Pseudomonas Aeruginosa Biofilm Formation in Different Environments

poster abstractVarious bacteria, such as the soil microbe Pseudomonas aeruginosa, form into strong structures to defend themselves from antibiotics and other harmful materials. These structures are called biofilms. The goal of this research is to isolate P. aeruginosa from several soil samples and determine whether they are able to form biofilms in those environments. Another goal of this research is to find out how different environmental factors affect the formation of Pseudomonas biofilms. We isolated P. aeruginosa from soil samples using Pseudomonas Isolation Agar plates. The colonies most similar to P. aeruginosa were picked, cultured, and tested by PCR in order to confirm that the strains were actually P. aeruginosa. Using these methods, so far we have collected 12 P. aeruginosa strains and we are collecting more strains from different soil samples. In future studies, we will determine whether these strains form biofilms in soil. We will also demonstrate the effect of magnesium on P. aeruginosa on biofilm formation. These studies will begin to investigate how altering environmental conditions can influence persistence of this bacterial pathogen in the soil. These studies can have broad implications for transmission of the bacterium from the environment to humans during disease

Utilizing the C2Maps Platform for Characterizing Drug-Protein Relations, Generating Mobile Games, and Constructing Integrated Pathway Models

poster abstractThe C2Maps platform is a collection of genome-wide data that display the connections between drugs, diseases and genes. The C2Maps is used as a tool to compare and extrapolate known map data into unknown areas. By using C2Maps, researchers can compare genetic, sequential and physical information about disease specific proteins. Manual curation is important for the C2Maps platform in order to validate the literature mining approach and to overcome high levels of data noise generated from molecular networks. Currently we are examining specific drug-protein relationships in several diseases. In this research, the C-Maps website is being used to manually curate abstracts about disease specific drugprotein relations and then it is determined whether a drug “Up Regulates”, “Down Regulates”, or “Indirectly” affects a specific protein. Presently, more than 2000 specific protein-drug relations have been examined through the platform. We theorize that new drug-protein relations will be discovered through curation efforts. To broaden the scope of curation data generated, a C2Maps mobile game is in the process of being developed. This game takes advantage of novel technology in mobile development to create a game that will allow several researchers to contribute to the curation process. The data generated from the manual curation approach can be used to validate various protein-drug relationships in pharmacology and can determine the best possible drugs targeting specific proteins in cancer. Optimal drugs and their respective targets for a specific disease can then be incorporated into an integrative pathway model to analyze the mechanism of the drug. Specific properties of the drug, including chemical structure, can then be examined to determine how a specific drug acts on particular target proteins

Optical coherence tomography enables imaging of tumor initiation in the TAg-RB mouse model of retinoblastoma

PURPOSE: Retinoblastoma is the most common primary intraocular malignancy in children. Although significant advances in treatment have decreased mortality in recent years, morbidity continues to be associated with these therapies, and therefore, there is a pressing need for new therapeutic options. Transgenic mouse models are popular for testing new therapeutics as well as studying the pathophysiology of retinoblastoma. The T-antigen retinoblastoma (TAg-RB) model has close molecular and histological resemblance to human retinoblastoma tumors; these mice inactivate pRB by retinal-specific expression of the Simian Virus 40 T-antigens. Here, we evaluated whether optical coherence tomography (OCT) imaging could be used to document tumor growth in the TAg-RB model from the earliest stages of tumor development. METHODS: The Micron III rodent imaging system was used to obtain fundus photographs and OCT images of both eyes of TAg-RB mice weekly from 2 to 12 weeks of age and at 16 and 20 weeks of age to document tumor development. Tumor morphology was confirmed with histological analysis. RESULTS: Before being visible on funduscopy, hyperreflective masses arising in the inner nuclear layer were evident at 2 weeks of age with OCT imaging. After most of these hyperreflective cell clusters disappeared around 4 weeks of age, the first tumors became visible on OCT and funduscopy by 6 weeks. The masses grew into discrete, discoid tumors, preferentially in the periphery, that developed more irregular morphology over time, eventually merging and displacing the inner retinal layers into the vitreous. CONCLUSIONS: OCT is a non-invasive imaging modality for tracking early TAg-RB tumor growth in vivo. Using OCT, we characterized TAg-positive cells as early as 2 weeks, corresponding to the earliest stages at which tumors are histologically evident, and well before they are evident with funduscopy. Tracking tumor growth from its earliest stages will allow better analysis of the efficacy of novel therapeutics and genetic factors tested in this powerful mouse model

Drug Discovery Through Drug Perturbation Pathway Modeling and Network Analysis

Due to intrinsic complex molecular interactions, the “one disease – one target – one drug” strategy for disease treatment is no longer the best option to treat cancers. To assess drug pharmacological effects, we assume that “ideal” drugs for a patient can treat or prevent the disease by modulating gene expression profiles of this patient to the similar level with those in healthy people. A new approach for drug-protein interactions curation, drug-drug similarity network comparison, and integrative pathway model construction and evaluation was introduced to determine optimal drugs for various cancers. Drug-protein interaction curation is conducted to discover novel drug-protein relationships and is categorized as: up regulated, down regulated, indirect up or down, ambiguous and unknown. The manual curation can be utilized for drug repurposing and examining drug mechanism on a pathway level. A drug-drug similarity network model is built by examining similar targets, therapeutic mechanisms, side effects, and chemical structures. Drug similarity analysis is useful for drug repositioning because similar drugs may have compatible therapeutic or toxic effects for a disease. Drug similarity networks are constructed and examined through a molecular network visualization platform. An integrative disease-specific pathway model is also built to gain a more holistic view of disease mechanisms by including every significant disease-specific protein. Including drugs on the pathway through target information can also offer a clear mechanism for the drug’s action. We also transform integrated pathways into network models and ranked drugs based on the network topological features of drug targets, drug-affecting genes/proteins, and curated disease-specific proteins. Combining our three approaches could potentially lead to advances in drug repurposing and repositioning

Kif14 overexpression accelerates murine retinoblastoma development

The mitotic kinesin KIF14 has an essential role in the recruitment of proteins required for the final stages of cytokinesis. Genomic gain and/or overexpression of KIF14 has been documented in retinoblastoma and a number of other cancers, such as breast, lung and ovarian carcinomas, strongly suggesting its role as an oncogene. Despite evidence of oncogenic properties in vitro and in xenografts, Kif14's role in tumor progression has not previously been studied in a transgenic cancer model. Using a novel Kif14 overexpressing, simian virus 40 large T-antigen retinoblastoma (TAg-RB) double transgenic mouse model, we aimed to determine Kif14's role in promoting retinal tumor formation. Tumor initiation and development in double transgenics and control TAg-RB littermates were documented in vivo over a time course by optical coherence tomography, with subsequent ex vivo quantification of tumor burden. Kif14 overexpression led to an accelerated initiation of tumor formation in the TAg-RB model and a significantly decreased tumor doubling time (1.8 vs. 2.9 weeks). Moreover, overall percentage tumor burden was also increased by Kif14 overexpression. These data provide the first evidence that Kif14 can promote tumor formation in susceptible cells in vivo

E-Mobility -- Advancements and Challenges

Mobile platforms cover a broad range of applications from small portable electric devices, drones, and robots to electric transportation, which influence the quality of modern life. The end-to-end energy systems of these platforms are moving toward more electrification. Despite their wide range of power ratings and diverse applications, the electrification of these systems shares several technical requirements. Electrified mobile energy systems have minimal or no access to the power grid, and thus, to achieve long operating time, ultrafast charging or charging during motion as well as advanced battery technologies are needed. Mobile platforms are space-, shape-, and weight-constrained, and therefore, their onboard energy technologies such as the power electronic converters and magnetic components must be compact and lightweight. These systems should also demonstrate improved efficiency and cost-effectiveness compared to traditional designs. This paper discusses some technical challenges that the industry currently faces moving toward more electrification of energy conversion systems in mobile platforms, herein referred to as E-Mobility, and reviews the recent advancements reported in literature

Synthesis and Biological Evaluation of Novel Homoisoflavonoids for Retinal Neovascularization

Eye diseases characterized by excessive angiogenesis such as wet age-related macular degeneration, proliferative diabetic retinopathy, and retinopathy of prematurity are major causes of blindness. Cremastranone is an antiangiogenic, naturally occurring homoisoflavanone with efficacy in retinal and choroidal neovascularization models and antiproliferative selectivity for endothelial cells over other cell types. We undertook a cell-based structure–activity relationship study to develop more potent cremastranone analogues, with improved antiproliferative selectivity for retinal endothelial cells. Phenylalanyl-incorporated homoisoflavonoids showed improved activity and remarkable selectivity for retinal microvascular endothelial cells. A lead compound inhibited angiogenesis in vitro without inducing apoptosis and had efficacy in the oxygen-induced retinopathy model in vivo

Bringing Attention to Lesser-known Bone Remodeling Pathways

Osteoporosis, a disease of low bone mass, places individuals at enhanced risk for fracture, disability, and death. In the USA, hospitalizations for osteoporotic fractures exceed those for heart attack, stroke, and breast cancer and, by 2025, the number of fractures due to osteoporosis is expected to rise to nearly three million in the USA alone. Pharmacological treatments for osteoporosis are aimed at stabilizing or increasing bone mass. However, there are significant drawbacks to current pharmacological options, particularly for long-term management of this chronic condition. Moreover, the drug development pipeline is relatively bereft of new strategies. Consequently, there is an urgent and unmet need for developing new strategies and targets for treating osteoporosis. Casual observation led us to hypothesize that much of the bone remodeling research literature focused on relatively few molecular pathways. This led us to perform bibliometric analyses to determine the relative popularity of bone remodeling pathways in publications and US National Institutes of Health funding of the last 10 years. In this review article, we discuss these findings and highlight several less-examined signaling pathways that may hold promise for future therapies

Ferrochelatase is a therapeutic target for ocular neovascularization

Ocular neovascularization underlies major blinding eye diseases such as “wet” age-related macular degeneration (AMD). Despite the successes of treatments targeting the vascular endothelial growth factor (VEGF) pathway, resistant and refractory patient populations necessitate discovery of new therapeutic targets. Using a forward chemical genetic approach, we identified the heme synthesis enzyme ferrochelatase (FECH) as necessary for angiogenesis in vitro and in vivo. FECH is overexpressed in wet AMD eyes and murine choroidal neovascularization; siRNA knockdown of Fech or partial loss of enzymatic function in the Fechm1Pas mouse model reduces choroidal neovascularization. FECH depletion modulates endothelial nitric oxide synthase function and VEGF receptor 2 levels. FECH is inhibited by the oral antifungal drug griseofulvin, and this compound ameliorates choroidal neovascularization in mice when delivered intravitreally or orally. Thus, FECH inhibition could be used therapeutically to block ocular neovascularization

Examining LRP4’s Capacity to Participate in Inhibitory WNT Signaling in Bone Cells

Context: In states of health, bone mass is sustained in a coordinated effort by osteoblasts, osteoclasts, and osteocytes. WNT signaling through low-density lipoprotein receptor related protein 5/6 (LRP5/LRP6) is one of the central signaling pathways that aids in controlling bone homeostasis. A prominent antagonist of the WNT signaling pathway is sclerostin. Low-density lipoprotein receptor related protein 4 (LRP4) is required to facilitate sclerostin-mediated inhibition of LRP5/LRP6. Clinically, mutations in LRP4 (R1170W and W1186S) which diminish its ability to bind sclerostin result in bone overgrowth. The mechanism by which LRP4 participates in this process is unknown. In vitro experiments suggest that LRP4 physically binds sclerostin and presents it directly to nearby LRP5/LRP6. Objective: Delineating the mechanistic function(s) of LRP4 is important because if LRP4 directly provides sclerostin to LRP5/LRP6 then interfering with this process represents a potential therapeutic intervention for promoting anabolic bone formation. Design: Genetic cloning is currently underway to fluorescently tag LPR5, LRP6, LRP4 and the LRP4 missense mutation (R1170W) for FRET/FLIM microscopy experiments. FRET/FLIM microscopy will be used to examine the live in vitro signaling dynamics of LRP4. Results: We expect to determine if LRP4 co-localizes with LRP5 or LRP6 in living bone cells, whether this co-localization is sclerostin dependent, and whether the LRP4 missense mutation affects this interaction. Our results will characterize the novel role LRP4 plays in sclerostin-mediated WNT inhibition and capacity to effect bone mass