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During this study, the trailer was managed according to industry practices (Transport Quality Assurance program of the National Pork Checkoff). Temperature was recorded inside the trailer to generate a 3D representation of the temperature profile. GPS coordinates were recorded for every minute of each trip. A weather data set of hourly outdoor temperatures that correspond to the geographical trailer location was generated using the GPS coordinates and the data from the nearest local NOAA weather station. The outdoor weather data was summarized into an overall weather distribution to represent the outdoor conditions over the complete study. This distribution will be compared to geographical distributions within the US, based on ASHRAE records, to represent the study applicability. Additionally, correlations were assessed between the inside and outside environmental data to identify trends, specifically during extreme hot and cold conditions. Results of this study have the potential to identify areas of management for reducing the number of dead or down pigs during and after transport. Analysis is ongoing and complete results will be presented.
EFFECT OF STRAWBERRY GUAVA ON SURVIVAL AND REPRODUCTION IN THE NATIVE HAWAIIAN
TERRESTRIAL LAND SNAIL LEPTACHATINA CEREALIS (PULMONATA: AMASTRIDAE)Kaile Costa, Daniel Chung.
University of Hawaii Kapiolani Community College, Honolulu, HI.
The endemic Hawaiian land snail Leptachatina cerealis, was fed differing diets of mamaki (Pipturus albidus) leaves, its preferred food in the wild, and strawberry guava leaves (Psidium cattleianum), an invasive that forms vast
THE ALLEVIATION OF PAIN ASSOCIATED WITH THE DISBUDDING OF CALVESAmanda Mathias1, Dan Stein1, John Gilliam2, Michelle Calvo-Lorenzo1.
Oklahoma State University, Stillwater, OK, 2Oklahoma State University Center for Veterinary Health Sciences, 1 Stillwater, OK.
Disbudding, the removal of horn buds on a calf, is a necessary management procedure because it eliminates costs associated with horns and improves safety. However, disbudding is known to cause pain and distress in calves. Extralabel use of some pharmaceuticals allows for some alleviation of pain and distress, but it is not the best method.
FDA-approved pharmaceuticals would be better options to alleviate pain, however none exist. This study evaluates the effectiveness of ethyl alcohol (EtOH) to produce a cornual nerve block, when the area surrounding the horn is numb. To test this, calves were given 1 of 3 treatments: 5 ml saline solution (SAL), 5 ml of 2% lidocaine (LID), a solution mixture of 2.5 ml of 2% lidocane and 2.5 ml of 75% EtOH (MIX), or 5 ml of 75% EtOH (ALC). The calves were needle pricked in the area surrounding the horn bud at 5, 10, 15, 20, 30, and 60 minutes after treatment on day 1 to determine onset of the cornual nerve block, and once daily on days 2-14 to test duration. The study found SAL calves exhibited “not blocked” behaviors at all time points; all other treatments exhibited initial “blocked” behaviors at 5 minutes post-treatment. The study also found that MIX and ALC treatments maintained a cornual nerve block for 2 weeks. Results indicate the potential of using EtOH as a form of pain relief for disbudding calves. Future research incorporating the removal of the horn buds after EtOH administration will further evaluate the effectiveness of EtOH as a cornual nerve block.
AGOUTIS ON THE ALERTSharonda Carson1, Enzo Aliaga-Rossel.
University of Central Oklahoma, Edmond, OK, 2San Andres University, La Paz, BO.
1 Agoutis (Dasyprocta punctata) are mammals that are a key component of the Neotropical forest as agents for seed dispersal, seed predators, and prey for several species. They spend the day defending their territory from invading agoutis, searching for food, looking for potential mates, and watching for predators. The objective of the research will be to study the alert behavior of agoutis in two different habitats in Las Cruces, Costa Rica. A group of agoutis from the secondary forest and from the garden will be studied to see which are more alerted. One group has grown accustomed to the presence of humans and hearing disturbances while the other group lives in a less disturbed area where there is less influence from people. To test how alert the agoutis can be in the presence of a predator or an invading agouti, a series of test calls from an ocelot and an invader will be sounded. The calls will start at a distance far away and will advance the longer it takes the agoutis to respond. Observation notes will be taken after every test call to note the agoutis’ reaction. With one group having fewer interactions versus another group that is used to everyday disturbances, the expected conclusion will be for the agoutis in the secondary forest to be more alerted than the agoutis in the garden. The overall goal for the research project will be to have a better understanding of the agoutis’ alert behavior and the contribution they make to their habitat.
15 UNDERGRADUATE POSTER ABSTRACTS
EXPRESSION OF WNT-SIGNALING TRANSCRIPTS AT SPECIFIC STAGES OF FOLLICLE DEVELOPMENT IN
BOVINE GRANULOSA CELLSAllison Potts1, Andrea Stapp1, Belinda Gomez1, Bailey Parker1, Craig Gifford1, Dennis Hallford2, Jennifer Gifford1.
Oklahoma State University, Stillwater, OK, 2New Mexico State University, Las Cruces, NM.
1 Follicular maturation is a dynamic process requiring input from pituitary gonadotropins and ovarian-derived factors.
Members of the wingless-type mammary tumor virus integration site (WNT) signaling pathway have been recognized as differentially expressed and hormonally regulated in rodent ovaries. However, the role and expression of WNTsignaling molecules in ovarian follicle development in cattle is unknown. Therefore, the objective of this study is to characterize components of the WNT-signaling pathway at specific stages of follicular development by real-time PCR.
To identify gene expression changes in bovine folliculogenesis, granulosa cells and follicular fluid were collected from ovary pairs containing a stage III CL. Granulosa cells were isolated from small (1 to 5 mm) and large (8 to 22 mm) follicles and the corresponding CL. Real-time PCR quantification of select WNT family members was evaluated at distinct stages of development. Compared to small follicle granulosa cells, expression of the WNT transcriptional cofactor CTNNB1 was similar in large, dominant follicles (P = 0.53) but was decreased in the CL (P 0.01).
Expression of WNT ligands also demonstrated stage-specific regulation as WNT2B was reduced in large dominant follicles (P = 0.10) but increased in CL (P = 0.03) compared to small follicles. A comparable pattern of expression was demonstrated for WNT5A as small follicles had greater expression compared to large follicles (P 0.01) and was similar to CL (P = 0.56). Results from the current experiment indicate WNT-signaling molecules may be inhibitory to follicle development and luteinization in cattle.
BIODISTRIBUTION IN TISSUE OF NOVEL DRUG MOLECULES VIA MALDI-IMAGING MASS SPECTROMETRYJoseph Wayne Fowler, Jonathan Waxer, Carly Ferguson, Richard Gatti.
University of California, Los Angeles, Los Angeles, CA.
Premature termination codons (PTC) are the result of mutations within an organism’s genome that can cause a wide variety of genetic disorders. One such genetic disorder is ataxia-telangiectasia (AT), which affects the brain causing malignant health defects in mammals due to an AT-mutated protein. Nonaminoglycoside compounds induce a read-through of PTCs, developing functional, AT-mutated proteins theoretically treating AT. Nonaminoglycosides have been found to treat PTC genetic disorders in vitro but have been untested in vivo. Two nonaminoglycosidic read-through compounds of interest (RTC 6 and 16) should theoretically be able to pass the blood-brain barrier to treat PTC disorders because of their small size and nonpolar characteristics. A total of 48 mice were tested following an intraperitoneal injection: 16 untreated, 16 RTC-6 treated, and 16 RTC-16 treated. Imaging mass spectrometry was used to identify the presence of RTC 6 and 16 in the liver, heart, kidneys, lungs, and brain of dosed mice. As expected, no traces of RTC 6 or 16 were found in the brains or livers of 8 control mice. RTC 16 was discovered in the brains of 3 treated mice. Although RTC 16 was located in the liver of 4 treated mice, no trace of RTC 16 was found in the brain of 8 treated mice. After complete tissue testing, the next step will be to test mouse models to obtain enough data on the stability of nonaminoglycoside compounds and to ultimately synthesize a capable drug to treat ataxiatelangiectasia.
CHARACTERIZATION OF THE ACYL-COENZYME A SYNTHETASE PA4198 IN PSEUDOMONAS AERUGINOSARachel Gomez, Debra Dunaway-Mariano.
University of New Mexico, Albuquerque, NM.
Pseudomonas aeruginosa is an opportunistic pathogen with antibiotic resistance and is a large contributor to hospital infections, including those affecting patients with cystic fibrosis. As a member of the Pseudomonadacae family, Pseuodmonas aeruginosa can use a variety of organic compounds such as fatty acids for energy. A key enzyme for metabolism of fatty acids are acyl-coenzyme A synthetases which activate fatty acids for further reactions. The focus of this project is to characterize the acyl-CoA synethetase PA4198 in terms of its biological function and range as well as its efficiency as an enzyme. This includes cloning, expression, purification and kinetic analysis of PA4198
EFFECTS OF SALICYLIC ACID, ACETYLSALICYLIC ACID, AND METHYL SALICYLATE ON
R-PHOSPHOFRUCTOKINASE-1-ENZYME ACTIVITY: COMPARISONS TO ASCORBYL FATTY ACID INHIBITIONSLouciana Sanchez1, Edward Alexander1, Percy Russell2, Alberto Palacios1, Rogelio Estrada1, Anita Williams2.
San Diego Mesa College, San Diego, CA, 2University of California, San Diego, La Jolla, CA.
1 Research has confirmed the benefit of aspirin and salicyclates in cancer prevention. One proposed mechanism for this action involves inhibition of the enzyme cycloxygenase (COX). Studies on fatty acid derivatives of ascorbic acid (AA) have shown them to have antimetastatic action on certain cancers. In vitro experiments have shown that these fatty acid derivatives inhibit the activity of the enzyme r-phosphofructokinase (r-PFK-1). Mechanisms for the fatty acid derivative inhibitions may involve antioxidation by the 2, 3-ene-diol group in ascorbic acid and allosteric effects of lipophilic side chains. The purpose of this research was to investigate the inhibition properties of salicyclic acid (SA), acetylsalicyclic acid (ASA), and methyl salicylate (MS) on r-PFK-1. Our hypothesis was, since these compounds are antioxidants, antioxidation by SA, ASA, and MS on r-PFK-1 should take place and yield results similar to those found for ascorbic acid derivatives: ascorbic acid 6-butyrate, ascorbic acid 6-palmitate, ascorbic acid 6-sterate, ascorbic acid 2, 6-dibutyrate, and ascorbic acid 2, 6-dipalmitate, (Ic50: 0.13, 0.021, 0.026, and 1.2 mM). SA, ASA, and MS were obtained commercially. The r-PFK-1 enzyme was purified employing centrifugation and ion-exchange filtration.
Inhibitions were carried out in buffered solutions of 95% ethanol. Our results show that SA, ASA, and MS show no significant inhibitions of r-PFK-1 when compared to the fatty acid derivatives of ascorbic acid. From these results, we conclude that the side chain length and lipophilic characteristics of the ascorbic acid derivatives may be the major contributors in the inhibition of r-PFK-1 rather than antioxidation.
DETECTING THE MECHANISM OF FEEDBACK INHIBITION OF TYROSINE HYDROXYLASE; DOPAMINE
BINDING TO GLUTAMATE RESIDUES IN THE REGULATORY DOMAINJessica Villacorta, Susan Colette Daubner.
St. Mary’s University, San Antonio, TX.
Tyrosine hydroxylase (TyrH) catalyzes the rate-limiting step of catecholamines synthesis by converting L-tyrosine to L-DOPA. The post-translational mechanisms that control TyrH activity include phosphorylation by protein kinases and dephosphorylation by phosphatases, feedback inhibition, and protein complexes. This research focuses on the structural changes of the regulatory domain by feedback inhibition caused by dopamine-binding inhibition. When dopamine binds to TyrH, the flexible loop of the regulatory domain changes conformation in order to close off the active site. We hypothesize that one or two glutamate residues found in the flexible loop are responsible for anchoring down dopamine and hence closing the R domain. To test our hypothesis, we replaced the glutamate residues found at positions 43 and 48 of the regulatory domain with a glutamine. Mutants were obtained by site-direct mutagenesis, and the mutated proteins were overexpressed in E. coli. Then the proteins were purified through standard chromatographic techniques. The Michaelis-Menten parameters were obtained in order to ensure that the catalytic domain was not affected by the mutation. For the E43Q mutant, the Km value for tetrahydropterin was 44.4 µM, the KM value for tyrosine was 49.23 µM, and the VMax value was 48.94 min-1. For the E48Q mutant, the KM value for tetrahydropterin was 40.06 µM; the KM value for tyrosine was 41.61 µM, and the VMax value was 53.64 min-1. Knowing that the mutant proteins are catalytic active allows us to proceed with studies of their interactions with dopamine.
17 UNDERGRADUATE POSTER ABSTRACTS
INVESTIGATION OF TRIAZABOROPYRIDINIUM (HPY) DYES FOR A CELL VIABILITY ASSAYAlan Yazzie, Jeffrey Arterburn.
New Mexico State University, Shiprock, NM.