The Immunomodulatory Role of Alcohol on HIV-Infected Monocyte-Derived Dendritic Cells

Alcohol is known to induce inflammation in the presence of the human immunodeficiency virus (HIV). In our previous studies, we revealed that alcohol induces cannabinoid receptors which play a role in the regulation of inflammatory cytokine production in monocyte-derived dendritic cells (MDDC). However, the ability of alcohol to alter MDDC function during HIV infection has not been clearly elucidated yet. To study the potential impact of alcohol on HIV-infected MDDC (confirmed by p24 ELISA), monocytes were isolated from commercially available buffy coats and cultured for 7 days with GM-CSF and IL-4. MDDC were infected with HIV- 1Ba-L and treated with different concentrations of alcohol (0.1% band 0.2%) for 4-7 days. MDDC phenotype, endocytosis, cytokine production, and ability to transmit HIV to T cells were analyzed. Uninfected CD4+ T cells were co-cultured for 7 days with either infected/treated MDDC or the supernatants from infected/treated MDDC. Inflammatory cytokine arrays were performed using supernatants from HIV-infected MDDC treated with alcohol. Results showed that HIV positive MDDC treated with alcohol had higher levels of infection compared to untreated HIV positive controls. CD4+ T cells exposed to HIV-infected MDDC acquired 100-fold higher levels of p24 compared to CD4+ T cells exposed to only supernatants. CD4+ T cells exposed to HIV-infected and alcohol-treated MDDC had higher levels of infection compared to controls. Cytokine array data show dysregulation of cytokine production by alcohol. In addition, MDDC phenotype and endocytic capacity were altered in the alcohol treated MDDC. Our results indicate a crucial role of MDDC in HIV transmission to T cells and provide insights into the inflammatory role alcohol exerts on dendritic cell function in the context of HIV infection. Supported by the National Institute on Alcohol Abuse and Alcoholism award R00AA021264, the National Institute on Drug Abuse award R01DA034547, and the Institute on NeuroImmune Pharmacology at FIU.

Economy of the island of eco-tourism: the role of shark-diving and shark harvesting

Beyond its importance in maintaining ecosystems, sharks provide services that play important socioeconomic roles. The rise in their exploitation as a tourism resource in recent years has highlighted economic potential of non-destructive uses of sharks and the extent of economic losses associated to declines in their population. In this work, we present estimates for use value of sharks in Fernando de Noronha Island - the only ecotouristic site offering shark diving experience in the Atlantic coast of South America. Through the Travel Cost Method we estimate the total touristic use value aggregated to Noronha Island by the travel cost was up to USD 312 million annually, of which USD 91.1 million are transferred to the local economy. Interviewing people from five different economic sectors, we show shark-diving contribute with USD 2.5 million per year to Noronha’s economy, representing 19% of the island’s GDP. Shark-diving provides USD 128.5 thousand of income to employed islanders, USD 72.6 thousand to government in taxes and USD 5.3 thousand to fishers due to the increase in fish consumption demanded by shark divers. We discover, though, that fishers who actually are still involved in shark fishing earn more by catching sharks than selling other fish for consumption by shark divers. We conclude, however, that the non-consumptive use of sharks is most likely to benefit large number of people by generating and money flow if compared to the shark fishing, providing economic arguments to promote the conservation of these species.

Role of ABL Family Kinases in Breast Cancer

The ABL family of non-receptor tyrosine kinases, ABL1 (also known as c-ABL) and ABL2 (also known as Arg), links diverse extracellular stimuli to signaling pathways that control cell growth, survival, adhesion, migration and invasion. ABL tyrosine kinases play an oncogenic role in human leukemias. However, the role of ABL kinases in solid tumors including breast cancer progression and metastasis is just emerging.

To evaluate whether ABL family kinases are involved in breast cancer development and metastasis, we first analyzed genomic data from large-scale screen of breast cancer patients. We found that ABL kinases are up-regulated in invasive breast cancer patients and high expression of ABL kinases correlates with poor prognosis and early metastasis. Using xenograft mouse models combined with genetic and pharmacological approaches, we demonstrated that ABL kinases are required for regulating breast cancer progression and metastasis to the bone. Using next generation sequencing and bioinformatics analysis, we uncovered a critical role for ABL kinases in promoting multiple oncogenic pathways including TAZ and STAT5 signaling networks and the epithelial to mesenchymal transition (EMT). These findings revealed a role for ABL kinases in regulating breast cancer tumorigenesis and bone metastasis and provide a rationale for targeting breast tumors with ABL-specific inhibitors.

Role of Vinculin in Regulating Force-Sensitive Dynamics of Adherens Junctions During Collective Cell Migration

Dynamic processes such as morphogenesis and tissue patterning require the precise control of many cellular processes, especially cell migration. Historically, these processes are thought to be mediated by genetic and biochemical signaling pathways. However, recent advances have unraveled a previously unappreciated role of mechanical forces in regulating these homeostatic processes in of multicellular systems. In multicellular systems cells adhere to both deformable extracellular matrix (ECM) and other cells, which are sources of applied forces and means of mechanical support. Cells detect and respond to these mechanical signals through a poorly understood process called mechanotransduction, which can have profound effects on processes such as cell migration. These effects are largely mediated by the sub cellular structures that link cells to the ECM, called focal adhesions (FAs), or cells to other cells, termed adherens junctions (AJs).

Overall this thesis is comprised of my work on identifying a novel force dependent function of vinculin, a protein which resides in both FAs and AJs - in dynamic process of collective migration. Using a collective migration assay as a model for collective cell behavior and a fluorescence resonance energy transfer (FRET) based molecular tension sensor for vinculin I demonstrated a spatial gradient of tension across vinculin in the direction of migration. To define this novel force-dependent role of vinculin in collective migration I took advantage of previously established shRNA based vinculin knock down Marin-Darby Canine Kidney (MDCK) epithelial cells.

The first part of my thesis comprises of my work demonstrating the mechanosensitive role of vinculin at AJ’s in collectively migrating cells. Using vinculin knockdown cells and vinculin mutants, which specifically disrupt vinculin’s ability to bind actin (VinI997A) or disrupt its ability to localize to AJs without affecting its localization at FAs (VinY822F), I establish a role of force across vinculin in E-cadherin internalization and clipping. Furthermore by measuring E-cadherin dynamics using fluorescence recovery after bleaching (FRAP) analysis I show that vinculin inhibition affects the turnover of E-cadherin at AJs. Together these data reveal a novel mechanosensitive role of vinculin in E-cadherin internalization and turnover in a migrating cell layer, which is contrary to the previously identified role of vinculin in potentiating E-cadherin junctions in a static monolayer.

For the last part of my thesis I designed a novel tension sensor to probe tension across N-cadherin (NTS). N-cadherin plays a critical role in cardiomyocytes, vascular smooth muscle cells, neurons and neural crest cells. Similar to E-cadherin, N-cadherin is also believed to bear tension and play a role in mechanotransduction pathways. To identify the role of tension across N-cadherin I designed a novel FRET-based molecular tension sensor for N-cadherin. I tested the ability of NTS to sense molecular tension in vascular smooth muscle cells, cardiomyocytes and cancer cells. Finally in collaboration with the Horwitz lab we have been able to show a role of tension across N-cadherin in synaptogenesis of neurons.

The role of inflammatory mediators in the overactive and bladder pain syndromes

The Overactive Bladder (OAB) and Bladder Pain Syndrome (BPS) are debilitating disorders for which the pathophysiological mechanisms are poorly understood. Injury or dysfunction of the protective urothelial barrier layer, specifically the proteoglycan composition and number, has been proposed as the primary pathological characteristic of BPS. For OAB, the myogenic theory with dysfunction of the muscarinic receptors is the most reiterated hypothesis. For both over activity of the inflammatory response has been posited to play a major role in these diseases. We hypothesise that BPS and OAB are peripheral sensory disorders, with an increase in inflammatory mediators, such as cytokines and chemokines, which are capable of activating, either directly or indirectly, sensory nerve activity causing the disease. The aim of the PhD is to identify potential new therapeutic targets for the treatment of BPS and OAB. We used medium throughput quantitative gene expression analysis of 96 inflammation associated mediators to measure gene expression levels in BPS and OAB bladder biopsies and compared them to control samples. Then we created a novel animal model of disease by specific proteoglycan deglycosylation of the bladder mucosal barrier, using the bacterial enzymes Chondroitinase ABC and Heparanase III. These enzymes specifically remove the glycosaminoglycan side chains from the urothelial proteoglycan molecules. We tested role of the identified mediators in this animal model. In addition, in order to determine on which patients peripheral treatment strategies may work, we assessed the effect of local anaesthetics on patients with bladder pain. Gene expression analysis did not reveal a difference in inflammatory genes in the OAB versus control biopsies. However, several genes were upregulated in BPS versus control samples, from which two genes, FGF7 and CLL21 were correlated with patient clinical phenotypes for ICS/PI symptom and problem indices respectively. In order to determine which patients are likely to respond to treatment, we sought to characterise the bladder pain in BPS patients. Using urodynamics and local anaesthetics, we differentiated patients with peripherally mediated pain and patients with central sensitisation of their pain. Finally to determine the role of these mediators in bladder pain, we created an animal model of disease, which specifically replicates the human pathology: namely disruption in the barrier proteoglycan molecules. CCL21 led to an increase in painrelated behaviour, while FGF7 attenuated this behaviour, as measured by cystometry, spinal c-fos expression and mechanical withdrawal threshold examination. In conclusion, we have identified CCL21 and FGF7 as potential targets for the treatment of BPS. Manipulation of these ligands or their receptors may prove to be valuable previously unexploited targets for the treatment of BPS.

The Moderating Role of Personality on Workplace Conflict and Outcomes

The purpose of this study was twofold. The first was to further clarify and expand or understanding of the relationship between interpersonal conflict, incivility, and their roles as stressors in the stressor-strain relationship. The second goal was to examine how neuroticism, extraversion, agreeableness, conscientiousness, trait anger, and sphere specific locus of control moderate the stressor-strain relationship between task conflict, relationship conflict, incivility and workplace and health outcomes. The results suggest that extraversion, neuroticism, conscientiousness, agreeableness, trait anger, and locus of control play significant roles in how workplace aggression affects individuals. These findings suggest that occupations that experience a high level of workplace aggression should consider incorporating these personality traits into their selection system as a way of limiting or reducing the effects workplace aggression can have on individual health, wellbeing, and job outcomes.

Flammability Standards – A Major Role in Flame Retardant Exposure? Televisions as a case study.

Flame retardants (FRs) are added to materials to enhance the fire safety level of readily combustible polymers. Although they have been purported to aid in preventing fires in some cases, they have also become a significant cause for concern given the vast data on environmental persistence and human and animal adverse health effects. Evidence since the 1980s has shown that Canadian, American and Europeans have detectable levels of FRs in their bodies. North Americans in particular have high levels of these chemicals due to stringent flammability standards and the higher use of polybrominated diphenyl ethers (PBDEs) in North America as opposed to Europe. FRs have been detected in household dust and some evidence suggests that TVs could be a significant source of exposure to FRs. It is imperative to re-visit the flammability standard (UL94V) that allows for FR use in TVs plastic materials by providing a risk versus benefit analysis to determine if this standard provides a fire safety benefit and if it plays a major role in FR exposure. This report first examined the history of televisions and the progression to the UL94V flammability test standard to understand why FRs were first added to polymers used in the manufacturing of TVs. It has been demonstrated to be due to fire hazards resulting from the use of plastic materials in cathode-ray tube (CRT) TVs that had an “instant-on” feature and high voltage and operating temperatures. In providing a risk versus benefit analysis, this paper presents the argument that 1) by providing a market survey the current flammability test standard (UL94V) is outdated and lacks relevance to current technology as flat, thin, energy efficient Liquid Crystal Displays (LCDs) dominate over traditionally used heavy, bulky and energy-intensive CRTs; 2) FRs do not impart fire safety benefits considering that there is a lack of valid fire safety concern, such as reduced internal and external ignition and fire hazard, and a lack of valid fire data and hazard for television fires in general and finally; 3) the standard is overly stringent as it does not consider the risk due to exposure to FRs in household dust due to the proliferation and greater use of televisions in households. Therefore, this report argues that the UL94V standard has become trapped in history and needs to be updated as it may play a major role in FR exposure.

Reform and Redesign: A Review of the Changing Role of the Public Purposes in R&D and Technology Institutes (Working Paper 17)

As the globalization of knowledge has taken hold over the past decade, and as governments around the world review their new roles in support of the production of knowledge, several factors have shaped the context in governments’ approach to public research. Arguably, none has been more affected by these pressures for reform than government scientific and technology laboratories or institutes. Often ignored in the re-shaping of national systems of innovation, these organizations play an important role in advancing national economic and social objectives. This paper, by reviewing examples of reforms underway in several countries, including Canada, France, Germany, the UK, Japan, USA and Latin America, will argue that government research and technology institutes — often historically surrogates for industrial research — are gradually re-defining their mandates to meet the new pressures of globalization as well as satisfying growing public demands for increased relevance and efficiency in responding to citizens’ and industry needs.

“It is through others that we become ourselves.” A study of Vygotskian play in Russian and Irish schools.

50 years after publishing his seminal work on play and its role in child development, Vygotskian theory is still highly influential in education, and particularly in early years. This paper presents two examples of full integration of Vygotskian principles into schools in two very different settings. Both report improvements in learning and in well-being, and exemplify the theory-practice-theory cycle, highlighting the development of new theoretical constructs arising out of putting theory firmly into practice. In both settings, the positive results have come from years of effort, in which school personnel who may have been skeptical at first, have been inspired by the impact of adopting Vygotskian play on the children they teach. The Northern Ireland study shows that at least some of the Golden Key principles (mixed-age play and enhanced home-school links) translate perfectly into very different cultural-historical contexts.

Asymmetric hydrogen flame in a heated micro-channel: role of Darrieus-Landau and thermal-diffusive instabilities

Present work examines numerically the asymmetric behavior of hydrogen/air flame in a micro-channel subjected to a non-uniform wall temperature distribution. A high resolution (with cell size of 25 μm × 25 μm) of two-dimensional transient Navier–Stokes simulation is conducted in the low-Mach number formulation using detailed chemistry evolving 9 chemical species and 21 elementary reactions. Firstly, effects of hydrodynamic and diffusive-thermal instabilities are studied by performing the computations for different Lewis numbers. Then, the effects of preferential diffusion of heat and mass transfer on the asymmetric behavior of the hydrogen flame are analyzed for different inlet velocities and equivalence ratios. Results show that for the flames in micro-channels, interactions between thermal diffusion and molecular diffusion play major role in evolution of a symmetric flame into an asymmetric one. Furthermore, the role of Darrieus–Landau instability found to be minor. It is also found that in symmetric flames, the Lewis number decreases behind the flame front. This is related to the curvature of flame which leads to the inclination of thermal and mass fluxes. The mass diffusion vectors point toward the walls and the thermal diffusion vectors point toward the centerline. Asymmetric flame is observed when the length of flame front is about 1.1–1.15 times of the channel width.

The Role of Morphogens in B Cell Development

Morphogens are signalling molecules that play a significant role in modulation of cell fate and development. Hedgehog proteins (Hh) are morphogens that have been shown to be involved in the development of immune cells. In this study, it is demonstrated that treatment of B cells with rShh, can increase B cell activation and also promote survival of B cells at 18hours post-stimulus. Also, at this time point, there was found to be an increase in secretion of antibody isotypes and IL-6. By 40hours post-stimulus, it was observed that the level of B cell activation was apparently arrested in treated B cells, whereas the level of activation continued to rise in untreated B cells. Interestingly, it was observed that there was an increase in the percentage of; CD23-CD25+ B cells when B cells were treated with rShh and this was accompanied by an increase in apoptosis. Consistent with this finding in relation to apoptosis, there was an increased expression of the pro-apoptotic protein Bnip3 in B cells treated with rShh by 40hours post-stimulus. It was observed that there were three subsets of B cells arising in our culture at 40hours, which were all found to possess different characteristics. It was demonstrated that treatment with rShh can increase B cell differentiation towards FO-I at 18hours post-stimulus. By 40hours post-stimulus, Hh signalling can divert differentiation away from the FO-I B cell towards the T2-MZP, which was accompanied by an increase in IL-10 secretion. Gene expression analysis revealed that Hh signalling could modulate a number of molecules involved in delivering the BCR signal into the cells such as Btk, Nfatc1 and Traf2. Additionally, deletion of Dhh, showed that there was a skewed peripheral B cell development in the Dhh-/- mice. Overall, our data demonstrate that Hh signalling can regulate the development of B cells in response to an activation stimulus by strengthening the BCR signalling pathway.

SMART social science? Examining the nature and role of social scientific expertise in institutional design

Daniel Bromley argues against Oran Young’s FIT model as a basis for environmental governance, on the grounds that humans cannot manage nature and that attempts to do so are based on a scientistic, modernist conceit. At issue is the role of natural and social scientists in adjudicating questions about what we ought to do to close governance gaps and address unsustainable behaviors. If Bromley is right, then the lessons of the American pragmatist tradition recommend against attempts to “fit” social institutions to the natural world. The first objective of this paper is to argue that Bromley’s view is not in keeping with the pragmatism of C. S. Peirce and John Dewey, which actually places a high value on natural and social scientific modes of inquiry in the service of social ends. I argue that Young’s proposal is in fact a development of the pragmatist idea that social institutions must be fit in the sense of fitness, i.e., resilient and able to navigate uncertainty. Social institutions must also evolve to accommodate the emerging values of the agents who operate within them. The second objective of this paper is to examine the role of social science expertise in the design of social policies. Governance institutions typically rely on the testimony of natural scientists, at least in part, to understand the natural systems they operate within. However, natural systems are also social systems, so it seems pertinent to ask whether there is a role for social systems experts to play in helping to design environmental governance institutions. I argue that social scientists can make a unique contribution as experts on social institutions, and as such, are necessary to bring about a transformation of the unsustainable institutions that are preventing us from achieving stated sustainable development goals.

The Role of Art in Computer Game Design

Computer games are significant since they embody our youngsters’ engagement with contemporary culture, including both play and education. These games rely heavily on visuals, systems of sign and expression based on concepts and principles of Art and Architecture. We are researching a new genre of computer games, ‘Educational Immersive Environments’ (EIEs) to provide educational materials suitable for the school classroom. Close collaboration with subject teachers is necessary, but we feel a specific need to engage with the practicing artist, the art theoretician and historian. Our EIEs are loaded with multimedia (but especially visual) signs which act to direct the learner and provide the ‘game-play’ experience forming semiotic systems. We suggest the hypothesis that computer games are a space of deconstruction and reconstruction (DeRe): When players enter the game their physical world and their culture is torn apart; they move in a semiotic system which serves to reconstruct an alternate reality where disbelief is suspended. The semiotic system draws heavily on visuals which direct the players’ interactions and produce motivating gameplay. These can establish a reconstructed culture and emerging game narrative. We have recently tested our hypothesis and have used this in developing design principles for computer game designers. Yet there are outstanding issues concerning the nature of the visuals used in computer games, and so questions for contemporary artists. Currently, the computer game industry employs artists in a ‘classical’ role in production of concept sketches, storyboards and 3D content. But this is based on a specification from the client which restricts the artist in intellectual freedom. Our DeRe hypothesis places the artist at the generative centre, to inform the game designer how art may inform our DeRe semiotic spaces. This must of course begin with the artists’ understanding of DeRe in this time when our ‘identities are becoming increasingly fractured, networked, virtualized and distributed’ We hope to persuade artists to engage with the medium of computer game technology to explore these issues. In particular, we pose several questions to the artist: (i) How can particular ‘periods’ in art history be used to inform the design of computer games? (ii) How can specific artistic elements or devices be used to design ‘signs’ to guide the player through the game? (iii) How can visual material be integrated with other semiotic strata such as text and audio?

The economic crisis as a game changer? Exploring the role of social construction in sustainability transitions

Continuing economic turbulence has fuelled debates about social and political reform as much as it has stimulated actions and initiatives aimed at a more fundamental transition of dominant economic systems. This paper takes a transition perspective to explore, from a Western European viewpoint, how the economic crisis is actually viewed through a variety of interpretations and responded to through a range of practices. We argue that framing societal phenomena such as the economic crisis as "symptoms of transition" through alternative narratives and actions can give rise to the potential for (seemingly) short-term pressures to become game changers. Game changers are then defined as the combination of: specific events, the subsequent or parallel framing of events in systemic terms by engaged societal actors, and (eventually) the emergence of (diverse) alternative narratives and practices (in response to the systemic framing of events). Game changers, when understood in these terms, help to orient, legitimize, guide, and accelerate deep changes in society. We conclude that such dynamics in which game changers gain momentum might also come to play a critical role in transitions. Therefore, we argue for developing a better understanding of and methodologies to further study the coevolutionary dynamics associated with game changers, as well as exploring the implications for governance.

Renal Drug Transporters – Clinical Significance and Role in Drug-Drug Interactions

Thesis (Ph.D.)--University of Washington, 2016-07