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Rise Sponsored Summer Symposium 2015 RISE SPONSORED SUMMER SYMPOSIUM 2015 Friday, September 4, 2015 9am-2:30pm Building 4, 2nd floor, Room 314 TIME ORAL PRESENTATIONS (RISE Intensive Students) 9-9:10a Opening Remarks: Dr. Jill Adler-Moore 9:10-9:30a Jade Lolarga Mentor: Dr. Jijun Hao Western University Presentation Title: Generation of Integration-Free Methods of Feline Induced Pluripotent Stem Cells 9:30-9:50a Alma Madrigal Mentor: Dr. Ansel Zhao Presentation Title: Understanding RGS2 function in Human Adipogenesis and Osteogenesis 9:50-10:10a Jennifer Rubio Mentor: Dr. Jill Adler-Moore Presentation Title: Evaluation of Lipidated and Non-Lipidated CDN Adjuvants in a gD3pep-HD and a gD3pep-Cys Liposomal Vaccine using an HSV-2 Intravaginal Mouse Challenge Model" 10:10-10:30a Hernán Reza Mentor: Dr. Jill Adler-Moore Presentation Title: Standardization of a murine model of pulmonary aspergillosis 10:30-10:50a Adam Marentes Mentor: Dr. Nancy Buckley Presentation Title: Role of cannabinoid-receptor 2 (CB2) on tissue fungal load in a systemic Candida albicans infection in mice 10:50-11:10a Jennifer Enriquez Mentor: Dr. Andrew Steele Presentation Title: The Dopamine Transporter (DAT) plays a role in high fat diet obesity but does not in circadian entrainment to feeding 11:10-11:30a Timothy Batz Mentor: Dr. Bharti Sharma Presentation Title: The evolutionary impact of a Pack-MULE element in tomato and its wild relatives 11:30-11:50a Hessam Ghanimi Mentor: Ángel A. Valdés Presentation Title: Phylogenetic Reconstruction of the Genus Berthella Using Molecular and Morphological Data 11:50a-12:10p Eric Breslau Mentor: Ángel A. Valdés Presentation Title: Species Delimitation and Phylogenetic Analysis of Dolabrifera brazieri. 12:10-1p LUNCH AND POSTER PRESENTATIONS (RISE Invitation Students) Melody Sycks Mentor: Dr. Andrew Steele Presentation Title: Analysis of nutrient absorption in obesity resistant mice by adiabatic bomb calorimetry Gabriela Garza-Vazquez Mentor: Dr. Andrew Steele Presentation Title: To binge or not to binge: Studying the effects of Dopamine Receptor 1 signaling on feeding in mice Natalie Eulogio Mentor: Dr. Marcia Murry Presentation Title: Can Algae save the Dairy Industry? William Morris Mentor: Dr. Jill Adler-Moore Presentation Title: Is two better than one? Investigating a novel Aspergillus vaccine in combination with anti-fungal therapy. Christian Garrido Mentor: Dr. Nipam Patel UC Berkley Presentation Title: Role of teashirt and tiptop in the regulation of Hox genes in the marine crustacean Parhyale hawaiensis ORAL PRESENTATIONS Continued (RISE Intensive Students) 1-1:20p Isis Janilkarn-Urena Mentor: Dr. Marcia Murry Presentation Title: Algal Bioremediation of Organic Wastes Coupled to Biomass Production for Feed and Biofuels 1:20-1:40p Danielle McHaskell Mentor: Dr. Jayson Smith Presentation Title: The quantitative effect of phlorotannins on the feeding preference of Aplysia vaccaria. 1:40-2p Pablo Unzueta Mentor: Dr. Francis Flores Presentation Title: A kinetic study of (3-oxo-3-phenyl-1-propen-1-yl)- Ferrocene: A chalcone derivative. 2-2:10p Closing Remarks: Dr. Jill Adler-Moore Generation of Integration – Free Methods of Feline Induced Pluripotent Stem Cells J Lolarga and Jijun Hao Department of College of Veterinary Medicine, Western University of Health Science, Pomona, CA Introduction: Hypertrophic Cardiomyopathy (HCM) is a very common type of heart disease found in adult cats caused by mutations in cardiac myosin binding-C (MYBPC3), which is inherited as an autosomal dominant trait. IPSCs have been used for differentiation into various tissue cells, which include cardiomyocytes. Therefore iPSCs provide a possible in vitro method for generating disease-specific iPSC-derived cardiomyocyte cells that could be used as a model for understanding the HCM disease to test and develop therapeutic drugs. IPSCs generation have been used in various species such as humans, mice, monkeys, and horses for disease modeling and drug examination. However, there are limited disease modeling and drug examination done in cats. Methods: Feline wild-type somatic cells were grown in 100mm tissue culture dish using Basic MEF Medium. Once the feline wild-type somatic cells were 90-100% confluent, the cells were plated in a 24 – well plate that contained 2.5x104 cells/ml. Day 2, the cells were transfected through Lipofection using two different vectors and plated with MEF Medium with no Penicillin Streptomycin. Day 3, the wells were washed with Phosphate Buffered Saline (PBS) and added MEF Media with Penicillin Streptomycin. Day 6-7, the cells were plated in a 6-well tissue culture dish coated with Geltrex with approximately 2.0x105 cells/ml. From Day 6-22 the cells were changed and grown in E8 Medium and changed on a daily basis. Day 22-27 colonies are ready to be harvested and observed for iPSC formation. Results: On Day 22, the feline wild-type somatic cells were observed and showed no signs of iPSCs formation. The following morphology of the possible feline wild-type iPSCs still showed the same morphology compared to the primary feline wild-type somatic cells that were plated in the 100mm tissue culture dish. During the growth period from Day 6 –Day 22, a suitable formation of a possible iPSCs colony was present in one of the wells. However as the growth period proceeded, there was no sign of colony formation in the well. When observed under the 10x microscope, possible single colonies of iPSCs were visible based on their circular morphology. However, these possible single iPSCs did not form any colonies, which is an important characteristic of iPSCs. Conclusions: The following results showed no signs of iPSCs colony formation based on the observation on Day 22. This could be due to the regents that were used for Lipofection, which had no effect on the feline wild-type somatic cells during transfection. In addition, future studies include repetition of the Lipofection transfection method with minimal changes to the protocol as well as testing other methods of transfection. Research supported by NIH RISE, Cal Poly Pomona, and Western University of Health Science. Understanding RGS2 Function in Human Adipogenesis and Osteogenesis A Madrigal, Yuanxiang Zhao Department of Biological Sciences, California State Polytechnic University, Pomona, CA Introduction: Human adipogenesis and osteogenesis define the commitment and differentiation processes by which mesenchymal stem cells (MSCs) mature into functional adipocytes or osteoblasts to form fat or bone, respectively. Between osteogenic and adipogenic lineage commitment, a theoretical inverse relationship exists, such that differentiation towards one lineage results at the expense of the other. This relationship is supported by cellular and clinical studies showing that bone mass and fat mass are closely regulated. At the clinical level, the majority of medical conditions associated with bone loss, including aging and osteoporosis, are accompanied by an increase in marrow fat content. Likewise, conditions associated with high bone mass are associated with decrease marrow fat mass. Because of this, identification of signaling pathways that are either pro-osteogenic/anti-adipogenic or both positive regulators of these two cellular processes have important applications in the improvement of human health and treatment of clinically associated diseases. In this study, we focus on understanding the role of members of the Regulator of G protein Signaling family, in particular regulator of G protein signaling 2 (RGS2), in adipose-derived human mesenchymal stem cells (ad-hMSCs) upon adipogenic and osteogenic induction. Methods: Adipose-derived hMSC (p4) were cultured in HycloneAdvanceSTEM Cell Media (CM). Ad-hMSCs were differentiated using adipogenic inducing media (AIM) consisting of 1uM or 0.2 uM DEX, 0.45uM IBMX, and 10ug/ml Insulin for 12 days or osteogenic inducing media (OIM) consisting of 1 uM or 0.2 uM DEX, 10mM glycophosphate, 0.05 mM ascorbic acid for 16-21 days. RGS2 expression pattern was characterized by RT-PCR during both adipogenic and osteogenic differentiation of hMSCs at 12h, 24h, 36h, 48h, 72h, 96h, 5D, 6D, 7D and 16D post media induction. siRNA mediated RGS2 silencing was used to determine RGS2 function. Ad- hMSCs were reverse transfected with siCON or siRGS2 siRNA and plated equally in 24 well plates two days prior to AIM/OIM induction. Adipogenesis was characterized by oilRedO staining and quantification, dapi and mature adipocyte counting, and RT-PCR of adipogenic markers. Osteogenesis was characterized by Alkaline Phosphatase (ALP) activity, AlizarinRed Staining and quantification, dapi counting, and RT-PCR of osteogenic markers. Results: Overall, RGS2 was up-regulated during both adipogenic and osteogenic induction but appear to oscillate in response to media change at 48h intervals. siRGS2 targeted silencing resulted in decrease adipogenic efficiency and adipocyte number in AIM treated siRGS2 samples compared to controls (siCON) (p<0.05), but did not have a significant effect on overall cell number (p>0.05). siRGS2 targeted knockdown in OIM treated samples resulted in a significant decrease in calcium deposits as evaluated by AlizarinRed (p<0.05), but did not show a significant difference in ALP activity at day 3, 7, 14 compared to controls (siCON). Conclusions siRNA targeted RGS2 expression knockdown resulted in a decrease in both adipogenic and osteogenic differentiation efficiency suggesting RGS2 is a positive regulator of both adipogenesis and osteogenesis. Evaluation of Lipidated and Non-Lipidated
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