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Wyoming INBRE Research Network Retreat

September 15-17, 2016 University of Wyoming/ National Park Service Research Station (AMK

Ranch)

Grand Teton National Park, WY

Program

Wyoming INBRE is supported by a grant from the National Institute of General Medical

Sciences (2P20GM103432) from the National Institutes of Health. Any opinions expressed or results presented are solely the responsibility of the authors and does not necessarily represent

the official views of the National Institutes of Health.

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Thursday September 15, 2016

7:00pm Welcome reception- Berol Lodge

Friday September 16, 2016 *Please note that all meals are provided for all meeting attendees at the conference site (Berol Lodge). To find the lodge follow the road heading roughly north to the top of the hill. Please park in the parking lots as you enter the Research Station.

7:00-8:50am Breakfast* All early morning (9-10:30am) speakers should load their PowerPoint presentations on the main computer in the Berol conference room before 8:50am.

9:00-9:10am Welcome/ Introductions: R. Scott Seville, Wyoming INBRE Program Director and PI, University of Wyoming at Casper

9:10-9:40am Wyoming INBRE Bioinformatics Core, University of Wyoming. How can

INBRE bioinformatics core best serve Wyoming institutions. Vikram Chhatre, Nicolas Blouin, and Naomi Ward

9:40-10:00am Wyoming Geographic Information Science Center (WyGISC), University

of Wyoming. Examples and Options: Using Scripting Languages to Facilitate Research and Learning (and Employment?). Shannon Albeke

10:00-10:10am OP1 (University of Wyoming at Casper) INBRE supported research at Casper

College and the University of Wyoming at Casper. Dagmara Motriuk-Smith

10:10-10:20am OP2. (University of Wyoming at Casper) Eimeria in Mediterranean geckos and ornate box turtles. Kristine Carmen, Catherine Kerr, Dagmara Motriuk-Smith, Chris T. McAllister, R. Scott Seville

10:20-10:30am OP3 (University of Wyoming at Casper) Recombinant Spider Silk-Like Protein Production for Biomaterial Generation. Tianna M. Aikey, Hunter McCurdy, Patrick Johnson and Florence Teulé-Finley

10:30-11:00am Break* All late morning (11am-12pm) speakers should load their PowerPoint

presentations on the main computer in the Berol conference room before 10:50am.

11:00-11:10am OP4 (University of Wyoming at Casper) Long-term fire succession as a framework for training undergraduate students in research. Hayley C. Lanier, Zachary P. Roehrs, Meredith A. Roehrs, and R. Scott Seville

11:10-11:20am OP5 (University of Wyoming at Casper) Mapping the dynamic ecology of ants (Formicidae) in post-burn Yellowstone. Isaac T. Andersson and Hayley C. Lanier

11:20-11:30am OP6 (University of Wyoming at Casper) Can sleepy bats think? Lewis Hein and Hayley C. Lanier

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11:30-11:40am OP7 (University of Wyoming at Casper) Does Raptor Distribution Differ Between Urban and Rural Habitats of Wyoming? Haley Tolbert, and Hayley C. Lanier

11:40-11:50am OP8 (Laramie County Community College) Occupancy Modeling to Examine Meso-Mammal Diversity and Abundance at an Urban-Rural Interface of Cheyenne, Wyoming. Devyan J. Paiz, Francis G. Schaffer, Ami L. Wangeline, Hayley C. Lanier, and Zachary P. Roehrs

11:50-12:00pm OP9. (Laramie County Community College) Ear and PIT tag loss rates in small mammals from the Greater Yellowstone Ecosystem. Molly E. Loetscher, Zachary P. Roehrs, Meredith A. Roehrs, Hayley C. Lanier, and R. Scott Seville

12:00-1:20pm Lunch* All early afternoon (1:20-3:30pm) speakers should load their PowerPoint presentations on the main computer in the Berol conference room before 1:15pm.

1:20-1:30pm OP10 (Laramie County Community College) Small mammal survey of the Cheyenne Business Park Natural Area, Cheyenne, Wyoming. Jessica M. Williams, Molly E. Loetscher, Meredith A. Roehrs, and Zachary P. Roehrs

1:30-1:40pm OP11 (Laramie County Community College) Bioinformatics Education with Coursera: Is it worth it? Suki Smaglik

1:40-1:50pm OP12 (Northwest College) From Antibiotics nouveau to Zonotrichia -INBRE Research at Northwest College. Eric C. Atkinson, Allan Childs, Elise Kimble, and Uko Udodong

1:50-2:00pm OP13 (Northwest College) Lichen the Odds: Search for Novel Antibiotics in Wyoming Lichen. Scott Chanthongthip, Elise Kimble, and Michael Cuddy.

2:00-2:10pm OP14 (Northwest College) Environmental sources of novel antibiotic-producing bacteria. Joel Hunt, Ryan Winchell, Elise Kimble, and Uko Udodong

2:20-2:30pm OP15 (Central Wyoming College/ University of Wyoming) Analysis of Effect of TRPV1 Activation on Triglyceride, Free Fatty Acid, and Cholesterol Levels in an Obese Mouse Model. Kaylan Schilling, Joy Watkins, Jana Favela, Asia Williams, Rachel Tighe, Rachel Graham, Anna Hepp, Padmamalini Baskaran, Steven McAllister, and Baskaran Thyagarajan

2:30-2:40pm OP16 (Central Wyoming College) Microplastic Pollution in the Snake River: A snapshot. Ellen Yeatman, and Kirsten Kapp

2:40-2:50pm OP17 (Western Wyoming Community College) INBRE Research at Western WY Community College Provides Student Opportunities. Bud Chew

2:50-3:00pm OP18 (Western Wyoming Community College) Pressure-Volume Loop Analysis of Rbm20 -/- Rats and DGAT 1 -/- Mice. Benjamin A. Sabat, Hanna Ahuja, Sarah Bailey, Gabriel Correia Rodrigues, and Bud Chew

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3:00-3:30pm Break* All late afternoon (after 3:30pm) speakers should load their PowerPoint presentations on the main computer in the Berol conference room before 3:25pm.

3:30-3:40pm OP19 (Western Wyoming Community College) A comparison of fire and pine beetle (Dendroctonus ponderosae) disturbances on seed banks in a forest ecosystem. Katheryn Thomas, Will Clark and Megan Lahti

3:40-3:50pm OP20 (University of Wyoming) How do intrinsically disordered hub proteins achieve broad binding specificity? Grant Bowman

3:50-4:00pm OP21 (University of Wyoming) Inhibition of UGCG Expression Via MicroRNA and SiRNA. Jesse Hinshaw and Anya Lyuksyutova

4:00-4:10pm OP22 (University of Wyoming) A regulatory genetic network in C. elegans contributes to epidermal structural integrity during development. Sarina Bernazzani, Melissa Kelley and David S. Fay

4:10-4:20pm OP23 (University of Wyoming) Conserved ankyrin repeat proteins and NIMA kinases regulate extracellular matrix remodeling and intracellular trafficking in Caenorhabditis elegans. Vladimir Lažetić and David S. Fay

4:20-4:30pm OP24 (University of Wyoming) Genetic Analysis in NimA-Related Kinase Pathways in C. elegans. Joseph Braveen, Vladimir Lažetic and David S. Fay

4:30-4:40pm OP25 (University of Wyoming) CARD9 Knockout Rescues Heart Function by Reducing Fibrosis and Hypertrophy in a Pressure-overload Model of Murine Heart Failure. Matthew Peterson, Samantha Haller, Kayla Wilson, Thomas D. Paul, and Guanglong He

4:40:4:50pm OP26 (Sheridan College)- INBRE Supported Undergraduate Research at Northern Wyoming Community College District (NWCCD). Ami Erickson

4:50-5:00pm OP27 (Central Wyoming College) A Plan for a Long Term Investigation of

Human Exposure to West Nile Virus in Fremont County, Wyoming. Joy Watkins, Kaylan Schilling, Adam Conner, Shanda Barlow, Grant Hosking, Kelvin Kinyatta with Steven McAllister

5:00-5:30pm Break*- Poster session preparation. Poster presenters should set up their posters in the Conference Room (Berol Lodge, CR) before 5:20pm.

5:30-7:30pm Poster reception (Berol lodge: Conference Room/CR) and BBQ dinner

P1 Native bee diversity in burned and non-burned areas of the Rocky Mountain Forest System and the importance post-fire succession in conservation. Kendra David and Hayley C. Lanier (University of Wyoming at Casper)

P2 Genetic analyses to determine road-barrier effects on small mammal populations. Laura M. Diesburg and Hayley C. Lanier (University of Wyoming at Casper)

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P3 Caulobacter PopZ forms an intrinsically disordered hub in organizing bacterial cell poles. Josh Holmes and Grant Bowman ( University of Wyoming)

7:30pm-8:30pm Keynote presentation

The Repairable Brain, Really? Dr. John Sladek, Professor Emeritus Department of Neurology, Pediatrics & Neuroscience, University of Colorado School of Medicine and Chair of the Wyoming INBRE External Advisory Committee Department of Neurology, Pediatrics & Neuroscience (Emeritus), University of Colorado School of Medicine, Research Complex 1 North, Room 7133, Mail Stop 8315 12800 E. 19th Ave. Aurora, CO 80045. Email: [email protected] Abstract. The human brain contains about a trillion neurons that are organized into specific and well defined neural circuits that control everything we do from walking to waking to creating music and being innovative. For over a century there was a consensus among scientists that the brain was incapable of making new nerve cells and regenerating new connections. We now know that neurogenesis occurs in the adult brain and that severed axons can grow, albeit not necessarily to restore proper function. Neurodegenerative disorders such as as Parkinson’s disease, ALS, Alzheimer’s disease and many others including stroke might be stopped or even reversed if new neurons could take the place of those lost during these progressive neurological disorders or after trauma such as spinal cord injury. Implantation of replacement cells of fetal or stem cell origin has been attempted clinically with some success and holds considerable promise for restoration of function. Another intriguing possibility is the stimulation of cell division in the adult, a conserved evolutionary mechanism for cell renewal that is common in some vertebrates. Professor Sladek will discuss these possibilities in the context of his pioneering research in a non-human primate model of Parkinson’s disease.

Saturday September 17, 2015

7:30am-8:50am Breakfast* All morning speakers should load their PowerPoint presentations on the main computer in the Berol conference room before 8:50am.

9-9:10am Wyoming INBRE Developmental Research Project Program, University of Wyoming. Update from the Director of the Developmental Research Project Program (DRPP). David S. Fay

9:10-9:45am An overview of the biomedical startup model; How venture capital and the digital age are impacting healthcare innovation. Gregory Fluet (Consultant at Sapient Capital Management, L.L.C.)

9:45-9:55am OP28 (University of Wyoming) MetabocinTM : A novel agent to ameliorate metabolic syndrome. Markert Laurel, Padmamalini Baskaran, and Baskaran Thyagarajan

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9:55-10:05pm OP29 (University of Wyoming) Sirtuin-1 and Regucalcin Emerge as a Potential Target to Defy Redox Stress–Induced Accelerated Aging. Kara Nazminia, Justine Frantz, Padmamalini Baskaran and Baskaran Thyagarajan

10:05-10:30am Break*

10:30-12:00pm INBRE Principal Investigators’ session

12:30pm- Lunch*

2pm- INBRE group activity (outdoors)

6-7:30pm Dinner* (Make your own pizza contest; by school)

Oral Presentations- Abstracts (Berol lodge Conference Room)

Presentations abstracts are listed as per the schedule. Oral Presentations marked as OP are limited to 10 minutes including questions. Friday September 16, 2016: How can INBRE bioinformatics core best serve Wyoming institutions Chhatre, Vikram, Nicolas Blouin, and Ward Naomi

Wyoming INBRE Bioinformatics Core, University of Wyoming, 1000 E. University Avenue, Laramie, WY 82071

E-mail: [email protected]

Abstract. We introduce services available through the INBRE Bioinformatics Core at the University of Wyoming. In addition to the UW campuses at Laramie and Casper, these services are available to research and teaching community at all seven community colleges across the state. Following its mandate of raising bioinformatics awareness, the Core has recently become fully functional through the appointment of two staff scientists. The scientists will engage in several activities: (1) collaborate with Wyoming researchers on bioinformatics projects, (2) disseminate beginner training on introduction to bioinformatics computing, (3) organize periodic workshops for faculty, staff and students on advanced bioinformatics analyses of next generation sequencing data, (3) provide general data analysis support for community members, and (4) assist Wyoming community in efficiently using the infrastructure available through the Advanced Research Computing Center (ARCC) at the University of Wyoming. In addition, the Core has made significant investment towards buying priority access to this computing infrastructure for the benefit of the INBRE funded researchers. The Core has an informative website accessible at http://inbre.wygisc.org. We invite the INBRE community to participate in this discussion on how the Core can best serve their bioinformatics needs.

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Examples and Options: Using Scripting Languages to Facilitate Research and Learning (and Employment?)

Albeke, Shannon

University of Wyoming, 1000 E. University Avenue, Laramie, WY 82071


Abstract. As data become more abundant and larger in size, scientists are increasing their reliance upon scripting languages to process and analyze these data. Writing code to customize workflow and automate processes can increase efficiency, minimize errors and allow researchers to analyze very large datasets. I will present two examples of research that used multiple scripting languages to create, process and analyze ecological data. Because increasing student exposure to these analytical tools is an important role that instructors can embrace to expand a highly marketable skillset, I will also present some possible tools to help facilitate bringing scripting into the classroom.

As data become more abundant and larger in size, scientists are increasing their reliance upon scripting languages to process and analyze these data. Writing code to customize workflow and automate processes can increase efficiency, minimize errors and allow researchers to analyze very large datasets. I will present two examples of research that used multiple scripting languages to create, process and analyze ecological data. Because increasing student exposure to these analytical tools is an important role that instructors can embrace to expand a highly marketable skillset, I will also present some possible tools to help facilitate bringing scripting into the classroom.

OP1: INBRE supported research at Casper College and the University of Wyoming at Casper

Motriuk-Smith, Dagmara

University of Wyoming at Casper, 125 College Drive, Casper, WY 82601


Abstract. The goal of the INBRE supported research is to train undergraduate students in conducting biomedical research. Nine students receiving the INBRE internship during the summer 2016 were engaged in diverse biomedical research projects ranging from studying antibiotic resistance genes in the environment, electrospinning spider silk nanofiber mats from recombinant spider silk-like proteins to morphology and phylogeny of parasites. In addition, numerous projects related to the mammalian and invertebrate habitat diversity and population genetics were conducted. Our students learned biological, molecular, and fieldwork techniques. They participated in writing and editing abstracts and papers, and in preparation of presentations for conferences. Casper College and University of Wyoming at Casper faculty members received three Wyoming INBRE Scaled Participatory Research and Education Model grants (SPREM), one Wyoming INBRE UW - Wyoming Community College Collaborative Grant award, two faculty members were connected to UW-Laramie faculty members via collaborative activities.

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OP2: Eimeria in Mediterranean geckos and ornate box turtles

Carmen, Kristine, Catherine Kerr, Dagmara Motriuk-Smith, Chris T. McAllister, and R. Scott Seville



Abstract. Oocysts from two different hosts, Hemidactylus turcicus (Mediterranean geckos) and Terrapene ornata (Ornate box turtles) were collected and analyzed. Samples were originally collected in Texas, Florida, Arkansas, and Mississippi and analyzed in Casper, Wyoming. Oocysts from the H. turcicus were identified as Acroeimeria lineri (Eimeria lineri) and Choleoeimeria turcicus (Eimeria turcicus). In five of the samples from H. turcicus, A. lineri was present. Overall, 97 oocysts were documented and measured. These oocysts had an average size of 23.95 x 18.00 µm with an average shape index (length/width) of 1.3. The sporocysts of this species measured 7.63 x 6.72 µm with an average shape index of 1.1. C. turcicus was also described. Sixty-six oocysts were photographed and measured. These oocysts had an average size 34.50 x 17.55 µm and an average shape index of 2.0. Sporocysts in these oocysts were 9.40 x 8.07 with a shape index of 1.2. DNA was extracted from these oocysts, but so far the results are inconclusive. A new species of Eimeria was described and documented from the T. ornata samples. Seventy oocysts were measured and had an average size of 21.02 × 14.11 µm, the shape index was 1.5. Sporocysts were ellipsoidal, 9.84 × 6.06 µm with a shape index of 1.6. Polar granules were visible in almost every oocyst, with some oocysts containing as many as 2 or 3. DNA was extracted from these samples and two unique rDNA 18S sequences were obtained. Construction of phylogenetic trees that define evolutionary relationships will be attempted in the future.

OP3: Recombinant Spider Silk-Like Protein Production for Biomaterial Generation 1Aikey, Tianna M., 1,2Hunter McCurdy, 2Patrick Johnson, and 1Florence Teulé-Finley 1University of Wyoming at Casper, 125 College Dr., Casper, WY 82601 2University of Wyoming, 1000 E. University Avenue, Laramie, WY 82071


Abstract. The unique and wide range of mechanical properties of spider silk fibers make spider silk proteins desirable for many potential biomedical applications. Some of these include their uses in the fabrication of scaffolds for tissue engineering, artificial ligaments or nanofiber mats for wound dressings. The dragline and flagelliform silks from orb weaver spiders are very strong and elastic respectively due to key structural amino acid sequences making up their silk protein compounds. The modular and repetitive sequence of these fibrous silk proteins allows for their recombinant production. In this study, a 60 kDa chimeric flagelliform-dragline spider silk-like protein (SSLP) was produced recombinantly in Escherichia coli. The clone cells were harvested after expression of the silk gene was induced using isopropyl b-D-1-thiogalactopyranoside (IPTG). Total protein content was recovered through standard E. coli cell lysis. After heat treatment of the total protein extract, the SSLPs were purified using immobilized metal (nickel)

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affinity chromatography (IMAC). SDS-PAGE/Coomassie and Western blot analyses were performed to confirm SSLP presence. The eluted SSLPs were then dialyzed prior to their lyophilization. The dry SSLPs constitute the raw material for nanofiber mat generation through electrospinning. These spider silk-based mats will be tested for cytotoxicity in vitro to evaluate their biocompatibility as potential wound dressings.

OP4: Long-term fire succession as a framework for training undergraduate students in research 1,2Lanier, Hayley C., 3Zachary P. Roehrs, 3Mereditn A. Roehrs, and 1,2R. Scott Seville 1University of Wyoming at Casper, 125 College Drive, Casper, WY 82601 2University of Wyoming, 1000 E. University Avenue, Laramie, WY 82071 3Laramie County Community College, 1400 E. College Dr., Cheyenne, WY 82007


Abstract. Although the interest in providing undergraduates with self-directed research experiences is growing, many impediments exist to allowing students the freedom of developing their own projects. Acquiring equipment, IACUC approvals and obtaining permits is challenging, particularly when students are given the latitude to develop a wide range of questions. As a result, many PIs opt for undergraduate assistants, rather than undergraduate investigators, to provide research experience to their students. While undergraduate assistant experiences are valuable, students may not always develop the critical thinking, problem solving, and science process skills they need to become independent scientists. We present data on using a long-term study of fire ecology to: 1) develop a wide range of student-driven questions in areas as diverse as ecology, evolution, and disease ecology; 2) create opportunities for hands-on teaching and learning of concepts related to the process of science; 3) provide opportunities for involving and training numerous undergraduates with varying goals; and 4) produce a more skilled and competitive biomedical student that will successfully transition to UW and/or continue to post-baccalaureate biomedical studies. The encompassing student as scientist model effectively lowers the ‘activation energy’ associated with developing student-directed projects, and provides a framework within which a variety of student generated questions can be pursued both in the lab and field.

OP5: Mapping the dynamic ecology of ants (Formicidae) in post-burn Yellowstone

Andersson, Isaac T. and Hayley C. Lanier



Abstract. During the last three decades, wildfires in the Western United States have been increasing in numbers and intensity. As wildfire season grows and the resulting burn areas cover more of the Western states, it has become imperative to understand the natural fire cycle and the factors involved with burn area recovery. Formicidae, or ants, are a key species in healthy ecosystems throughout the world. Ants aerate the soil and influence the chemical balance, they are important predators and prey species, and they also consume and recycle large amounts of organic materials. Because of their critical role in ecosystem functioning, it is important to understand how ant populations are shaped by the recovery and maturation of

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previously burned areas. Limitations on ant populations in post-burn areas may be related to the abundance of food (seed bearing plants and invertebrates) relative to adjacent mature pine forests (bottom-up controls). Alternatively, top-down effects from predators, such as spiders and shrews, may be the driving factor for ant populations. For this study we used the pitfall samples of invertebrates collected in 2014 and 2015 as part of a long-term study of the 1988 Yellowstone Fires. Invertebrates were sorted to order and, in some cases, family. We compared models of ant abundance relative to explanatory variables that were bottom-up (vegetation characteristics, downed trees, and abundance of invertebrate prey), top-down (shrew and spider captures), or both to determine which model best explained the observed abundance data. Ants are small, silent, relentless shapers of the Western landscape, and by understanding better how they are impacted by post-fire succession of the forest cycle will help us to better understand the interdependence and change in this regions.

OP6: Can sleepy bats think?

Hein, Lewis and Hayley C. Lanier



Abstract. Sleep is a nearly universal phenomenon in the animal kingdom, but is still poorly understood in many animals. In part because it is a cryptic behavior little is known about how, where, or why many animals sleep. My REU project with Dr. Barrett Klein at the Smithsonian Tropical Research Institute focused on the "why" of sleep, searching for functional benefits, in particular benefits associated with learning, shared across distant lineages. My work was on the Jamaican fruit bat (Artibeus jamaicensis). We examined sleep in the context of learning ability, with simple maze trials for spatial learning and the influence of sleep restriction on a bat’s ability to complete these tasks. Our results tentatively showed that extended loss of sleep loss impairs learning ability and memory consolidation. These findings, coupled with the fact that species depend on learning ability for their everyday survival, provide useful information for conservation in continually noisy, well-lit human landscapes as well as reinforce the fundamental importance of sleep across animals.

OP7: Does Raptor Distribution Differ Between Urban And Rural Habitats of Wyoming?

Tolbert, Haley, and Hayley C. Lanier

University of Wyoming at Casper, 125 College Dr., Casper, WY 82601


Abstract. Raptors are iconic apex predators of the prairie and grassland habitats of Wyoming. However, due to extensive invasion of their habitat by urbanization, they may be facing a greater threat to their survival. This study seeks to see if (1) raptor density differs between urban and rural habitats, and (2) if this difference is due to increased food abundance in human-modified habitats. Over the summer of 2016, I used distance sampling techniques and roadkill surveys to examine these questions, focusing on raptors that thrive in prairie, grassland, and riparian habitats. The results indicated that raptors are not adapting to human environments, with 1.9 raptors per km2 in rural habitats vs.0.7 raptors per km2 in urban areas. An AIC comparison of separate urban and rural models versus a combined model strongly supported modeling the two habitats separately. There was also a significant difference between roadkill in different habitats with on average 1 roadkill per km in rural habitats and 0.4 roadkill per km in

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urban zones. While raptors and our prey proxy roadkill were slightly correlated, this correlation was toward a larger abundance of both in rural areas. This suggests that even in a state with a small human footprint, urbanization may be negatively affecting important apex predators.

OP8: Occupancy Modeling to Examine Meso-Mammal Diversity and Abundance at an Urban-Rural Interface of Cheyenne, Wyoming 1Paiz, Devyan J., 1Francis G. Schaffer, 1Ami L. Wangeline, 2Hayley C. Lanier, 1Zachary P. Roehrs 1Laramie County Community College, 1400 E. College Dr., Cheyenne, WY 82007 2University of Wyoming at Casper, 125 College Dr., Casper, WY 82601


Abstract. Camera traps are a passive surveying method used to inventory and gather natural history and ecological information on vertebrates. In this study, camera traps were used to obtain occupancy estimates for meso-mammals in the Cheyenne Business Park Natural Area (CBPNA), an industrial complex on the edge of Cheyenne, Wyoming. Eight camera traps were deployed sampling forty three 100×100 m survey grids, with each grid surveyed by one camera for 7 days, totaling 301 trap days. Grids were assigned to 1 of 3 habitats (prairie, riparian, or woodland). Occupancy data were analyzed using a single-season, single-species model across the CBPNA. In preliminary results riparian habitats hosted a more diverse meso-mammalian community in comparison to prairie habitats. At present, 4 mammal species (Neovison vison, Ondatra zibethicus, Procyon lotor, Sylvilagus floridanus) have been detected on the CBPNA and occupancy has been estimated. Neither N. vison nor P. lotor are well documented historically within Laramie County. While all of these species are known to be present in Laramie County, none have been documented on the CBPNA and this is the first study to estimate their abundance in southeastern Wyoming. Species expected, but not detected may have been due to anthropogenic or seasonal effects requiring further research. Results presented consist of winter data, but are part of ongoing research to elucidate meso-mammal population dynamics in southeastern Wyoming.

OP9: Ear and PIT tag loss rates in small mammals from the Greater Yellowstone Ecosystem 1Loetscher, Molly E., 1Zachary P. Roehrs, 1Meredith A. Roehrs, 2Hayley C. Lanier Hayley C., and 2R. Scott Seville 1Laramie County Community College, 1400 E. College Dr., Cheyenne, WY 82007 2University of Wyoming at Casper, 125 College Dr., Casper, WY 82601

Email: [email protected]

Abstract. When conducting capture-recapture research, study individuals must be marked and retain those marks between capture periods. This individual identification is necessary to track movements and estimate population demographic parameters (e.g. estimate population size). Tag loss is problematic in these studies as it leads to an inability to track individuals and an overestimation of population size. Ear tag loss is well documented in the literature, but loss

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rates are species and habitat specific. Few studies have examined loss rates of passive integrated transponder (PIT) tags, and no studies have document loss rates for either of these tags in the small mammal species in this region. Data collected for a long-term project in the Greater Yellowstone Ecosystem allowed for the comparison of loss rates for ear and PIT tags. The results of this study demonstrate that ear tags are commonly lost (26%) after 1 week. PIT tags are less likely to be lost (10%), but when lost, it is usually within the first 1–2 days. Based on our observations this early PIT tag loss is likely due to miss injection of the PIT tag or the PIT tag working its way back out the injection site. In most cases PIT tags remained in animals, with individuals being recaptured more than a month later. We recommend that ear tags for short-term (1–5 day) and PIT tags for longer term (1–2 months minimum) identification should be used to ensure mark retention over the course of a capture-recapture study.

OP10: Small mammal survey of the Cheyenne Business Park Natural Area, Cheyenne, Wyoming

Williams, Jessica M., Molly E. Loetscher, Meredith A. Roehrs, and Zachary P. Roehrs

Laramie County Community College, 1400 E. College Dr., Cheyenne, WY 82007


Abstract. Embedded in an industrial district and centered on the hundred year flood plain of Dry Creek is the Cheyenne Business Park Natural Area (CBPNA), Cheyenne, Wyoming. This flood plain prairie is managed by the Laramie County Conservation District for wildlife, recreation and education. Over the past year we have conducted a small mammal census of the CBPNA, including 4,050 trap nights. Sherman live traps were set in transects with 50 traps per line, and baited with rolled oats and peanut butter. This effort has resulted in the capture of 269 small mammals for a 6.6% trap success rate. Small mammals captured included 4 confirmed species: Reithrodontomys megalotis (western harvest mouse), Microtus ochrogaster (prairie vole), Peromyscus maniculatus (North American deermouse), Mus musculus (house mouse) and a possible second species of Microtus sp. (vole). Of the species caught, the most common was the western harvest mouse with 62% of captures followed by the voles, at 26% of captures. There was a difference between habitats in which these two most abundant species were caught. While harvest mice were found in all habitats on the CBPNA, voles were found primarily in dense vegetation less prone to flooding. Although, only the prairie vole has been recorded in this region, some of the captured voles lacked the yellowish orange underbelly wash, indicative of this species. Further study of teeth and skulls of these voles is being conducted for final identification. Deer mice (7%) and house mice (6%) were also captured in disturbed habitats. Deer mice were more common in disturbed prairie areas and house mice in flood prone areas or along the railroad tracks. Increasing human use of the CBPNA will require more intensive management of this area. This research provides a base line for future management and education on the CBPNA.

OP11: Bioinformatics Education with Coursera: Is it worth it?

Smaglik, Suki

Laramie County Community College, 1400 E. College Dr., Cheyenne, WY 82007


Abstract. Thanks to a SPREM Sabbatical sub-award I was able to enroll for certificates in bioinformatics training, through Coursera, while I was on sabbatical from CWC and continuing

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today. Two Bioinformatics Specialties, Genomic Data Science (Johns Hopkins - JHU) and Bioinformatics (UCSD), each consisting of 6 or 7 multi-week courses and a capstone project. So far I have earned 2 certificates for Genomic Data Science courses, with plenty more to learn! The quality of delivery varies between institutions. The JHU course is well organized, with plenty of practice, quizzes, a challenging final problem and an engaged teaching assistant. The UCSD Bioinformatics course is a bit out of date, with the lectures and the teaching assistant being of very little help. The instructors do their best to add quite a bit of humor, though. It comes with an interesting interactive textbook, which includes both practice and graded problems. The quiz questions cover rather obscure, lightly covered topics. While assured that we needed neither biology nor computing skills, this specialization is really created for folks who have both. I decided to take a step back and enroll (non-cert) in their beginner version: Biology Meets Programming: Bioinformatics for Beginners, which goes more slowly through the first several chapters of the book. I also found the course Bioinformatics: Introduction and Methods, from Peking University (taught in English) useful in understanding the history and evolution of bioinformatics, as well as current methods. This course assigns benchmark papers to read and discuss, and the lectures are clear and very understandable. My take-away message is that I am glad to have a miniscule understanding of the concepts and methods behind turning big data sets into simple diagrams, but I will likely leave this type of data processing to the experts, and enjoy interpreting the results.

OP12: From Antibiotics nouveau to Zonotrichia -INBRE Research at Northwest College

Atkinson, Eric C., Allan Childs, Elise Kimble, and Uko Udodong

Northwest College, 231 West 6th Street, Powell, WY 82435


Abstract. At NWC, INBRE supported undergraduate research, exploration, and mentoring span topics including novel antibiotic research, characterization of bactericidal compounds, rotifer endocrine disruption, and avian disease. Our broadening trend continued in FY 2015/2016. We have worked to meet the following program goals: 1) enhance opportunities for WY community college undergraduates to better understand (and ultimately participate in) the field of biomedical research; and, 2) develop a pipeline of students with an interest in biomedical science who would then go on to complete their baccalaureate degrees and/or graduate degrees at UW. With significant support from our Executive Dean we initiated a 2-credit class (BIOL 2465-Research Problems in Biology) to partner with our INBRE program providing transcript documentation of research participation. Six students graduated with AS degrees, 3 of which will be transferring to UW in Life Sciences (generally, Zoo/Phys) while the remaining student transferred to UM to MSU. Five transfer students applied for INBRE Transition Fellowships (one wishing to delay until 2017), one was awarded. Another INBRE student transferred to UW (Spring 2016) in Chemical Engineering and will back-transfer credits to attain his AS. Our 4 supported and 3 informal faculty mentors have guided 17 students within our program during 2015/2016. Recently, previous participants have been accepted to the School of Pharmacy (UW), WAMI (UW), and the Peace Corps. In addition to WY INBRE events, 4 students attended professional meetings, 1 presenting a poster at an international conference in Canada. Additionally, we were awarded SPREM support to remodel a computer lab turning it into our new Molecular Techniques Lab.

OP13: Lichen the Odds: Search for Novel Antibiotics in Wyoming Lichen

Chanthongthip, Scott, Elise Kimble, and Michael Cuddy

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Abstract. Currently, an increasing concern exists about antibiotic resistance in pathogenic and potentially pathogenic bacteria. As a result, many researchers are searching for novel antibiotics from environmental sources (i.e. other bacteria, animals, lichen, etc.). Our research focuses on some of the native lichen species (Xanthoparmelia chlorochroa and Caloplaca citrina) in Wyoming and their ability to inhibit pathogens. To test the lichens’ secondary metabolites’ ability to inhibit potential infection we used three pathogens: Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa via the Kirby-Bauer disc diffusion protocol. With a dosage amount of 50 µL reconstituted extract, results showed zones of inhibition by X. chlorochroa on S. aureus (zone of 9.5 mm) and Pseudomonas (zone of 4.0 mm), whereas C. citrina extract inhibited only S. aureus (zone of 5.9 mm). This demonstrates significant implications for the future of lichen-derived antibiotics and their efficacy against S. aureus and Pseudomonas aeruginosa. The antibiotic-producing lichens’ secondary metabolites will be analyzed through nuclear magnetic resonance (NMR) and liquid chromatography/mass spectrometry (LC/MS) to identify specific compounds that inhibit pathogens.

OP14: Environmental sources of novel antibiotic-producing bacteria

Hunt, Joel, Ryan Winchell, Elise Kimball, and Uko Udodong



Abstract. The ability of pathogenic bacteria to be selected for antibiotic-resistant traits, as demonstrated by strains of methicillin-resistant Staphylococcus aureus and carbepenem-resistant Enterobacteriaceae, is concerning in the medical field. Several reasons, including economic factors, have led to the paucity of new antibiotics, which nonetheless remain an important treatment option for health care providers. We have focused our investigation of possible antibiotic-producing microbes on local environmental sources, including cave soil, deer rumen, marsh mud and water, a thermal spring, and two compost piles. All of these sources have hosted inhibitory microbes, as shown by testing environmentally acquired bacteria on lawns of three pathogens – E. coli, P. aeruginosa, and S. aureus. Identification of the environmentally acquired bacteria is ongoing using PCR amplification of the 16S ribosomal RNA gene.

OP15: Analysis of Effect of TRPV1 Activation on Triglyceride, Free Fatty Acid, and Cholesterol Levels in an Obese Mouse Model 1Schilling, Kaylan, 1Joy Watkins, 1Jana Favela, 1Asia Williams, 1Rachel Tighe, 1Rachel Graham, 1Anna Hepp, 2Baskaran Padmamalini, 1Steven McAllister, and 2Baskaran Thyagarajan 1Central Wyoming College, 2660 Peck Avenue, Riverton, WY 82501 2School of Pharmacy, University of Wyoming, 1000 E. University Avenue, Laramie, WY 82071


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Abstract. Metabolic syndrome is comprised of the conditions of obesity, impaired glucose metabolism, dyslipidemia, and cardiovascular complications. It can lead to stroke, a major cause of death worldwide. Previous research suggests that transient receptor potential vanilloid subfamily 1 (TRPV1) protein expressed in adipose tissue and vascular smooth muscle cells is a potential target to treat dyslipidemia, fatty liver and hypertension. Research from Baskilab demonstrates that TRPV1 protein is expressed in mammalian adipose and liver tissues and that activation of TRPV1 by capsaicin prevented high fat diet (HFD)-induced obesity, non-alcoholic fatty liver disease and hypertension, without modifying the quantity of food or water consumed by mice. In our collaborative research with Baskilab, we measured triglyceride, free fatty acid, and cholesterols levels in the liver of normal chow or HFD (capsaicin; 0.01%)-fed wild type and TRPV1-/- mice. We evaluated the hypothesis that HFD-feeding increased lipid levels and that TRPV1 activation by capsaicin prevented this. Our data provides new evidence for the role of TRPV1 activation in regulating lipid metabolism, suggests that TRPV1 regulates de novo lipogenesis and lipolysis, and that activation of TRPV1 is a novel strategy to prevent hyperlipidemia and associated complications in diet-induced obesity.

OP16: Microplastic Pollution in the Snake River: A snapshot

Yeatman, Ellen, and Kirsten Kapp

Central Wyoming College, 240 S. Glenwood St. #124, P.O. Box 4795, Jackson, WY 83001


Abstract. Over the past 40 years, world production of plastic resins increased some 25-fold creating a waste stream comprised of 60-80% plastics. Microplastics, tiny fragments of plastic invisible to the naked eye usually less than 5mm in size, are receiving increasing attention from the scientific community as an important and potentially detrimental environmental toxin due to this growth of the plastic manufacturing industry. Microplastics, once in the aquatic environment adsorb potentially harmful toxins and leach out additives. When organisms ingest microplastics, not only are they harmed by simply ingesting them, but they also get exposed to high levels of toxins. Of the few freshwater studies addressing microplastics, high concentrations of microplastics in the Great Lakes, Lake Geneva and Lake Hovsgol, a remote lake in the mountainous region of Mongolia, were revealed. To understand their potential environmental impacts, monitoring for the presence and abundance of them is necessary. This study examines the Snake River for the presence of microplastics from the headwaters in YNP to its confluence with the Columbia River. Grab samples (~1,750 mL each) and volume reduced samples (using a 100 micron plankton net) were collected approximately every 50 river miles along the Snake River, a 1,400 mile stretch of river. Grab samples are processed through 0.45µm filters using vacuum filtration and volume reduced samples are exposed to wet peroxide oxidation prior to vacuum filtration. Both samples are then visually inspected under stereoscopes, with a fluorescent adapter. Any suspect particles will be analysed using Raman spectrometry to confirm the type of polymer that it is.

OP17: INBRE Research at Western WY Community College Provides Student Opportunities

Chew, Bud

Western Wyoming Community College, 2500 College Drive, Rock Springs, WY 82901


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Abstract. INBRE research at WWCC is designed to expose promising freshman and sophomore students to undergraduate research, to encourage students to consider a career that includes research, to enhance students’ opportunities in a competitive world, and to further scientific knowledge through presentation and publication of research work. Currently, 9 students participate in INBRE funded research; topics include wildlife biology/ecology, synthesis of novel organic molecules, and cardiovascular physiology. Three faculty members have established INBRE research programs; two additional faculty members are beginning their involvement with the program. WWCC currently has 6 INBRE Transition Fellows at the University of Wyoming (2 seniors, 4 juniors. It is fair to say that research at WWCC would not be possible without the WY INBRE grant. The opportunities presented by INBRE have allowed the college to recruit and retain better faculty members; students involved in INBRE research at WWCC become better, more confident students, while not only developing enhanced lab/field skills, but also responsibility, perseverance, and leadership skills.

OP18: Pressure-Volume Loop Analysis of Rbm20 -/- Rats and DGAT 1 -/- Mice

Sabat, Benjamin A., Hanna Ahuja, Sarah Bailey, Gabriel Correia Rodrigues, and Bud Chew


Abstract. WWCC cardiovascular research progressed on projects in 2015-2016. Assessment of Cardiac Function of Rbm 20 -/- Rats Using Pressure-Volume Loop Analysis in 3m, 6m, and 9m Old Rats, will soon be submitted for publication. RNA binding motif 20 (Rbm20), a muscle splicing factor, is responsible for post-translational splicing of titin in a sarcomere, and heart diastolic and systolic properties. Rbm20 -/- (KO) rats have impaired cardiomyocyte relaxation and increased compliance of the ventricular walls, pathologies similar to mutant humans. KO rats begin to die suddenly at 5m of age; afflicted humans die in their twenties. We hypothesized that KO rats, when compared to Rbm20 +/+ (WT) rats, would not exhibit cardiac dysfunction at 3m of age, but would by 6m, leading to heart failure. Results indicate a trend for cardiac output to be reduced at 3m, which became significant (p<0.05) at 6m. Those KO surviving to 9m had nearly normal cardiac outputs. All differences in cardiac output can be attributed to stroke volume. We conclude that cardiac dysfunction in KO rats begins very early in life, but surprisingly, not all KO rats go into heart failure. Project 2, Assessment of Cardiac Function in DGAT 1-/- Mice Fed a High Fat Diet, generated pilot data. This project required modifying PV Loop techniques for mice. Diacylglycerol acyltransferase 1 (DGAT 1) is a key enzyme in conversion of fatty acids into triglycerides in intestinal absorptive cells; when absent, an animal cannot store fat, so blocking DGAT 1 is a potential therapeutic target for obesity. However, little is known about the health of DGAT -/- (KO) mice. We hypothesized that KO mice would have normal cardiac function after feeding a high fat diet for 6m. Preliminary results suggest that, when normalized for body mass, cardiac output is not different than DGAT +/+ (WT).

OP19: A comparison of fire and pine beetle (Dendroctonus ponderosae) disturbances on seed banks in a forest ecosystem

Thomas, Katheryn, Will Clark, and Megan Lahti

Western Wyoming Community College, 2500 College Drive, Rock Springs, WY 82901


Abstract. Fire and insect are two major influences on forest ecosystems in North America (Perera et al., 2011) and forests have some adaptations to these events, such as serotinous cones in pine trees (Pinus spp.). Recent pine beetle (Dendroctonus ponderosae) outbreaks in

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response to climate change have led to rapid and extreme disturbances throughout much of the western United States in the last 10 years (Perera et al., 2011). While fire and insects are considered natural disturbances in forest ecosystems, the rapid and profound outbreak of pine beetles is considered unnatural. Seed banks are known to be important reserves for re-establishing forest ecosystems after a disturbance (e.g. Ferrandis et al., 1996). Upon preliminary research, we have not found any publications quantifying the effects of pine beetles on seed banks or germination rates. Our research investigates the effects of fire and pine beetle on seed bank (diversity and density) and germination rate in a pine forest habitat. We will use ANOVA tests to determine whether the effects of disturbance by pine beetles is similar to the effects of fire disturbance. This information will have great implications for better understanding the response to and recovery from pine bark disturbance, particularly for restoration ecology and forest management.

OP20: How do intrinsically disordered hub proteins achieve broad binding specificity?

Bowman, Grant



Abstract. When multiple intracellular signaling pathways intersect, complexity is often streamlined by filtering these pathways through a single hub with the capacity to bind many different proteins. For many hubs, protein binding occurs through an intrinsically disordered domain. How do intrinsically disordered proteins achieve specific binding to a broad range of proteins with structurally divergent binding interfaces? The goal of this project is to use a combination of genetic, biochemical, and biophysical approaches to address this question. The focus of these studies is a bacterial hub protein called PopZ, but the fundamental advancements in our understanding of this type of binding interface will be directly relevant to other intrinsically disordered hub proteins, including several that are widely studied in cancer biology.

OP21: Inhibition of UGCG Expression Via MicroRNA and SiRNA

Hinshaw, Jesse, and Anya Lyuksyutova



Abstract. Metabolic syndrome occurs in 24%-27% of Americans and is associated with type 2 diabetes mellitus and atherosclerotic cardiovascular disease (ASCVD). Nonalcoholic steatohepatitis (NASH) occurs in 33% of metabolic syndrome patients and is a condition in which the liver becomes inflamed and damaged due to its high fat content. This condition can lead to fibrosis and cirrhosis of the liver, becoming life threatening. Few methods of treating NASH exist, however one potential avenue of treatment is inhibition via microRNAs or targeted siRNAs of the gene UGCG for glucosyl ceramide synthase (GCS). High levels of UGCG are associated with development of NASH, making it a prime target for inhibition. My project has focused on creating DNA constructs using an episomal vector (EEV) with GFP fused to the UGCG 3’ untranslated region (UTR) from both mice and humans. Additional constructs will be required that contain the short sequence that encodes miR-200, a microRNA that has a binding site in the 3’UTR of UGCG or three siRNA that have been designed to target the 3’UTR of UGCG. These constructs are then transfected into HEK

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(Human Embryonic Kidney) cells to determine if direct interaction of microRNA with the 3’UTR of UGCG is responsible for the reduction in expression of UGCG based on the known presence of microRNA binding sites present on the 3’ UTR. This will be evaluated using microscopy to determine if GFP levels are down-regulated after the transfection of microRNAs. The miR-200 or siRNA construct can then be transfected into mice with metabolic syndrome and NASH to monitor the effects of this microRNA in a model mammalian system. The completion of this project will yield important information regarding the inhibition of UGCG in mammalian cells and may lead to treatments for NASH and metabolic syndrome patients.

OP22: A regulatory genetic network in C. elegans contributes to epidermal structural integrity during development

Bernazzani, Sarina, Melissa Kelley, and David S. Fay



Abstract. During development, biomechanical forces contour the body and provide shape to internal organs. We previously demonstrated that a regulatory network involving the conserved proteins SYM-3/FAM102A, SYM-4/WDR44, and MEC-8/RBPMS are required to maintain epidermal integrity in response to biomechanical forces in C. elegans embryos. Embryos and larvae that are doubly mutant for mec-8; sym-3 or mec-8; sym-4 have defective anterior morphology, called the Pharynx Ingressed, or Pin phenotype. We showed that MEC-8 regulates the alternative splicing of multiple structural proteins including a fibrillin-like protein, FBN-1, which acts specifically in epidermal cells and is a component of the apical extracellular matrix (aECM or sheath). FBN-1 functions to withstand a variety of biomechanical forces acting on the epidermis, including an internally directed pulling force exerted by the pharynx as it undergoes stretch and elongation. Our current studies indicate that FBN-1 may attach directly to epidermal cells via integrins, which have not previously been reported to act within the epidermal aECM of embryos. Consistent with this, mutation of RGD (integrin-binding) sites in FBN-1 leads to molting defects. In addition, mutations in fbn-1, sym-3 and sym-4 can enhance the “notched-head” phenotype (seen in integrin mutants) and inhibition of integrin subunits enhances Pin in sym-3 and sym-4 mutant backgrounds.

Our studies further indicate that SYM-3 and SYM-4 act in a parallel pathway to MEC-8 and may promote the trafficking of structurally important proteins in epidermal cells. Functional SYM-3 and SYM-4 fluorescent reporters localize to vesicle-like structures at or near the plasma membrane in epidermal cells. Furthermore, SYM-3 and SYM-4 act in a common genetic pathway with the RAB-11 GTPase, a known regulator of endocytic recycling and exocytosis, which has been shown to regulate integrin localization in other systems. In other studies, we have identified novel components of the aECM that function with FBN-1 to stabilize epidermal architecture during embryogenesis. Taken together, we have identified a network of genes that are required to maintain epidermal architecture in response to a variety of biomechanical forces during embryogenesis.

OP23: Conserved ankyrin repeat proteins and NIMA kinases regulate extracellular matrix remodeling and intracellular trafficking in Caenorhabditis elegans

Lažetić, Vladimir and David S. Fay


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Abstract. Molting is an essential developmental process in nematodes during which the epidermal apical extracellular matrix, the cuticle, is remodeled to accommodate further growth. Using genetic approaches, we identified a requirement for three conserved ankyrin repeat rich proteins, MLT-2/ANKS6, MLT-3/ANKS3 and MLT-4/INVS, in Caenorhabditis elegans molting. Loss of mlt gene function results leads to severe defects in the ability of larvae to shed old cuticle and results in developmental arrest. Genetic analyses demonstrate that MLT proteins functionally cooperate with the conserved NIMA kinase family members, NEKL-2/NEK8 and NEKL-3/NEK6/NEK7 to promote cuticle shedding. MLT and NEKL proteins are specifically required within the hyp7 epidermal syncytium and fluorescently tagged mlt and nekl alleles are expressed in puncta within this tissue. Expression studies further show that NEKL-2–MLT-2–MLT-4 and NEKL-3–MLT-3 colocalize within largely distinct assemblies of apical foci. MLT-2 and MLT-4 are required for the normal accumulation of NEKL-2 at the hyp7–seam-cell boundary and loss of mlt-2 causes abnormal nuclear accumulation of NEKL-2. Correspondingly, MLT-3, which binds directly to NEKL-3, prevents NEKL-3 nuclear localization, supporting the model that MLT proteins may serve as molecular scaffolds for NEKL kinases. Our studies additionally show that the NEKL-MLT network regulates early steps in clathrin-mediated endocytosis at the apical surface of hyp7, which may in part account for molting defects observed in nekl and mlt mutants. Taken together, our studies have identified a conserved NEKL-MLT protein network that regulates remodeling of the apical extracellular matrix and intracellular trafficking, functions that may be conserved across species.

OP24: Genetic Analysis in NimA-Related Kinase Pathways in C. elegans

Joseph, Braveen, Vladimir Lažetic, and David S. Fay



Abstract. The family of NimA-related kinases (NEKs) are highly conserved in eukaryotes where they have been implicated in variety of diseases and cellular functions. Nevertheless, the targets of NEKs, as well as the components of NEK signaling pathways, remain largely unknown. The C. elegans genome encodes for 4 NEK family members (NEKL-1/NEK9, NEKL-2/NEK8, NEKL-3/NEK6/7, and NEKL-4/NEK10). We recently reported a novel function for NEKL-2 and NEKL-3 in regulating the completion of C. elegans molting during larval development. Our published and recent studies indicate that incomplete molting stems, at least in part, from defects in endocytosis in the apical epidermis of C. elegans larvae, thus implicating NEKs in processes associated with intracellular trafficking. To gain further insight into the molecular, cellular, and developmental functions of NEK kinases, we have undertaken a number of approaches including genetic analysis. Using CRIPSR/Cas9 methods we generated a range of loss-of-function (LOF) alleles of nekl kinases. We find that weak hypomorphic alleles in nekl-2 and nekl-3, which display no defects as single mutants, are synthetically lethal, leading to near uniform larval arrest in nekl-2; nekl-3 double mutants. Using a semi-clonal F1 screen, we were successful in identifying ~12 suppressor mutations of varying strengths and we will describe progress on their molecular identification using whole genome sequencing. In addition, we are following up on preliminary data linking NEKL-2/NEK8 to the C. elegans bicaudal ortholog, BICD-1. Bicaudal proteins functions as molecular tethers that couple cargos, including intracellular vesicles, to microtubule motor proteins, such as dynein. Regulation of BICD-1 by NEKL-2 may thus account for defects in endocytosis and molting that occur in nekl-2 mutant strains.

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OP25: CARD9 Knockout Rescues Heart Function by Reducing Fibrosis and Hypertrophy in a Pressure-overload Model of Murine Heart Failure

Peterson, Matthew, Samantha Haller, Kayla Wilson, Paul D. Thomas, and Guanglong He



Abstract. Introduction. Hypertension can lead to left ventricular hypertrophy, hypertensive heart disease, and heart failure. The progressions of each are tied closely to elevated systemic and local inflammation. Caspase Recruitment Domain-containing Protein 9 (CARD9), a cytosolic protein expressed in macrophages and neutrophils, mediates secretion of inflammatory cytokines by these cells, which infiltrate the myocardium after injury and perpetuate a sustained inflammatory response. We reported rescued myocardial function by knockout of CARD9 (CARD9-\-) in high-fat diet feeding. Macrophage deletion has been reported to ameliorate the progression of TAC-induced left ventricular hypertrophy and fibrosis. Combined, these findings suggest that CARD9 may be a valuable target in a TAC model of heart failure.

Hypothesis. We hypothesized that knockout of CARD9 would rescue progression of TAC pressure-overload induced heart failure.

Methods. C57BL/6 wild-type (WT) and CARD9-\- mice were assigned to thoracic aortic constriction (TAC) or to sham (CONT). To assess heart function, fractional shortening (FS%), was measured by echocardiogram 1, 2, and 3 months post-surgery.

Results. FS% was compromised in WT-TAC compared to WT-CONT at 1 (35.2 vs. 46.7; p<0.0001), 2 (31.0 vs. 46.2; p<0.0001), and 3 months (30.8 vs. 45.2; p<.01), but only significantly so in CARD9-\-TAC at 2 months (38.6 vs. 45.5; p<0.05). At 3 months, mice were sacrificed. Heart weight/tibia length was elevated in WT-TAC compared to WT-CONT (13.57 vs. 7.65; p<0.0001), but not in CARD9-\-TAC. Histological results of Masson’s trichrome stain suggested hypertrophy in both TAC groups, but revealed less fibrosis in CARD9-\-TAC than in WT-TAC.

Conclusion. CARD9 is a novel target for TAC-induced cardiac dysfunction in part through reductions in fibrosis and hypertrophy. Further study is needed to investigate signaling transduction involved, as well as the extent to which CARD9-\- may be protective in order to determine if it is a potential target for intervention in hypertensive heart disease.

OP26: INBRE Supported Undergraduate Research at Northern Wyoming Community College District (NWCCD) Erickson, Ami Sheridan College, 3059 Coffeen Ave, Sheridan, WY 82801 Email: [email protected] Abstract. Two campuses with diverse faculty expertise and interests are able to involve undergraduate students in research projects due largely to the funding support from the INBRE CC Network grant. Northern Wyoming Community College district includes both Gillette and Sheridan College, and faculty on both campuses have utilized funding to encourage undergraduate research. Recent projects include “REDOX Reactions of Biometals in Simulated

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Body Fluid and Human Plasma” and “Studying Grapevine Cellular and Physiological Response to Abiotic Stress,” which is also a collaborative project with UW’s Dr. Sadanand Dhekney and partially funded by an INBRE CC collaborative grant. Current progress and results from these projects, as well as discussion of possible future projects, strategies and challenges will be presented. OP27: A Plan for a Long Term Investigation of Human Exposure to West Nile Virus in Fremont County, Wyoming

Watkins Joy, Kaylan Schilling, Adam Conner, Shanda Barlow, Grant Hosking, Kelvin Kinyatta with Steven McAllister

Central Wyoming College, 2660 Peck Avenue, Riverton, WY 82501


Abstract. West Nile virus (WNV) was originally discovered in Uganda in 1937. WNV belongs to the Flaviviridae family and is in the same genus as Dengue Fever, Tick Borne Encephalitis, Zika virus, and Yellow Fever. Individuals contracting the virus may be asymptomatic, experience mild symptoms of fever, malaise, or develop a severe disabling illness such as meningitis, encephalitis, or polio like paralysis. There is currently no human vaccine available for WNV. WNV was first detected in the United States in 1999. The virus rapidly migrated across the contiguous 48 states over the course of ten years. WNV last reached epidemic proportions in Wyoming in 2007 when 185 humans were diagnosed with WNV, including two fatalities. The majority of WNV infections of that year were found in Fremont County with 118 infected. Our previous serosurveys conducted in 2011 and 2012 indicated the majority of the population in Fremont County had not been exposed to WNV. We also identified three subjects with abnormally high levels of IgM antibodies five years after initial infection. Our plan is to conduct additional surveys over multiple years to continue to monitor levels of exposure to WNV and to investigate our intriguing observation.

Saturday September 17, 2016: Update from the Director of the Developmental Research Projects Program (DRPP)

Fay, David S.



Abstract. The major goals of the Developmental Research Projects Program (DRPP) are to promote high-quality health-related research and to strengthen the funding, training, infrastructure, and stature of the biomedical sciences at UW and partner institutions throughout the state. As such, DRPP funds are viewed as investments, with expected returns of high-quality publications, extramural grants, and meaningful educational experiences for graduate and undergraduate students. In order to accomplish these goals the DRPP seeks to attract the deepest pool of applications possible by drawing from three broad and inclusive areas of biomedical research: (1) cardiometabolic syndrome; (2) technology for chronic disease research and therapeutics; and (3) genetic and genomic analysis in health-related research. Several distinct funding mechanisms provide different levels of support to investigators including Thematic awards, which allow for a maximum of $75K/year for three years, and Pilot Project

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awards, which provide $25K/year for two years. Both Thematic and Pilot proposals are sent out for external review and must demonstrate clear potential to become independent extramurally funded projects. In addition, the DRPP funds Collaborative awards, which promote scientific partnerships with faculty at UW and community colleges, and graduate student fellowships, which support Ph.D. research in the biomedical sciences at UW. Currently the DRPP supports nine Thematic, five Pilot, and four Collaborative projects in addition to 24 graduate students.

The DRPP also recognizes the importance of mentoring faculty who receive Thematic and Pilot awards, most of whom are non-tenured Assistant Professors. To address this critical area, the DRPP has recently launched a new set of initiatives. These include practical mentoring exercises, such as the chalk talk series, at which INBRE faculty present their plans for grant submissions and research. In addition, the DRPP underwriting valuable services including professional scientific editing and pre-review of grants and manuscripts. The purpose of these initiatives to help INBRE PIs be as productive and successful as possible and to give INBRE PIs a competitive advantage as they transition to independence.

An overview of the biomedical startup model; How venture capital and the digital age are impacting healthcare innovation.

Fluet, Gregory

Consultant at Sapient Capital Management, L.L.C

Abstract. Going from an idea to a billion dollar company (or at least something more than what was invested). A brief history of healthcare innovation, venture capital and how biomedical research can evolve into a business. In addition to the traditional startup model, we will discuss how the digital age is impacting capital formation for new healthcare startups. In other words, how Aristotle, Thomas Aquinas, the age of exploration and the Dutch East India Trading Co., led to surgical robots, cardiac stents, Stemcentrx, Theranos and FitBit. (All in 40 mins.)

OP28: MetabocinTM : A novel agent to ameliorate metabolic syndrome

Markert, Laurel, Baskaran Padmamalini, and Baskaran Thyagarajan



Abstract. Obesity foreshadows metabolic diseases. An imbalance between energy intake and expenditure causes obesity. Currently, one third of the US population is obese and the growing adult and childhood obesity necessitates the development of a novel strategy to counter obesity and its comorbidities. Preliminary research from our laboratory suggest that supplementation of MetabocinTM, an agent that specifically activates the transient receptor potential vanilloid subfamily 1 (TRPV1) in high fat diet counters diet-induced obesity. MetabocinTM prevented weight gain caused by feeding high fat diet in the wild type mice. MetabocinTM neither altered the food or water intake in wild type or TRPV1-/- mice nor protected TRPV1-/- mice (that genetically lack TRPV1 protein) from the effects of high fat diet. MetabocinTM also significantly suppressed impairment of glucose handling, hyperglycemia, insulin resistance, hypertension and hypercholesterolemia associated with high fat diet-induced obesity. High fat diet

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significantly decreased the left ventricular internal diameter (diastolic) and fractional shortening and dietary MetabocinTM antagonized these effects in the wild type mice. Analyses of mechanisms involved in the effect of MetabocinTM suggested that the drug upregulated the expression and activity of cellular metabolic sensor sirtuin-1. Our data suggest that MetabocinTM antagonizes metabolic syndrome by activating TRPV1 expressed in adipose tissues. Further studies are underway in evaluating the signaling pathways by which TRPV1 activation regulates sirtuin-1 expression and activity.

OP29: Sirtuin-1 and Regucalcin Emerge as a Potential Target to Defy Redox Stress–Induced Accelerated Aging

Nazminia, Kara, Justine Frantz, Baskaran Padmamalini, and Baskaran Thyagarajan



Abstract. Accelerated aging occurs during space flights due to high-energy radiation induced-oxidative stress. Recent research suggests that oxidative stress downregulates the expression of cellular proteins, which regulate cellular senescence and aging processes. Senescence marker protein-30 (SMP-30; regucalcin) and sirtuin-1 (SiRT-1; a protein that acts as a cellular sensor for energy and regulates aging) are important biomarkers, which are down regulated upon oxidative stress. We hypothesized that SMP-30/SiRT-1 overexpression/activation attenuates redox stress, and oxidative stress mediated aging process. To mimic the effects of oxidative stress caused by high-energy radiation, we used hydrogen peroxide (H2O2; an oxidizing agent) for our experiments. Our preliminary data suggest that H2O2 treatment suppressed the expression of senescence markers SiRT-1, SMP30 and p53 in transient receptor potential vanilloid subfamily 1 (TRPV1) expressing human embryonic kidney 293 (HEK293) cells and that overexpression of SMP-30 protected the expression of SiRT-1 and p53 from the effect of H2O2. Effect of capsaicin (TRPV1 agonist) and resveratrol (SiRT-1 activator) on H2O2-induced effect on SMP-30 and SiRT-1 expression in HEKTRPV1 cells will be evaluated.

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Poster Abstracts (Berol lodge Conference Room)

P1: Native bee diversity in burned and non-burned areas of the Rocky Mountain Forest System and the importance post-fire succession in conservation

David, Kendra, and Hayley C. Lanier



Abstract. Fire management is an important factor to native bee conservation. Burned dead trees are important nesting sites for cavity dwelling bees, and bare ground that can result from fires is important to ground dwelling bees Allowing natural fires to occur in our Rocky Mountain Forests may be an important factor in increasing native bee populations, as larger populations of native bees have been found in early successional stages of post-fire forests in other forest systems This information may be critical to future fire management practices in order to conserve native bees. This work focused on the two recent fire disturbances on Casper Mountain (2006 and 2012) that allowed for a broad focus on responses in the native bee populations throughout the Rocky Mountain Forest. It also allowed for comparison of native bee populations in Rocky Mountain Forest ecosystems with different post-fire succession times. Because fire opens up more areas to flowering plants, I hypothesize that more native bees will be found in early post-fire succession (East Casper Mountain) than later post-fire succession (West Casper Mountain) or non-burned (control) areas. I tested this hypothesis by collecting bees across burned and adjacent control areas over the summer of 2016. The total number of bees collected from Casper Mountain was 4690, with most belonging to Halictus (sweat bees), Ceratina (small mining bees), Lasioglossum (sweat bees), and Bombus (bumble bees). Overall, bees were more abundant in burn areas than in the control area (p = 4.248e-06) and exhibited higher family-level diversity in burned areas (p = 8.885e-05). The local impact of higher bee diversity also appeared extended to adjacent plots, with control plots near the most recent burn more closely resembling the burn in terms of abundance and diversity. This is important information for fire management practices in regards to native bee conservation. P2: Genetic analyses to determine road-barrier effects on small mammal populations

Diesburg, Laura M. and Hayley C. Lanier



Abstract. Roadways are viewed as an absolute necessity all over the world. They provide a means to allow us to travel quickly and easily across distant regions. Unfortunately, the impact these roadways have on wildlife such as small mammals, specifically their impact on gene flow, is generally poorly known. We used genetic analyses to look at the impact of the John D. Rockefeller Jr. Memorial Parkway in Teton County, Wyoming, on the long-term movements of Red-backed Voles (Myodes gapperi) and Deer Mice (Peromyscus maniculatus). Both Red-Backed Voles and Deer Mice were analyzed in order to get a comparison between two species with different dispersal capabilities and habitat preferences. We extracted DNA from 52 Red-

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backed Voles and 6 Deer Mice to examine the impacts on population structure. Although the final sample sizes were small (cytochrome b was sequenced for 14 and 6 individuals, respectively), the results are suggestive of a decrease in gene flow across the road. Future analyses will focus on using microsatellite data and additional Deer Mice samples to provide a detailed look at variation.

P3: Caulobacter PopZ forms an intrinsically disordered hub in organizing bacterial cell poles

Holmes, Josh, and Grant Bowman



Abstract. Despite their relative simplicity, bacteria have complex anatomy at the sub-cellular level. At the cell poles of Caulobacter crescentus, a 177 amino acid protein called PopZ self-assembles into three-dimensional polymeric superstructures. Remarkably, we find that this assemblage interacts directly with at least eight heterologous proteins, which are involved in cell cycle regulation and chromosome segregation. The binding determinants within PopZ include 24 amino acids at the N-terminus, a 32 amino acid region near the C-terminal homo-oligomeric assembly domain, and portions of an intervening linker region. Together, the N-terminal 133 amino acids of PopZ are sufficient for interacting with all heterologous binding partners, even in the absence of homo-oligomeric assembly. Structural analysis of this region revealed that it is intrinsically disordered, like p53 and other hub proteins that organize complex signaling networks in eukaryotic cells. Through live cell photobleaching, we find rapid binding kinetics between PopZ and its partners, suggesting that many pole-localized proteins become concentrated at cell poles through rapid cycles of binding and unbinding within the PopZ scaffold. We conclude that some bacteria, like their eukaryotic counterparts, use intrinsically disordered hub proteins for network assembly and sub-cellular organization.

Acknowledgements

Wyoming INBRE is supported in part by a grant from the National Institute of General Medical Sciences (2P20GM103432) from the National Institutes of Health. Any opinions expressed or results presented are solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Special thanks to the administration and staff of the University of Wyoming/ National Park Service Research Station for their assistance in organizing and running the conference.

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