- IUPUI Research Day 2016
ItemSelective Plane Illumination Microscopy, A New Imaging Modality Available at the Indiana Center for Biological Microscopy(Office of the Vice Chancellor for Research, 2016-04-08) Winfree, Seth; Smith, Nathaniel; Dunn, Ken; Kamocka, Malgorzata; Molitoris, BruceMicroscopy is a primary tool for studying 3D tissue models. Microscopy provides the only means of distinguishing the behaviors of individual cells in a heterogeneous context that obscures biochemical assays. SPIM (Selective Plane Illumination Microscopy) is new approach that is ideally suited to the unique problems involved in high-resolution imaging of 3D tissue models. In the simplest form of SPIM, a cylindrical lens is used to generate a thin lightsheet (1-10 microns) that illuminates a sample. An imaging objective lens, placed orthogonal to this lightsheet is used to collect an image of fluorescence that is selectively excited in this single illuminated plane. The sample is then rotated, and the process is repeated until a multiview dataset of the entire sample is collected. These cross-section images are then assembled to give a complete 3D image of the sample. This approach offers several advantages over conventional methods of imaging thick tissues. First, SPIM provides superior axial resolution for large field-of-view images, deconvolved SPIM volumes have isotropic 3D resolution. Second, SPIM is a “gentle” imaging approach and is better suited to imaging living tissues than either confocal or multiphoton microscopy, supporting studies of cell migration, development, signaling and physiology. Third, imaging speeds can be 30 to 200 fold faster than scanning confocal or multiphoton systems, enabling resolution of dynamic events, and rapid collection of large image datasets. We describe the assembly and customization of an OpenSPIM based lightsheet microscope (IU OpenSPIM) as a platform for developing new imaging technologies. To this end we have implemented software and hardware for multi-channel laser control and temperature and perfusion control. We present examples of highresolution, live and high speed imaging, demonstrating these capabilities. The IU OpenSPIM is a centerpiece in the development of new software for 3D tissue cytometry and a novel screening platform. ItemA Characterization of Different Spark Regimes for Ignition Delay Comparison with Conventional Spark Plugs(Office of the Vice Chancellor for Research, 2016-04-08) Wozniak, Zachary M.; Burton, Jesse C.; Hedrick, Cameron J.; Deng, Qiuyu; Robinson, Daniel W.The introduction of plasma into combustion and ignition processes has continuously proved to be advantageous when compared to the conventional spark ignition in a wide range of categories. From the capability to ignite leaner mixtures and improve fuel economy to an effective reduction of hazardous emissions and ignition delay, the benefits of integrating non-equilibrium plasma can be utilized for numerous applications including hot jet ignition. Detailed design specifications for the electrode configuration, circuit schematic, and combustion rig are developed and presented. Using a CCD camera and high performance oscilloscope, this paper aims to identify, characterize, and compare the different effects of frequency and pulse width of a driver circuit on the plasma sparks quantitatively in terms of the current, voltage, and energy attributes. Four different plasma regimes are investigated with frequencies ranging from 5.44 Hz to 95.46 kHz and pulse energies ranging from 168 μJ to 14.42 J. The maximum voltage and current characteristics of the plasmas indicate a glow discharge referencing previous experiments. Future work is laid out for a comparison of the ignition progression between a non-thermal plasma system and a traditional spark with using Schlieren imaging. ItemPersonal Sentiment and Marketing of Electronic Cigarettes Among Twitter Users(Office of the Vice Chancellor for Research, 2016-04-08) Lomax, Victoria; Wendling, Brooke; Wright, EmilieThe use of electronic cigarettes (e-cigarettes) is growing in the United States and there is increasing controversial dialogue surrounding e-cigarettes on social media, including Twitter. With the recent spike in popularity, we conducted a systematic review of the literature to: a.) examine what Twitter users are exposed to regarding e-cigarettes, and b) identify potential ramifications for this exposure. Using predesignated inclusion and exclusion criteria, relevant articles were located using PubMed, EMBASE, EBSCOhost, and CINAHL Complete and reviewing reference lists of relevant articles. Full text, English language, peer-reviewed articles relevant to e-cigarette dialogue on Twitter were reviewed. Of the twelve studies, seven met the inclusion criteria. From our analysis of the content, two key themes were found: marketing (predominant theme) and positive personal sentiment regarding e-cigarette use. Also, within our review, common ramifications for increased marketing and positive sentiment were identified. First, the rise in marketing reaching vulnerable populations, specifically adolescents and young adults, may contribute to the growing use of e-cigarettes and influence positive perceptions of these smoking behaviors. Second, there is controversial information shared regarding the health effects of e-cigarette use. This is an emerging topic and there is relatively scant literature available related to e-cigarette dialogue on Twitter. As a result of our review, we recommend Twitter as a platform for methodically analyzing social media trends and informing health care providers of current issues regarding e-cigarettes. Although more research on the health risks of e-cigarettes is required, there is the need for the current health information on e-cigarettes to be disseminated through Twitter. Health care providers also need to discuss e-cigarette use with patients in the clinical settings. Continued surveillance of e-cigarette use and marketing, as well as examination of the necessity for marketing regulations are important as e-cigarette use becomes more prevalent. ItemUnderstanding Microbubble Coalescence Using High-Speed Imaging and Lattice Boltzmann Method Simulation(Office of the Vice Chancellor for Research, 2016-04-08) Zhou, Shuyi; Cao, Yuanzhi; Chen, Rou; Chen, Chuanyi; Yu, Huidan (Whitney); Zhu, Likun; Sun, TaoMicrobubble coalescence is one of the important research areas of bubble dynamics. The purpose of this research is to seek deeper understanding and relative mathematical relation on microbubble coalescence. To fulfill that, we conducted both experiments and simulations. For the part of experiment, we fabricated a microfluidic gas generator with better performance leading corresponding fluidic chemical reaction. After that we utilized ultrafast synchrotron X-ray imaging facility at the Advanced Photon Source of Argonne National Laboratory to capture the gas generating and microbubble merging phenomena using high speed imaging. These experiments show how the microbubbles with the same ratio contact and merge in the reaction channel and different concentration of reactants. As for the part of simulation, we lead the simulation basing on lattice Boltzmann method to simulate microbubble coalescence in water with unequal diameter ratio. Focuses are on the effects of size inequality of parent bubbles on the coalescence geometry and time. The “coalescence preference” of coalesced bubble closer to the larger parent bubble is well captured. A power-law relation between the preferential relative distance and size inequality is consistent to the recent experimental observations. Meanwhile, the coalescence time also exhibits power-law scaling, indicating that unequal bubbles coalesce faster than equal bubbles. ItemThe translational regulator dFMRP interacts with epidermal growth factor receptor to regulate apoptosis in Drosophila(Office of the Vice Chancellor for Research, 2016-04-08) Zic, Jessica Z.; Sherwood, Jacqueline E.; Tessier, Charles R.Posttranscriptional gene regulation is required for all aspects of cellular and tissue development and is a major mechanism underlying many diseases ranging from neurological disorders to cancer. The translational repressor fragile x mental retardation protein (FMRP) is ubiquitously expressed throughout development but is silenced in Fragile X Syndrome, an autism spectrum disorder. Interestingly, high levels of FMRP have recently been identified in human metastatic breast cancer. FMRP overexpression in these patients is directly correlated with increased lung metastasis suggesting a direct role for translational regulation both in cell proliferation and in invasive cell migration. Interestingly, however, FMRP can promote both proliferation and apoptosis. To dissect FMRP’s role in cancer development and progression, we are exploiting the powerful genetic system of Drosophila. Drosophila is an excellent model organism for human diseases associated with FMRP due to the strong evolutionary conservation of the fragile x mental retardation gene 1 which encodes this protein. dFMRP was overexpressed in the Drosophila imaginal wing disc, an epithelial tissue model. Contrary to a role in proliferation, overexpression of dFMRP leads to obvious cell loss in the adult wing and an increase in apoptotic markers. Using a combinatorial genetic screen, we have identified genes which are able to suppress this apoptotic phenotype and thus may be important for FMRP-‐dependent tumorigenesis. Our focus is now on the epidermal growth factor receptor (EGFR) signaling pathway since blocking this mechanism is able to completely rescue the dFMRP-‐overexpression wing defects. Clonal analysis reveals that dFMRP overexpressing cells survive their dFMRP-induced apoptotic programming when co-‐expressing a dominant negative form of EGFR. Additional clonal analyses are being used to explore the potential significance of this survival on tumor formation and metastasis. Item(Dis)Agreement in Parent-Child Perceptions of Injustice and Their Relationship to Pain Outcomes(Office of the Vice Chancellor for Research, 2016-04-08) Wuest, David G.; Miller, Megan M.; Scott, Eric. L.; Trost, Zina; Hirsh, Adam T.Perceiving one’s pain as unjust and thinking about pain in a catastrophic manner are linked to worse outcomes in children with chronic pain. Dyads where the child catastrophized more than the parent experienced particularly poor outcomes in previous research. We investigated the concordance between parent and child injustice perceptions and its relationship to pain outcomes. 139 patients (age=15.4±2.1; 71.9% female) attending the pain clinic at Riley Children’s Hospital completed measures of perceived injustice, pain, and QOL. Parents completed a measure of perceived injustice about their child’s pain. Parent-child dyads were categorized into one of four groups based on concordance of injustice perceptions: (1) concordant high, (2) concordant low, (3) discordant high parent (P) – low child (C), and (4) discordant low P – high C. Parent injustice perceptions were significantly higher than child perceptions (t(138)=5.80, p<.001, d=.50). ANOVAs identified significant group differences for pain intensity (F(3,138)=2.80, p<.05, η2=.06) and QOL (F(3,138)=15.11, p<.01, η2=.25). For pain intensity, discordant low P – high C dyads reported the highest pain, and significantly higher pain than discordant high P – low C dyads (mean difference [MD]=1.94, p<.05). Concordant high dyads reported the second highest pain. A similar pattern emerged for QOL. Discordant low P – high C dyads reported the worst QOL, and significantly worse QOL than concordant high dyads (MD=-10.22, p<.01), concordant low dyads (MD=-23.70, p<.01), and discordant high P – low C dyads (MD=-28.97, p<.01). Concordant high dyads reported the second worse QOL. Overall, dyads where the child endorsed high injustice perceptions, regardless of parental perceptions, experienced worse pain and QOL, with the worst outcomes observed for discordant dyads (low P – high C). Children in low P – high C dyads may feel invalidated and, thus, use maladaptive strategies in an attempt to communicate the severity of their pain. Research is needed to identify the mechanisms underlying these relationships. ItemInternational Factors Impacting Corporate Social Responsibility at STEM Organizations(Office of the Vice Chancellor for Research, 2016-04-08) Whinery, Tiffany E.Corporate Social Responsibility (CSR) has gained significant attention over the past decade and continues to grow as a rising global effort in STEM organizations across multiple industries. Many countries have a longstanding history of practicing CSR and have adopted or adapted the Western model of corporate volunteerism. However, little research has been conducted to determine cultural factors that impact this practice such as volunteer motivation, satisfaction, and project types. This study explores the CSR efforts of, Eli Lilly and Company (Lilly), whose annual day of service provides employees across the globe the opportunity to serve various organizations and their communities through diverse projects. This research aims to determine the influence of culture dimensions and societal norms on multiple aspects of Lilly’s volunteer efforts such as the acceptance of the practice and perspectives of volunteers. ItemImproving bone properties and fracture susceptibility: experimental models of genetic manipulation, pharmacologic intervention, and cellular perturbation reveal new approaches for improving bone health(Office of the Vice Chancellor for Research, 2016-04-08) Witcher, Phillip C.; Lee, Joowon; Assaf, Noor; Mertz, Susan; Singh, Karan; Thompson, William R.; Robling, Alexander G.Bone, a crucial support structure in the human body, is often taken for granted for its lightweight properties and unparalleled strength. Skeletal fracture is a major clinical condition affecting millions of Americans, which results from abnormal aging, hormonal imbalance, genetic conditions, and lifestyle choices (e.g., exercise). Because fractures are caused by a number of different factors, reducing fracture incidence requires a multifactorial approach to unraveling the underlying biology of bone metabolism, in order to discover new ways to improve bone properties and prevent fractures. We have taken such an approach by conducting (1) genetic manipulation experiments in mice, where genes predicted to be involved in bone mass regulation were mutated; (2) pharmacologic experiments to quantify the dose-response effect of an agent that inhibits bone loss, and (3) cell culture experiments, aimed at revealing molecular pathways activated by mechanical stimulation. METHODS: Mice with mutations in two genes, likely to regulate bone mass (SOST, DKK1) were generated and subjected to in vivo dual energy x-ray absorptiometry (DEXA) scans at 6-wk old. Whole body scans were analyzed for bone mineral density (BMD) using Lunar Piximus II v2.10 software. Mice (6-wk) were also dosed (0, 1, 10, 100, or 1000 mg/kg) with daily alendronate HCl, a bisphosphonate that inhibits osteoclast activity. Six wks later, the mice were sacrificed, and the femurs were dissected and sectioned for histological analysis of bone formation parameters, including mineralizing surface (MS/BS), mineral apposition rate (MAR), and bone formation rate (BFR/BS). To understand the cellular signaling events in response to mechanical loading, bone marrow mesenchymal stem cells (MSCs) were treated with 10, 20, 30, or 40μM PF7408671, an S6 kinase inhibitor. Cells then were subject to 100 cycles of biaxial mechanical strain (2%, 10 cycles/min). Protein lysates were separated by electrophoresis and probed for phosphorylation of Rictor and Akt by Western blot. RESULTS: Mice harboring mutations in either the SOST gene or the DKK1gene exhibited significantly increased BMD compared to wild-type control mice, though the SOST mutation had a stronger effect on BMD than DKK1. Mice with compound mutations (SOST and DKK1 mutations) had significantly greater BMD than mice with either single mutation, suggesting that inhibition of SOST and DKK1 might be an effective means to increase bone mass in patients susceptible to fracture. Mice treated with high-dose alendronate (100 or 1,000 mg/kg) exhibited significant decreases in bone formation parameters (MS/BS, MAR, and BFR/BS) compared to untreated (0 mg) mice, suggesting that while this compound might be beneficial for inhibiting bone loss, it also inhibits bone formation. The signaling hub, mTORC2, is a critical regulator of mechanical force in MSC progenitors. Our data demonstrate that S6 kinase is an upstream activator of mTORC2 in response to mechanical strain. CONCLUSION: Our experiments suggest that genetic manipulation of mice reveal viable protein targets (e.g., SOST, DKK1) that could ultimately be manipulated pharmacologically to improve bone mass. We also found that an FDA-approved class of drugs inhibits bone formation even at very low doses, suggesting that additional pro-anabolic compounds might benefit patients taking bisphosphonates. On a cell signaling level, we found that the mTORC2 pathway shows considerable promise for pharmacologic manipulation to simulate the effects of exercise. Taken together, these experiments highlight the utility of a broad approach to solving bone metabolism challenges that can affect fracture susceptibility. ItemIdentifying Metabolic Pathways Producing Alkamides in Echinacea purpurea(Office of the Vice Chancellor for Research, 2016-04-08) Williams, Jermell; Teitgen, Alicen; Minto, RobertEchinacea purpurea is a widely used herbal supplement that is frequently taken to relieve cold symptoms. Alkamides are a secondary metabolite found throughout the Echinacea genus that contain fatty acid chains incorporated into amides and are believed to be the bioactive agent in Echinacea. Our goal is to identify and understand the specific metabolic processes by which E. purpurea produces alkamides. In our experiment, Echinacea seedlings were grown to where the first true leaf emerged and unfurled which is when alkamide production is known to be most active. Alkamides were then extracted and taken to the GC/MS and LC/MS for analysis. Extracted alkamides were analyzed by triplequadrupole chromatography to investigate 13C labeling by glucose. Solid phase extractions were also performed to better observe fragmentation patterns. Fatty acids were also extracted to determine if fatty acids and alkamides were affected the same way by light or the lack of light, which would indicate that they are being synthesized in the same place. It was determined that neither compound experienced a synthesis decrease in the dark significant enough to support a model where acyl chains are newly created in the chloroplasts. Therefore alkamides are more likely to be made in the mitochondria. We are currently in the process of examining the spectra in order to determine the structures of the alkamides as well as any metabolic relationships. ItemThe NeoWarm biomedical device: Assessment of feasibility and cultural acceptability, identification of potential barriers and challenges, and stakeholder mapping(Office of the Vice Chancellor for Research, 2016-04-08) Watts, Thomasina; Siddiki, Furhan; Savita, AakashIntroduction: Across the globe, approximately 4 million newborns die each year; complications from hypothermia underlie many of these deaths. Regions with fewer resources for neonatal care have higher rates of hypothermiarelated death. Kangaroo Mother Care (KMC) is the practice of prolonged skintoskin contact to prevent hypothermia among small and premature infants. KMC is cost effective, and proven to reduce hypothermia; however, KMC programs are often discontinued or fail to expand. A built prototype of a biomedical device, called NeoWarm, has been developed to augment KMC initiatives. Identification of potential barriers and facilitators to adoption the NeoWarm technology is urgently needed. Methods: In order to assess the feasibility of NeoWarm, and to identify current barriers to implementation of KMC and NeoWarm, a comprehensive literature review was conducted. Key barriers and facilitators to existing KMC programs in subSaharan Africa, Asia, and Latin America were identified. Stakeholder mapping and analysis in relation to the NeoWarm device for three “target countries” within each of these global regions was performed. Potential stakeholders were identified and categorically ranked in terms of influence and relevance. Results: Three key barriers to KMC programs were identified. These included: unacceptability among male stakeholders; lack of support from health care providers and insufficient health infrastructure, leading to fears of tuberculosis and other infections spreading in crowded KMC wards. Comprehensive stakeholder mapping for Kenya, India, and Guatemala revealed a complex web of potential influencers and regulatory processes for adoption of NeoWarm technology. Conclusion: The NeoWarm device may support increased acceptance of KMC among male stakeholders and some health care providers; however, the concerns regarding spread of tuberculosis among KMC motherbaby pairs was an unexpected finding, which will significantly inform subsequent NeoWarm development and testing. Stakeholder mapping and analysis revealed many potential NeoWarm partners within each region whom had not been previously identified.