VictorJuiz/thesis huggingface dataset:
Targeted Drug Delivery to Breast Cancer using Polymeric Nanoparticle Micelles
Keywords: Drug delivery
Cancer
Chemotherapy
Anti-cancer therapy
Polymeric nanoparticles
Nanoparticle micelles
Pharmacokinetics
Biodistribution
Live cell binding
Tumour xenograft models
Tumour pathophysiology
Orthotopic tumour model
Nanoparticle targeting
Passive targeting
Active targeting
Formulations
Antibody targeting
Breast cancer

Broad distribution and activity limit the utility of anti-cancer compounds by causing unacceptable systemic toxicity and narrow therapeutic indices. To improve tumour accumulation, drug-loaded macromolecular assemblies have been designed to replace conventional surfactant-based formulations. Their nanoscale size enhances tumour accumulation via hyperpermeable vasculature and reduced lymphatic drainage. Incorporating targeting ligands introduces cell specificity through receptor-specific binding and uptake, enabling drugs to reach intracellular targets. In this work, the targeting properties of polymer nanoparticle micelles of poly(2-methyl-2-carboxytrimethylene carbonate-co-D,L-lactide)-graft-poly(ethylene glycol)-furan (poly(TMCC-co-LA)-g-PEG) were verified using in vitro and in vivo models of breast cancer. To select a relevant mouse model, the vascular and lymphovascular properties of two tumour xenograft models were compared. Greater accumulation of a model nanocarrier was observed in orthotopic mammary fat pad (MFP) tumours than size matched ectopic subcutaneous tumours, suggesting that the organ environment influenced the underlying pathophysiology. Immunostaining revealed greater vascular thickness, density and size, and thinner basement membranes in MFP tumours, likely contributing to greater blood perfusion and vascular permeability. Based on these observations, MFP tumour-bearing mice were used to characterize the pharmacokinetics and biodistribution of a taxol drug, docetaxel, encapsulated in poly(TMCC-co-LA)-g-PEG nanoparticles. The nanoparticle formulation demonstrated longer docetaxel circulation in plasma compared to the conventional surfactant-based formulation. As a result, greater docetaxel retention was uniquely measured in tumour tissue, extending exposure of tumour cells to the active compound and suggesting potential for increased anti-cancer efficacy. Furthermore, active targeting of antibody-modified nanoparticles to live cells was shown to be selective and receptor-specific. Binding isotherms were used to quantify the impact of antibody density on binding strength. The equilibrium binding constant increased linearly with the average number of antibodies per particle, which is consistent with a single antibody-antigen interaction per particle. This mechanistic understanding enables binding behaviour to be adjusted in a predictive manner and guides rational nanoparticle design. These studies validate poly(TMCC-co-LA)-g-PEG nanoparticles as a platform for targeted delivery to cancer on both a tissue and cellular level, forming a compelling justification for further pre-clinical evaluation of this system for safety and efficacy in vivo.
The Effects of Up-regulated Happiness on Others’ Prosocial Behavior: The Role of Cultural Thinking Style
Keywords: social effects of emotion
prosocial behavior
inauthentic displays of happiness
analytical-holistic thinking
culture
trust

Past research has suggested that up-regulating displays of happiness leads to positive social outcomes. In my dissertation research, I suggest that whether the up-regulation of displays of happiness leads to positive outcomes depends on observers’ culture. I developed and tested a model of how one aspect of observers’ culture, analytic-holistic thinking, influences the relation between expressers’ displays of happiness and observers’ prosocial behavior. In two studies, I examined how observers’ analytical-holistic thinking moderates the effect of expressers’ inauthentic displays of happiness (regulated through surface acting) and expressers’ authentic displays of happiness (regulated through deep acting) on observers’ prosocial behavior and trust inferences. In a laboratory experiment (Study 1), observers’ analytical-holistic thinking moderated the effect of project leaders’ inauthentic displays of happiness (vs. neutral emotional displays) on observers’ trust in the expresser, but not on observers’ prosocial behavior (time taken to edit a cover letter). In a field fundraising experiment (Study 2), observers’ analytical-holistic thinking moderated the effect of fundraisers’ inauthentic displays of happiness (vs. authentic displays of happiness) on observers’ trust in fundraisers, intentions to volunteer for the charity organization, and monetary donations to the charity organization. Further, observers’ trust in fundraisers mediated the moderating effect of observers’ analytical-holistic thinking on the relation between fundraisers’ inauthentic displays of happiness and observers’ intention to volunteer. Contributions to the literature on the social effects of emotions and to the literature on fundraising and helping as well as practical implications are discussed.
Electrode/Organic Interfaces in Organic Optoelectronics
Keywords: organic light emitting diodes
charge injection
organic electronics
electrodes

Organic semiconductors have the advantage over traditional inorganic semiconductors, such as Si or GaAs, in that they do not require perfect single crystal films to operate in real devices. Complicated multi-layer structures with nanometer scale thicknesses can thus be easily fabricated from organic materials using low-cost roll-to-roll manufacturing techniques. However, the discrete nature of organic semiconductors also implies that they typically contain almost no intrinsic charge carriers (i.e., electrons or holes), and thus act as insulators until electrical charges are injected into them. In electrical device applications this means that all of the holes and electrons within a device must be injected from the anode and cathode respectively. As a result, device stability, performance, and lifetime are greatly influenced by the interface between the organic materials and the electrode contacts. Despite the fundamental importance of the electrode/organic contacts, much of the basic physical understanding of these interfaces remains unclear. As a result, the current design of state-of-the-art organic optoelectronic devices tends to be based on trial and error experimentation, resulting in overly complicated structures that are less than optimal. In the present thesis, various electrode/organic interfaces relevant to device applications are studied using a variety of different techniques, including photoelectron spectroscopy and the iii temperature dependent current-voltage characteristics of single carrier devices. The fundamental understanding gleaned from these studies has been used to develop new strategies for controlling the energy-level alignment at electrode/organic interfaces. A universal method for tuning the work function of electrode materials using a halogenated organic solvent and UV light has been developed. Application of this technique in organic light emitting diodes enabled the first highly simplified two-layer device with a state-of-the-art record breaking efficiency.
Synthetic Application of Amphoteric Aziridine Aldehydes and alpha-Boryl Aldehydes
Keywords: nitrogen-containing heterocyclic compounds
organoboron

A range of N-H alkynylaziridines were prepared from amphoteric unprotected aziridine aldehydes without protecting-group manipulation. Unprotected alpha -amino allenes can be obtained from these strained propargyl amines via a 9-BBN mediated hydride transfer. Further transformation of alpha -amino allenes to 2,4,6-trisubstituted pyridines was realized. We also developed another class of amphoteric molecules – alpha-boryl aldehydes, equipped with the tetrahedral MIDA boryl group. A wide range of boryl-substituted building blocks or functionalized boronic acid derivatives have been accessed from these bench-stable alpha-borylcarbonyl compounds. Further chemoselective transformations of these alpha-boryl aldehyde derived building blocks have been conducted, where alph-boryl isocyanates, alpha-aminoboronic acids, acylboronates, and borylated heterocycles were achieved through the decarboxylative functionalization of alpha-borylcarboxylic acids.
The Interaction of Ice Sheets with the Ocean and Atmosphere
Keywords: Kalman smoother
tip jet
ice sheets
sea level fingerprints

A rapidly melting ice sheet produces a distinctive geometry of sea level (SL) change. Thus, a network of SL observations may, in principle, be used to infer sources of meltwater flux. We outline a new method, based on a Kalman smoother, for using tide gauge observations to estimate the individual sources of global SL change. The Kalman smoother technique iteratively calculates the maximum likelihood estimate of Greenland and West Antarctic ice sheet melt rates at each time step, and it allows for data gaps while also permitting the estimation of non-linear trends. We have also implemented a fixed multi-model Kalman filter that allows us to rigorously account for additional contributions to SL changes, such as glacial isostatic adjustment and thermal expansion. We report on a series of detection experiments based on synthetic SL data that explore the feasibility of extracting source information from SL records before applying the new methodology to historical tide gauge records. In the historical tide gauge study we infer a global mean SL rise of ~1.5 ± 0.5 mm/yr up to 1970, followed by an acceleration to a rate of ~2.0 ± 0.5 mm/yr in 2008. In addition to its connection to SL, Greenland and its large ice sheet act as a barrier to storm systems traversing the North Atlantic. As a result of the interaction with Greenland, low-pressure systems located in the Irminger Sea, between Iceland and Greenland, often produce strong low-level winds. Through a combination of modeling and the analysis of rare in-situ observations, we explore the evolution of a lee cyclone that resulted in three high-speed-wind events in November 2004. Understanding Greenland’s role in these events is critical in our understanding of local weather in this region.
Global Proteomic Detection of Native, Stable, Soluble Human Protein Complexes
Keywords: protein
proteomics
HeLa cells
293 cells
HPLC
interactome
proteome
cofractionation
mass spectrometry
human
protein complex
chromatography

Protein complexes are critical to virtually every biological process performed by living organisms. The cellular “interactome”, or set of physical protein-protein interactions, is of particular interest, but no comprehensive study of human multi-protein complexes has yet been reported. In this Thesis, I describe the development of a novel high-throughput profiling method, which I term Fractionomic Profiling-Mass Spectrometry (or FP-MS), in which biochemical fractionation using non-denaturing high performance liquid chromatography (HPLC), as an alternative to affinity purification (e.g. TAP tagging) or immuno-precipitation, is coupled with tandem mass spectrometry-based protein identification for the global detection of stably-associated protein complexes in mammalian cells or tissues. Using a cell culture model system, I document proof-of-principle experiments confirming the suitability of this method for monitoring large numbers of soluble, stable protein complexes from either crude protein extracts or enriched sub-cellular compartments. Next, I document how, using orthogonal functional genomics information generated in collaboration with computational biology groups as filters, we applied FP-MS co-fractionation profiling to construct a high-quality map of 622 predicted unique soluble human protein complexes that could be biochemically enriched from HeLa and HEK293 nuclear and cytoplasmic extracts. Our network is enriched in assemblies consisting of human disease-linked proteins and contains hundreds of putative new components and novel complexes, many of which are broadly evolutionarily conserved. This study revealed unexpected biological associations, such as the GNL3, FTSJ3, and MKI67IP factors involved in 60S ribosome assembly. It is my expectation that this first systematic, experimentally-derived atlas of putative human protein complexes will constitute a starting point for more in depth, hypothesis-driven functional investigations of basic human molecular and cellular biology. I also note that my generic FP-MS screening approach can, and is currently, being applied by other members of the Emili laboratory to examine the global interactomes of other mammalian cell lines, tissues, sub-cellular compartments, and diverse model organisms, which should expand our understanding of proteome adaptations and association networks associated with cell physiology, animal development and molecular evolution.
Heterarchy and Hierarchy in the Formation and Dissolution of Complex Hunter-gatherer Communities on the Northern Plateau
Keywords: Complex Hunter-Gatherers
Mid-Fraser Archaeology
Heterarchy
Social Organization

This research explores the changing nature of social organization associated with the growth and breakup of large nucleated hunter-gatherer winter settlements in the Mid-Fraser region of south-central British Columbia, ca. 2000-300 cal. B.P. It uses hierarchy and heterarchy as overarching conceptual frameworks for theorizing and evaluating structures of social and political organization. Regional radiocarbon data were used to examine issues of demography and to evaluate the role of scalar stress in producing social change in these burgeoning communities. In order to explore aspects of economic practice and wealth distribution over time artifacts, fauna, and features from sixteen different housepits from five different village sites near the present-day town of Lillooet, British Columbia were analyzed. Results suggest that the villages formed around 1800 cal. B.P. and attained peak population ca. 1200 cal. B.P. The onset of the Medieval Climatic Anomaly at that time altered resource conditions, resulting in greater reliance on mammalian rather than riverine resources. Increased pressure on these resources led to the incorporation of greater amounts of small bodied mammals after 1000 cal. B.P. Apparent declining numbers of houses within large villages after 1200 cal. B.P. suggest that village abandonment began at this time, with individual families likely settling in dispersed villages. The large villages were totally abandoned by 900-800 cal. B.P. Lack of evidence for wealth differentiation in these contexts suggest that social hierarchy based on control over access to resources never emerged in the large villages and that more egalitarian conditions prevailed. Heterarchical structures that allow for shifting balance of power between bands and individual families is argued to have characterized the shift between population aggregation and dispersal.
The Role of Endoplasmic Reticulum Aminopeptidase 1 (ERAP1) in the Pathogenesis of Ankylosing Spondylitis
Keywords: Spondyloarthritis
HLA-B27
Endoplasmic Reticulum
Antigen presentation

Ankylosing spondylitis (AS) is associated with HLA-B*2704 and B*2705 but not with HLA-B*2706 and B*2709. Genome wide studies recently identified ERAP1 as an important genetic association in AS and could be the missing link in the pathogenesis of AS. I studied the implications of the two known actions of ERAP1 on AS pathogenesis. For assessing the peptide trimming function, surface HLA-B27 and MHC-I free heavy chain (FHC) expression on peripheral blood mononuclear cells of AS patients were studied. Subsequently, in an in vitro system of C1R cells expressing different AS-associated and AS-neutral HLA-B27 subtypes, I studied the effect of ERAP1 suppression on HLA-B27 and FHC expression. To assess the cytokine receptor shedding function, I studied serum cytokine receptor level variation with ERAP1 polymorphisms and its relationship to disease activity in AS patients. Finally, I studied the effect of variants of ERAP1 and other members of the antigen presentation machinery on radiographic severity in AS patients. AS patients with the major allele of the ERAP1 rs27044 polymorphism had higher FHC expression on monocytes. In C1R cells ERAP1 suppression led to an increase in intracellular FHC (IC-FHC) and B27-peptide complexes identified by a special MARB4 antibody, but only in C1R cells expressing the AS-associated subtypes HLA-B*2704 and B*2705. ERAP1 variants had no effect on serum cytokine receptor levels. Baseline radiographic severity was associated with ERAP1 polymorphism in univariate analysis only. LMP2 variants were associated with baseline radiographic severity in multivariate analysis. ERAP1 affects peptide presentation and FHC formation by HLA-B27 and could be the missing link in the pathogenesis of AS. ERAP1 through its differential HLA-B27 subtype interaction could explain why certain subtypes of HLA-B27 are associated with AS while others are not. Larger studies are required to look closely at the effect of ERAP1 on radiographic severity and progression in AS.
The Role of 53BP1 and its Phosphorylation in the DNA Damage Response
Keywords: DNA Damage
Genetic Instability
Cancer
Ionizing radiation
53BP1
Hypoxia
Cell cycle
Radiosensitivity

The tumour suppressor p53-binding protein 1 (53BP1) is phosphorylated following DNA double strand breaks (DSBs); however, little is understood about the upstream signaling pathways that control this phosphorylation. Additionally, it is not known how these processes combine with 53BP1 to control the survival of cells following DNA damage such as that imparted by ionizing radiation (IR), which is the basis of radiotherapy. In this thesis, I have shown that 53BP1 is phosphorylated specifically in S-phase cells, but not relocalized to intranuclear foci, in response to severe oxygen stress. This occurs with only partial dependence on the ATM kinase (Chapter 2). Following IR, I find that both ATM and DNA-PKcs contribute to intranuclear phosphorylated 53BP1 foci, but that this phosphorylation is independent of proximal signaling molecules that control the localization of 53BP1 to initial DSBs (Chapter 3). Furthermore, I show that 53BP1 loss confers sensitivity to IR and this can be further augmented by inhibition of ATM and DNA-PKcs kinases suggesting that there are both 53BP1-dependent and -independent pathways of survival from IR (Chapter 4). These findings may have important implications for molecular pathology and personalized medicine as 53BP1 has recently been found to be activated or lost in subsets of human tumours. I have collaborated to initiate the development of a novel system to interrogate the implications of 53BP1 loss as traditional siRNA approaches in human cancer cells were not feasible (Chapter 5 and Appendix 2). This system can be used in vivo as tumour xenografts to further understand how 53BP1 and the tumour microenvironment interact endogenously and in response to IR. I also present the possibility and proof of concept for the use of 53BP1 as a biomarker in primary human prostate cancer tissue where little is known about 53BP1 biology (Chapter 5).
Molecular Mechanisms of Glycine Primed NMDA Receptor Internalization
Keywords: NMDA receptor
Neuroscience
Glycine
Endocytosis

N-Methyl-D-aspartate receptors (NMDARs) are a principal subtype of excitatory ligandgated ion channel with prominent roles in physiology and disease in the mammalian central nervous system (CNS). Activation of NMDARs requires binding of both glutamate and glycine. Apart from its co-agonist action, glycine can also prime NMDARs for subsequent internalization upon binding of both glutamate and glycine. However, the molecular basis responsible for mediating and regulating glycine priming and NMDAR endocytosis is largely unknown. In my thesis, I discovered the principle that although NMDAR gating and priming share a common requirement for glycine binding, the molecular constraints for gating are distinct from those for priming through two mutations of the glycine binding site in GluN1 subunit of the NMDAR that, while maintaining gating of NMDARs, eliminate glycine priming of the receptors. One of the molecular signatures of glycine priming is recruitment of the endocytic adaptor protein AP-2. I have characterized the two regions in GluN2 subunits required for enhanced AP-2 association. This unexpected result suggests binding of glycine initiates a conformational change transmitted from GluN1 to GluN2 allowing for docking of endocytic machinery. Furthermore, I have discovered that naturally occurring splice variants of GluN1 subunit, containing a 21 amino acid sequence in the N-terminus domain (N1) cassette, abrogate glycine stimulated AP-2 recruitment and glycine-primed NMDAR internalization. These findings imply that there are distinct populations of native NMDARs in the CNS – those lacking N1 that show glycine-primed internalization and those containing N1 that are not primable. Collectively, my thesis work demonstrates a dramatic all-or-none priming effect with splice variants of NMDARs, a highly unexpected discovery providing novel insight into the molecular mechanisms and physiological role of glycine priming. Ultimately, elucidating principles and mechanisms of glycine priming lay the foundation for new types therapeutic approaches for CNS disorders, approaches without the deleterious consequences of directly blocking NMDARs.
The Mortality Cost of Undertriage of Major Trauma in Ontario
Keywords: Injury
Trauma system
Trauma

Introduction: Multiple studies suggest that severely injured patients treated at trauma centers have a lower mortality compared to patients treated at non-trauma centers. In many trauma systems, a significant proportion of patients continue to be transported from the scene to a non-trauma center (undertriaged); only a fraction of these patients are subsequently transferred to trauma center care. Although previous analyses have attempted to examine the mortality associated with transfer and with undertriage, these studies were not population-based, and therefore potentially underestimated the mortality cost of undertriage at the system level. Methods: In this dissertation, we developed an algorithm to convert ICD-10 diagnosis codes to Injury Severity Score. This algorithm allowed us to utilize population-based data to examine the outcomes of all severely injured patients surviving to reach an emergency department in Ontario. We examined whether, among severely injured patients, transfer from a non-trauma center to a trauma center is associated with increased mortality compared to direct transport from the scene. In addition, we used an instrumental variable analysis to produce a population-based estimate of the mortality cost of undertriage in a subset of patients injured in motor vehicle collisions. Results: Patients requiring transfer to trauma center care have significantly higher mortality at 30 days than patients transported directly from the scene of injury (Odds ratio 1.24; 95% CI, 1.10-1.40). Among patients involved in motor vehicle collisions, only 45% were transported directly to a trauma center. In this subset of patients, those triaged directly to a trauma center had significantly lower mortality at 24 hours (Odds ratio 0.58, 95% CI 0.41-0.84) and 48 hours (Odds ratio 0.68, 95% CI 0.48-0.96) compared to undertriaged patients. There was a trend towards decreased mortality among patients triaged to a trauma center at 7 days and 30 days. Conclusions: Undertriage and transfer after major trauma are associated with substantial increase in mortality compared to direct transport to a trauma center. These data suggest a need to design strategies to improve access to trauma center care in Ontario.