing an additional mechanism to explain the distinct contributions of RET isoforms to cellular processes. Finally, we investigated the impact of altered sub-cellular localization in the context of thyroid carcinoma. Activation of RET has been implicated in a number of thyroid tumours that differ in their inherent oncogenicity. We observed that altered subcellular localization of oncogenic forms of RET, RET/PTCs, enhance their oncogenicity. Interestingly, RET/PTC tumours are indolent and rarely metastasize compared to other RET-mediated forms of cancer. Further investigation revealed that RET/PTC oncogenes are expressed off relatively weak promoters, resulting in quantitatively less RET/PTC oncoprotein expression in these tumours compared to mutant RET expression in more aggressive cancers. Together, our results represent the first in-depth study of the trafficking properties of RET and indicate the importance of proper sub-cellular localization and trafficking in the maintenance of normal cell metabolism. / Thesis (Ph.D, Pathology & Molecular Medicine) -- Queen's University, 2009-11-19 22:51:47.38

No description available.

The trafficking of illicit drugs by sea has become an industry comprised of many individual enterprises of variform size and organization. Seizure statistics for the 1980s indicate that 70% of the total quantity of drugs intercepted in the trafficking stage were interdicted in the maritime sector or attributed to having been transported by sea. More significantly, it appears that only between 8 - 12% of the total volume of drugs trafficked are intercepted. The use of the seaborne modes of transport is the result of planetary geography which made the maritime medium one of only two ways by which drugs may enter several states. In response, varying sophisticated counter-trafficking offensives, policies and strategies have been implemented and contemplated in select geographical regions - examples being the Caribbean and Pacific Basins. However, the importation of illicit substances to the primary consuming states has not been curbed and indications are that the overall flow of drugs remains unimpeded. This thesis focuses on the maritime trade in illicit drugs during the 1980s by providing both qualitative and quantitative analyses of the activity. Specifically, the theme addressed is the question of why there is so little success in combating the maritime drug trade. Embraced by the study are the various geographical, physical, technical and socio-political elements supportive of the trade. Among the pertinent topics revealed are the flow structure to the trade, the categories of drugs transported, the classes of vessels utilized, the methods of concealment and deception employed, the involvement of organized crime, the contributing geographical elements and the unique variations to specific routes as determined by destination and region. Additionally, the international law suppressing the maritime trade in illicit drugs is examined. To lend completeness to the study a brief review of the historical dimension to the smuggling of drugs by sea is included along with analysis of drug production and consumption. Because the threat of drugs is perceived to be greatest, albeit wrongly, among the developed states this thesis tackles the subject from the perspective of the coastal member states of O.E.C.D. Lastly, recommendations and innovations to old strategies are proffered specifically as they apply to the maritime component of the illicit drug trade.

Integral membrane proteins are targeted to discrete compartments through the action of a number of transport pathways. The post-translational modification of cargo with ubiquitin is a key regulator of protein sorting. Ubiquitinated cargo are bound by specific cargo sorting machinery and directed towards the appropriate destination. Therefore, the identification and characterization of the proteins involved in cargo ubiquitination is critical to understanding the regulation of protein sorting. In the work presented here, we characterize the role of the E3 ubiquitin ligases, RNF126 and Rabring7, in two distinct membrane trafficking pathways. First, we show that RNF126 and Rabring7 are involved in the ligand induced downregulation of cell surface receptors. RNF126 and Rabring7 associate with the EGFR, amongst other RTKs, and promotes its ubiquitination. RNF126 and Rabring7 are required for the efficient sorting of the EGFR through the late endocytic compartment. We also show that the depletion of Rabring7 attenuates the degradation of MET and that both RNF126 and Rabring7 regulate the sorting of CXCR4 from an early endocytic compartment. In addition, the depletion of RNF126 or Rabring7 destabilizes ESCRT-II and reduces the number of multivesicular bodies formed after EGF stimulation. Second, we found that RNF126 regulates the sorting of the CI-MPR. In cells transiently depleted of RNF126, the CI-MPR is dispersed into a transferrin receptor positive endocytic compartment. This effect is specific to the CI-MPR as other cargos that are sorted between the endosome at the Golgi remain unaffected. We found that RNF126 physically associates with the clathrin adaptor GGA3 and promotes its ubiquitination, suggesting that RNF126 regulates GGA3 mediated CI-MPR sorting. Together, this work furthers our understanding regarding the role of ubiquitin in membrane traffic.

In this thesis, I examine how health stakeholders in British Columbia think and talk about human trafficking. I interrogate the health stakeholders’ speech as a site where broad societal discourses associated with human trafficking manifest. Using critical race theory, interlocking analysis, and a Foucauldian discourse analysis approach, I critically deconstruct health stakeholders’ understandings of human trafficking and persons who are trafficked. I pay particular attention to the discursive strategies the health stakeholders employ to construct the subjectivities of both persons who are trafficked and themselves in human trafficking discourse. I argue that these meaning-making processes and the uncritical reproduction of dominant human trafficking discourse in the health sector at least, in part, account for the lack of development and implementation of provincial human trafficking-specific policy and services to date. Given this absence, this thesis encourages health stakeholders to create evidence-based initiatives to address human trafficking and the health needs of persons who are trafficked. / Graduate / 0452 / aclancey@hotmail.com

No description available.

Anion Exchanger 1, AE1, is a membrane glycoprotein that functions as a dimer in the red blood cells (RBC) as well the kidney. It functions to exchange Cl- for HCO3- in an electroneutral manner, with the RBC AE1 having an additional function in maintaining its biconcave shape. Mutations in AE1 can cause Hereditary Spherocytosis (HS) and distal Renal Tubular Acidosis (dRTA). A mutation, C479W, has been discovered in Edmonton that caused the rare incidence of HS and dRTA in young patient who is heterozygous for C479W and G701D, a recessive dRTA mutation. Expression in MDCK cells has demonstrated that C479W AE1 is retained intracellularly, is misfolded, but can dimerize. C479W AE1s trafficking to the plasma membrane is not rescued by interaction with WT protein, or the small molecules glycerol and DMSO, or by reduced temperature. C479W AE1 also has an increased interaction with the ER chaperone protein, Calnexin.