PROCEEDINGS 91 ANNUAL MEETING 35STProceedings of the 91 Annual MeetingstVirginia Academy of ScienceVirginia Polytechnic Institute and State UniversityMay 22-24, 2013Aeronautical and Aerospace SciencesEVOLUTION OF U.S. MILITARY AIRCRAFT. M. Leroy Spearman & Robert W.Heath, Langley Research Center, Hampton, VA & Independent Contractor, NewportNews, VA. The airplane was accepted as a part of the military system in August 1907with the establishment of an Aeronautical Division of the U.S. Army Signal Corps Anairplane was procured from the Wright Brothers and flight training began at FortMeyers, VA. In August 1909 an improved Wright Flyer was formally accepted andwas identified as `Airplane No. 1`.Another U.S, designer was becoming well knownand in 1911 a Curtiss airplane was the second airplane to be sold to the U.S. Army. Thefirst appropriation for military aeronautics was in the 1912 War Department budgetfor the sum of $125.000 and orders were placed three Wright airplanes and two Curtissairplanes. European countries, on the brink of World War I, were making progress inthe development of military aircraft. The U.S. entered the war in1917 but had nomilitary aircraft. U.S. volunteer pilots did gain experience while flying British andFrench aircraft and the U.S. industry gained experience by manufacturing someEuropean airplanes. Following the war, the U.S. acquired a number of British andFrench airplanes for the Army Air Service. Native aircraft were soon to appear,primarily trainer airplanes and early Martin bombers. Concern that the U.S. waslagging in aeronautical research lead to the establishment of the National AdvisoryCommittee for Aeronautics (NACA) An aeronautical laboratory was constructed atLangley Field, VA and, since 1920, the laboratory has contributed to the technologicalgrowth of U.S. military aircraft as they evolved from the classic propeller–drivenfabric-covered biplane to the all-metal monoplanes In the 1940`s jet-propulsion wasintroduced and the flight envelop was changed to include transonic and supersonicflight More recently some military aircraft designs have employed stealth technologythat is intended to make the aircraft less visible to most tracking systems – exampleis the F22 Raptor stationed at Langley Field, VA.DELINEATION OF AERODYNAMIC PROBLEMS WITH WIND TUNNEL TESTS. M. Leroy Spearman, Langley Research Center, Hampton, VA. The use of windtunnels in the development of new aircraft and missile concepts is well known. However the wind tunnel is also useful in establishing the characteristics of existingaircraft or missiles. As flight speeds increased compressibility of the air became amajor problem. Regions of sonic flow caused some separated flow that reduced thecontrol effectiveness. Wind tunnel tests indicated that blunting the trailing edge of flap-type controls to reduce the surface slope would eliminate the separated flow and restoreVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss136 VIRGINIA JOURNAL OF SCIENCEthe control effectiveness. Blunted trailing edges. were applied to the ailerons and thehorizontal tail of several high-speed aircraft and the roll control and pitch control wasrestored. Further research lead to the development of the all-moving tail that could bemoved symmetrically for pitch control and differentially for roll control. In1949Convair produced the delta-winged F-102 supersonic aircraft that had difficulty inachieving the desired supersonic speeds. Production was halted and NACA-Langleyundertook an investigation of the concept. A major revision involved the applicationof the transonic area rule that resulted in sliming the body in the region of the wingattachment- the coke bottle` contour. The transonic drag rise was reduced, supersonicspeed was achieved and production was resumed. Wind tunnel tests in conjunctionwith analytical results can be useful in determining the performance characteristics ofexisting aircraft. One example is the Soviet MiG-25 airplane first seen in a1967airshow. The airplane was said to be capable of Mach number 3 flight. There wassome doubt of this performance in the Western World Estimates of the overallplanform and profile shape could be gotten from airshow photographs. A study wasundertaken at NASA-Langley to determine the cross-sectional shape that would berequired to meet the speed. A wind tunnel model was built and tests verified the speed.In 1976 a MiG-25 defected to Japan and a detailed examination of the airplaneindicated that the NASA-Langley model was essentially correct.VARIATIONS OF VARIABLE GEOMETRYAIRCRAFT. M. Leroy Spearman,Langley Research Center, Hampton, VA. The geometric shape of an aircraft usuallyconsists of a fuselage, wing and tails and a propulsion system. The fuselage providesthe space required for the cargo, equipment and crew. The wing provides the liftrequired to sustain flight. The tails provide for stability and control The propulsionsystem provides for forward movement of the aircraft. The shape and arrangement ofthese components are designed in such a way as to assure that the missionrequirements for the aircraft are met Thus for different mission requirements adifferent geometric shape is required. This lead to the concept of a single aircrafthaving variable geometry that would be suitable for multiple missions An example ofsuch an aircraft is the General Dynamics F-111 in which the wing panels can be set ata low sweep position for take-off and landing and low-speed flight – for high-speedflight the wing panels are rotated back to a high sweep angle. With the F-111 the entirewing panels are moved using inboard pivots. Some research has been done withoutboard pivots in which only a portion of the wing panel is swept. And some researchhas been done with only a single pivot wherein the entire wing is rotated into what isknown as a skewed wing. Several aircraft having variable sweep wings have beenproduced in the U.S., and in several European countries and in the Soviet Union.Agriculture, Forestry, and AquacultureDOES PROXIMITY TO COMPOST PILES AFFECT FLY POPULATIONS IN HIGHTUNNEL TOMATO? Mark Kraemer, Walter Mallory & Steven Pao, AgriculturalResearch Station, Virginia State University, Petersburg, VA 23806. Compost iscommonly used in organic vegetable production but it is not known if nearby compostpiles pose a health threat. This study evaluated fly populations in two high tunnels,Virginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 37STwith and without compost. Both tunnels had 3 rows of bush tomatoes. The treatmenttunnel had tubs of manure and straw placed in each corner. Flies were monitored overa period of 10 weeks using 15 x 25 cm yellow sticky cards, one in each quadrant ofeach tunnel. The control tunnel was 5 m distant to the treatment tunnel. Four types offilth fly were found in the tunnels, plus hover flies and Tachinid parasitoids. The lesserhouse fly (Fannia canicularis) was the most numerous and increased over time. Otherflies were the common house fly (Musca domestica), stable fly (Stomoxys calcitrans),and black garbage fly (Hydrotaea aenescens). There were no significant differencesbetween filth fly populations between tunnels. Thus, a distance of greater than 5 mfrom compost piles is required to reduce filth fly populations.EFFICACY TESTS OF VERNONIA LEAF EXTRACT AND FRACTIONS TOCONTROL SMALL RUMINANTS GRASTROINTESTINAL PARASITES (GIN). F. Djibodé Favi, A. B. Yousuf, S. A. Wildeus, W. H. Mallory & M. E. Kraemer,Agriculture Research Station, Virginia State University, Petersburg VA 23806. Gastrointestinal nematodes (GIN), were the greatest danger to the small ruminantindustry worldwide and Haemonchus contortus (Rudolphi) Cobb is one of the mostpathogenic to ruminants. It piercing the abomasum causes a number of significantcomplications that can lead to death. Leave from vernonia varieties were used forthousand years in the old world to cure GIN in animal and human. Tested extracts weremostly inconclusive in 3600 documented records. The objective was to test fractionsof leaf extract to determine efficacy conditions of Vernonia galamensis (Gilbert) fromAfrica. Aqueous and oily fractions from leave extract were parted to obtain concentratefractions. Fractions were applied to larva in 12- cell plate (10 larva/µl) and larvalmortality was accessed for three days. The controls were 30% solvent and commercialdewormers Cydectin. Eggs filtered from feces of infested animal were diluted to 50eggs/100µl were tested in 12-cell plate. Hatching were monitored from 1-3 days. Cydectin kills L3 larva within 1-3 days post treatment (dose dependent). Egg leaks andthe shell has a round hole. Fraction A kill larva between 1-2 days post treatment. Larva vomit struggle and die. Egg shows atypical frozen yolk. Fraction A was selectedfor further studies. TEMPERATURE EXTREMES OF THE BROWN MARMORATED STINK BUG(Halyomorpha halys). John D. Aigner, Thomas P. Kuhar, Ashley K. Lohr, &Christopher R. Philips, Virginia Tech, Department of Entomology, 216 Price Hall,Blacksburg, VA 24061. Today’s global economy continues to grow, increasing thechances that invasive species become established in the United States. Brownmarmorated stink bug (BMSB), Halyomorpha halys (Stål), is a major agricultural pestthat was accidentally introduced in Pennsylvania and is rapidly spreading throughoutthe North America. This project was designed to better understand the bug’sphysiology and its ability to withstand a range of temperatures so that we can betterunderstand the potential climatic and geographic limitations of the insect. This wasdone by monitoring the lethal high temperature of BMSB and by identifying thesupercooling points (SCP’s) associated with BMSB at several intervals throughout theyear. Field-collected and lab-reared BMSB egg masses, nymphs, and adults wereexposed to temperatures ranging from 35 to 45 C (95 to 113 F) for four hours or untilo oall bugs died, whichever was shorter. Temperatures above 41 C (104 F) resulted ino oVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss138 VIRGINIA JOURNAL OF SCIENCEsignificant mortality of all stages of BMSB. All life stages of BMSB survivedexposures to temperatures below 40 C (104 F). Supercooling points were determinedo oto range from -8°C (17.6°F) to -22°C (-7.6°F) using field collected BMSB. TheseSCP’s change throughout the year however, no discernible pattern was recognized.These SCP’s have been monitored from December of 2011 through April 2013.OMNIVORY AND PREDATOR FUNCTION IN AGRICULTURAL ECOSYSTEMS. C. R. Philips, T. P. Kuhar & D. A. Herbert, Dept. of Entomology, Virginia Tech,Blacksburg, VA 24061. Understanding how buckwheat (Fagopyrum esculentumMoench) companion plantings impact natural enemy abundance, and pest suppressionin adjacent crops, may provide alternative control methods thereby slowing the rate ofinsect resistance to pesticides and reducing pesticide exposure to the applicator andsurrounding habitats. Buckwheat has been widely used as a companion planting onvegetable farms, vineyards, and orchards to supply nectar and pollen and encouragearthropod natural enemy populations. However, scientific data demonstrating the truebiological control benefit of such companion plantings are scarce. The primaryobjective of this project was to determine the impact of buckwheat companion plantingon lepidopteran pest abundance, parasitoids abundance, parasitism, and the predatorand parasitoid communities in collards. Overall abundance of pest caterpillars did notvary with distance and imported cabbageworm was the most abundant species.Parasitism of imported cabbageworm larvae did not differ with distance from availablenectar sources. Abundance of parasitoids was not significantly different with distance,and the average rate of parasitism was 50%. Even at low numbers, it is likely thatparasitoids play an important role in lepidopteran population dynamics. It also appears,however, that generalist predators may also be playing an important role. Futureresearch will investigate the biology, habitat use, and predation impacts of some ofthese predators.MONOSEX CULTURE OF FRESHWATER SHRIMP (Macrobrachium rosenbergii). Brian Nerrie, Virginia Cooperative Extension, Virginia State Univ., Petersburg VA23806. In Virginia sustainable production of freshwater shrimp (Macrobrachiumrosenbergii) has been shown to be an excellent agriculture diversification in thetobacco growing region, especially for limited-size farms. Research and demonstratedfarming success in Israel, India and Bangladesh indicated improved yields for all-maleshrimp populations. Research in Mississippi and Kentucky confirmed higher yields. Benefits and disadvantages of all-male or all-female monosex populations arediscussed. Adoption of all-male production practices in Virginia would result inenhanced per acre production levels and larger individual animal size. However,multiple factors need to be considered by the private sector. The factors impacting onprofits are: availability of all-male juveniles, stress from time and distance to stocking,survival rate, higher juvenile costs and whether consumer preference for higher pricesfor larger shrimp exceeds a larger quantity of smaller shrimp of the same value. Profitability will be the major determinant if monosex culture is adopted.DISEASES OF CHANNEL CATFISH REARED IN CAGES IN VIRGINIA. DavidCrosby, Virginia Cooperative Extension, Virginia State University, PO Box 9081,Petersburg, VA 23806. The main production method for growing catfish in VirginiaVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 39STis using cages in farm ponds. Most catfish fingerlings used for stocking cages comesfrom out of state commercial farms. After the initial stocking, parasitic and bacterialoutbreaks can occur up to three days later. Aeromonas hydrophila and Flavobacteriumcolumnare are the most common bacterial pathogens that cage producers willencounter. These pathogens are quite common in the aquatic environment and are theleading causes of disease problems in cages. Columnaris infections can occur withinone day after stocking catfish into cages. A highly pathogenic strain of Aeromonashydrophila that originated in Alabama has been diagnosed from catfish stocked incages in Virginia. A study conducted at Virginia State University (VSU) found thatcatfish fingerlings imported from commercial farms usually had external parasites.Trichodina was found on catfish from four of the six commercial catfish fingerlingshipments to VSU from 2007 – 2009. All shipments had Henneguya, the causativeagent of proliferative gill disease. Trichodina is the most common parasite on catfishin cages that causes mortalities. In February of 2013, winter saprolegnasis or winter killoccurred in a caged catfish operation resulting in the high mortalities. This incidentoccurred in a pond with very low alkalinity, less than 17 ppm. Anytime a fish farmeris stocking catfish into cages from a commercial fish farm may increase the potentialrisk of a disease outbreak.CAN STANDARD SOIL TESTS PREDICT ORGANIC BLUEBERRYPRODUCTION IN PLOTS WITH DIFFERENT SOIL AMENDMENTTREATMENTS? Roman J. Miller. Dept. of Biol. Eastern Mennonite University,Harrisonburg, Virginia 22802. What best predicts blueberry productivity: Soil tests?Foliar analysis? Plant growth? This question was examined in four blueberry plots withdifferent mulches incorporated into their soil: organic horse manure (OHM), organicsheep manure (OSM), organic pine straw (OPS), and organic Planter Choice (OPC).Soil tests revealed normal or high nutrient levels in all plots. pH values were nearoptimal in OHM and OPC, but too high in OSM and too low in OPS. Soil respiration,foliar nutrient and chlorophyll values were within expected ranges or optimal in allplots. Growth parameters such as bushiness for Duke was greatest in OPS; whileBluecrop bushiness was greatest in OSM. Average plant height for OPS and OPCbushes was 50% greater than OSM bushes and 100% greater than OHM bushes. Berryproductivity in the OPS and OPC plots doubled the OSM plot and exceeded the OHMplot threefold! While soil profiles and foliar analysis both failed to adequately predictberry productivity, simple bush height in unpruned young blueberry bushes emergedas the single best predictor of blueberry productivity. (Supported by USDA SpecialtyCrop Grant #2011-546)PostersANALYSIS OF ANTHOCYANINS AND TOTAL PHENOLIC COMPOUNDS INBLUEBERRIES: METHOD DEVELOPMENT AND PRELIMINARY RESULTS. A. Kniss, E. Harnish, S. Cessna & R. Miller, Departments of Biology and Chemistry,Eastern Mennonite University, Harrisonburg, VA 22802. Anthocyanidins and otherphenolic compounds are known to have antioxidant capacity. Blueberries have beenreported to be one of the richest sources of antioxidants among common fresh fruits.Interest in the micronutrients of blueberries has grown with increasing evidence ofVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss140 VIRGINIA JOURNAL OF SCIENCEpossible health benefits. In order to compare the antioxidant profile of differentvarieties of high bush blueberries growing in differing conditions, severalmethodologies were tested. We found that two different UV-Visible spectroscopy-based measures of antioxidants were manageable for testing large numbers of berries. These are the FRAP assay (ferric ion reducing antioxidant power) and the Folin-Ciocalteau assay of total phenolic compounds. Our initial findings with these twoassays indicate that Duke and Jersey varietals are lower in antioxidants than Blue Cropand Blue Gold. Several methods for HPLC quantification of acid hydrolyzedanthocyanidins were also tested; here we report the most successful extraction,hydrolysis and chromotography method. These three measures provide quantitativeanalysis of antioxidants in blueberry cultivars. Each method was developed to the pointof functional use, and will be used to determine health of organically grown blueberriesand compare them to conventionally grown blueberries. (Supported by USDASpecialty Crop Grant #2011-546)ABUNDANCE OF APPETITE REGULATORY FACTOR mRNA IS INFLUENCEDBY 3-HOUR FEED WITHDRAWAL IN CHICKENS SELECTED FOR HIGH ANDLOW BODY WEIGHT. W. Zhang, L. A. Nelson, P. B. Siegel. M. A. Cline, & E. R.Gilbert, Department of Animal and Poultry Sciences, Virginia Tech, Blacksburg, VA24061. Long-term (55+ generations) selection for low (LWS) and high (HWS) bodyweight in chickens has resulted in a 10-fold difference in body weight at selection ageand correlated responses in appetite and body composition. Some of the LWS areanorexic, while HWS are hyperphagic and obese. We hypothesized that differences inappetite regulatory factor mRNA between the lines would be accentuated by foodwithdrawal. Five-day old male LWS and HWS chicks (n=24) were randomly dividedinto 2 treatments; 1) continuous access to a starter diet or 2) 3 h feed withdrawal, afterwhich hypothalamus was collected. The mRNA abundance of AGRP, NPY, NPYR1,MC4, GLUT1, and FOXO1 was measured by real time PCR. Data were analyzed byANOVA using the Glimmix procedure (SAS 9.3). For AGRP, NPY and NPYR1, therewas greater (P<0.05) abundance in LWS than HWS, whereas for FOXO1 and MC4there was greater (P < 0.05) expression in HWS. Food withdrawal affectedhypothalamic NPY mRNA abundance, where NPY mRNA was greater in fed LWSchicks as compared with fed HWS (P<0.05) and fasted LWS chicks (P<0.05). Inconclusion, there was greater mRNA abundance of food intake-stimulatory factors inLWS and greater abundance of satiety-inducing factors in HWS. These results suggestthat differences in food intake between LWS and HWS are associated with differencesin the appetite circuitry of the hypothalamus and that these differences are accentuatedunder certain feeding conditions.Astronomy, Mathematics, and PhysicsAN INEXPENSIVE RADIO TELESCOPE IN A COLLEGE PHYSICS LAB, ANUPDATE. C. Crook & T.C. Mosca III, Dept. of Chem. and Phys., & Dept. of Math, Rappahannock Community College, Warsaw VA 22572. An amateur radiotelescopeon the Glenns campus of Rappahannock Community College was used by first-yearphysics students to conduct original research on solar radio frequency emissions. MostVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 41STnotably, RF signals were recorded that fit the characteristic profile of solar flareemissions that approximately coincided with a flare confirmed by NASA and otherradio observatories. Slight anomalies in the time that this flare was recorded werediscussed but not resolved. The project also provided a logical context for theapplication of physics theories that are not directly observable, such as energytransmission by magnetohydrodynamic waves through the solar plasma.FLUCTUATIONS IN A THERMOELECTRIC AND INFRARED POWERGENERATION. H. S. Mann , A. E.Masters & G. X. Scarel , James Madison1 2 1 1University, Dept. of Physics and Astronomy, Harrisonburg, VA 22807, & Custom2Thermoelectric, Bishopville, Maryland 21813. In recent years, the interest inalternative energies has risen greatly. Infrared radiation represents a significant portionof the solar spectrum which is worth exploiting. Capturing infrared radiation andconverting it into usable electricity has been demonstrated using power generators.However there is evidence that power generators interact differently with heat orinfrared radiation. This work addresses the fluctuations in voltage observed in a powergenerator interacting with heat in an open and isolated environment. The results arethen compared with the fluctuations in voltage generated by infrared radiation on apower generator in an open and isolated environment. The findings from thesemeasurements contribute to further understanding the distinction between infrared andthermoelectric power generation, as well as the fundamental physics of the powergenerators involved.A WISE SEARCH FOR EXCESS MID-INGRARED EMISSION FROM 100,000NEARBY M DEARFS IN THE SUPERBLINK PROPER MOTION SURVEY. C. A.O’Donnell , S. Lepine & B. Rojas-Ayala , Department of Astronomy, University1,2 2,3 2 1of Virginia, Charlottesville VA 22904, American Museum of Natural History, New2York NY 10024, & City University of New York, New York NY 10075. We searched3the Wide-field Infrared Survey Explorer (WISE) catalog to identify mid-infraredcounterparts of 100,000 M dwarfs within 100 parsecs of the Sun selected from theSUPERBLINK proper motion survey. An M dwarf with infrared excess can be a signof dust or planetary material that is re-emitting absorbed optical light; both possibilitieswould presumably increase a system's overall infrared flux compared to the expectedflux of the star alone. Additionally, such systems are likely to be very young. TheWISE survey covered 4 bands centered at 3.4, 4.6, 12, and 22 micrometers (W1, W2,W3, and W4 respectively). Since almost all of the M dwarfs did not have reliable W4magnitudes, we focused on the W1-W3 color and found that the vast majority of Mdwarfs had W1-W3 < 0.5. After systematic examination of all stars with redder colors(i.e., W1-W3 > 0.5), including visual verification of the WISE images, we haveidentified 172 M dwarfs with clear infrared excesses (0.5 < W1-W3 < 2.48) and 223additional candidates with possible but weaker infrared excesses (0.5 < W1-W3 <1.27). All these systems, especially the 172 high-excess systems, should be highpriority targets for follow-up observations, including exoplanet survey programs.Finally, we identified 5 stars with particularly high infrared excesses (1.0 < W1-W3 <2.48), and their spectral energy distributions were consistent with a dust model. Futureobservations will confirm the source of the excess emission.Virginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss142 VIRGINIA JOURNAL OF SCIENCETHE SIGNATURE OF RADIATIVE POLARITONS: DIELECTRIC FUNCTIONAND SIMULATED SPECTRA. Yosyp Schwab & Giovanna Scarel, James MadisonUniversity, Harrisonburg, VA 22807. Surface phonon polaritons have recentlyreceived a lot of attention for their role in heat transfer in nano-structured materials.However, much less attention has been given to radiative polaritons and their vital rolein determining the optical properties of insulating oxides and in infrared (IR) powergeneration. Here it is shown through experiment and simulation that radiativepolaritons radiate in the far-IR/microwave frequency region. In order to obtain thesimulated results, the classic dielectric function for insulating oxides was modified. Acontribution including the radiative polaritons was added, leading to an excellentagreement with the experimental spectra. This is the first time that the location of thefrequency range of radiative fields has been experimentally shown, as well as theexistence of the radiative fields themselves.INSTRUCTIONAL INSTRUMENTATION LAB EXPERIMENTS AT JAMESMADISON UNIVERSITY. Joseph D. Rudmin, & Anne Henriksen, Dept. ofIntegrated Science and Technology, James Madison Univ., Harrisonburg, VA 22807. James Madison University college students experience a series of laboratory activitiesillustrating critical concerns in designing laboratory experiments, and gain experiencewith common representative measurement instruments. The laboratory experiencesmaximize clarity of illustration of concepts, within a very modest laboratory budget. The laboratory experiences build on each other. National Instruments compact DAQhardware and LabView software are used for the most rapid data acquisition thatstudents collect.MATTER-ANTIMATTER ASYMMETRY AND NEW STATES OF MATTER WITHTHE BELLE EXPERIMENT. Leo Piilonen, Dept. of Physics, Virginia PolytechnicInst. and State Univ., Blacksburg VA 24061-0435. I introduce the broad topic ofelementary particle physics and how it helps us to understand the nature and workingsof the universe at its most fundamental level. I then illustrate this topic through thestudies of the difference in behavior between matter and antimatter and the search fornew states of matter as probed in the Belle experiment and its successor, Belle II,operating at the High Energy Accelerator Research Organization (KEK) in Japan.NUMERICAL MODELING OF THE CLIMATE CHANGE IMPACT ON RAREGENOTYPES IN NATURE. Charles L. Armstrong, Iordanka N. Panayotova, & LisaHorth, Dept. of Mathematics & Statistics & Dept. of Biological Sciences, OldDominion Univ., Norfolk, VA 23529. This study uses computer models created usingMATLAB software to analyze the effects of climate change on rare genotypes that areactivated by temperature-sensitive enzymes. The primary focus of this study is theeffect that these changes would have on the black-spotted mosquitofish (Gambusiaholbrooki). We also analyze the effects that these temperature increases and resultingdecreases in melanic mosquitofish inheritance will have on the overall mosquitofishpopulation. For this study we used a model previously published by Dr. Panayotovasimulates the life cycle of this fish. For this study we have created models that accountfor rising temperatures, yearly temperature fluctuations, the relationship between risingtemperatures and melanic inheritance, and different patterns of temperature increases.Virginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 43STOur results indicate that increasing temperatures will be devastating for the melanicmosquitofish population, resulting in extinction within 1000 years. Despite thesefindings, our simulations also indicate that if the temperature stabilizes before themelanic mosquitofish becomes extinct the melanic population will also stabilize nearthe frequencies present at the time the temperature stabilizes. (Supported by OldDominion University Spring 2013 Undergraduate Research Grant.)Biologywith Microbiology and Molecular BiologyG EN O M E M U T AT IO N M APPER: NEX T -G EN B IO IN FO R M AT ICSAPPLICATIONS FOR VIRAL PATHOGENS DETECTION ESPECIALLY HUMANADENOVIRUSES. Amirhossein Shamsaddini, Donald Seto, & Ancha Baranova,School of Systems Biology, College of Science, George Mason University. To date,applying high-resolution genomics and bioinformatics approaches has yielded in-depthand better views of the natural variation of Viral Pathogens. Emerging next-generationsequencing technologies have revolutionized the collection of genomic data forapplications in bioforensics, biosurveillance, and for use in clinical settings. However,to make the most of these new data, new methodology needs to be developed that canaccommodate large volumes of genetic data in a computationally efficient manner. Wepresent Two Different Applications of Computational Framework to analyzeSequences for rapid species identification and Evolution Patterns Analysis. Softwarefor our approaches is available at http//:ssb.gmu.edu/gmm.TRACKING BROWN ADIPOSE TISSUE IN VISCERAL FAT TISSUE OFPATIENTS WITH OBESITY. E. Dadkhah , R. Mehta , K. Doyle , A. Baranova,2 1, 2 1, 2 1,A. Birerdinc & Z. Younossi , Betty and Guy Beatty Center for Integrated2 1, 2 1 , 2 , 3Research, Inova Health System, Falls Church VA, Center for the Study of Chronic2Metabolic Diseases, School of Systems Biology, George Mason University, Fairfax,VA, & Center for Liver Diseases, Inova Fairfax Hospital. The Prevalence of obesity3is going to increase and is estimated to reach 42% of the population by 2030. Severalfactors are related to obesity. One of the attractive subjects of researches which is eventrying to use as a therapy for obesity these days is brown adipose tissue (BAT). Theimbalance between BAT and white adipose tissue (WAT) can induce obesity. BATpromotes energy burning that avoids obesity. In this study, the proportion of BAT toWAT DNA was studied in visceral adipose tissue to find out any relation between theamount of BAT and BMI. Ninety eight fresh fat tissues were collected from obesepatients at Inova Fairfax Hospital after collecting consent forms. PCR was performedwith 15ng of extracted DNA. As BAT differs from WAT by the large amount ofmitochondria, the ratio of mitochondrial DNA to genomic DNA considered as the ratioof BAT to WAT in the fat tissue. Two Cyclin B1 primers (CCNB1a & CCNB1C) andtwo mitochondrial primers (M12 & M13) were selected as genomic and mitochondrialDNA indicators respectively. C(t) values were normalized using CCNBb (IPC) forgenomic and M14 for mitochondrial DNA. Technique optimization efforts are ongoing.Virginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss144 VIRGINIA JOURNAL OF SCIENCEIN V IV O FLORESCENT IM AG IN G O F HO LLO W PO LY M ERICNANOCAPSULES LOADED WITH ALEXAFLOUR 750. Venkata SureshPatthipati , Christopher Osgood , James Swanson , Ramjee Balasubramanian , Kalpana1 1 1 2Mahadevan , & Sangbum Han , Department of Biological Sciences, ODU, Norfolk,2 2 1VA, Department of Chemistry and Biochemistry, ODU, Norfolk, VA. Nanoparticle2based imaging strategies hold significant promise in addressing current diagnostic andtherapeutic challenges. However, previous nanomaterials failed to clear effectivelyfrom the circulation via the renal route. This work demonstrates the use of highlyfluorescent, water-soluble, resorcinarene nanocapsules (122 nm) and their effectivenessin biomedical imaging. These hollow polymeric nanocapsules loaded with AlexaFluor 750 fluoresce brightly in the urinary bladder of mice under the in vivo fluorescentimager indicating that their major route of clearance is through the kidney. TEManalysis of the urine recovered from the mice showed the presence of intactnanocapsules. Such florescent nanocapsules can be very effective in visualizing theparts of the urinary tract. They are particularly useful in abdomino-pelvic surgeriesranging from caesarian delivery to laparoscopy. The use of these nanocapsules are notrestricted to the imaging as they could be loaded with a host of molecules making themefficient drug delivery vehicles in treating cancer and other pathologies.THE DIFFRENTIAL ROLE OF CBP UBIQUITIN LIGASE ACTIVITIES IN p53REGULATION. Oluwatoyin E. Akande & Steven R. Grossman, Dept. ofMicrobiology & Immunology, Virginia Commonwealth Univ., Richmond, VA 23298. The acetyltransferase CREB-Binding Protein (CBP) is a known transcriptional co-activator involved in p53 regulation and has been shown to encode cytoplasmic, butnot nuclear, E3 autoubiquitination and p53-directed E4 ubiquitin ligase activities. Inthis work, we sought to determine the regulation of differential ubiquitin ligaseactivities between nuclear and cytoplasmic CBP. Understanding how CBP regulatesp53 ubiquitination and stability may lead to potential therapeutics that can modulatep53 stability in cancer cells. We show that a nuclear interacting factor repressescytoplasmic CBP E3 (autoubiquitination) ligase activity in unstressed U2OS cells. Weincubated immunoprecipitated cytoplasmic CBP from U2OS cells with either nuclearlysates or CBP- immunodepleted nuclear fraction. Our results showed reduced CBP E3autoubiquitination in the mixtures when compared to purified cytoplasmic CBP alone.We further showed that DNA damage by doxorubicin (dox) in U2OS cells activatedE3 ligase activity in the nuclear fraction. These observations may provide a possibleexplanation for the inactive nuclear E3 ubiquitin ligase activity but active cytoplasmicE3 and p53-directed E4 ubiquitin ligase activities of CBP in unstressed cells. The doxdependent activation of nuclear CBP E3 autoubiquitination may result from doxdependent inactivation or re-localization of the nuclear inhibitor to other cellularcompartments. Further work will identify the nuclear interacting factor and in parallel,determine if the differential ubiquitin ligase activity observed between cytoplasmic andnuclear CBP is otherwise dependent on post-translational modifications of CBP itself.ENRICHMENT OF CAPTIVE SQUIRREL MONKEYS. LaCheryl A. Ball & Eric L.Walters, Department of Biology, Old Dominion University, Norfolk, VA 23529. Foodenrichment is a technique used by the zoo industry to promote overall wellness ofanimals in captivity. I measured responses of captive, Saimiri sciureus squirrelVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 45STmonkeys to food enrichment at the Virginia Zoo (Norfolk, VA). The research involveddetermining pre-treatment activity levels in order to test the effect of food enrichmenton post-treatment activity levels. I hypothesized that foraging and active behaviorswould increase as follows: baseline 104M ). We have prepared cryptands with association constants in the range of 10 to 10-1 4 6M with 4,4-bipyridinium compounds (“paraquats” or “viologens”) and have-1developed several efficient ways to functionalize the most powerful versions of thesehosts, the pyridino cryptands. These functionalized hosts now enable new avenues tocreate supramolecular polymeric structures beginning with both functionalizedpolymers as well as properly functionalized monomeric species. Combined withmodern polymerization methodologies these systems enabled our recent work directedtoward formation of star, block, graft and brush copolymers and also supramolecularpolymers. These procedures will be discussed along with characterization of theresultant novel materials.Virginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss170 VIRGINIA JOURNAL OF SCIENCEDESIGN AND SYNTHESIS OF ROMP IMIDAZOLIUM POLYMERS FOR USE ASACTUATORS. T. L. Price Jr. , U. H. Choi , D. Wang , Arunachalam Murugan , D.1 2 3 1V. Schoonover , M. Zhang , R. H. Colby , J. R. Heflin , R. B. Moore & H. W.1 1 2 3 1Gibson , Dept. of Chem., Virginia Tech, Blacksburg, VA, 24061, Dept. of Materials1 1 2Science & Engineering, Penn State University, University Park, PA, 16802, & Dept.3of Phys., Virginia Tech, Blacksburg, VA, 24061. Previous work conducted by ourgroup has established that properly designed poly(meth)acrylates containingimidazolium pendant groups will exhibit high single ion conductivities (~10 S/cm). 5Polymers of this type are known to be capable of electromechanical actuation. In thispresentation we will describe our design criteria for poly(norbornene)s containingimidazolium pendant units and discuss advantages of the system; additionally, singleion conductivities (~10 S/cm) and preliminary studies of their actuation behavior will5be presented. (Supported by: U.S. Army Research Office grant number W911NF-07-1-0452: Ionic Liquids in Electro-Active Devices (ILEAD) MURI).DESIGN OF MULTICHROMOPHORIC SYSTEMS FOR MOLECULARELECTRONICS. E. M. N. Ndip, N. Pandey & K. Cole, Dept. of Chem. HamptonUniversity, Hampton, VA 23668. This work is part of our ongoing research on thedesign, synthesis, and laser spectroscopy of multichromophoric (aryl styrylheterocyclic (thiophene, furan, and pyrrole) ð-centered compounds) systems formolecular electronics. Theoretical studies have elucidated the effects of tuning onseveral electronic properties and determined their suitability as materials for molecularelectronic devices. Ground state geometries of various derivatives were optimizedusing both semi empirical (PM3) and DFT-B3LYP with the 6-31G** basis set. Thelinear absorption spectra, and energy gaps were computed for the various molecularsystems in vacuum at both the semi empirical (ZINDO-CI) and ab initio (TDDFT)methods on the optimized ground state geometries. The computed maximumwavelengths for representative systems are in good agreement with the experimentaldata (405-438nm) obtained from fluorescence experiments cited in the literature. Thedata also shows reasonable agreement between the semi-empirical and ab intiocalculated ?. Band gaps determined from the TDDFT/B3LYP calculations are moreconsistent with the experimental values for the thiophene derivatives. (Supported byNSF CREST (HRD-0734635, HRD-0630372, and ESI-0426328/002))ELECTROCHEMICAL INTERACTIONS OF ACTIVATED CARBONS WITHAQUEOUS AND BIOLOGICAL MEDIA: AN ACID-BASE MODEL. M. M.Goldin , G. R. Garaeva , & N. V. Sklifosovky , Dept. of Biol. & Chem., Liberty1 2 2 1University, Lynchburg, VA 24502 & Institute for Emergency Medicine. The2adsorption of metal ions and organic compounds on activated carbons was measured2 4in 0.10 M Na SO against Ag/AgCl. Activated carbon samples caused a drift in the pHof distilled water, 0.10 M sodium sulfate, and other solutions in which they wereimmersed. For carbons with positive OCP, the pH drifted toward acidic values, whilefor negative-OCP carbons, toward basic values. If the carbon/electrolyte heterogeneousequilibrium is considered as an Usanovich acid-base pair, the electron acceptor is anacid, and the electron donor is a base. The “acidic” positively charged carbon immersedin a solution acidifies it, while the “basic” negatively charged carbon makes it morebasic, which is in agreement with empirical data. Identical unmodified AKU and AG-3Virginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 71STactivated carbon samples were immersed in phosphate buffer solutions; the acidicsolution caused a positive shift in the OCP of carbon, while the basic solution produceda negative OCP shift, also consistent with the Usanovich model. Finally, unmodifiedand oxidized SKT-6A carbon, as well as an iodide-doped polypyrrole (PPy) compositebased on SKT-6A and pure chloride-doped PPy, were titrated by the Boehm methodto determine surface functionalization. The amount of carboxyl groups on the surfaceof these materials gives a strong linear correlation with OCP and overall functionalgroup data corroborates the increase in Brønsted-Lowry acidic groups on carbonsurface with increasing potential. This study is supported by funds from the Center forResearch and Scholarship Fund of Liberty University.ç –DEAROMATIZATION: MODIFICATIONS OF ANILINE AND 2-2(DIMETHYLAMINO) PYRIDINE. J. A. Pienkos , A. Knisely , B. Liebov , V.1 1 1Teran , V. E. Zottig , M. Sabat & W. H. Myers , Dept. of Chem., University of1 1 2 3 1Virginia, Charlottesville VA 22903, Nanoscale Materials Characterization Facility,2Dept. of Materials Science and Engineering, University of Virginia, Charlottesville VA22903, & Dept. of Chem., University of Richmond, Richmond VA 23173. Organic3scaffolding with a transition metal fragment allows for rapid access to new compounds.3ç -coordination of an arene to a ð-basic {TpW(NO)(PMe )} fragmentrenders the2aromatic molecule “dearomatized.” A dearomatizedarene exhibitsreactivity that isdifferent than the reactivity of the free, unbound ligand.Ligand exchange between3{TpW(NO)(PMe )(ç -benzene)} and 2-(dimethylamino) pyridineorN,N-dimethyl2anilinederivatives, in the presence of acid, affords new ç complexed cationic species.2These species can react with various electrophiles, generating allyls, whichcanreactwith aromatic molecules to form a carbon-carbon sp -sp center. Additionally, the3 2ç -coordinated N,N-dimethyl anilinedimethyl aniline derivative can be cyclopropanated2and further modified through ring opening reactions. Liberation of the boundligandsaffords novel á,â-unsaturated cyclohexenones.S T E P : S O L A R T H E R M A L E LE C T R O C H E M IC A L P R O C E S S , ACOMPREHENSIVE PROCESS FOR UTILIZING RENEWABLE ENERGY TOREDUCE ANTHROPOGENIC CARBON DIOXIDE EMISSIONS. Stuart Licht,Baochen Cui, Baohui Wang, Jason Lau & Jessica Stuart, Dept. of Chem., The GeorgeWashington Univ., Washington, DC. 20052. STEP is a new process that converts andstores solar energy. STEP has been shown experimentally to function at much higherefficiencies then photovoltaic solar cells and to support current densities that are 1 to3 orders of magnitude greater than those achieved by photoelectrochemical watersplitting or photoelectrochemical solar cells. STEP utilizes a portion of sunlight todrive photovoltaic charge transfer, while the remainder of the sunlight is applied assolar thermal energy to heat and decrease the energy of enodothermic electrolysisreactions. By taking the reactants to a higher temperature, the voltage needed to drivethe endothermic electrolysis reaction decreases improving the efficiency of thereaction. A secondary advantage of going to these higher temperatures is the ability touse molten salts as the medium for electrolysis. Molten salts can support very highmolecular concentrations at the electrode surface allowing for higher current densitieswith greater efficiencies. Select molten salts can support very high voltages, areresistant to poisoning and can act as a thermal buffer to maintain the high temperatureVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss172 VIRGINIA JOURNAL OF SCIENCEneeded for the reaction. STEP has been shown to be a suitable alternative for a varietyof carbon dioxide releasing processes such as the production of iron, cement, hydrogen,desalinization, fuels, and wastewater treatment giving STEP the potential to be acomprehensive approach for reducing anthropogenic carbon dioxide emissions.PostersCHEMO- AND REGIOSELECTIVE DIBORATION OF ALLENES. Xi Guo,Amanda Nelson & Webster L. Santos, Dept. of Chem., Virginia Polytechnic Instituteand State University, Blacksburg, VA, 24061. Organoboron compounds are importantsynthetic intermediates in organic synthesis. Metal-catalyzed diboration reaction withcommercially available diboron reagents is a traditional method to introduce boronmoiety. However, most of the previous work was focused on symmetrical diboronreagents such as bis(pinacolato)diboron and bis(catecholato)diboron, a differentiallyprotected diboron reagent is employed in this work. Diboration of different allenes withit provides the borylated product chemo- and regioselectively. This work is funded byDepartment of Chemistry, Virginia Polytechnic Institute and State University and ACSPetroleum Research Fund.DESIGN, SYNTHESIS, AND BIOLOGICAL EVALUATION OF NOVELINHIBITORS OF SPHINGOSINE KINASE 2. Emily A. Morris , Neeraj N.1Patwardhan , Mithun R. Raje , Yugesh Kharel , Kevin Lynch & Webster L. Santos ,1 1 2 2 1Dept. of Chem., Virginia Tech, Blacksburg, Virginia 24061 & Dept. of1 2Pharmacology, University of Virginia, Charlottesville, Virginia 22908. Sphingosinekinase (SphK) has become a prevalent target for many disease states in recent years. SphK phosphorylates sphingosine to sphingosine 1-phosphate (S1P). SphK can befound in two isoforms, SphK1 and SphK2, and differ in their cellular location. Inmouse studies, inhibition of SphK2 showed an increase in the circulating levels of S1Pin blood. This increase of S1P has been shown to be anti-fibrotic, as it leads to adecrease in the extracellular-matrix formation. Our lead molecule, SLR080811, isishown to have a K of 1 µM and 5-fold selectivity for SphK2 overSphK1.SLR080811’s structural motif contains a tail group, linker, and head group. Inthis poster, we will review the structural modifications that have been performed to thelinker and head group regions of SLR080811 in an attempt to improve inhibition ofSphK2.MUTAGENIC ANALYSIS OF BACTERIAL BETA-GLUCURONIDASE. SkyeHickling, Gina Burchett & Kimberly Lane, Dept. of Chem., Radford University,Radford, VA 24142. Camptothecin-11 (CPT-11) is used for chemotherapy in cancerpatients. It gets converted to the topoisomerase inhibitor SN-38. SN-38 is metabolizedin the liver to SN-38-glucuronide. Bacterial â-glucuronidase hydrolyzes theglucuronide re-releasing SN-38 into the intestines. This reformed SN-38 is toxic to thebody and â-glucuronidase is the key to the toxicity of the drug CPT-11. The use ofinhibitors of E. coli â-glucuronidase decreases the amount of damage to intestinal cells. Deficiencies in the human form of â-glucuronidase may lead to the onset of Slysyndrome, a lethal disease with no current treatment. Previous studies has shown thata stable dimer of the enzyme may exist. This current study has been using WinCootVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 73STand ArgusLab to develop mutations in silico of the active site in â-glucuronidase andsubunit interfaces. Active site mutations were performed at residues that have shownto be of some importance through the enzyme related to â-glucuronidase called â-galactasidase. These active site mutations were developed at the following residues:H296, E297, S360, F365, N412, E413, Y468, and E504, D163, V274, Q507, F281,K277. The subunit interface mutations were performed at the following residues toreduce electrostatic interactions: E6, R71, K13, D16, D319, R43, D77, T7, D477,K157, Y517, D53, R10, K77, N308, E523, H514, K576, and S579. In the future, MC-PRO by Schrodinger will be used to determine feasible mutations for laboratorytesting. Through these calculations, mutations will be developed and studied for theinteractions between E. coli â-glucuronidase and ligands in hopes of learning moreabout the enzyme mechanism and dimer stability.Computer ScienceNONTRADITIONAL APPLICATIONS OF AUTOMATA THEORY. BruceChittenden, Department of Computer Science , Hampton University, Hampton,Virginia 23668. Automata theory is typically taught as a mathematical theory ofcomputation with applications in compiler construction, specifically in the area oflexical analysis. Typically computer science students do not encounter automatatheory outside of a course on programming languages or compiler construction, and aretherefore left with the impression that automata theory is a fairly abstractmathematically theory which only applies in these areas. I would like to put forwardthe argument that we should expand how we teach automata theory to include severalpractical applications such as the specification and verification of protocols. Automatatheory provides a rich set of abstractions and tools that every student should learn toapply to many situations that they will encounter in the development of complexsoftware. I have found that automata theory is well suited for the specification andverification of many types of protocols. Any software situation where communicationsis occurring and state is maintained over a period of time lends itself to a softwaredesign and implementation based upon automata theory. The finite state machine isspecified symbolically, that is the inputs, outputs, states, and transitions are defined interms of symbols easily understood by humans but difficult in that form, to be useddirectly by a computer. An Analyzer Program can easily be created that processes thedescription of the Finite-State Machine and generates two tables of integers, outputsand next state, representing the machine behavior. These tables can be used directlyby the computer. The software implementation becomes completely table driven.Using todays programming languages such as C, C++, or Java, this is easilyimplemented using a switch statement. The significant benefit of this approach is thatit pushes the debugging of the protocol from coding and testing phase back into thedesign phase, therefore reducing the time to deliver working software.OPEN Q UEST ION S REGARDING UPPER BOUND ON M ATRISMULTIPLICATION OF O(n ) FOR w < 2.374. S. V. Providence, Department ofwComputer Science, Hampton University. Matrix multiplication is a fast algorithm thatcan be distributed and has fast performance on modern computer systems. A lowerVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss174 VIRGINIA JOURNAL OF SCIENCEbound of W(n ) for this algorithm holds even for sparse matrices. The best known lower2bounds are c × n , for constants c b 3. Strassen suggests matrix multiplication requires2strictly more than quadratic time. Several categories for finding an upper bound to thisalgorithm persist: bilinear algorithms, approximate algorithms and Schönhagestheorem, ultra fast solutions to tensor products, and Coppersmith-Winograd algorithm.We examine these approaches with regard to numerical accuracy and stability whileconsidering the current open questions and a possible choice for solution.A FLUID-BASED STOCHASTIC APPROACH FOR ANALYZING ANDMODELING NETWORK TRAFFIC. Yen-Hung Hu, Department of ComputerScience, Hampton University, Hampton, VA 23668. In this paper, a fluid-basedstochastic approach for modeling normal and malicious network activities isinvestigated and developed. To achieve the goal of this paper, several existing rawnetwork traffic traces are analyzed first. And then a fluid-based stochastic networkmodel for simulating normal and malicious network traffics according to characteristicsof such raw traffic traces is investigated and developed. Since observing that themajority of the collected network traffic traces demonstrates self-similar behavior, thispaper investigates this self-similar characteristic and adopts it as a key factor ofidentifying malicious activities. Several simulation results are included and the rawtraffic traces shown in this paper are gathered by the Cooperative Association forInternet Data Analysis (CADIA).EducationTHE WAY FORWARD: UNDERSTANDING OUR ECOLOGICAL PLACE. EmmaD. Burnett , Alana S. Burton & Eugene G. Maurakis , Department of Biology,1 2 1 1University of Richmond, Richmond VA 23173, Department of Geology, College of2William and Mary, Williamsburg VA 23185. This documentary film explores themodern relationship between Americans and the natural world by examining theperspectives of individuals with unique personal and professional connections to theenvironment. Interviews conducted with these individuals illuminate numerous facetsof our relationship with the environment, from a farmer’s reasons for investing in thelong-term health of the environment, to a scientist’s perspective on the role ofskepticism in the pursuit of truth and the biggest challenges to educating the public. Some of the general questions addressed include the implications of our attitudetowards the environment on our health and the health of future generations, challengesto the pursuit of general knowledge and understanding, including polarized viewpointsand special interest groups, and potential steps in overcoming these challenges. Through the examination of these topics by interview subjects from differentbackgrounds and occupations, the film presents various ways of thinking about theenvironmental problems that we face and highlights the importance of individual actionand constructive dialogue.ENVIRONMENTAL ISSUES AND AMERICAN ATTITUDES. Alana S. Burton¹,Eugene G. Maurakis², and David V. Grimes³, College of William and Mary, Dept. of1 2Biology, University of Richmond, & Old Dominion Appalachian Trail Club. In July3Virginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 75ST2012 the “Survey on Human- Environment Interaction” was launched usingSurveyMonkey. The survey aimed to explore how humans view their relationship tothe natural world as well as how demographics influence beliefs about environmentalissues. In September, with 383 respondents, the survey was closed for analysis. Over90% of respondents agreed that there is a direct link between human health andenvironmental health and that environmental degradation is disturbing; therefore, thisconsensus may serve as a useful backdrop to environmental education. Age, educationlevel, and political affiliation were found to play an important role in how humans viewenvironmental issues; therefore, these characteristics may be useful in the creation ofeducational programs.CURRICULUM MATERIALS AS A XOMPANION TO A LARGE MUSEUMEXIBITION ON HUMAN HEALTH AND WELLNESS. David B. Hagan, ScienceMuseum of Virginia, 2500 W. Broad Street, Richmond, VA 23220. The ScienceMuseum of Virginia has a statewide mission for informal science education. It hasspent 14 months in research, planning and design for an 8,000 square foot interactiveexhibit gallery focused on the human body and mind. It also produced a five-minuteeducation curriculum video on the human body, its systems, chemistry, and physiology.Research on the function and visitor learning was conducted on the interactive exhibitsusing formative evaluation. Student groups (elementary through high school) wereobserved engaging with a selected set from each of the three main categories ofexhibits, (Body, Mind and Play). The results of these evaluation cycles were used toimplement modifications to the interactive exhibit units. In the Body section theexhibits included: an isometric bench press, a test of strength in pushing a fixed bar ina standing position; a test of balance on a 12-foor tightrope six inches above the floor;a test of reaction time in response to visual stimuli; a test of the visitor’s ability toassume a yoga position and hold it for ten seconds; a test of the visitor’s flexibility asmeasured by their ability to fit into a small space; and three exhibits focused on concepts related to nutrition, diet and food preparation in the Kitchen Theater. Visitorexperiences in the Mind section include psychology-related engagements: a self-reported measure of happiness; two different short-term memory tests; a challenge ofthe visitor’s capability to cooperate in a group to succeed at a timed challenge; ademonstration of eye-tracking sensors showing where the visitor’s gaze was focusedon an image; and a Sleep Theater experience that takes the visitor on a narrated journeythrough a typical night’s sleep showing the sleep cycles and levels of sleep. The Playsection of the exhibit includes: playing percussion instruments, attempting to matchand hold a given note as a test of perfect pitch; and an interactive overview of pastimes,hobbies and sports. The exhibit has been tested and is in the process of installation.Environmental SciencePLANTS AS INDICATORS OF DISTURBANCE AT THE ABRAMS CREEKWETLANDS, A PROTECTED CALCAREOUS ECOSYSTEM IN WINCHESTERAND FREDERICK COUNTY, VIRGINIA. J. T. Walker, V. Thomas, W. S. Bousquet,D. F. Cooper, B. Ridgeway, S. Dieudonné, S. Kochman, J. Miller & G. Ayers. Environmental Studies Program, Shenandoah University, Winchester, Virginia 22601. Virginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss176 VIRGINIA JOURNAL OF SCIENCEA 1998 study by Shenandoah University (SU) supplemented investigations by theVirginia Natural Heritage Program in documenting the ecological significance of theAbrams Creek Wetlands. In the spring and fall of 2012 SU researchers resurveyed fourcommunities from the 1998 study. Using the relevé (Braun-Blanquet) method,community plots were described by their vegetation and physical characteristics. Foreach community, researchers compared the changes in relative cover by species from1998 to 2012. Indicator plant species were selected by three criteria: large (>3 s.d.)change in relative cover, rarity in Virginia, and low coefficient of conservatism. Factors potentially responsible for changes in vegetation were then identified. Whilethe analysis is preliminary, it provides opportunities to better understand wetlandecology, improve wetland management, and protect local and regional biologicaldiversity.STORMWATER MITIGATION CHOICES BY THE GENERAL PUBLIC. Shane H.Abinette & Eugene G. Maurakis , Biology Dept., Virginia Commonwealth1 2 & 3 1University, Science Museum of Virginia, & Biology Dept., University of Richmond. 2 3Objectives were to educate and survey visitors in the Rainkeepers exhibition at theScience Museum of Virginia regarding proper stormwater mitigation practices. Surveydata were gathered from visitors’ voting responses in interactive panels that asked 11questions regarding the user’s habits on littering, car washing, oil recycling and othertopics from December 2012 –April 2013. Results showed a high visitor willingness touse pervious concrete and rain barrels but a lack of actual use of these materials. Highlevels of participation in litter and pet waste pick up, and reduced use of lawn pesticidewere also found. A high willingness was also detected for recycling motor oil andconverting lawns to native vegetation. For all questions, a majority of respondentsclaimed they already did or were willing to practice good stormwater managementtechniques. Funded by a National Fish and Wildlife Foundation grant.LAND-USE PROBLEMS FOR LOCAL AIR QUALITY AND BIOL:OGICALCONSERVATION, A SIMPLE SOLUTION PROPOSED. Richard S. Groover,Department of Environmental Science and Policy, George Mason University, FairfaxVA 22030-4444. Local air quality and biological conservation is connected to land-usedecisions and resulting changes. The citizens and the governmental officials often maynot understand the importance of air quality or biodiversity conservation, andecosystem connectivity with respect to their land-use decisions, but they need tobecome aware of such. Individuals or developers who wish to maximize their use ofland and resources often have less concern for conservation. This presentation exploresthe current situation local habitat destruction and how these losses impact air quality,local biota in Virginia, how local governments are structured, local governmentdecision-making processes, and what conservation supporting tools exist for them touse. Two win/win strategies for achieving better success with local air quality andbiological conservation can improve local land-use decisions. One of the strategies isthe development of an innovative proffer and mitigation approach to increase trees andimprove carbon sequestration on land proposed for development. It includes an in-perpetuity protection of set-a-side forest areas. The second strategy involves an in-perpetuity protection of wildlife travel corridors, within the associated land beingVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 77STrezoned. Both strategies would be utilized during the zoning change phase for propertyto be developed.PostersA COMPARATIVE ANALYSIS OF THE HEALTH OF THE NI RIVER ANDMASSAPONAX CREEK IN SPOTSYLVANIA COUNTY. Dr. M. L. Bass, E. E.Stewart, A. T. Elliott & H. E. Richters, University of Mary Washington. The mainfocus of this research project was to determine the health of both the Ni River andMassaponax Creek in Spotsylvania County, VA. The two streams provide a gooddemonstration of the effects of urban development. There are four sample sites locatedon Massaponax Creek and three sites on the Ni River. At each site, macrobenthicsamples were taken as bio-indicators of pollution. These samples were corroborated bywater chemistry analyses, including conductivity, dissolved oxygen levels, fecalcoliform, alkalinity, calcium hardness, phosphate and nitrate concentrations and totaldissolved solids. All samples were taken within each season. An examination of themacrobenthic results indicate that throughout the summer, the %EPT of the Ni Riverwas approximately 90% at most sites. The low was of 82.8% at the McEwan Farmsite. At Massaponax Creek, the summer's %EPT ranged from 97.3% at the Wetlandto a low of 62.7% at Route 208. These ranges are caused by variations in waterchemistry. During autumn and winter, the %EPT's steadily fell, as the organismscompleted their aquatic life cycles. It can also be noted that one particular site onMassaponax was afflicted by a high CFU count, as well as high spikes in alkalinity,phosphate and hardness in the winter. It has been determined that the Ni River ishealthier than Massaponax Creek. This is represented by the higher EPT percentagesand diversity of macrobenthic individuals within the Ni River. However, MassaponaxCreek has maintained good health despite increased development within its watershed.Medical SciencesTHIOREDOXIN-INTERACTING PROTEIN MEDIATES HCYS-INDUCED NLRP3INFLAMMASOME ACTIVATION IN MOUSE PODOCYTES. Justine M. Abais,Krishna Boini, Min Xia, & Pin-Lan Li, Department of Pharmacology & Toxicology,Virginia Commonwealth University, Richmond VA 23298. Our recent studies havedemonstrated that NADPH oxidase-derived reactive oxygen species (ROS) activatesNLRP3 inflammasomes causing homocysteine (Hcys)-induced podocyte andglomerular injury, however the precise mechanism regarding how ROS activates theinflammasome is still unknown. The current study explored whether thioredoxin-interacting protein (TXNIP) mediates Hcys-induced NLRP3 inflammasome activationin podocytes. TXNIP, the regulatory inhibitor of the antioxidant thioredoxin (TRX),is thought to dissociate from TRX in response to elevated levels of oxidative stress tobind to inflammasome protein NLRP3 and activate the inflammasome complex. Genetic or pharmacologic inhibition of TXNIP by small interfering RNA or verapamilprevented Hcys-induced NLRP3 inflammasome formation and activation both in vitroand in vivo by reducing colocalization of NLRP3 with ASC or caspase-1, blockingHcys-induced coimmunoprecipitation of TXNIP with NLRP3, and diminishingVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss178 VIRGINIA JOURNAL OF SCIENCEcaspase-1 activity and IL-1â production. In addition, TXNIP inhibition protectedpodocytes from Hcys-induced injury as demonstrated by normal expression levels ofpodocyte markers podocin and desmin, and preservation of glomerular function byreduced proteinuria and albuminuria in vivo. These results concluded that TXNIPbinding to inflammasome protein NLRP3 is a key signaling mechanism for Hcys-induced NLRP3 inflammasome formation and activation, and subsequent glomerularinjury (supported by NIH grants DK54927 and 1F31AG043289).THE BALANCE OF T-LYMPHOCYTE SPECIFYING TRANSCRIPTIONFACTORS AND THE CYOTOKINE EXPRESSION IN OBESE PATIENTS WITHNAFLD. M. Keaton , R. Mehta , A. Neupane , & A. Baranova , Betty and Guy2 1, 2 2 1, 2 1Beatty Center for Integrated Research, Inova, Falls Church VA 22042 & School of2Systems Biology, George Mason Univ., Fairfax VA 22030. The accumulation ofimmune cells within visceral adipose tissue (VAT) in subjects with NAFLD has beenpoorly understood. The immune response in obesity is reported to involve the shift inh h hratio of subsets of T cells (T 1, T 2, T 17) within VAT through the transcriptionfactors, TBX21, GATA3 and RORã, respectively. The aim was to assess the ratio ofT-cell lineage transcription factors and T-cell associated cytokines in obese patientswith NAFLD. Forty-five biopsy-proven NAFLD patients’ VAT samples werecollected. RNA was extracted, and then converted, to cDNA. RT-PCR profiling wasperformed using validated primers for T cell lineage specifying markers (CD3E -CD4+ cell marker; TBX21 - Th1; GATA3 - Th2; FOXP3 - Treg). ACTB was used fornormalization. Group comparisons for ratios like TBX21/CD3E and TBX21/GATA3were carried out amongst all possible cohorts. There were consistently higher ratios ofGATA3/CD3E compared to TBX21/CD3E and FOXP3/CD3E within each histologicalcohort related to grade of steatosis, histologic NASH, and stage of fibrosis. Groupcomparison of cytokines showed TGF-B1 (FC=0.13, p=0.04) and IL-1B (FC=0.79,p=0.03) were down-regulated in patients with NASH. IL-1B (FC=0.25, p=0.007) wasdown-regulated in patients with advanced hepatic fibrosis. The immune imbalance thatis seen in obese NAFLD patients does not seem to depend on the expression oftranscription factors involved in T cell lineage determination in the adipose tissue.PRECLINICAL ASSESSMENT AND SPECTROSCOPIC CHARACTERIZATIONOF THE ABUSED SYNTHETIC CANNABINOID, CP47,497. K. L. Samano , L. E.1Wise , J. L. Poklis , A. Poklis & A. H. Lichtman , Dept. of Pharmacology &1 1 1,2,3 1 1Toxicology, Dept. of Forensic Science & Dept. of Pathology, Virginia2 3Commonwealth University, Richmond, VA, 23298. The objective of the present studywas to characterize the pharmacological consequences of the abused syntheticcannabinoid compound, CP47,497. Cannabimimetic effects were assessed using thewell-established tetrad model that is sensitive to the chief psychoactive compoundpresent in marijuana, THC, and consists of four outcome measures: catalepsy,antinociception (tail flick latency), hypothermia, and locomotor activity. CP47,4971 was significantly more potent than THC in each measure. The CB receptor antagonist,rimonabant, reversed all pharmacological effects of CP47,497. Consistent with these1findings, CB (-/-) mice were resistant to the cannabimimetic effects of CP47,497.Immediately following behavioral testing, mice were humanely euthanized and bloodand tissue were harvested for CP47,497 quantification. Samples are currently beingVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 79STanalyzed on an Applied Biosystems Liquid Chromatograph/Tandem MassSpectrometer (LC/MS/MS) interface utilizing electrospray ionization and selective ionmonitoring, using acetonitrile liquid-liquid extraction. Additionally, ongoing research1is examining tolerance and dependence liability, as well as CB receptor adaptationsfollowing repeated administration of CP47,497. In conclusion, the results of the present1study indicate that acute administration of CP47,497 elicits markedly more potent CBreceptor mediated effects (i.e. 5-9 fold) than THC.PRESERVATION OF T CELL FUNCTION AS A NECESSARY COMPONENT INCHEMOTHERAPEUTIC STRATEGIES IN TREATMENT OF MELANOMATUMORS. Se W. Jeong & Timothy N. J. Bullock, Dept. of Pathology, University ofVirginia, Charlottesville, VA 22908. Metastatic melanoma has a poor prognosis aspatients are only expected to live six to twelve months after diagnosis. PLX4720 is aspecific inhibitor that targets a B-Raf gene mutation that is characterized by a V to Esubstitution at position 600. This has been highly anticipated as more than half of thepatients present this mutation. While PLX4720 has had dramatic short-term success,the tumors escaped the B-Raf inhibition through other proliferative pathways.Therefore, six additional inhibitors were identified to have synergistic cytotoxicity withPLX4720 against melanoma tumors. This project probed how these combinations ofchemotherapies, that have proven synthetic lethality against melanoma, impacted uponT cell function and cytokine expression. Ultimately, the goal was to define the leastd e t r im e n ta l c o m b in a t io n s o f d ru g s to T c e l l s fo r c o m b in e dchemotherapy/immunotherapy approaches. Although most inhibitor combinations werenot deleterious, only one combination, PLX4720 and Masitinib, a platelet-derivedgrowth factor receptor inhibitor, was detrimental to T cell effector function. Therefore,our data indicates that it is possible to select multi-model therapy using combinationsof chemotherapies and immunotherapy.hOCTS: NOVEL TARGET FOR ANTIDEPRESSANT THERAPY. Kavita Iyer ,1Xiaolei Pan , Osama Alwassil , Douglas Sweet , & Malgorzata Dukat , Department2 1 2 1 1of Medicinal Chemistry & Department of Pharmaceutics, School of Pharmacy,2Virginia Commonwealth University, Richmond VA 23298. Drugs that targetnorepinephrine and serotonin transporters (NET and SERT, respectively) arecommonly utilized to treat depression, however they have certain drawbacks associatedwith them. There is an acknowledged need to identify novel antidepressants. Humanorganic cation transporters (hOCTs) are proteins that modify the transport ofendogenous neurotransmitters such as serotonin and norepinephrine. Recently,antidepressants have been shown to interact with hOCTs and may be a potential targetfor drug development. Our laboratory has synthesized a series of quinazoline analogs3 that are constrained analogs of m-chlorophenylguanidine (MD-354), a 5-HT receptorpartial agonist. Two quinazoline analogs (KEO-099 and KEO-147) were found to be35-HT receptor antagonists and tested positive in the mouse tail- suspension test.3However, the two analogs differed in their ability to bind to 5-HT receptors as well asNET and SERT. It was concluded that neither NET nor SERT are able to adequatelyexplain the antidepressant effects of the quinazoline analogs. This led us to examinethese analogs at hOCTs to determine if their action at hOCTs could contribute to theirVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss180 VIRGINIA JOURNAL OF SCIENCEantidepressant effects. Results from the preliminary tests indicate that quinazolinesinhibit the transport of 1-methyl-4-phenylpyridine (MPP ), a model substrate for+hOCTs. Moreover, we have established a rudimentary structure-activity relationship(SAR), which identifies substituents seemingly important for both binding as well asselectivity for hOCT subtypes.NLRPs IN VISCERAL ADIPOSE TISSUE OF MORBIDLY OBESE PATIENTSWITH NON-ALCOHOLIC FATTY LIVER DISEASE (NAFLD). Arpan Neupane ,2Rohini Mehta , Ancha Baranova , Arian Afendy , Zachary Goodman , & Zobair1,2 1,2 1 1Younossi , Betty and Guy Beatty Center for Integrated Research, Inova Health System,1 1Falls Church, VA & School of Systems Biology, George Mason University, Fairfax,2VA. Inflammasome-associated Nucleotide-binding oligomerization domain, Leucine-rich Repeat and Pyrin domain containing (NLRP) family proteins serve as intracellularsensors of PAMPs (toxins, whole pathogens, etc.) and DAMPs (dsDNA, cellulardebris, etc.). Inflammasome mediated activation of caspase-1 leads to cleavage of pro-IL1B and pro-IL18 and in turn induce expression of a number of cytokine genestriggering a cascade of inflammatory and immune responses. Obesity is a state of low-grade chronic inflammation and inflammation has been shown to play an important rolein obesity-associated non-alcoholic fatty liver disease (NAFLD). NAFLD is stronglyassociated with visceral obesity. In this study we investigated the expression of 14NLRPs genes in visceral adipose tissue of morbidly obese subjects (N=46) withNAFLD. Expression of 11 NLRPs members was not detected (Ct $ 35). Interestingly,NLRP1 and NLRP3 (the most well studied NLRPs) were found be downregulated inmorbidly obese subjects with severe NAFLD. This study for the first time determinesthe expression of NLRPs in visceral adipose tissue of morbidly obese subjects withNAFLD. The results show downregulation of inflammasome sensors with increasingseverity of obesity-associated NAFLD. Further studies are needed to dissect themechanism of NLRP downregulation.COMBINED ADMINISTRATION OF THE SELECTIVE FAAH INHIBITOR PF-3845 AND COX INHIBITOR DICLOFENAC: INVESTIGATION IN A MURINEMODEL OF NERUPPATHIC PAIN. T. W. Grim , S. O’Neal , S. J. Kinsey , J. M.1 1 2Pearson & A. H. Lichtman , Department of Pharmacology and Toxicology, Virginia3 1 1Commonwealth University, Richmond, VA, Department of Psychology, West Virginia2University, Morgantown, VA, & Ironwood Pharmaceutics, Inc. Cambridge, MA. 3Fatty acid amide hydrolase (FAAH), the primary degradative enzyme for theendogenous cannabinoid anandamide, offers a novel target to attenuate neuropathicpain, a condition refractory to existing analgesics. In the present study, we employedthe chronic constriction injury model of neuropathic pain (CCI) to assess whethercombined administration of the FAAH inhibitor, PF-3845, and the nonselectivecyclooxygenase inhibitor (COX) inhibitor, diclofenac, would produce augmentedantinociceptive effects. Dose response studies were conducted for each compound andsubthreshold doses were given in combination to test for augmented antinociceptiveeffects in the CCI model. The co-administration of 5 mg/kg PF-3845 and 30 mg/kgdiclofenac produced significant antinociception compared with either drug given alone.Metabolomic analysis found that PF-3845 elevated brain anandamide levels andVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 81STdiclofenac reduced prostaglandin brain levels, regardless of whether the compoundswere given separately or in combination.PHOSPHORYLATION OF RAPTOR FOLLOWING PEMETREXED ACTIVATIONOF AMPK OVERCOMES TSC2 DOWN REGULATION AND mTORC1ACTIVITY. Stuti Agarwal, Catherine Bell, & Richard Moran, Deptartment ofPharmacology & Toxicology, Virginia Commonwealth University, Richmond VA23298. Pemetrexed is a highly unusual antifolate in that it has significant therapeuticutility against non-small cell lung cancers (NSCLC) and mesothelioma. We haveshown that pemetrexed has a second mechanism, in addition to inhibition ofthymidylate synthase, which leads to robust activation of AMP-dependent kinase(AMPK), and have suggested that this effect is causal of the activity of this drugagainst lung cancers. The most frequently mutated gene in NSCLC is p53 and, yet, theeffects of pemetrexed are retained in p53-null carcinoma cells. Like K-Ras mutationand EGFR alterations, p53 loss causes hyperactivity of mTORC1; this is due primarilyto loss of TSC2 function. The effects of pemetrexed also result in a functional loss oftranscriptional activation by p53, with resultant loss of TSC2 function and concomitantexpansion of RhebGTP. However, the robust stimulation of AMPK phosphorylationof Raptor in pemetrexed-treated cells overwhelms these effects and causes suppressionof mTORC1.IL28B GENOTYPE ASSOCIATIONS WITH METABOLIC SYNDROME. LeahByars , Ancha Baranova , James M. Estep & Zobair Younossi , Betty and Guy2 1, 2 1,3 1, 2 , 3 1Beatty Center for Integrated Research, Inova Health System, Falls Church VA, Center2for the Study of Chronic Metabolic Diseases, School of Systems Biology, GeorgeMason University, Fairfax, VA & Center for Liver Diseases, Inova Fairfax Hospital. 3Hepatitis C Virus (HCV) is a viral infectious disease that affects an estimated 130-170million people. In a number of recent studies, the IL28B gene has been shown to playan important role in the outcome of HCV treatment. Metabolic syndrome (MetS) is agroup of medical disorders usually associated with obesity. Recent, but not conclusive,evidence suggests that IL28B genotypes may be associated with metabolic confoundersof HCV. In HCV patients, there is a significant correlation between the presence of CCgenotype and several metabolic factors. In non-HCV patients with NAFLD, no suchcorrelation was found. The metabolic profiles of the CC and Non-CC groups ofNAFLD patients were also not different from each other. The beneficial effects of CCgenotype on metabolic profiles may only be conferred after incidental infection withHCV, or after the treatment with antiviral therapy. IL28B genotyping in larger groupsof non-HCV patients is warranted to determine the true effect of IL28B genotypes onmetabolic outcomes.ACTION OF DECONSTRUCTED MDPV ANALOGS AT hDAT. F. T. Sakloth , R.1Kolanos , E. Solis , L. De Felice & R. A. Glennon , Department of Medicinal1 2 2 1 1Chemistry, Virginia Commonwealth University, Richmond VA 23298 & Department2of Physiology and Biophysics, Virginia Commonwealth University, Richmond VA980551. The number of cases reported on the abuse of psychoactive “bath salts” hasintensified in the past few years. These “synthetic cathinones” are â-keto analogs ofamphetamine and were scheduled as of October 21, 2011. MethyenedioxypyrovaleroneVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss182 VIRGINIA JOURNAL OF SCIENCE(MDPV), a major constituent of bath salts, is the first synthetic cathinone known to actas a dopamine re-uptake inhibitor, i.e. to show a hyperpolarizing current. The purposeof this project was to understand which structural features contribute to itshyperpolarizing action, and determine what is it about MDPV that converts it from adepolarizing to a hyperpolarizing agent? MDPV was deconstructed into seven analogs,making only one structural change at a time. All the analogs, as with MDPV, showeda hyperpolarizing current in electrophysiological studies using frog oocytes transfected5 0 with hDAT. The IC values were measured and they ranged over a 200-fold span.Amongst the structural features that contribute to affinity, the tertiary amine and/or theextended side chain are optimal for affinity, but the extended side chain seems criticalfor affinity. However, the carbonyl oxygen atom and the methylenedioxy ring are notmajor contributors to affinity and not required for the hyperpolarizing action. Inaddition to understanding the structural contribution of MDPV to its hyperpolarizingaction we are the first to report on the mechanism of newly confiscated MDPV analogs.[Supported by NIH grant DA 5R01DA033930.]ENDOCANNABINOID METABOLIC INHIBITORS FOR THE TREATMENT OFAFFECTIVE ASPECTS OF MORPHINE WITHDRAWAL. T. F. Gamage & A. H.Lichtman, Dept. of Pharmacology and Toxicology, Virginia CommonwealthUniversity, Richmond VA 23298. Inhibition of the endocannabinoid catabolicenzymes fatty acid amide hydrolase (FAAH) with PF3845 or monoacylglycerol lipase(MAGL) with JZL184 reduces morphine withdrawal signs. It is uncertain if these anti-withdrawal effects extend to aspects related to the affective components of withdrawal.In the present study, we tested whether FAAH or MAGL inhibition would block theacquisition of conditioned place avoidance (CPA) to morphine withdrawal. Whilemorphine (30 mg/kg) blocked acquisition of CPA and reduced jumping, neitherJZL184 (4 or 40 mg/kg), PF3845 (1, 3, or 10 mg/kg), nor clonidine (0.03-1 mg/kg)affected acquisition of naloxone-precipitated morphine withdrawal-induced CPA.Additionally, delta-9-tetrahydrocannabinol (1, 3, or 10 mg/kg) did not affectwithdrawal CPA. However, both JZL184 and THC significantly reduced thepercentage of mice that exhibited jumping behavior. These data suggest that whileinhibition of endocannabinoid metabolism reduces overt behavioral withdrawal signs,it is ineffective at reducing the aversive aspects of withdrawal. Thus, these drugs onlyreduce a subset of aspects related to opioid withdrawal. Future studies will examine theeffects of combination treatment of dual FAAH/MAGL inhibition on acquisition ofwithdrawal CPA as well as other affective-related withdrawal behaviors.PostersADIPONECTIN PREDICTOR OF LONG-TERM SUSCEPTIBILITY TO OBESITY. E. Jenkie , J. Bahamonde , B. Brenseke , M. Friedman , L. Mitchell & R. Prater ,1 2 2 1 1 1,2EVCOM; VMRCVM. In 2009-2010, 35% of adult Americans were obese and at high1 2risk for metabolic disease due to sedentary behavior and chronic overindulgence of fatand carbohydrates. Adiponectin is a protein hormone secreted by adipose tissue and isinversely correlated with body fat percentage in adults. It aids in the suppression ofmetabolic derangements that may result in obesity, type 2 diabetes, atherosclerosis andmetabolic syndrome. Our laboratory proposed that maternal malnutrition, representedVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 83STas high fat diet (HFD), high sugar diet (HSD) or ethanol consumption (EtOH)throughout pregnancy, would elevate long-term risk of obesity in mouse offspring andthat adiponectin in offspring from dams exposed to these diets could be measured inorder to predict long-term susceptibility to obesity. Liver adiponectin was measured incontrol, HFD, HSD and EtOH groups at postnatal day 1, 21 and 42 in C57BL/6 miceusing a mouse adiponectin ELISA. Serum insulin levels were measured, andhistopathology reports and x-ray microtomography images were obtained for diseasedetection. Adiponectin decreased with age in all groups. The treatment groups hadlower adiponectin levels (HFD group was lowest), compared to controls. Additionally,HFD offspring were found to have higher insulin levels than controls, and their femursdisplayed fat accumulation and lesions resembling osteoporosis. Findings suggest thatprenatal malnutrition can lead to diminished adiponectin levels and that measuringadiponectin may help to predict future risk of obesity and related disorders. Thisbiomarker may then be manipulated in novel ways (e.g., nutritional interventions) inorder to improve long-term health.THE EFFECTS OF GLIAL CELL MODULATORS ON METHAMPHETAMINE(METH) INTRAVENOUS SELF-ADMINISTRATION AND FOOD-MAINTAINEDBEHAVIOR IN RATS. S. E. Snider , E. S. Hendrick & P. M.1 1Beardsley , Department of Pharmacology and Toxicology, Institute for Drug and1,2,3 1 2Alcohol Studies, Center for Biomarker Research and Personalized Medicine, School3of Pharmacy, Virginia Commonwealth University, Richmond Virginia. Methamphetamine (METH) administration activates glial cells in the CNS andincreases pro-inflammatory cytokine production and release. Glial cell activation andpro-inflammation have been linked to drug abuse-related behavior. Ibudilast (AV411;3-isobutyryl-2-isopropylpyrazolo-[1,5-a]pyridine), which inhibits phosphodiesterase(PDE) and glial-mediated pro-inflammatory activity, AV1013, an amino analog ofibudilast with similar glial but limited PDE activity, and minocycline, a tetracyclinederivative with similar glial attenuating capabilities, all attenuate METH (0.03mg/kg/inf) self-administration. The effects of ibudilast (10 mg/kg) and minocycline (30& 60 mg/kg) on food-maintained responding were also examined using a behavioraleconomic approach. Demand curves for both METH (0.03 mg/kg/inf) and food-maintained behavior were obtained by increasing the FR value daily using thefollowing progression: 1, 3, 6, 9, 13, 19, 26, 35, 47, 62, 82, 108. After matching theessential values (á levels) of METH and food pellets, twice daily ibudilast or oncedaily minocycline, was administered for three days under FR1 conditions. Ibudilast andminocycline both significantly (p<0.05) reduced both METH- and food-maintainedresponding. These results indicate that ibudilast and minocycline have complex effectspotentially able to attenuate behavior maintained by multiple events.BLOCKADE OF THE NMDA RECEPTOR ION CHANNEL ENHANCES THEDISCRIMINATIVE STIMULUS EFFECTS OF NITROUS OXIDE. Kellianne J.Richardson & Keith L. Shelton, Department of Pharmacology & Toxicology, VirginiaCommonwealth University, Richmond VA 23284-2006. The abuse-related CNS2 2effects of nitrous oxide (N O) are poorly understood. In vitro data suggests that N OA alters the function of NMDA and GABA receptors, amongst others. Our goal was toVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss184 VIRGINIA JOURNAL OF SCIENCEassess the neurotransmitter systems responsible for producing the intoxicating effects2of N O using drug discrimination. Twenty-four mice were trained to discriminate 102 2 2 min of exposure to 60% N O/40% O versus 100% O in daily 5 min operant sessions.2Mice acquired the discrimination in an average of 38 days. N O producedconcentration-dependent substitution for the training concentration. Full substitution2 2required 7 min of 60% N O exposure but the offset of stimulus effects N O followingthe cessation of exposure was more rapid. In substitution studies the NMDA channel2blocker MK-801 produced up to 49% N O lever-selection at 0.56 mg/kg. While MK-801 failed to produce more than partial substitution, an intermediate dose of 0.172mg/kg MK-801 significantly shifted the N O concentration-effect curve to the left. TheAcompetitive NMDA antagonist CGS 19755 and the positive GABA modulator2midazolam failed to substitute for N O. The stimulus effects of nitrous oxide alsopartially overlap with volatile solvents and anesthetics. Taken together these data2suggest the subjective stimulus properties of N O may be partially mediated by NMDAantagonism. Other receptor systems not yet examined are probably also involved.Supported by NIDA grant RO1-DA020553.THE EFFECTS OF WESTERN-STYLE DIET ON ATTENTIONAL SET SHIFTINGPERFORMANCE. S. Marwitz, L. Woodie & S. Blythe, Dept. of Biol., Washington& Lee University, Lexington, VA 24450. The rate of obesity in the human populationhas risen to alarming levels in recent years. Obesity has been linked with a number ofmaladies such as insulin resistance, leptin resistance and diabetes. However, there isnow growing evidence that obesity may even be linked to hippocampal damage. Weconducted a pilot study to find if an attentional set shifting task could detect damagedone to the hippocampus from a high fat diet. We used two groups of rats, onemaintained on a high fat diet (n=19) and the other maintained on a control diet (n=19).The diets continued for 11 weeks when we performed the attentional set shifting task.Before the attention task the rats were maintained on limited versions of theirrespective diet so that they reached 85% of their body weight. In the task the rats werepresented with two identical glass cups filled with a different material and scent. Ratswere required to remember which scent or material was baited, with tasks increasingin difficulty. We found that diet did not have a significant effect on performance in thetask (p=0.177, p=0.751). However, there was significant decline in performance as thetasks progressed and became more difficult (p=0.052, p=0.033). With this data andfurther research into other attention tasks, we hope to establish an appropriate test toshed light on the effects of a high fat diet on the brain. (Supported by: Howard HughesMedical Institute Fund, Levy Foundation Grant, The Virginia Academy of Sciencesand Washington & Lee University.)STATISTICAL ANALYSIS OF ENVIRONMENTAL RISK FACTORS OF BREASTCANCER. Amirhossein Shamsaddini , Ancha Baranova , Aybike Birerdinc ,1,2 1,2 1,2School of Systems Biology, College of Science, George Mason University & Betty1 2and Guy Beatty Obesity and Liver Program, Inova Health System, Falls Church, VA. Breast cancer is the most frequently diagnosed cancer among women, accounting forabout 30% of all new cancer cases each year. In the early 1990s, it was suggested thatexposure to some environmental chemicals such as organochlorine compounds (OC)may play a causal role in the etiology of breast cancer through estrogen-relatedVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 85STpathways. We used 1999–2009 National Health and Nutrition Examination Survey datato examine associations between serum concentrations of OC pesticides and breastcancers. Statistical analysis (Logistic Regression) was performed with SPSS v.20 onthese databases after carefully screening for any inconsistency. Although further studyis necessary to confirm these findings, these statistical results suggest that OC pesticideexposures may have a significant effect on breast cancer risk.HUMAN LEUKOCYTE ANTIGEN (HLA) CLASS I COMPLEX: CLINICALSIGNIFICANCE AS AN INDEPENDENT PREDICTOR OF NON-ALCOHOLICFATTY LIVER DISEASE. Ali Moosvi , Azza Karrar , Zachary Goodman , Ancha1,3 2,3 2Baranova & Zobair Younossi , Center for the Study of Chronic Metabolic1,2,3 1,2 1Diseases, George Mason University, Fairfax, VA, Betty and Guy Beatty Center for2Integrated Research, Inova Health System, Falls Church, VA, & School of Systems3Biology, College of Science, George Mason University, Fairfax, VA. The overall aimof our study is to assess the association between the Human Leukocyte Antigen (HLA)Class I Antigens polymorphism and hepatic steatosis, which is an independentpredictor of cardiovascular disease. Patients with biopsy-proven Non-Alcoholic FattyLiver Disease (NAFLD) (n =199) were genotyped with the PCR Sequence SpecificOligonucleotides (PCR-SSO) for the HLA-A,-B, and –C. NAFLD patients weregrouped to steatohepatitis (NASH) and Non-NASH NAFLD. Univariate andmultivariate analyses were performed to determine significant correlations. HLA –A11, -B51 were found to be strongly associated to high-grade hepatic steatosis. Inmultivariate analysis approach, our data demonstrated an independent associationbetween HLA-A11 and lower risk of high-grade hepatic steatosis, in addition to anindependent association between HLA-A31, -B64, -B57 and histologic NASH. Notabledifferences were seen within the HLA-C loci, where Cw7 correlated to a protectiveeffect against Advanced Fibrosis, while Cw8 had a predisposing effect. Identified riskhaplotypes will aid in management of NAFLD by stratifying patients into groups withincreased risk in developing not only NASH or Advanced Fibrosis, but alsocardiovascular diseases.MATRICELLULAR PROTEINS AND THEIR SPLICE VARIANTS IN OBESENAFLD PATIENTS. Z. Asif , R. Mehta & A. Baranova , Center for the Study1,3 1 , 2 1,2 1of Chronic Metabolic Diseases, George Mason University, Fairfax, VA, Betty and2Guy Beatty Center for Integrated Research, Inova Health System, Falls Church, VA,& Biology Department, College of Science, George Mason University, Fairfax, VA. 3Matricellular proteins (MPs) are extracellular proteins that modulate cell-matrixinteractions and cell function. MPs are doing this by remodeling the extracellularmatrix. Obesity is a chronic disease that has reached epidemic proportions worldwide.Obesity is accompanied by excessive storage of fat in adipose tissue and ectopic siteslike liver. Obesity is associated with many chronic diseases, notably nonalcoholic fattyliver disease (NAFLD). During obesity, the adipose tissue undergoes extensiveremodeling (hyperplasia and hypertrophy) to accommodate excessive fat deposition.This extracellular matrix remodeling is, in turn, associated with altered expression ofMPs. Some of the MPs have alternate isoforms whose expression and roles have notbeen examined in the context of obesity and associated NAFLD. These isoforms maybe released into circulation and may affect distant sites such as liver. The aim was toVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss186 VIRGINIA JOURNAL OF SCIENCEconduct a literature survey and shortlist MPs implicated in obesity and fatty acidmetabolism. A literature survey was done on MPs using PUBMED and GoogleScholar. MPs of interest were shortlisted and primers were designed for the differentvariants. Among the MPs, primers were designed for osteopontin (OPN), angiopoietin-like 4 (ANGPTL4), and autotaxin (ATX). Their expression in obese patients withNAFLD was detected. Future work: To design primers specific to each mRNAisoform and to detect their expression in obese patients with NAFLD.THE RELATIONSHIP BETWEEN DEPRESSION AND THE FATTY LIVERINDEX. Nayeem Hossain , Michael Estep , & Ancha Baranova , Betty and Guy1,2 1,2 ,3 1,2 1Beatty Center for Integrated Research, Inova Health System, Falls Church, VA, Center2for the Study of Chronic Metabolic Diseases, School of Systems Biology, GeorgeMason University, Fairfax, VA, & Center for Liver Diseases, Inova Fairfax Hospital,3Falls Church, VA. The fatty liver index (FLI) uses various chemical and physicalmeasurements. This study observed the effect of depression’s chemical environmenton these values, focusing on triglyceride level and body mass index (BMI). Mann-Whitney tests compared triglyceride levels and BMI of those with depression to thosenot depressed in the overall population; subpopulations diagnosed with NASH werecompared for further analysis. Triglyceride levels of depressed individuals weresignificantly higher than those not depressed in the overall population (P < 0.0232).The triglyceride value of individuals with both NASH and depression was notsignificantly higher (P < 0.1738) than those with only NASH. The average BMI wasnot significantly higher in depressed individuals than those not depressed in bothoverall population (P < 0.3371) and NASH subpopulation (P < 0.4122). Elevatedtriglyceride levels cause the FLI of depressed individuals without NASH to be higherthan those of individuals without depression or NASH. Greater waist circumference,attributed to elevated triglyceride levels, further affects FLI values.THE GENE EXPRESSION PATTERNS IN THE ADIPOSE TISSUE OFMETABOLIC SYNDROME PATIENTS IN THE CONTEXT OF GENDER. AshleyGreer , Brianda Beverley , Micheal Estep , Betty and Guy Beatty Center for1,2 1 ,2 ,3 1,2 ,3 1Integrated Research, Inova Health System, Falls Church VA, CSCMD, GMU, Fairfax,2VA, & NVCC, Annandale, VA. Adipose tissue metabolism differs between men and3women. Study of these differences may shed light into pathologies such as metabolicsyndromes that affect men and women differently. The aim of this study is to examinemRNA and miRNA expression from visceral adipose tissue and circulating orexigenichormones with respect to gender and obesity associated disease. Visceral adipose tissue(VAT) was collected from biopsy-proven NAFLD patients who were undergoingbariatric surgery (N=24, Male=6, Female=18). MiRNA expression data from the VATof NAFLD patients were obtained. Differences in circulating concentration orexpression was considered significant between men and women if the absolute valueof the change was >1.5 and a P-value of <0.05 was obtained. Regarding the cohort inwhich miRNA expression was measured, of the males in this group, 33% of patientshave a diagnosis of diabetes, and 50% have a diagnosis of non-alcoholic steatohepatitis(NASH). Many of the differentially expressed miRNA correlate significantly withclinical and laboratory data in women. The variation in the expression patterns betweenmen and women support that there is a gender-dependent difference in morbid obesity.Virginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 87STFurther analysis on the expressions and functions of these molecules is necessary.Further study of this gene could express a difference in growth regulation in men andwomen. Better understanding of the functions for these molecules may help to developgender-specific screenings or treatments for morbidly obese men and women.PARADOXICAL EFFECTS OF MORPHINE WITHDRAWAL IN MOUSEMODELS OF ANXIETY-LIKE BEHAVIOR. J. P. Crowley III, T. F. Gamage, & A.H. Lichtman, Dept. of Pharmacology and Toxicology, Virginia CommonwealthUniversity, Richmond VA 23298. Opioid dependence presents a serious threat tosociety and affects approximately 2.4 million individuals in the United States.Cessation of drug taking leads to a withdrawal syndrome consisting of somatic signs,flu-like symptoms, and anxiety, which contribute to relapse. Typical models of anxietyhave a lack of face validity in terms of expression of anxiety-like behavior duringopioid withdrawal. It’s been shown that mice going through morphine withdrawaldisplay an increased open-arm time in the elevated plus maze, a behavior typicallyinterpreted as anxiolytic. We assessed morphine withdrawal in two assays typicallyused to assess anxiety, the light/dark box (LD box), a conflict task similar to the EPM,and novelty-induced hypophagia (NIH), which measures consumption of palatable foodin the home cage and a novel environment. In this assay a decrease in consumptionand/or increase in latency is interpreted as anxiogenic behavior. Male ICR mice wereimplanted with either 75 mg morphine or placebo pellets and tested 48 h later receivingeither naloxone (0.03-0.1 mg/kg) to precipitate withdrawal or saline. Morphinewithdrawal increased the time spent in the light side of the LD box and reduced thelatency to consume food in the NIH test – effects typically interpreted as anxiolytic.The increased time in the light side might reflect a shift in defensive behaviors towardescaping. Additionally, naloxone may be blocking morphine’s suppressive effects onfood consumption thus manifesting as a decrease in latency to consume. Further studieswill test the effects of withdrawal in assays with greater face validity for escapebehavior.LIPID, GLUCOSE AND LIVER ENZYMES PROFILING OF OBESE NAFLDPATIENTS. Sweta Sedhai , Katherine Doyle , Amir Shamsad , Aybike Birerdinc1.2 1,2 1,2 1, & Ancha Baranova , Betty and Guy Beatty Center for Integrated Research, Inova2 1,2 1Health System, Falls Church, VA & School of Systems Biology, College of Science,2George Mason University, Fairfax, VA. Insulin resistance is a major and very commonissue in chronic diseases such as obesity, type 2 diabetes, cancer and cardiovasculardiseases. It is also a key feature of Nonalcoholic Fatty Liver Disease (NAFLD). Themain aim of this study is to quantify the insulin levels as well as glucose, ALT andAST levels in the serum of obese NAFLD patients. The data obtained in thisexperiment will be used in future studies for sample stratification. The Cholestech LDXdiagnostic tool was used in this experiment. Lipid Profile GLU and ALT.AST(GPT.GOT) cassettes were placed in the Alere Cholestech LDX system. Thecorrelation analysis was performed for 70 serum samples of obese NAFLD patients inwhich 54 serum samples were obtained from ELISA technique and 16 serum sampleswere done in Cholestech LDX system. The significant values for TRG was 0.001 withcorrelation coefficient (0.397), HDL was 0.018 with correlation coefficient (-2.90),LDL was 0.107 with correlation coefficient (0.210), Non-HDL was 0.020 withVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss188 VIRGINIA JOURNAL OF SCIENCEcorrelation coefficient (.285), TC/HDL was 0.021 with correlation coefficient (0.283)and GLU was 0.00 with correlation coefficient (0.499). The data obtained throughcorrelation analysis and linear regression supports the main idea of this study. Theincreased level of IL-8, IL-6, TNFa and C-peptide insulin are found in the patients withhigher glucose, ALT and AST levels.Natural History and BiodiversityFREE-LIVING SOIL NEMATODE POPULATION DIVERSITY DYNAMICS ATAN ASIMINA TILOBA SITE IN VIRGINIA. Sarah R. Marzec & Theresa M. Grana,Department of Biological Sciences, Univ. of Mary Washington, Fredericksburg VA,22401. Nematodes are microscopic roundworms that are highly successful in manyenvironments. The model organism Caenorhabditis elegans is specifically a free-livingnematode which can be found in soil but has been mostly isolated from anthropogenichabitats. Little is known about the environmental factors that affect C. elegans andwhere it is proliferate in nature. Information on ecological factors affectingCaenorhabditis species will be useful in identifying selective pressures that caninfluence genomic changes. The goals of this study are to find C. elegans and otherCaenorhabditis species and shed light on relationships between the ecological factorsand proliferating populations of nematodes. An Asimina tiloba site provides a naturalVirginian habitat with a food source for Caenorhabditis species. This site is sampledevery two weeks and any relevant ecological factors are recorded for the duration ofa year. Nematodes are isolated from each soil sample and are separated based onmorphology. The life stages of the nematodes from each sample are recorded and thenthe nematodes are sequenced for species identification. At present, five samples havebeen collected and several strains of nematodes have been isolated among and withinthe samples. All nematodes have shown to be in the dauer life-stage, a non-feedingmigratory stage, showing that there are no established colonies. This is most likely dueto the cold weather and we expect future results taken between the months of Maythrough October to show proliferating populations.INVENTORY AND CONSERVATION ASSESSMENT OF THE MOTH FAUNA OFVIRGINIA (LEPIDOPTERA). Steven M. Roble, Virginia Department of Conservationand Recreation, Division of Natural Heritage, Richmond, VA 23219. The insect orderLepidoptera includes the butterflies, skippers, and moths. Virginia’s butterfly andskipper fauna of some 170 species has been well-studied for more than a century andcontinues to attract interest from biologists and amateur naturalists alike. In contrast,the much larger moth fauna of the Commonwealth remains poorly studied with theexception of a few pest species such as the Gypsy moth (Lymantria dispar). No formalcompilation of the Virginia moth fauna exists, although survey efforts by staff of theVirginia Department of Conservation and Recreation, Division of Natural Heritage(DCR-DNH), during the past quarter century have begun to elucidate the composition,distribution, and conservation status of the “macro-moth” component of this fauna.Nearly 1,200 species of macromoths have been documented in Virginia, with acomparable number of “micro-moths” also expected to occur in the state, for a totalVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 89STfauna of about 2,400-2,500 species. A number of these species have not been formallydescribed in the scientific literature. Currently, the DCR-DNH rare animal list includes95 species of moths (92 macro-moths and 3 micro-moths), 11 of which have not beenrecorded in the state in more than 50 years. None of these species is formally listed asthreatened or endangered in Virginia. The DCR-DNH animal “watchlist” contains 123additional species of macro-moths that may warrant conservation attention. Withfunding via a State Wildlife Grant from the Virginia Department of Game and InlandFisheries and the U.S. Fish and Wildlife Service, DCR-DNH has recently developeda web-based atlas of Virginia’s rare Lepidoptera (butterflies, skippers, and moths) andOdonata (dragonflies and damselflies) that will debut in June 2013 athttp://www.vararespecies.org.DEVELOPMENTA TOLERANCE TO NICOTINE DIFFERS BETWEENINCIPIENT SPECIES OF A PARADITIC WASP WITH RESPECT TO HOSTFOOD-PLANT. Justin P. Bredlau & Karen M. Kester, Dept. of Biology, VirginiaCommonwealth University, Richmond VA 23284. Nicotine evolved as a defenseagainst insect herbivores. In response, tobacco-feeding insects as well as parasiticwasps that attack and develop within tobacco-feeding insects evolved counter defensesto nicotine. We tested the hypothesis that two incipient species of the braconid wasp,Cotesia congregata Say, and their reciprocal hybrids would differ in developmentalresponses to nicotine dosage level in the host diet. “MsT wasps” originated from thesolanaceous specialist, Manduca sexta L. (“tobacco hornworm”) on tobacco, and “CcCwasps” from the catalpa specialist, Ceratomia catalpae Boisduval (“catalpa sphinx”)on catalpa. Reciprocal crosses (MsT% x CcC&, CcC% x Ms&) were established bypairing unmated males and females. Females were permitted a single oviposition intoa 3 -instar larva of M . sexta. Parasitized caterpillars were fed on a laboratory diet withrd0%, 0.1%, or 0.3% (wet weight) nicotine until wasp larvae egressed from the host andspun cocoons. For each hornworm host, we counted the number of egressed andunegressed larvae, cocoons, and emergent adult wasps. Results demonstrate that MsTwasps are highly adapted to nicotine and responded to nicotine in a dosage-dependentmanner. In contrast, exposure to 0.1% nicotine resulted in high mortality of CcC wasps.Responses of the hybrid crosses varied with respect to maternal wasp type (MsT orCcC) and sex. For example, most haploid males with CcC mothers failed to egress,whereas hybrid females from either cross did, suggesting that nicotine tolerance is adominant heritable trait or suite of traits.HABITAT CHARACTERISTICS INFLUENCING HIBERNATION SITESELECTION BY WOOD TURTLES (GLYPTEMYS INSCULPTA LE CONTE), ATHREATENED SPECIES IN THE SHENANDOAH VALLEY, VIRGINIA. J. A.Miller & W. S. Bousquet, Envtl. Studies Program, Shenandoah Univ., Winchester VA22601 & T. S. B. Akre, Dept. of Bio. Longwood Univ., Farmville VA 23909. InNovember 2012, researchers captured hibernating wood turtles, Glyptemys insculptaLe Conte, and recorded habitat data twice along two stream reaches of a tributary tothe Shenandoah River. Hand capture techniques followed the standardized protocolused by the Wood Turtle Study Group, which required three researchers to search forturtles in 50-meter segments of 1.0-Kilometer stream reaches. Stream habitat variablesfor each 50-meter segment and descriptive data for each turtle captured were recorded.Virginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss190 VIRGINIA JOURNAL OF SCIENCEMost turtles occurred within 5 to 10 meters of the stream banks, demonstratingpreference for shallow, slow-moving stream segments with abundant leaf packs.Statistical analysis revealed a positive correlation between the number of root massesand the number of wood turtles (r = 0.268). Stream depth and number of turtlesshowed a negative correlation (r = -0.215). Root masses had a stronger correlation atthe first site (r = 0.428) than at the second site (r = -0.029). Although the first site andoverall creek data showed low correlations between stream width and number of turtlesfound, the second site exhibited a high correlation (r = 0.5). Further study at these andother sites in the Shenandoah Valley region will help identify the stream variables thathave the greatest influence on wood turtle hibernation site selection, allowing prioritiesto be set for stream protection and rehabilitation.DEPOSITIONAL HISTORY OF THE CARMEL CHURCH BONEBED, AMIOCENE (14 Ma) MARINE VERTEBRATE SITE IN CAROLINE COUNTY,VIRGINIA. Alton C. Dooley, Jr., VA Museum of Natural History, Martinsville, VA24112. The Carmel Church Quarry in eastern Virginia includes at least eightdisconformity-bound Tertiary marine units. One of these units contains a dense fossilvertebrate bonebed, consisting primarily of cetaceans, chondrichthyans, osteichthyans,and turtles, with smaller numbers of other marine and terrestrial mammals. Previousstudies of terrestrial mammals and diatoms indicate that the bonebed correlates to theuppermost part (Bed 15) of the Plum Point Member of the Calvert Formation, with anage of 13.7-14.7 Ma (middle Miocene), and that it disconformably overlies the earlyEocene Nanjemoy Formation. Yet the bonebed contains large numbers of reworkedfossils, including taxa with little or no temporal overlap, such as Carcharoclesmegalodon, Carcharocles cf. C. angustidens, and Otodus obliquus. Most reworkedfossils in the bonebed show extensive evidence of abrasion, but some Eocene taxa(such as O. obliquus) are occasionally found in pristine condition. The presence ofthese specimens suggests that the Nanjemoy-Calvert disconformity included at leasttwo transgressions between the late Eocene and middle Miocene that were completelyremoved by erosion prior to deposition of the Calvert Formation, and that pristineEocene fossils have been reworked from the Nanjemoy into the Calvert throughbioturbation.PostersHABITAT USE AND SWIMMING BEHAVIOR OF LONGNOSE DACE(RHINICHTHYS CATARACTAE) FROM A MERCURY CONTAMINATED RIVER. Kimberly J. Bolyard & Joelle E. Bennett, Dept. of Biol., Bridgewater College,Bridgewater VA 22812. Mercury in the environment has the potential to negativelyaffect the behavior of aquatic organisms through impacts on the nervous system andother physiological functions. We investigated the habitat use and swimmingperformance of longnose dace (Rhinichthys cataractae) from a river with mercurycontamination and from a non-contaminated river in the Shenandoah Valley. Laboratory studies showed that dace from the non-mercury river spent significantlymore time in areas containing large sticks than in areas with pine brush, pallet wood,or nothing. Dace from the mercury contaminated river did not spend significantly moretime in any particular section of the test pool. In a second test, dace from the mercuryVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 91STcontaminated river spent more time swimming in a current over a five minute testperiod than did dace from the uncontaminated river but the difference was notsignificant. Finally, tested fish from the uncontaminated river were significantly longerthan fish from the mercury contaminated river though they were not significantlyheavier. As a result of spending less time in cover and possibly more time swimmingin a current, longnose dace in mercury contaminated rivers may experience higher riskof predation than dace in uncontaminated rivers.DESCRIBING NEMATODE DIVERSITY. Jessica A. Dochney, Thy N. Mai, &Theresa M. Grana, Department of Biological Sciences, University of MaryWashington, Fredericksburg, VA 22401. Nematodes play important roles in nutrientcycling and decomposition. The nematode phylum is one of the most diverse andabundant groups of animals, with a projected 1 million species. However, fewer then29,000 nematode species have been described and only a few species have been studiedin laboratories. The nematode, Caenorhabditis elegans has been well studied as agenetic and developmental model organism. Such studies have contributed to ourunderstanding of basic cell biology, genetics, and neurobiology. Our goal is to find anddescribe new species of nematodes, particularly those that could build upon ourunderstanding of Caenorhabditids. Since 2005, the number Caenorhabditds known toresearchers has doubled from 22 to nearly 50. A representative of one of these newspecies was found on the UMW campus on the northwest side of Seacobeck Hall. Ourwork is part of the formal description of this new species, which will help otherresearchers in understanding the life history of Caenorhabditids. To describe the newspecies, we are taking high-resolution images, measuring identifying body features, andcomparing these data to described species.LEARNED RESPONSES TO HERBIVORE-INDUCED PHYTOCHEMICALS INTHE PARASITIC WASP, COTESIA CONGREGATA (SAY) (HYMENOPTERA:BRACONIDAE). Christopher Crockett & Karen M. Kester, Dept. of Biology, VirginiaCommonwealth University, Richmond VA 23284. Volatile phytochemicals, producedin response to herbivore feeding, aid parasitic wasps in the location of suitableherbivore hosts for reproduction. Several studies have demonstrated that wasps learnthese host-induced volatiles (HIVs) in association with oviposition (“ovipositionallearning”) but whether wasps also can learn these cues within a critical periodfollowing adult emergence (“ post-emergence learning”) is not known. We tested thehypothesis that Cotesia congregata, an endoparasioid of the tobacco hornworm(Manduca sexta L.) learns to recognize HIVs from tomato (var. ‘Castlemart’) throughpost-emergence learning. Inexpensive two- and four-way choice olfactometers werebuilt and evaluated for use with this species, and a behavioral assay was developed. Totest our hypothesis, newly-emerged wasps were given a post-emergence experiencewith leaves from a plant on which a hornworm had fed (“induced leaves”) or a controlplant (“non-induced leaves”). After a refractory period of 48 hours, orientationpreferences of wasps to induced or non-induced leaves were assayed using a two-wayT-tube olfactometer. More wasps experienced with non-induced leaves chose volatilesfrom non-induced leaves (X2 = 3.92, df=1, N = 50, p < .0477), and more waspsexperienced with induced leaves chose volatiles from induced leaves (X2 = 6.48, df=1,N = 50, p < .0109). Results suggest that parasitic wasps can learn HIVs though post-Virginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss192 VIRGINIA JOURNAL OF SCIENCEemergence experience. Continuing work will compare wasp responses to ‘Castlemart’mutants with two levels of HIV silencing.MIGRATION DYNAMICS OF NORTHERN SAW-WHET OWLS IN THE INNERPIEDMONT OF CENTRAL VIRGINIA. Esther Stine & Gene D. Sattler, Dept. ofBiology & Chemistry, Liberty Univ., Lynchburg, VA 24502. Northern Saw-whetOwls (Aegolius acadicus) were mist netted during fall migration in Campbell County,Virginia in 2002-2007 and in 2012 using an audiolure of the male advertisement call. Birds were aged using feather molt criteria and sexed using a wing-mass discriminatefunction. The number of owl captured ranged from a low of 7 to a high of 101. Themajority of birds were encountered during the first two weeks of November, with amedian capture date of November 9 . In most years adults predominated, making upthat least 68% of captures. However, both 2007 and 2012 were invasion years in whichhigher than normal numbers were netted and hatch year birds predominated (59% inboth years). Females predominated in each year, making up at least 61% of captures. The inner Piedmont of central Virginia was established as an important corridor forNorthern Saw-whet Owl migration, and capture rates exceeding those found on theCoastal Plain where the species’ migration in Virginia has previously beeninvestigated.AGE AND PALEO-ENVIRONMENTAL SIGNIFICANCE OF UPPER PALEOZOICOSTRACODES FROM THE APPALACHIAN AND EASTERN ILLINOIS BASINS. Shelby C. Sanders & Neil E. Tibert , Department of Earth and Environmental Sciences,University of Mary Washington, Fredericksburg, VA 22401. Ostracodes recoveredfrom eastern North American coal measures in the Appalachian and Illinois Basinshave the potential to resolve the temporal and spatial relationships of the stratadeposited during the Pennsylvanian-Permian boundary interval. The Greene Formationof the Dunkard Group is the youngest Paleozoic stratigraphic unit in the AppalachianBasin that contains plant fossils of uncertain age. Fossil ostracodes from the GIL 30core in western Kentucky (Illinois Basin) are comparable to the nonmarine ostracodeassemblages from the Dunkard and can be used to constrain the age of the youngestsedimentary rocks in the Appalachian Basin. The GIL30 core was recovered from adown-faulted block located near Sturgis, KY and contains strata deposited during thelatest Pennsylvanian and earliest Permian. We observed three ostracode assemblagesin the GIL 30 core: (A) a marine assemblage dominated by Hollinella cushmani; (B)a brackish assemblage dominated by Geisina upsoni, Cavellina nebrascensis, andVelatomorpha fittsi; and (C) a freshwater assemblage dominated by Whipplellacuneiformis and Haworthina bulletta. The freshwater assemblage in the GIL 30 can becorrelated with confidence to the uppermost units of the Greene Formation (e.g.,Windy Gap Limestone) of the Dunkard Group in the stratotype area of northeasternWest Virginia and southwestern Pennsylvania. These findings confirm an EarlyPermian age for at least the uppermost Dunkard Group strata.TURTLES OF THE FREDERICKSBURG CANAL: INTRODUCED SPECIES ANDESTIMATES OF POPULATION SIZES. Y. Takeda & W. Wieland, Department ofBiological Sciences, Univ. of Mary Washington. The yellow-bellied slider(Trachemys scripta scripta), a non-native to the Fredericksburg area, were found in theVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 93STFredericksburg canal in the spring of 2011. The objectives for summer 2012 was todetermine if this species has established a population, and in addition which species ofturtles live in the canal and obtain life history information on these species, includingestimates of population size. Hoop nets were placed along a 150 m stretch of the canalin the general area of where the single specimen of the yellow-bellied slider was found. Length and weight were taken for each individual and we attempted to determine ageby counting the ridges on the plastron. Population size was determined using openpopulation mark-recapture models. Turtles captured in the canal were red-belliedcooters (Pseudemys rubriventris), painted turtles (Chrysemys picta), eastern snappingturtles (Chelydra serpentina serpentina), stinkpots (Sternotherus odoratus), and thered-eared slider (Trachemys scripta elegans). The yellow-bellied slider was not found. The single specimen found in 2011 was likely introduced as these are common in thepet trade. However, the red-eared slider is also a non-native. The population sizeestimates of species for which we had a sufficient sample size are given by the Jolly-Seber model (open population) for the area of focus (roughly 150m). Estimates includepopulation size and 95% confidence interval: red-bellied cooters, 35 (24-74); paintedturtles, 9 (9-12); red-eared slider, 23 (10-114). Age determination proved to beproblematic. Estimates for growth rates were determined for red-bellied cooters. However, due to small sample size and the short time span over which the study wasconducted (29 capture days) these estimates are suspect.PsychologySTORIES OF BYSTANDER INTERVENTION FROM ACROSS THE GLOBE. S.Y.Teie, P. Randall, R. Wallace & B. Rivero, Center for Applied Behavior Systems,Department of Psychology, Virginia Tech, Blacksburg VA 24060. Actively Caring forPeople (AC4P) wristbands are used to recognize others for performing prosocialbehavior. In 2011, the website AC4P.org was launched to provide a public space forindividuals to share their experiences around AC4P wristband recognition. 74.4% ofwristbands were passed by beneficiaries of kind acts who reciprocated with thewristband (Reciprocators), and 24.6% of wristbands were passed by observers of kindacts who intervened with the wristband (Bystanders). The qualitative analysis of AC4Pstories revealed a differentiation between stories about wristbands passed byReciprocators and those passed by Bystanders. Future research will explore genderinteractions using larger sample sizes.A BEHAVIOR-BASED INTERVENTION TO INCREASE PROSOCIALRECOGNITION IN AN ORGANIZATION. K. M. Pacque, S. M. McCarty, S.Butterworth & C. Holmes, Department of Psychology, Va. Polytechnic Inst. & StateUniv., Blacksburg VA 24061. Today’s fraternal culture needs more caring,compassionate, and interpersonal relationships. News reports document problems frombinge drinking and sexual assault charges, to the death of organization members fromhazing. 73 students in a Greek fraternity received an Actively Caring for People(AC4P) intervention and used Twitter to recognize prosocial behavior. Students self-reported frequency data and relational data on behaviors and relational ties,respectively. Social network analysis (SNA) and multiple regression analyses assessedVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss194 VIRGINIA JOURNAL OF SCIENCEinfluence ties, power ties, influence centrality, and power centrality on performance ofrecognition. Additionally, in- and out-degree ties of Aristotle’s three variations offriendship predicted organization members’ sense of organizational belonging to thefraternity. In general, multiple regression models with SNA measures predicted AC4P-related behaviors better than frequency ties alone. Only friendships based on“goodness” predicted students’ sense of belonging in their organization.A BULLYING-PREVENTION INTERVENTION FOR MIDDLE SCHOOLS:PROMOTING AND REWARDING PROSOCIAL BEHAVIOR TO REDUCEAGGRESSION. B. Tarzia, L. Anderson, G. Yam & M. Armstrong, Center for AppliedBehavior Systems, Department of Psychology, Virginia Tech. This paper discusses thedevelopment of an Actively Caring for People (AC4P) approach to reduce bullying inmiddle schools. An intervention teaching character strengths enabling students todemonstrate discretionary, prosocial behavior is likely to have positive outcomes. Anassessment was conducted to evaluate the relationship between identified AC4Pcharacter strengths and positive outcomes across two schools in Southwest, VA. Students were asked to score their subjective well-being, hope, gratitude, self-efficacyand self-esteem, as well as their involvement in bully-victim behavior anddiscretionary, prosocial behavior (i.e. AC4P behavior). Using regression analyses,students’ self-reported scores in hope, gratitude and self-efficacy were analyzed. Hopeand gratitude were significant predictors in whether or not they participated in bullyingand/or victimizing behavior. In regards to AC4P behavior, hope was a significantpredictor for both performing and receiving AC4P behavior. A one-way ANOVA wasrun with Tukey’s post hoc breaking down students’ bullying behavior by category (i.e.uninvolved students, bullies-only, victims-only and bully-victims). Students’ self-reported “hope” scores were significantly different in all three bully-behaviorcategories (i.e. bullies-only, victims-only and bully-victims) from uninvolved students.THE EFFECT OF NEGATIVE AFFECT ON VISUAL SEARCH PERFORMANCEWITH AND WITHOUT AUTOMATION. Clinton Carter, Rachel R. Phillips, &Poornima Madhavan, Department of Psychology, Old Dominion University, NorfolkVA 23529. To examine the impact of affective valence on threat detectionperformance with and without automation under different levels of uncertainty,participants completed a visual search task in which they had to identify foe presenceor absence in synthetic aperture radar images. Participants consisted of 67 students (23males, 44 females) between the ages of 18 and 51 (M = 21.27, SD = 5.84) from a largesoutheastern university. Participants were randomly assigned to one of two affectivevalence conditions (negative or neutral) and one of two automation conditions (noautomation or perfect automation) and completed the visual search task with both easyand difficult images. A series of 2 automation x 2 affective valence x 2 task difficultymixed factorial ANOVAs were conducted for the dependent variables of sensitivity,response bias, and confidence. Results revealed that there was a significant interactionbetween affective valence and automation for response bias. Specifically, in thenegative condition, response biases were similar between those who received theassistance of the automation and those who did not. However, in the neutral condition,those who received the automation were more liberal than those who did not receivethe automation. These participants also reported the highest confidence and were partVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 95STof the most sensitive group. These findings suggest that participants in the negativecondition may have utilized the aid less than those in the neutral condition. Thisindicates that the potential emotional experiences of an operator should be consideredwhen determining when and where to integrate automated decision aids.HOW THE FRAMING OF METAPHORS AFFECTS PUBLIC POLICY TOWARDTHE MENTALLY ILL. Victoria E. Bennett & Della N. Gibson, Department ofPsychology, University of Mary Washington, Fredericksburg, VA 22401. Metaphorshave been shown to influence people’s decisions about campaigns, ads, and politics.Specifically, studies have looked at how metaphors have influenced attitudes towardcrime and obesity. We examined the effect of blaming and victimization metaphors onpublic policy attitudes toward the mentally ill. It was hypothesized that victimizationmetaphors, as compared to blaming metaphors, would lead to increased support ofoutpatient treatment, increased support of government funding, and increased supportof rights and opportunities for the mentally ill. The results showed no significantdifferences between the three conditions. Additionally, metaphors may not influencepeople with strongly held beliefs toward an issue.A CONCEPTUAL MODEL FOR BYSTANDER INTERVENTION: EXPANDINGPROSOCIAL BYSTANDER BEHAVIOR. M. Sihalath, B. Wright, P. Pierucci & R.Cobb-Ozanne, Center for Applied Behavior Systems, Department of Psychology,Virginia Tech, Blacksburg VA 24060. The frequency of bullying and interpersonalviolence remains high throughout our culture. In these situations where intervention isneeded, bystanders play a vital role in potentially reducing interpersonal violence.Bystanders in emergency or violent situations choose whether or not to intervene forthe prevention of harm and/or injury. This extends the bystander intervention modelfrom the prevention of interpersonal harm to the promotion of prosocial behavior.Participants were given green wristbands to recognize desirable prosocial behavior andwere measured on the four factors from the Latané and Darley model (i.e., “notice”,“interpret”, “responsibility”, and “skills”). In regards to predicting prosocialrecognition, the results suggest “responsibility” and ability to “interpret” weresignificant predictors. In regards to predicting intention to intervene, the data suggests“notice” to be a significant predictor.StatisticsSTATISTICAL PREDICTION FOR VIRGINIA LYME DISEASE EMERGENCEBASED ON SPATIAL-TEMPORAL COUNTS DATA. Yuanyuan Duan , Jie Li , Yili1 1Hong , Korine N. Kolivras , James B. Campbell , Stephen P. Prisley & David N.1 2 2 3Gaines , Department of Statistics, Virginia Tech, Blacksburg, VA 24061 , Department4 1 2of Geography, Virginia Tech, Blacksburg, VA 24061, Department of Forest Resources3and Environmental Conservation, Virginia Tech, Blacksburg, VA 24061 & Virginia4Department of Health, Richmond, Virginia 23219. The emergence of infectiousdiseases over the past several decades has highlighted the need to better understand andprepare for epidemics as endemic infectious diseases. These diseases are usuallyexpanding their geographic range and are recorded over multiple time periods, makingVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss196 VIRGINIA JOURNAL OF SCIENCEthe analysis and prediction more complicated. This study is based on areal (census tractlevel) counts data of lyme disease cases in Virginia from 2003 to 2010. Ourvisualization of incident rate using kernel smoothing showed the spatial-temporalemergence pattern of Lyme disease. We built a spatial Poisson regression model withrandom effect to incorporate spatial-temporal correlations. The random effects weremodeled by Conditional Auto-Regressive (CAR) model. We used Markov chain MonteCarlo (MCMC) algorithm to fit the model in a Bayesian framework. Using this model,we analyzed 5 year sum counts from 2006 to 2010 to identify significantlandscape/demographic variables for different eco-regions. We also analyzed yearlycounts data and predict 2011’s count based on previous years. This study was fundedby NSF-BCS-GSS Grant.PRODUCT COMPONENT GENEALOGY MODELING AND WARRANTYRETURN PREDICTION. Caleb King , Yili Hong & William Q. Meeker ,1 1 2Department of Statistics, Virginia Tech, Blacksburg, VA 24061 & Department of1 2Statistics, Iowa State University, Ames IA 50011. Many industrial products consist ofmultiple components that are necessary for maintaining their integrity. There is alreadyan abundance of literature on modeling the lifetime of such components throughcompeting risks models. However, none of these models take into consideration thefact that many of these components are part of a specific generation of components andthat each of these components may go through several generations throughout a periodof observation. We believe that by incorporating this extra information, which isreadily available in most product warranty databases, better accuracy can be achievedin predicting time to failure thus yielding more reliable warranty return information.We present methods for estimating parameters for this new generational model as wellas provide a comparison with existing methods through the use of simulation. Ourresults indicate that the generational model outperforms the existing methods inpredicting warranty returns.DEGRADATION DATA ANALYSIS USING NONLINEAR MODELS VIA SHAPE-RESTRICTED SPLINES. Zhibing Xu & Yili Hong, Department of Statistics,Virginia Tech, Blacksburg, VA 24061. The traditional reliability analysis is based onthe failure-to-time data. However, many products are designed with high reliability andlow risk for failures. It is hard to collect enough failure-to-time data to give a preciseestimation for the highly reliable products in a limit time, even under acceleratedconditions. Comparing to the traditional life time data, degradation data not only canprovide enough information in a short time, but also give a better estimation andprediction of the life time of products. With the improvement of technology, life-affecting environmental variables are recorded as well as the degradation measure overtime. For example, the large amount of ultraviolet exposure will accelerate thedegradation of the polymer coating. Thus, it is important to incorporate theenvironmental variables, also called dynamic covariates, into the degradation pathmodel. Existing research suggests a linear mixed effect model with splines to combinethe dynamic covariates. With unspecific effect function for the dynamic covariates,shape restricted splines method is used. However, the linear model may not widelyused in real applications, because most of the degradation data follow a nonlinear trend,especially in the pharmacokinetic research. In this paper, we propose a nonlinear mixedVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 97STeffect model combing with dynamic covariates to model the degradation data. Shaperestricted splines are used in the proposed model and a modified alternating algorithmis developed. The performance of the algorithm is evaluated by simulations. Anoutdoor weathering dataset is used for illustration of the proposed method.DECISION TREES WITH HIGH DIMENSIONAL DATA AND EXPLICITCOVARIATE SELECTION. Lucas Roberts, Department of Statistics, Virginia Tech,Blacksburg, VA 24061. In this talk, we survey the decision tree literature from botha model based and a heuristic perspective. Primarily, these methods have beensuccessfully deployed in cases where prediction is of interest. When we are mainlyinterested in inference, which in this context means identifying the most usefulcovariates, these methods can exhibit poor behavior. These inferential problems areexacerbated in high dimensional spaces. After reviewing some general covariatefiltering approaches, we present our method, which does covariate filtering anddecision tree search jointly. We state necessary and sufficient conditions ensuringconsistency of the tree classifier and we show the benefits of our approach throughsimulated and real examples. We also give a rule for ranking and selecting covariatesusing output from our model. We compare our covariate ranking and selectionprocedure to current common approaches.CLUSTER-BASED PROFILE MONITORING IN PHASE I ANALYSIS. YajuanChen, Jeffery B. Birch & William Woodall, Department of Statistics, VirginiaPolytechnic Inst. & State Univ., Blacksburg VA 24061. An innovative profilemonitoring methodology is introduced for Phase I analysis. The proposed technique,referred to as cluster-based profile monitoring, incorporates a cluster analysis phase toaid in determining the possible existence of “profiles” in the historical data set (HDS)resulting from an out-of-control process. To cluster the profiles, the proposed methodfirst replaces the data from each sampled unit with an estimated profile, using someappropriate regression method, and clusters the profiles based on their estimatedparameters. This cluster phase then yields an initial main cluster which contains at leasthalf the profiles. The initial estimated population average (PA) profile is obtained byfitting a linear mixed model to those profiles in the main cluster. Profiles, determinedusing the Hotelling’s T2 statistic, that are not contained in the initial main cluster areiteratively added to the main cluster and the mixed model is used to update theestimated parameters for the PA profile. Those profiles contained in the final maincluster are considered as resulting from the in-control process while those not includedare considered as resulting from an out-of-control process. A simulated example andMonte Carlo results demonstrate the performance advantage of this proposed methodover a current non-cluster-based method with respect to more accurate estimates of thePA parameters.ESTIMATION AND EVALUATION OF OPTIMAL THRESHOLDS FOR THEBELIEVE THE POSITIVE SEQUENTIAL TESTING STRATEGY. Amber R. Wilk& Donna K. McClish, Department of Biostatistics, Virginia CommonwealthUniversity, Richmond VA 23219. Many continuous medical tests rely on a thresholdfor diagnosis. A sequential testing strategy, called Believe the Positive, classifies apatient positive if either the first test is greater than a threshold è1 or negative on theVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss198 VIRGINIA JOURNAL OF SCIENCEfirst test and greater than è2 on the second test. A threshold pair è = (è1, è2) wasdefined as optimal if it maximized GYI = Sensitivity (Se) + r*(Specificity (Sp) – 1).Of interest is to determine if the optimal threshold pair estimates are “good” whencalculated from a sample. Formulas were derived to estimate è* assuming tests followa binormal distribution, using the Newton-Raphson algorithm. A simulation study wasperformed assessing bias, root mean square error (RMSE), percentage of under/overestimation of Se/Sp, and coverage of simultaneous confidence intervals (SCI)/ellipsesfor sets of population parameters and sample sizes. Bootstrapping was used to estimatethe variance of each optimal threshold pair. When the area under the curve (AUC)between the two tests is equal, the median RMSE and median bias in Se and p appearsto decrease. The coverage of the SCI and confidence ellipses improves as sample size,AUC, and the ratio of standard deviations of each group increases. This also holds truefor percentage of overestimation of Se and Sp except when the allocation betweengroups favors those without disease. This method is an improvement over an empiricalestimate.LIKELIHOOD BASED FORECASTING OF LOCALLY CHANGING VARIANCESTRUCTURED TIME SERIES DATA. Rajan Lamichhane & Norou Diawara, Dept.of Mathematics and Statistics, Old Dominion University, Norfolk VA 23529. In thistalk, we assume several non-linear time series models with additive noise components,and the model fitting is proposed in two stages. The first stage identifies the densityratio using all the clusters information, without specifying any prior knowledge of theunderlying distribution function. In the second stage, we partition the time series intoconsecutive non-overlapping intervals of quasi stationary increments where thecoefficients shift from one stable regression relationship to a different one usingbreakpoints detection algorithm. These breakpoints are estimated by minimizing thelikelihood from the residuals. We develop a method based on finite mixture ofGaussian distributions to forecast this type of partitioned data. The Expectation-Maximization (EM) algorithm, with initial values obtained from the empiricalestimates; give the estimates of mixture distribution. We propose likelihood basedmethod to identify the most probable value from a pool of past values for theforecasting.ESTIMATING EXPLAINED VARIATION FOR AN UNDERLYING LINEARMODEL USING LOGISTIC REGRESSION. D. Sharma, Department of Mathematicsand Statistics, James Madison University, Harrisonburg, VA 22807. The coefficientof determinant, also known as the R statistic, is used to assess the extent of the2strength of the relationship between a response and explanatory variables in a linearregression model. It measures the proportion of variation in the response variableexplained by a set of independent variables. In many real life events, interest lies onmodeling the relationship between a continuous response variable and a set ofpredictors. But in practice, the continuous dependent variable of interest may not beobservable and is usually represented by its binary proxy. In such situations, logisticregression is a popular choice. There are many R type statistics proposed to measure2of explained variation for logistic regression. The pseudo R measure ( stands out2because of its intuitive interpretation and independence on the proportion of successin the sample. It, however, severely underestimates the proportion of explainedVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 99STvariation (P ) in the variable underlying the binary indicator of event occurrence. In2this research we present a method for estimating the explained variation for theunderlying linear model using the pseudo R statistics obtained from a logistic2regression analysis.STRUCTURED FUNCTIONAL ADDITIVE REGRESSION IN REPRODUCINGKERNEL HILBERT SPACES. Hongxiao Zhu , F. Yao & H. H. Zhang ,1 2 3Department of Statistics, Virginia Tech, Blacksburg, VA 24061, Dept. of Statistics,1 2University of Toronto, & University of Arizona. Functional additive models (FAMs)3provide a flexible yet simple framework for regressions involving functional predictors.The utilization of data-driven basis in an additive rather than linear structure naturallyextends the classical functional linear model. However, the critical issue of selectingnonlinear additive components has been less studied. In this work, we propose a newregularization framework for the structure estimation in the context of ReproducingKernel Hilbert Spaces. The proposed approach takes advantage of the functionalprincipal components which greatly facilitates the implementation and the theoreticalanalysis. The selection and estimation are achieved by penalized least squares using apenalty which encourages the sparse structure of the additive components. Theoreticalproperties such as the rate of convergence are investigated. The empirical performanceis demonstrated through simulation studies and a real data application.DICTIONARY LEARNING FOR GENETIC DATA IN RELATED INDIVIDUALS. Xiaowei Wu & H Zhu, Department of Statistics, Virginia Tech, Blacksburg, VA24061. Dictionary learning has been widely used for sparse representation and datarestoration in computer vision and image analysis. However, its application instatistical genetics has not been fully explored. We consider the problem of learningthe sparsity structure of genetic data in related individuals. We propose anonparametric Bayesian method based on a latent beta process factor analysis model.The method can be used in association study with related individuals for multiplepurposes, for example, genotype imputation, estimation of allele frequency or kinshipcoefficients. Simulation study shows good performance and advantages of the proposedmethod in comparison with some existing methods. We also apply our method to theanalysis of the GAW 18 data set.EVALUATING THE HEALTH EFFECT(S) OF ENVIRONMENTAL CHEMICALSAND NUTRITION: CONSIDERATIONS FOR HIGHLY CORRELATED DATA. Chris Gennings, Virginia Commonwealth University, Richmond VA 23298. Biomonitoring of environmental chemicals in human tissues and fluids has shown thatall people, not just those working in or living near major pollution sources, carry a“body burden” of synthetic chemicals in their blood, fat, mother’s milk, semen, urineand breath. Despite increasing public concern regarding the potential health effects ofpervasive and persistent environmental exposures, there are few, if any,recommendations for life style changes to mitigate such effects. Our research focus isa holistic evaluation of the impact of environmental chemicals on health effectsincluding metabolic endpoints (e.g., obesity, hyperlipidemia) and hepatotoxicity andthe role nutrition may play in potentially mitigating the impact of chemical exposures.As such, we are not focused on single chemicals or nutrients – instead, we are focusedVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1100 VIRGINIA JOURNAL OF SCIENCEon their potential combined effects. Standard statistical methods suffer frommulticollinearity effects due to the complex correlation structure among thecomponents. We have developed novel statistical methods to determine empiricalweights that identify subsets of components most associated with a response variable.The results are weighted indices of body burden and nutrients – i.e., simple tounderstand and visualize indices that may be easily combined with other behavioraldata for additional analyses. Characterization of the approach will be described throughsimulation studies. The methods will be demonstrated using NHANES data.Structural Biology, Biochemistry, and BiophysicsUBIQUITIN MODULATES TOLLIP’S LIPID-MEDIATED ENDOSOMALMEMBRANE BINDING. S. Mitra, A. Traughber, S. Gomez, & D. G. S. Capelluto.Department of Biological Sciences, Virginia Tech, Blacksburg, VA 24061. Ubiquitylation is a highly controlled post-translational modification of proteins, inwhich proteins are conjugated either with monoubiquitin or polyubiquitin chains.Ubiquitin modifications on target proteins are recognized by ubiquitin-bindingdomains, which are found in several effector proteins. In this study, we describe for thefirst time how ubiquitin controls the function of the Toll-interacting protein (Tollip),which is an effector protein in the innate immune signalling pathway and an adaptorprotein for endosomal trafficking. We have demonstrated that the central C2 domainof Tollip preferentially interacts with phosphoinositides. Remarkably, we haveobserved an ubiquitin dose-dependent inhibition of binding of Tollip tophosphoinositides and it does so specifically by blocking Tollip C2 domain-phosphoinositide interactions. This led us to discover that the Tollip C2 domain is anovel ubiquitin-binding domain. In addition, we have biophysically characterized theassociation of the Tollip CUE domain to ubiquitin and compared it with Tollip C2domain-ubiquitin binding. We have also found that ubiquitin binding to dimeric TollipCUE domain induces a drastic conformational change in the protein, leading to theformation of a heterodimeric Tollip CUE-ubiquitin complex. These data suggest thatubiquitin binding to the Tollip C2 and CUE domain and ubiquitin-mediateddissociation of CUE dimer reduces the affinity of Tollip protein for endosomalphosphoinositides, allowing Tollip’s cytoplasmic sequestration for its cytosoliccommitments.M ECHANISTIC STUDIES OF A FLAVIN DEPENDENT LYSINEMONOOXYGENASE FROM NOCARDIA FARCINICA. R.M. Robinson, P.Rodriguez, N. Keul, & P. Sobrado, Department of Biochemistry, Virginia Tech,Blacksburg VA 24061. The kinetic and chemical mechanism of the lysinemonooxygenase Nocobactin G (NbtG) from Nocardia farcinica was investigated usingboth steady-state and rapid reaction kinetics. NbtG hydroxylates both L- and D-lysine,a unique characteristic among members of the N-hydroxylating monooxygenases(NMOs), which are typically selective for the L-stereoisomer of their respectivesubstrate. NbtG is mostly uncoupled (~75%) and produces superoxide and hydrogen2peroxide as oxidative by-products. O consumption increases from 0.581 s in the-1absence of lysine to 1.09 s and 3.03 s when saturating concentrations of L- and D--1 -1Virginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 101STcat mlysine are present, respectively. NbtG has a ~8-fold higher k /K value for L-lysinesuggesting it is the preferred substrate. Reduction of NbtG by NADPH is independentof both lysine stereoisomers and occurs in two exponential phases with rates of 1.00s and 0.119 s for the first and second phase, respectively. Primary kinetic isotope-1 -1redeffect studies with pro-(R)-NADPD indicate a k value of ~1.5 where both phases ofDreduction are isotopically sensitive. The rate of flavin oxidation is affected by lysineand increases from 3.1 x 10 M s in the absence of lysine to 3.9 x 10 M s and 5.43 -1 -1 3 -1 -1x 10 M s when saturating concentrations of L- and D-lysine are present, respectively.3 -1 -1catSolvent kinetic isotope studies yield a k value of ~2.4. These results indicate theD2Oactive site of NbtG is loosely stereospecific for lysine and hydride transfer is onlypartially rate-limiting, but a chemical step that is influenced by proton exchange withsolvent is rate-limiting. This work was supported by a grant from the National ScienceFoundation MCB-1021384.A CONSERVATIVE ISOLEUCINE TO LEUCINE MUTATION CAUSES MAJORREARRANGEMENTS AND COLD-SENSITIVITY IN KLENTAQ DNAPOLYMERASE. E. Y. Wu , E. P. Kornberg , B. Zielinski , W. M. Barnes , & M. B.1 1 1 2Kermekchiev , Dept. of Biology, University of Richmond, Richmond, VA 23173,3 1Dept. of Biochemistry and Molecular Biophysics, Washington University School of2Medicine, St. Louis, MO 63110, & DNA Polymerase Technology, Inc., St. Louis, MO363104. Assembly of PCRs at room temperature can lead to low yields or unintentionalproducts due to mispriming. A conservative mutation of isoleucine 707 to leucine inDNA polymerase I from Thermus aquaticus substantially decreases its activity at roomtemperature without compromising its ability to amplify DNA. To understand whysuch a small change located 24Å from the active site can have a large impact onenzyme activity at low temperature, we solved the X-ray crystal structure of the largefragment of Taq DNA polymerase containing the cold-sensitive mutation (Cs3CKlenTaq) in the closed, ternary (E-DNA-ddNTP) and open, binary (E-DNA)complexes. The Cs3C KlenTaq ternary complex was identical to the wild-type exceptfor the mutation and a rotamer change in a nearby phenylalanine 749, suggesting thatthe enzyme should remain active. However, soaking out the nucleotide substrate at lowtemperature results in an altered binary complex made possible by the rotamer changeat F749 near the tip of the polymerase O-helix. Surprisingly, two adenosines in the 5´-template overhang fill the vacated active site by stacking with the primer strand,thereby blocking the active site at low temperature. These results explain the coldsensitivity phenotype of the I707L mutation in KlenTaq and serve as an example of alarge conformational change affected by a conservative mutation.EVOLUTION AND NETWORK ANALYSIS OF LIPOCALIN PROTEINS USINGCOMPUTATIONAL APPROACHES. Nardos F. Sori & Lesley H. Greene. Dept. ofChemistry and Biochemistry, Old Dominion University, Norfolk Va. 23529. Thelipocalins are a functionally diverse and divergent superfamily of proteins that sharea common three- dimensional structure which consists of an antiparallel â-barrel anda C-terminal á-helix. Lipocalins are found in various organisms with a wide range offunctions such as pheromone activity, lipid transport and coloration. Even thoughlipocalins are found widely in nature, the first indication of its existence in plants wasobserved in late 20 century by the sequence analysis of Violaxanthin de-epoxidasethVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1102 VIRGINIA JOURNAL OF SCIENCEand Zeaxanthin epoxidase, which were shown to be lipocalin-like proteins. Recentlylipocalins from Wheat and Arabidopsis were identified as true lipocalins through theelucidation of three structurally conserved regions. The study of these lipocalins is vitalas these proteins are believed to help plants tolerate oxidative stress and extremeconditions which broadens our understanding of plant sustainability in differentenvironments. Protein structure homology modeling was used to construct apreliminary structure of the temperature induced wheat lipocalin. Furthermore,conservation of amino acids through evolution was analyzed by using modifiedShannon entropy parameter with 20 divergent lipocalins. It revealed that three positionshave conservation that have greater than 0.45 which indicates high conservation.THERMOTOLERANCE IN MAMMALIAN CELLS. Ovidiu Lipan, Dept. of Phys.,Univ. of Richmond, Richmond VA, 23173. The heat-shock response network controlsthe adaptation and survival of the cell against environmental stress. A key element ofthe heat-shock network is the heat-shock transcription factor-1 (HSF1), which istransiently activated by elevated temperatures. The HSF1 homotrimers bind to the heatshock element on the DNA and control the expression of the hsp70 gene. The Hsp70proteins protect cells from thermal stress. The modulation of the activity of the hsp70-promoter by the intensity of the input stress is thus critical for cell's survival. Thepromoter transient activity starts from a basal level and rapidly increases once the stressis applied, reaches a maximum level and attenuates slowly back to the basal level. Themolecular construct used as a measure of the response to thermal stress is a Hsp70-GFPfusion gene transfected in Chinese hamster ovary (CHO) cells. The time profile of theGFP protein depends on the transient activity, of the heat shock system. The GFP timeprofile is recorded using flow cytometry measurements, which allows a quantitativemeasurement of the fluorescence of a large number of cells (10 ). The GFP responses4to one and two heat shocks were measured for 261 conditions of different temperaturesand durations. We found that: (i) the response of the cell to two consecutive shocksdepends on the order of the input shocks. This phenomenon is thermotolerance withoutrecovery time in between shocks; (ii) the responses may be classified as mild or severe,depending on the temperature level and the duration of the heat shock and (iii) theresponse is highly sensitive to small variations in temperature. INHIBITION OF SIDEROPHORE BIOSYNTHESIS BY TARGETING A.FUMIGATUS ORNITHINE HYDROXYLASE: A STRUCTURE-BASED VIRTUALSCREENING STUDY. S. Badieyan & P. Sobrado , Department of Biochemistry,1 1,2 1Virginia Tech, Blacksburg, VA 24061 & Virginia Tech Center for Drug Discovery,2Virginia Tech, Blacksburg, VA 24061. Siderophore A (SidA) from Aspergillusfumigatus is a flavin-dependent monooxygenase that catalyses hydroxylation of L-ornithine. N5-hydroxyl-ornithine is subsequently incorporated into hydroxamate-containing siderophores. The pathogenicity of A. fumigatus in mammals is dependenton the availability and function of siderophores. Thus, inhibition of the siderophorebiosynthetic pathway may significantly affect the virulence of this fungus. SidA playsa central role in this pathway and is considered the key target for inhibitory study.Availability of several high-quality structures of SidA in complex with differentligands allowed the in-silico screening of a large library of drugs, natural products, andsynthetic compounds. The screening results were sorted by the absolute and normalizedVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 103STbinding energies and then filtered based on the formation of at least one hydrogen bondto the part of the ornithine binding site with several hydrogen bond donors. The top hitshave average molecular weights of 200 Da and ÄG bindings of -7.5 kcal/mol. They arehighly soluble nitrogen- or oxygen-rich compounds that make several hydrogen bondswith residues in the active site of SidA. This study was supported by NSF grant MCB1021384 and by the Virginia Tech Biodesign and Bioprocessing Center.A NOVEL TANK-BINDING KINASE 1 (TBK1) SUBSTRATE AND ITSENDOGENOUS CONTROL OF A CRITICAL CATALYTIC HUB IN THE TYPE-IINTERFERON RESPONSE. James D. Marion, Jr. & Jessica K. Bell. VirginiaCommonwealth University, Richmond, VA 23298. To defend against pathogenchallenge, multi-cellular organisms mount an immune response that recognizes,sequesters and eradicates invading infectious agents. In response to receptor-mediatedpathogen detection, several signaling pathways converge to activate TBK1, a kinasewhich phosphorylates substrates that lead to the production of pro-inflammatorycytokines and type-I interferons. Increasingly, TBK1 dysregulation has been linked toautoimmune disorders and cancers, heightening the need to understand its regulatorycontrols. Here, we describe the mechanism by which Suppressor of IKKe (SIKE)inhibits the TBK1-mediated phosphorylation of IRF3, a transcription factor essentialto type-I interferon production. Kinetic analysis revealed that SIKE not only inhibitsIRF3 phosphorylation, but is also a high-affinity, TBK1 substrate. Further analysisdetermined that TBK1 phosphorylation of IRF3 and SIKE displayed negativecooperativity. Biophysical experiments revealed that TBK1 directly phosphorylatedSIKE on 6 C-terminal residues that mediated TBK1:SIKE associations and exhibitedstriking homology to the IRF3 phosphorylation motif. Taken together, our findingsdemonstrate that SIKE functions as a TBK1 substrate that regulates this criticalcatalytic hub, not through direct repression of activity, but by redirection of catalysisthrough substrate affinity.MOLECULAR MECHANISM OF MEMBRANE TARGETING BY ENDOSOMALADAPTOR PROTEINS. Shuyan Xiao & Daniel G. S. Capelluto, Protein SignalingDomains laboratory, Department of Biological Sciences, Virginia Tech, BlacksburgVA, 24061. The Toll-interacting protein (Tollip) is an effector protein in the innateimmune signaling pathway and an adaptor protein for endosomal trafficking. Tollipmediates protein sorting by association with Tom1, polyubiquitinated cargo, andclathrin. Tollip is modular in architecture with an N-terminal Tom1-binding domain(TBD), a central C2 domain, followed by a C-terminal CUE domain. Tom1 is amember of a protein family characterized by the presence of the VHS(Vps27/Hrs/STAM) domain at the N terminus, and a central GAT domain. In thisstudy, we identified that Tollip-TBD directly binds to Tom1-GAT with a very highaffinity. The solution structure of the complex formed by the Tollip-TBD and Tom1-GAT domain reveals that Tollip-TBD undergoes a coil/helix folding transition uponbinding to GAT domain, forming an anti-parallel â-sheet followed by a short á helixstructure. The amphipathic helix á1 and á2 of GAT play a key role in the interactingwith TBD. We also demonstrated that Tom1 GAT binds to Tollip TBD and ubiquitinin a mutually exclusive manner. Overall, our study provides the structural andmolecular basis of Tollip-TBD and Tom1-GAT binding interactions, which will aid inVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1104 VIRGINIA JOURNAL OF SCIENCEunderstanding the role of Tollip in endosomal trafficking. This work was supported bythe American Heart Association.STRUCTURAL BASIS OF PHOSPHATIDYLINOSITOL 4,5-BISPHOSPHATE(PIP2) RECOGNITION BY THE TIRAP PIP2-BINDING MOTIF. Xiaolin Zhao,Shuyan Xiao, Ayana Stukes & Daniel G. S. Capelluto, Department of BiologicalSciences, Virginia Tech., Blacksburg VA. 24061. TIRAP is an adaptor moleculeessential to the TLR2 and TLR4 signaling pathways. TIRAP contains a TIR domain,which is responsible for association with another adaptor protein, MyD88; and a N-terminal PIP2 binding domain (PBD) that binds to plasma membrane PIP2. Theassociation of MyD88 and TLR is stabilized by the binding of TIRAP to the plasmamembrane. Remarkably, a minimal region (residues 15-35) within PBD is sufficientto associate to the plasma membrane. We show that this region, that we called thePIP2-binding motif (PBM), adopts a helical conformation in the presence of themembrane mimic dodecylphosphocholine micelles and, more importantly, binds PIP2.NMR data indicates that TIRAP PBM binds PIP2 in a fast exchange regime, suggestinga moderate affinity for lipid binding. According to the TIRAP PBM resonanceassignments, we found that conserved residues Lys16, Leu18, Lys20, Lys31, andLys32 play important roles in association with PIP2. We are in the process to preciselyidentify the PIP2-binding site in TIRAP PBM and to obtain the solution structure of thepeptide to understand the progressive mechanism and regulation of membrane insertionof the protein.PROBING CONFORMATIONAL CHANGES WITH MULTIWAVELENGTHCOLLISIONAL QUENCHING. Ellis Bell & Chun Li, Laboratory for StructuralBiology, Biophysics & Bioinformatics, Dept. of Chemistry, University of Richmond,Richmond VA 23173. To probe the local environments of tryptophans in proteins wehave developed a novel analysis of fluorescence collisional quenching using multipleemission wavelengths, combined with differently charged quenchers (acrylamide:neutral, Iodide: negative & Cesium: positive). With N Acetyl tryptophan, similar Stern-Volmer constants are obtained at all wavelengths, as expected. With glutamatedehydrogenase, (4 tryptophans per subunit), 4 distinct families of constants areobtained with the highest occurring at long wavelengths and progressively decreasingas the wavelength band analyzed decreases showing this approach separates the effectson individual tryptophans in the protein. We have explored nucleotide effects onaccessibility and charge environment of the 4 tryptophans. Results indicate that overall,nucleotide ligands NADH, NADPH and ADP significantly lower accessibilityparticularly in the region of subunit interfaces while GTP has less effect and in the caseof W72 and W281 increases accessibility. Acrylamide quenching gives Stern-Volmerplots concave upwards, which become linear when lifetime measurements are used.With NAD(P)H, the charged quenchers, Iodide and Cesium (but not the neutralAcrylamide) give negative slopes which appear to result from differential effects ontryptophan-NAD(P)H resonance energy transfer. Funded in part by NSF Grant MCB0448905 to EB.SYNTHETIC BIOLOGY: SYNTHETIC CIRCUITS AS A TOOL FORBIMOLECULAR SYSTEMS EXPLORATION. Warren C. Ruder, Dept. of BiologicalVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 105STSystems Engineering, Virginia Tech, Blacksburg VA, 24061. Professor Ruder joinedVirginia Tech’s Biological Systems Engineering department as an assistant professorin 2012. His expertise in synthetic biology, cellular biomechanics, and lab-on-a-chipsystems complements Virginia Tech’s strengths in biomolecular, biomedical, andenvironmental engineering. Most recently, he spent two and a half years in thesynthetic biology laboratory of James Collins, a Howard Hughes Medical Investigatorand NAE and IOM member based at Boston University and the Harvard Wyss Institutefor Biologically-Inspired Engineering. Professor Ruder began his academic careerstudying civil and environmental engineering at MIT and graduated with an S.B. in2002 with a focus in mechanics. He spent two years as a research assistant at HarvardMedical School studying cell calcium and cAMP signaling systems. He then spent fouryears at Carnegie Mellon University, where he received an M.S. in MechanicalEngineering and a Ph.D. in Biomedical Engineering. His thesis work focused on thedesign of micromechanical systems to explore cell biomechanics.PostersWHERE DID THE SYNUCLEINS COME FROM? A COMPUTATIONAL LOOKINTO THE PAST. Agatha Munyanyi, John Bedford & Lesley H. Greene, Departmentof Chemistry & Biochemistry, Old Dominion University, Norfolk VA 23529. á-, â-and ã-Synucleins have been described as physiologically unstructured proteinsprimarily expressed in the brain tissue of vertebrates. Very recently á-synuclein hasbeen shown to form a helical tetramer which is resistant to fibril formation underphysiological conditions. This folded form of á-synuclein may be its natural state. Abioinformatics investigation was conducted to construct a synuclein superfamily tofacilitate understanding their physiological structure, function and evolutionary history.The results identified similarity to an endoglucanase enzyme from bacteriumAcetobacter pomorum , a CRE-DUR-1 protein from a nematode and a putative proteinfrom the Tasmanian Devil. The Tasmanian Devil protein in turn identified acytochrome c from the bacterium vibrio cholera. In conclusion however, the synucleinsmost surprisingly seem to be orphan proteins in the vertebrate line.MOLECULAR DOCKING AND VIRTUAL SCREENING OF PPAR-ä AGONISTS. K. E. Ascetta, S. N. Lewis & D. R. Bevan, Department of Biochemistry, Virginia Tech,Blacksburg VA 24061. Peroxisome proliferator-activated receptor subtype delta(PPAR-ä) is a transcription factor that causes increased fatty acid transport andoxidation in white adipose tissue and skeletal muscle. It also plays a role incarbohydrate catabolism and fat synthesis in the liver. All of these processes requireactivation by agonists, which can be developed as treatments for metabolic disorderslike type II diabetes and heart disease. One way to identify viable PPAR-ä agonists isthrough virtual screening. Screening requires a 3D model that best represents thereceptor. In this study eighteen ligand-bound PPAR-ä crystal structures were testedwith docking to determine which ones could serve as representative models. The fitnessof these structures was evaluated by measuring root mean squared deviation (RMSD)and assessing protein-ligand interactions that match crystal structure references. Thefour fittest models were then used to screen a small group of published known PPAR-äagonists that are not present in crystal structures. Here, only protein-ligand interactionsVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1106 VIRGINIA JOURNAL OF SCIENCEwere used as measures for successful docking. Of the fittest models, 3SP9 and 1GWXfaired the best in re-docking and cross-docking, while 3DY6 and 3SP9 most accuratelypredicted the protein-ligand interactions of the non- crystalized agonists. Presently,3SP9 appears to be the model best fit for virtual screening to identify PPAR-ä agonists.VIRGINIA TECH CENTER FOR DRUG DISCOVERY SCREENINGLABORATORY. P. Sobrado , N. J. Vogelaar & D. G. I. Kingston , Department of1 1 2 1Biochemistry, Department of Chemistry, & Virginia Tech Center for Drug2 1,2Discovery, Virginia Tech, Blacksburg, VA 24061. A new high-throughput screeninglaboratory has been established at Virginia Tech and is available for use by thescientific community. The facility instrumentation for microplate preparation includesa versatile liquid dispenser and a robot capable of high-accuracy plate-to-platetransfers. Detection of assay results can be done with UV-Vis absorbance,fluorescence, time-resolved fluorescence, glow luminescence, or through use of aUPLC. The facility has over 35,000 compounds available for screening use. Theapplication of high-throughput screening methodologies was demonstrated in an assaydesigned to identify inhibitors of SidA (AfSidA) from Aspergillus fumigatus. In apreliminary screening of 640 compounds from the ChemBridge Diverset compoundlibrary, N-(5-chloro-2-methylphenyl)-N-[(4-methylphenyl)sulfonyl]glycine wasidentified as a potential inhibitor and found to have a SidA-binding constant of 14 ?M.Further screening of the ChemBridge library is underway. The Virginia Tech Centerfor Drug Discovery is supported by the Fralin Life Science Institute, the Virginia TechCollege of Science, and the Virginia Tech College of Agricultural and Life Sciences.CHARACTERIZATION OF A FLAVIN-DEPENDENT –MONOOXYGENASEFROM CUPRIAVIDUS TAIWANENSIS. Aaron. Gringer , Isabel Da Fonseca & Pablo1 1Sobrado , Department of Biochemistry, Virginia Tech Center for Drug Discovery1,2,3 1 2& Fralin Life Science Institute, Virginia Tech, Blacksburg, VA 24061. Microbial iron3acquisition depends on complex steps and many of which are not shared by highereukaryotes. Inhibition of siderophore biosynthesis; a specific set of microbial reactionscritical for iron acquisition and storage, is therefore a potential antimicrobial strategy.One essential enzyme for this process is a flavin-dependent N-monooxygenase (NMO).Seeking for more insights of how this enzyme class works, we cloned, purified andinitially characterized the L-ornithine-5-monooxygenase from Cupriavidus taiwanensis.The codon optimized gene was cloned into pVP56K and expressed in E. coliBL21(DE3). The recombinant target protein with MBP fusion was purified usingcataffinity column and digested to remove MBP by treatment with Tev protease. The kMand K values with L-ornithine are 0.382 ± 0.009 s and 55 ± 7 µM, and with NADPH-1are 0.30 ± 0.04 s and 46 ± 2 µM, respectively. Supported by NSF grant MCB-11021384.COMPUTATIONALLY CHARACTERIZING THE STRUCTURAL DYNAMICSAND INTERACTIONS OF IRISIN D. R. Maddox, A. M. Brown, & D. R. Bevan,Department of Biochemistry, Virginia Tech, Blacksburg VA 24061. Obesity is anepidemic in today’s society with many negative effects, including type II diabetes,heart disease, and stroke. Due to this public health problem, the study of adipose tissueis central to the understanding of metabolic abnormalities associated with theVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 107STdevelopment of obesity. Irisin is a protein cleaved from the fibronectin type III domaincontaining 5 (FNDC5) protein in the skeletal muscle. It affects white adipose tissue byacting as a metabolic mediator. This effect is known as the “browning” of white fatwhere white adipose tissue is physiologically converted into brown adipose tissue. Weapplied computational techniques to investigate the characteristics of irisin by studyingproteins homologous to irisin. Homology modeling with Modeller, DaliServer/LOMETS structure searches, and molecular dynamics simulations performedwith GROMACS have been utilized to study the way irisin acts in comparison to thewell-known homologous proteins. As a preliminary result, irisin acts much like theneural cell adhesion molecule 2 (NCAM2), which is thought to bind to the alpha-2,8-polysialic acid surface receptor in neural cell membranes. NCAM2 shares secondaryand tertiary structure similarity to irisin, which suggests the proteins may share surfacereceptor types. Exploring the structural characteristics of the NCAM2 receptor mayimprove understanding of the role of irisin in metabolism.TOWARDS THE STRUCTURAL AND FUNCTIONAL CHARACTERIZATION OFTHE HFQ HOMOLOG FROM AQUIFEX AEOLICUS. Kimberly Stanek, JenniferPatterson, & Cameron Mura, Department of Chemistry, University of Virginia,Charlottesville, VA 22904. Bacterial survival requires careful regulation of mRNAturnover. The host factor ‘Hfq’ is a bacterial RNA chaperone that functions in mRNAturnover and related pathways, by modulating mRNA-sRNA interactions. As thebacterial branch of the 'Sm' superfamily, Hfq homologs tend to assemble into cyclicoligomers that associate with RNA. An Hfq has been identified in the genome of thedeep-branching thermophile Aquifex aeolicus (Aae), but little is known about itsstructure or activity. Earlier work has demonstrated that Aae Hfq cannot complementHfq-deletion strains of Salmonella enterica, implying that different species havedistinct RNA-binding profiles in vivo. To elucidate the structure and function of AaeHfq, the protein has been over-expressed, purified, and crystallized; Aae Hfq oligomershave also been studied by cross-linking. In addition, Aae Hfq has been shown to co-purify with small endogenous RNAs. The binding of Aae Hfq to FAM-U6 and FAM-A18 has been studied via fluorescence polarization assays and binding constants havebeen determined.COMPARISON OF ATOMISTIC MOLECULAR MECHANICS FORCE FIELDSFOR SIMULATING THE ALZHEIMER'S AMYLOID â-PEPTIDE. S. R. Gerben, J.A. Lemkul, & D. R. Bevan, Department of Biochemistry, Virginia Polytechnic Instituteand State University, Blacksburg, Virginia 24061-0308. Macromolecular functionarises from structure, and many diseases are associated with misfolding of proteins.Molecular simulation methods can augment experimental techniques to understandmisfolding and aggregation pathways with atomistic resolution, but the quality of thesepredictions is a function of the parameters used for the simulation. There are manybiomolecular force fields available, but most are validated using stably foldedstructures. Here, we present the results of molecular dynamics simulations on theamyloid b-peptide (Ab), whose misfolding and aggregation give rise to the symptomsof Alzheimer’s disease. Because of the link between secondary structure changes andpathology, being able to accurately model the structure of Aâ would greatly improveour understanding of the disease, an outcome with implications for studying otherVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1108 VIRGINIA JOURNAL OF SCIENCEprotein misfolding disorders. To this end, we compared five popular atomistic forcefields (AMBER03, CHARMM27, GROMOS96 53A6, GROMOS96 54A7, and OPLS-AA) to determine which could best model the structure of Ab. By comparing secondarystructure and calculated NMR shifts to available experimental data, we conclude thatAMBER03 and CHARMM27 over-stabilize helical forms of Ab, whereas OPLS-AA,GROMOS96 53A6, and GROMOS96 54A7 produce very similar results that agreewell with experimental observables.CHARACTERIZATION OF UDP-ARABINOPYRANOSE AS A SUBSTRATE OFEUKARYOTIC UDP-GALACTOPYRANOSE MUTASES. William T. Ryan, KarinaKizjakina & Pablo Sobrado, Department of Biochemistry, Virginia Tech., BlacksburgVA 24061. UDP-Arabinofuranose is a precursor for L-arabinofuranose, an importantconstituent in plant cell walls. In plants, UDP-arabinopyranose (UDP-Arap) isconverted into UDP-arabinofuranose (UDP-Araf) by the enzyme UDP-arabinopyranosemutase (UAM). UDP-galactopyranose mutases (UGMs) interconvert UDP-galactopyranose (UDP-Galp) into UDP-galactofuranose (UDP-Galf), a reactionimportant for virulence in the human pathogen Aspergillus fumigatus. The substratesfor both classes of mutases are very close in their chemical composition, yet theseenzymes perform their function through seemingly different mechanisms. While UGMfamily requires a flavin cofactor for catalysis, UAM does not require this cofactor for activity. Herein, we investigate the activity of eukaryotic UGMs with UDP-Arap/f asa substrate. It was determined that A. fumigatus UGM was able to utilize UDP-Arafcatonly when the flavin was in the reduced state. A k value of 0.013 s was determined.-1mThis is ~5000-fold lower than with UDP-Galf. The K value for UDP-Araf was 65 ìM,which is 1.72-fold lower than for UDP-Galf. Fluorescence polarization binding assaydrevealed that the K value for UDP-Arap was 347 ìM and 1179 ìM in the reduced andoxidized state, respectively. Supported by NIH grant R01 GM094469.SOLUTION CONDITIONS AND OXIDATION STATE OF METHIONINE-35AFFECT THE AGGREGATION PROPERTIES OF THE AMYLOID â-PEPTIDE (1-40). A. M. Brown, J. A. Lemkul, N. Schaum, & D. R. Bevan, Department ofBiochemistry, Virginia Tech, Blacksburg VA 24061. The amyloid â-peptide (Aâ) isa 40-42 residue peptide that is the principal toxic species in Alzheimer’s disease (AD).With AD being the primary causes of senile dementia in the United States,understanding the mechanism by which Aâ aggregates is essential in therapeuticdesign. The conversion of Aâ from a helical configuration in the membrane to a â-strand configuration is essential for aggregation. The oxidation of methionine-35(Met35) to the sulfoxide form (Met35 ) has been identified as potential modulator ofoxAâ aggregation. Experimental studies disagree on the role Met35 plays in Aâox neurotoxicity, which may be due to inconsistent solution conditions (pH, buffer,temperature). We applied atomistic molecular dynamics (MD) simulations as a means40to probe the dynamics of the 40-residue alloform of Aâ (Aâ ) containing Met35 orMet35 . We found that Met35 oxidation decreases the â-strand content of the C-oxterminal hydrophobic region (residues 29-40), with a specific effect on the secondarystructure of residues 33-35, thus potentially impeding aggregation. Further, there is animportant interplay between oxidation state and solution conditions, with pH and saltconcentration augmenting the effects of oxidation. The results presented here serve toVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 109STrationalize the conflicting results seen in experimental studies and provide a40fundamental biophysical characterization of Aâ dynamics in both reduced andoxidized forms. Ultimately, understanding the characteristics of Aâ that are important, for aggregation, such as the oxidation state of Met35 can provide insight into thebiochemical mechanism of AD.CHARACTERIZATION OF AN IMP CYCLOHYDROLASE IN HALOFERAXVOLCANII. Michelle L. Pasier & Cathy A. Sarisky, Dept. of Chem., Roanoke College,Salem VA 24153. Purine biosynthesis is a fundamental metabolic pathway present inessentially all life forms. The domain Archaea, unlike bacteria and eukaryotes, does notuniversally conserve enzymes across this pathway. Archaeal purine biosyntheticenzymes are also not necessarily homologous to non-archaeal enzymes. The finalreaction in de novo pathway is the conversion of 5-formamidoimidazole-4-carboxamide ribotide (FAICAR) to inosine-5'-monophosphate dehydrogenase (IMP).In Haloferax volcanii, a halophilic archaeon, this reaction is catalyzed by an IMPcyclohydrolase. The genome of H. volcanii is sequenced, but the characterization ofits genes is not yet completed. The gene HVO_0011 encodes a protein with a highamino acid sequence similarity to TK0430’s, an archaeal IMP cyclohydrolase.HVO_0011 was inserted into E. coli cells in the vector pMal-c5e. These cells wereinduced to express the protein of interest. This protein was extracted from the inducedcells by sonication and purification. The protein of interest was then separated from E.coli proteins using size exclusion and placed in a buffer simulating H. volcanii cellularconditions. An assay of the protein of interest and FAICAR was shown to produce IMPby its presence in the assay solution by HPLC chromatography. This confirms theprotein encoded by HVO_0011 as an IMP cyclohydrolase.DISABELED-2 (DAB-2) MODULATES PLATELET- CANCER CELLINTERACTIONS THROUGH ITS SULFATIDE BINDING DOMAIN. K. J.Andreano, X. Fu, D. G. S. Capelluto, C. V. Finkielstein. Dept. of Biol. Sci., VirginiaTech, Blacksburg, VA 24060. Platelets’ role in cancer progression and metastasis hasbeen attributed to platelet-mediated enhancement of tumor cell survival, extravasationand angiogenesis. Correlations exist between the tumor cells’ ability to aggregateplatelets in vitro and their metastatic potential in vivo that it is manifested as ahypercoagulable state. We have identified a tumor suppressor (Dab-2) that is releasedupon platelet activation and that modulates the extent of blood clotting. Our resultsshow Dab-2 is released from platelet alpha-granules to the membrane surface whereit binds to the integrin receptor, inhibiting platelet aggregation. Dab-2 binding tointegrins is modulated by sulfatides, glycosphingolipids that accumulate in plateletmembrane and effectively compete for Dab-2. We found that the N-terminal region ofDab-2 binds to sulfatides through two conserved sulfatide-binding sites. Uponactivation, sulfatides protect Dab-2 from thrombin cleavage, facilitate its internalizationand modulate the surface expression of P-selectin, a coagulation protein needed forstabilization of platelet aggregates. P-selectin mediates tumor cell adhesion to vascularendothelial cells and the interaction between activated platelets and cancer cells(emboli) during metastasis. Our data show that Dab-2/sulfatide recognition influencesthe stability of platelet aggregates heterotypically with cancer cells through sulfatidebinding and, indirectly, by controlling P-selectin levels. Thus, we hypothesize thatVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1110 VIRGINIA JOURNAL OF SCIENCEmanipulation of Dab-2 function in response of platelet activation will impact emboliformation offering an alternative route for therapeutic strategies aimed to controlmetastatic processes.THE ROLE OF ENTROPY IN THE REGULATION OF 3-PHOSPHOGLYCERATEDEHYDROGENASE. C. Meehan, A. Magnuson, C. Roach & E. Bell, Laboratory forStructural Biology, Biophysics & Bioinformatics, Dept. of Chemistry, University ofRichmond, Richmond VA 23173. Proteins can take on an array of conformationsdetermined by regional flexibility of the protein. This may play critical roles incatalysis and allosteric regulation. The relationship between structure and function wasstudied to determine the contributions particular amino acids in 3phospho-glyceratedehydrogenase (3pGDH), may play in flexibility and function of the protein. Basedupon available crystal structures a number of amino acids were identified by analysisof the temperature factors that may show altered flexibility in response to serinebinding. Addtionally, in conjunction with analysis of the temperature factors of boundcofactor (NAD(H)) we observe that two of the 4 central loops in the tetramer structureare more flexible than the other two and that this correlates with increased affinity forthe cofactor. This creates an entropy sink that can be used to harness flexibility changesfor catalysis and allosteric regulation. Various point mutations of specific amino acidresidues involved in this “entropy sink” were created and the expressed proteins werepurified using Ni-NTA affinity chromatography. Preliminary results suggest significanteffects on activity. Our observations suggest the presence of an entropy sink, whichcontributes to cofactor binding cooperation and possibly catalysis. This work issupported by NSF Grant MCB 0448905 to EB.ANS AS A PROBE OF LIGAND INDUCED CHANGES IN OLIGOMERICDEHYDROGENASES. G. Kingdom , S-H. Kim , C. Guzman , C. Metz and E. Bell ,1 2 1 1 1Laboratory for Structural Biology, Biophysics & Bioinformatics, Dept. of Chemistry,1University of Richmond, Richmond VA 23173 & Maggie Walker Governor’s School,2Richmond, VA 23220. 8-anilino-1-naphthalenesulfonic acid (ANS) fluorescence inthe presence or absence of 3 oligomeric dehydrogenases has been used to assessligand-induced changes in conformational plasticity. We have explored cofactor andcarboxylic acid induced changes in various dimeric malate dehydrogenases includingporcine mitochondrial and cytosolic forms and watermelon glyoxysomal forms. With3-phosphoglycerate dehydrogenase, a tetrameric enzyme found in higher eukaryotesincluding humans and in prokaryotes such as E. coli, we have compared the E coli andhuman forms using NSD mutants (containing the cofactor and substrate bindingdomains and the critical cofactor domain dimerization interface where a conservedtryptophan crosses the interface). Human NSD enhances ANS fluorescence more thanE coli NSD suggesting increased conformational plasticity. Alpha-ketoglutarateappears to enhance indicators of conformational flexibility. With hexameric glutamatedehydrogenase, the effects of the substrate Glutamate and a non catalytically activeanalog glutarate on ANS fluorescence suggest that, while glutarate has little effect onthe overall conformational plasticity of the enzyme, reduced cofactors result in agreatly enhanced blue shift in ANS fluorescence, suggesting enhanced binding tonewly exposed hydrophobic regions of the protein. Funded in part by NSF Grant MCB0448905 to EB.Virginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 111STTHE ROLE OF CONFORMATIONAL FLEXIBILITY IN NITRIC OXIDESYNTHASE. Rachel Jones , & Ellis Bell , Clover Hill High School, Midlothian VA,1 2 123112, Laboratory for Structural Biology, Biophysics & Bioinformatics, Dept. of2Chemistry, University of Richmond, Richmond VA 23173. Nitric Oxide Synthase(NOS) plays a critical role in a variety of signaling pathways and responses to cellularchallenge. Although the enzyme is well studied, the role of conformational flexibilityin Nitric Oxide Synthase (NOS) activity has received little attention. NOS exists inthree isoforms, inducible Nitric Oxide Synthase (iNOS), associated with malignantdisease; neuronal (nNOS), and endothelial (eNOS). Each consists of the N-terminaloxygenase domain which binds Arginine and NADPH and the C-terminal reductasedomain that binds flavin adenine dinucleotide, and calmodulin. To understand the roleof conformational flexibility, experiments using 8-anilino-1-naphthalenesulfonic acid(ANS) fluorescence, and multi-wavelength collisional quenching with differentlycharged quenchers were conducted in the presence/absence of Arginine to determinehow exposed hydrophobic regions of the protein were changed in iNOS. The datasuggests that Arginine increases the number of ANS binding sites per protein molecule;implying an increase in iNOS non-polar regions. The collisional quenching datasuggests that Arginine increases the exposure of non-polar regions, suggesting thatArginine increases the flexibility of the protein. Analysis of the amino acid sequenceof iNOS suggests that these changes in flexibility could be related to the regulation ofdisordered regions of the protein. A comparison of the effects of a number ofinhibitors of iNOS on both ANS fluorescence or the multi-wavelength collisionalquenching suggest different structural basis for their inhibitory effects.2EXPLORING THE STRUCTURAL BASIS FOR PIP SENSITIVITY OFINWARDLY RECTIFYING POTASSIUM CHANNELS. Trevor Larry, KalenHendra, Arco Paul & Linda M. Boland, University of Richmond, Department ofBiology, Richmond, VA 23114. All vertebrate inwardly rectifying potassium (Kir)2channels are activated by phosphatidyl-inositol 4,5-bisphosphate (PIP ). Dynamic2changes in PIP concentrations impact how Kir channels control the cellular restingmembrane potential. This project investigates a novel Kir channel (AqKir) that wascloned from the marine sponge Amphimedon queenslandica. Study of the spongechannels may provide insight into key changes in structure and function that occurred2during animal evolution. To probe the requirement for PIP in the maintenance of2AqKir, we reduced PIP concentrations in three ways: (1) application of wortmannin(at mM concentrations) to inhibit the kinase that phosphorylates PI to PIP; (2)activation of a co-expressed muscarinic acetylcholine receptor to activate a G protein-coupled phospholipase C; (3) activation of a co-expressed voltage–sensing 2phosphatase (CiVSP). The results confirm that PIP regulates AqKir but the sponge2channel requires much less PIP to remain active when compared to vertebratehomologs. We used mutagenesis to identify two key residues that are important for2higher sensitivity to PIP of the vertebrate Kir channels.TO TRANSLATE OR NOT TO TRANSLATE: HOW DED1, A DEAD-BOXATPASE, REGULATES MRNA TRANSLATION. Angie Hilliker, Dept. of Biology,University of Richmond, Richmond, VA 23173. Translation regulation is important,especially during stress, in neurons, and in early development, where many mRNAsVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1112 VIRGINIA JOURNAL OF SCIENCEare stored in translationally repressed states. These repressed mRNAs can be returnedto translation, but this process is poorly understood. We have previously shownevidence that the RNA helicase, Ded1, performs two sequential functions to return arepressed mRNA back to translation in an ATP-dependent manner. First, Ded1promotes the formation of a “pre-48S mRNP” that includes the initiation factors eIF4Fand Pab1. Then, Ded1 hydrolyzes ATP, allowing the mRNA to become translated. We hypothesize that Ded1 acts as an ATP-dependent switch to return mRNAs back tothe translating pool. We show that Ded1 interacts directly with eIF4G, while othershave shown that the mammalian ortholog of Ded1, DDX3, interacts with both eIF4Eand Pab1. We find that Ded1 helps recruit eIF4E and Pab1 to the m7G cap in vitro. We are currently testing the roles of polyA binding protein and the polyA tail inhelping Ded1’s first role in forming the pre-48S intermediate. Additionally, geneticscreens have revealed factors that may modulate Ded1’s first or second function viapost-translational modifications. These studies will help us understand how Ded1 isregulated, which will illuminate how mRNAs return to from a repressed state back intotranslation.INDUCTION OF APOPTOSIS BY NONSTRUCTURAL PROTEINS OF THESINDBIS VIRUS IN XENOPUS LAEVIS EMBRYOS. Kaitlyn Childs, JacobGraham, Kevin M. Myles, & Carla V. Finkielstein, Virginia Polytechnic Institute andState University, Blacksburg VA 24060. Viral infections are of interest as is relatedto virotherapy and host-virus interaction studies for mechanism construction that mayresult in knockout therapies to reduce or diminish virulence. Sindbis virus, anAlphavirus of the Togaviridae family, is transmitted by mosquitos; the endogenousstrain is opportunistic, infecting immunocompromised individuals, elders and youngchildren. The viral genome is divided into nonstructural (ns) and structural openreading frames. The nonstructural region encodes a polyprotein consisting of nsP1,nsP2, nsP3, and nsP4. In this study, we have found that the nonstructural protein nsp3has the ability to induce apoptosis in an heterologous system and that this property isrestricted to its N-terminus domain. Interestingly, whereas the nonstructuralpolyprotein is able to trigger apoptosis in Xenopus embryos when a polycistronicmRNA is injected in one-cell stage, neither nsp1, 2 nor 4 was able to accomplish thisform of cell death on their own. Apoptosis was confirmed by measuring caspase-3activity, visualization of membrane blebbing, and cleavage of cyclin A2, a knowncaspase substrate in Xenopus early embryogenesis. Next, we investigated whether nsp3pro-apoptotic activity was inhibited by interacting with anti-apoptotic members of theBcl-2 family. Thus, Xenopus laevis embryos were injected with various nsp3 constructsin one-cell, collected at different times before MBT and will be analyzed for bindingby immunoprecitation. We hope to elucidate the mechanism behind apoptosis inductionof the Sindbis virus in Xenopus laevis embryos in this way.DOWN-REGULATION OF THE CIRCADIAN FACTOR PERIOD 2 BY THEONCOGENIC E3 LIGASE Mdm2: EVIDENCE OF THE PRESENCE OFCIRCADIAN AND CELL CYCLE CROSSTALK MECHANISMS. Jing-Jing Liu,Tetsuya Gotoh, Marian Vila-Caballer, Carlo S. Santos, Jianhua Yang & Carla V.Finkielstein, Integrated Cellular Responses Laboratory, Department of BiologicalSciences, Virginia Tech, Blacksburg, VA. The circadian rhythm and cell cycle are theVirginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1PROCEEDINGS 91 ANNUAL MEETING 113STtwo main oscillatory systems in cells. How cells sense time and decide what is the besttime for growing, proliferating or apoptosis? One possibility is that there are crosstalksbetween these two systems. Based on the fact that Period 2 (Per2) also plays essentialrole in DNA damage response, Per2 is supposed to connect circadian rhythm and cellcycle, which makes Per2 work as a tumor suppressor. We found Per2 regulating p53pathway but little is known about how Per2 itself is regulated. One interesting findingis that independent of transcriptional regulation, overexpressed Per2 protein alsooscillates; this implies posttranslational modifications are essential for sustaining Per2protein oscillation. Per2 binds to Mdm2, a well-known E3 ubiquitin ligase, both invitro and in vivo. Mdm2 induces Per2 ubiquitination in vitro, but further experimentsare needed to verify Mdm2 is an E3 ligase for Per2 in vivo.USE OF ELECTROSTATIC POTENTIALS TO EVALUATE PPARá FOR DRUGDISCOVERY. F. Buzzalino, N. Lewis & D. Bevan, Dept. of Biochemistry, VirginiaTech 24060. It is estimated that in the United States, approximately 600,000 deaths peryear are attributed to cardiovascular disease. Peroxisome proliferator activatingreceptors (PPARs), transcription factors that pertain to the nuclear receptor family, playan important role in maintaining metabolism. PPARá, particularly responsible for thecatabolism of fatty acids, resides in the liver, heart, kidneys, and skeletal muscle. Inrecent years Fibrates, synthetic ligands used to treat hyperlipidemia via specificactivation of PPARá, have demonstrated effectiveness in reducing cardiovasculardisease. Our goal is to identify unknown ligands that interact with PPARá in a similarmanner as fibrates. Prior to performing studies to identify unknown ligands, anappropriate PPARá structure must be chosen. In our studies we have analyzed thebinding of 11 PPARá crystal structures with the 11 crystalized agonists via dockingexperiments (re-docking, and cross-docking). Our analysis of the residue interactionsbetween the 11 PPARá structures and the agonists reveal that four residues – His440,Tyr314, Tyr464, Ser280 – are associated with agonist binding. Docking of 544compounds with three PPARá for the presence of the four residues was performed. ThePPARá structure 2P54 demonstrated to be representative of a structure that can bindvarying compounds. Our results lead us to believe that the most appropriate PPARástructure for docking of unknown ligands is 2P54. Identification of PPARá selectiveligands may help produce fibrates in hopes of reducing the number of casualtiesassociated with cardiovascular disease.Virginia Journal of Science, Vol. 64, No. 1, 2013 https://digitalcommons.odu.edu/vjs/vol64/iss1

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