Indiana University-Purdue University Indianapolis (IUPUI) / Drug addiction is a behavioral disorder characterized by impulsive behavior and continued intake of drug in the face of adverse consequences. Millions of people suffer the financial and social consequences of addiction, and yet many of the current therapies for addiction treatment have limited efficacy. Therefore, there is a critical need to characterize the neurobiological substrates of addiction in order to formulate better treatment options. In the first chapter, the striatal dopamine system is interrogated with [11C]raclopride PET to assess differences between chronic cannabis users and healthy controls. The results of this chapter indicate that chronic cannabis use is not associated with a reduction in striatal D2/D3 receptor availability, unlike many other drugs of abuse. Additionally, recent cannabis consumption in chronic users was negatively correlated with D2/D3 receptor availability. Chapter 2 describes a retrospective analysis in which striatal D2/D3 receptor availability is compared between three groups of alcohol-drinking and tobacco-smoking subjects: nontreatment-seeking alcoholic smokers, social-drinking smokers, and social-drinking non-smokers. Results showed that smokers had reduced D2/D3 receptor availability throughout the striatum, independent of drinking status. The results of the first two chapters suggest that some combustion product of marijuana and tobacco smoke may have an effect on striatal dopamine concentration. Furthermore, they serve to highlight the effectiveness of using baseline PET imaging to characterize dopamine dysfunction in addictions. The final chapter explores the use of [18F]fallypride PET in a proof-of-concept study to determine whether changes in cortical dopamine can be detected during a response inhibition task. We were able to detect several cortical regions of significant dopamine changes in response to the task, and the amount of change in three regions was significantly associated with task performance. Overall, the results of Chapter 3 validate the use of [18F]fallypride PET to detect cortical dopamine changes during a impulse control task. In summary, the results reported in the current document demonstrate the effectiveness of PET imaging as a tool for probing resting and activated dopamine systems in addiction. Future studies will expand on these results, and incorporate additional methods to further elucidate the neurobiology of addiction.

Indiana University-Purdue University Indianapolis (IUPUI) / Signal transducer and activator of transcription 3 (STAT3) is constitutively activated in malignant tumors, and its activation is associated with high histological grade and advanced cancer stage. STAT3 has been shown to play important roles in multiple aspects of cancer aggressiveness including proliferation, survival, self-renewal, migration, invasion, angiogenesis and immune response by regulating the expression of diverse downstream target genes. Thus, inhibiting STAT3 promises to be an attractive strategy for treatment of advanced tumors with metastatic potential. We firstly identified a STAT3 inhibitor, inS3-54, by targeting the DNA-binding site of STAT3 using an in-silico screening approach; however, inS3-54 was finally found not to be appropriate for further studies because of low specificity on STAT3 and poor absorption in mice. To develop an effective and specific STAT3 inhibitor, we identified 89 analogues for the structure-activity relationship analysis. By using hematopoietic progenitor cells isolated from wild-type and STAT3 conditional knockout mice, further studies showed that three analogues (A18, A26 and A69) only inhibited STAT3-dependent colony formation of hematopoietic progenitor cells, indicating a higher selectivity for STAT3 than their parental compound, inS3-54. These compounds were found to (1) inhibit STAT3-specific DNA binding activity; (2) bind to STAT3 protein; (3) suppress proliferation of cancer cells harboring aberrant STAT3 signaling; (4) inhibit migration and invasion of cancer cells and (5) inhibit STAT3-dependent expression of downstream targets by blocking the binding of STAT3 to the promoter regions of responsive genes in cells. In addition, A18 can reduce tumor growth in a mouse xenograft model of lung cancer with little effect on body weight. Taken together, we conclude that it is feasible to inhibit STAT3 by targeting its DNA-binding domain for discovery of anticancer therapeutics.

Indiana University-Purdue University Indianapolis (IUPUI) / Sequestration at the aggresome and degradation through autophagy are two approaches by which a cell can counteract the toxic effect of misfolded proteins. Tuberous sclerosis (TS) and cancer cells can become dependent on autophagy for survival due to the high demand for protein synthesis, thus making protein clearance a potential therapeutic target. Because of its histone deacetylase (HDAC) inhibitory activity, we hypothesized that 4-phenylbutyrate (4-PBA) inhibits HDAC6 and aggresome formation to induce TS cell death. We found that 4-PBA treatment increases cell death and reduces bortezomib-induced aggresome formation. To link these results with HDAC inhibition we used two other HDAC inhibitors, trichostatin A (TSA) and tubastatin, and found that they also reduce bortezomib-induced protein aggregation. Because tubulin is a target of HDAC6, we next measured the effect of the HDAC inhibitors and 4-PBA treatment on tubulin acetylation. As expected, tubastatin increased tubulin acetylation but surprisingly TSA and 4-PBA did not. Because 4-PBA did not significantly inhibit HDAC6, we next hypothesized that 4-PBA was alternatively inducing autophagy and increasing aggresome clearance. Surprisingly, autophagy inhibition did not prevent the 4-PBA-induced reduction in protein aggregation. In conclusion, we found 4-PBA to induce cell death and reduce aggresome levels in TS cells, but we found no link between these phenomena. We next hypothesized that loss of the Ral guanine nucleotide exchange factor Rgl2 induces cell death via autophagy inhibition in pancreatic adenocarcinoma (PDAC) cells. KRas is mutationally activated in over 90% of PDACs and directly activates Rgl2. Rgl2 activates RalB, a known regulator of autophagy, and Rgl2 has been shown to promote PDAC cell survival. We first confirmed that loss of Rgl2 does increase cell death in PDAC cells. Initial experiments using doubly tagged fluorescent p62 and LC3 (autophagy markers) suggested that loss of Rgl2 inhibited autophagosome accumulation, but after developing a more sophisticated quantitation method we found loss of Rgl2 to have no effect. We also measured endogenous LC3 levels, and these experiments confirmed loss of Rgl2 to have no effect on autophagy levels. Therefore, loss of Rgl2 increases cell death in PDAC cells, but does not have a significant effect on autophagy.

Indiana University-Purdue University Indianapolis (IUPUI) / Aromatase inhibitors (AIs) are the current standard of care for the treatment of hormone receptor positive breast cancer in postmenopausal women. Nearly one-half of patients receiving AI therapy develop musculoskeletal toxicity that is characterized by joint and/or muscle pain and approximately one-fourth of patients discontinue their therapy as a result of musculoskeletal pain. Since there are no effective strategies for prevention or treatment, insight into the mechanisms of AI-induced pain is critical to improve treatment. However, there are few studies of AI effects in animal models of nociception. To determine whether AIs produce hypersensitivity in animal models of pain, I examined the effects of AI administration on mechanical, thermal, and chemical sensitivity in rats. The results demonstrate that (1) repeated injection of 5 mg/kg letrozole in male rats produces mechanical, but not thermal, hypersensitivity that extinguishes when drug dosing is stopped; (2) administering a single dose of 1 or 5 mg/kg letrozole in ovariectomized (OVX) rats also induces mechanical hypersensitivity, without altering thermal sensitivity and (3) a single dose of 5 mg/kg letrozole or daily dosing of letrozole or exemestane in male rats augments flinching behavior induced by intraplantar ATP injection. To determine whether the effects of AIs on nociceptive behaviors are mediated by activation or sensitization of peptidergic sensory neurons, I determined whether letrozole exposure alters release of calcitonin gene-related peptide (CGRP) from isolated rat sensory neurons and from sensory nerve endings in rat spinal cord slices. No changes in basal, capsaicin-evoked or high extracellular potassium-evoked CGRP release were observed in sensory neuronal cultures acutely or chronically exposed to letrozole. Furthermore, letrozole exposure did not alter the ability of ATP to augment CGRP release from sensory neurons in culture. Finally, chronic letrozole treatment did not augment neuropeptide release from spinal cord slices. Taken together, these results do not support altered release of this neuropeptide into the spinal cord as mediator of letrozole-induced mechanical hypersensitivity and suggest the involvement of other mechanisms. Results from this dissertation provide a new experimental model for AI-induced hypersensitivity that could be beneficial in delineating mechanisms mediating pain during AI therapy.

Indiana University-Purdue University Indianapolis (IUPUI) / In recent decades, Petri nets (PNs) have been used to model traffic networks for different purposes, such as signal phase control, routing, and traffic flow estimation, etc. Because of the complex nature of traffic networks where both discrete and continuous dynamics come into play, the Hybrid Petri net (HPN) model becomes an important tool for the modeling and analysis of traffic networks. In Chapter 1 a brief historical summery about traffic systems control and then related work is mentioned followed by the major contributions in this research. Chapter 2 provides a theoretical background on Petri nets. In Chapter 3, we develop a HPN model for a single signalized intersection first, then we extend this model to study a simple traffic network that consists of two successive intersections. Time delays between different points of network are also considered in order to make the model suitable for analysis and simulation. In addition to HPN models, we also consider discrete Petri nets where their modeling simplicity enables the characterization of the occurrences of all events in the system. This discrete PN is particularly useful to give a higher-level representation of the traffic network and study its event occurrences and correlations. In Chapter 4, we build a discrete PN model to represent a traffic network with two successive intersections. However, we find that the model leads to unbounded places which cannot accurately reflect the dynamics of the traffic in terms of event occurrences. Hence, we introduce the Modified Binary Petri nets (MBPN) to overcome the limitation and resolve the confliction problem when we design our controllers. This MBPN model is a powerful tool and can be useful for the modeling and analysis of many other applications in traffic networks. Chapter 5 gives a summary for each chapter, provides conclusion and discusses future work for both discrete and hybrid Petri nets.

Indiana University-Purdue University Indianapolis (IUPUI) / Intrinsically disordered proteins are proteins characterized by lack of stable tertiary structures under physiological conditions. Evidence shows that disordered proteins are not only highly involved in protein interactions, but also have the capability to associate with more than one partner. Short disordered protein fragments, called “molecular recognition features” (MoRFs), were hypothesized to facilitate the binding diversity of highly-connected proteins termed “hubs”. MoRFs often couple folding with binding while forming interaction complexes. Two protein disorder mechanisms were proposed to facilitate multiple partner binding and enable hub proteins to bind to multiple partners: 1. One region of disorder could bind to many different partners (one-to-many binding), so the hub protein itself uses disorder for multiple partner binding; and 2. Many different regions of disorder could bind to a single partner (many-to-one binding), so the hub protein is structured but binds to many disordered partners via interaction with disorder. Thousands of MoRF-partner protein complexes were collected from Protein Data Bank in this study, including 321 one-to-many binding examples and 514 many-to-one binding examples. The conformational flexibility of MoRFs was observed at atomic resolution to help the MoRFs to adapt themselves to various binding surfaces of partners or to enable different MoRFs with non-identical sequences to associate with one specific binding pocket. Strikingly, in one-to-many binding, post-translational modification, alternative splicing and partner topology were revealed to play key roles for partner selection of these fuzzy complexes. On the other hand, three distinct binding profiles were identified in the collected many-to-one dataset: similar, intersecting and independent. For the similar binding profile, the distinct MoRFs interact with almost identical binding sites on the same partner. The MoRFs can also interact with a partially the same but partially different binding site, giving the intersecting binding profile. Finally, the MoRFs can interact with completely different binding sites, thus giving the independent binding profile. In conclusion, we suggest that protein disorder with post-translational modifications and alternative splicing are all working together to rewire the protein interaction networks.

Indiana University-Purdue University Indianapolis (IUPUI) / Nurse-physician collaboration has received much attention over the past decade in the USA. The release of three reports from the Institute of Medicine implicated poor communication and collaboration among nurses and physicians as a major contributing factor to the incidence of sentinel events and medical errors. Despite the growing awareness of the imperative related to collaboration between nurses and physicians to ensure patient safety, the problem of poor nurse-physician collaboration remains endemic throughout the country. Indian nurses, along with many other internationally educated nurses, comprise 12-15.2% of the nursing workforce in the USA. Little is known about how Indian nurses culture potentially influences their ability to effectively collaborate with physicians to ensure patient safety. The purpose of this study is to understand Indian nurses’ attitudes and perceptions about nurse-physician collaboration. Hermeneutic interpretive phenomenology as influenced by the work of Martin Heidegger guided this study through the use of interviews via Skype. The overall experience of the Indian nurses was of one experiencing a dramatic positive change in nurse-physician collaboration in the USA as compared to India. Four themes emerged describing this phenomenon: Respect/feeling heard, Being Trusted, Assurance of Accountability, and Finding Freedom. Indian nurses practicing in the USA find a freedom that empowers them to collaborate with physicians for patient safety. They, as all nurses may, benefit from continuing educational opportunities that demonstrate ways to collaborate more fully.

Indiana University-Purdue University Indianapolis (IUPUI) / In 2008, grassroots NGOs formed 13 alliances in response to the need for emergency relief and post-disaster recovery after the Sichuan Earthquake that occurred in West China and killed approximately 87,000 people. These alliances served to raise and deliver relief materials, train and supervise volunteers, promote information sharing, and assist victims with mental health and livelihood recovery. However, all alliances were terminated within less than four years. Although plenty of scholarship discusses how corporate alliances evolve or fail, few studies focus on interorganizational collaboration among nonprofits. To explore how NGOs developed collective actions in China’s adverse sociopolitical environment, the author performed three years of observation in four coalitions and interviewed 60 alliance leaders, employees, and volunteers. This paper identifies four types of termination these NGO alliances experienced: three of them failed at their very births, five self-disbanded shortly after the end of emergency aid, three dissolved due to failed institutionalization, and the remaining two evolved into independent organizations. Tracking their life cycles, this study finds four main factors accountable for their terminations: political pressure, funding shortage, short-term orientation, and leadership failure. In particular, the repressive NGO regulation regime and limited funding sources fundamentally restricted all alliances’ capacity and sustainability. Further, the transient nature of disaster relief efforts and the conflict between disaster management and planned work areas contributed to the short-term orientation among alliance members and, thus, led to the closure of some alliances shortly after they provided emergency relief. In addition, though generally exempt from internal rivalry that often undermines inter-firm partnerships, NGO alliances of all types were confronted with leadership challenges—partner misfits concerning resources, strategy, and mission; flawed governing structures, and undesired individual leadership. The four factors interplayed and led to alliance dissolution through different combinations. The paper points out that, in addition to environmental uncertainty, leadership failure has become a major challenge for nonprofit collaborations.

Indiana University-Purdue University Indianapolis (IUPUI) / This study examined the roles of commitment and attributions in uninvolved partner responses to imagined sexual infidelity. Undergraduate students (N = 298) in dating relationships participated in a hypothetical sexual infidelity scenario in which they imagined their romantic partner engaged in sexual intercourse with someone else. Measured-variable path analysis was used to evaluate the predictive ability of commitment and attributions on negative emotional responses and predicted relationship continuation. The hypothesized conceptual model demonstrated poor fit to sample data. Through exploratory model building, an alternative model was generated that demonstrated good fit to sample data. A subset of commitment, investment, predicted negative affect. In addition, attributions predicted predictions of relationship continuation. Negative emotional responses were highly endorsed on a validated measure for emotional responses, the PANAS-X (Watson & Clark, 1994). Further, study findings highlight the importance of the use of a compliance check in assessing successful participant completion of imagined infidelity scenario. Unique study contributions include directions for further conceptual model development for this area of research as well as support for the use of compliance checks and careful selection of infidelity scenario.

Indiana University-Purdue University Indianapolis (IUPUI) / Madeline Heilman’s (1983) Lack of Fit Model, which postulates why discrimination occurs in the selection of sex-typed jobs, has been applied to the interaction of applicant attractiveness. Yet recent research suggests that other appearance variables, namely sex-typed facial features, may be associated with perceptions of fit. Building upon Heilman’s 1983 model, the current study evaluated how sex-typed facial features relate to applicant selection for sex-typed fields. Undergraduate students were recruited for participation during the spring academic semester (n = 413) and data were analyzed using a 2x2x2 ANOVA. Results indicated that selection is significantly impacted by the three-way interaction of applicant sex, facial feature-type, and sex type of the applying field. Further, masculine-featured females and feminine-featured males were significantly less favored for selection within the feminine sex-typed field. Implications of these findings and the differential evaluation of male and female applicants in a feminine field are discussed.