Spring 1999 Volume 3, Issue 1

Abstracts of Graduate student presentations at the Third Annual Graduate Student Research Symposium held April 8, 1999 in Keathley University Center on the campus of Middle Tennessee State University. Sponsored by the Middle Tennessee State University College of Graduate Studies and Sigma Xi.

SEROTYPE-DEPENDENT INDUCTION OF TUMOR NECROSIS FACTOR-ALPHA AND REPLICATION PATTERNS IN HUMAN MONOCYTES BY REOVIRUS. Chad Brooks and Anthony Farone. Department of Biology.

LEGIONELLA MULTIPLICATION IN AMOEBAE SUSPENDED IN SPRING WATER WITH EXOGENOUS BACTERIA. Tamara Dodson and Anthony Newsome. Department of Biology.

IMMUNOGOLD IDENTIFICATION OF LEGIONELLA SPECIES IN BIOFILMS. Michele L. Summy and Anthony Newsome. Department of Biology.

INTEGRATING VRML, JAVA, AND HTML IN A WEB-BASED TOOL FOR COMPUTER LITERACY. Warren Patterson and Judy Hankins. Department of Computer Science.

FROM MICROPROCESSOR TO MICROCONTROLLER THE EVOLUTION OF A DIGITAL READOUT SYSTEM. Preston Baxter and Saleh Sbenaty. Industrial Studies.

THE RELATIONSHIP OF RELIGIOUS SELF-DISCREPANCIES TO DEPRESSION AND ANXIETY. Clinton Craun and Tom Brinthaupt. Department of Psychology.

COMPETITIVENESS IN SAME-SEX FRIENDSHIPS. Stacia Brantley and Tom Brinthaupt. Department of Psychology.

METHOD DEVELOPMENT FOR ANALYZING THE AQUEOUS HYDROLYSIS OF CHLORPYRIFOS. Meredith Rogers and John DiVincenzo. Department of Chemistry.

TRANSITION STATE ANALYSIS OF THYMIDINE PHOSPHORYLASE. Mansoureh Rezaee and Paul Kline. Department of Chemistry.

ALDOL CONDENSATION PRODUCTS PRODUCED DURING THE STATIC DRYING OF ACETONE OVER 4A MOLECULAR SIEVE. Xiaoming Want and Martin V. Stewart. Department of Chemistry.

SYNTHESIS OF D-ALA-D-ALA PEPTIDOMIMETICS AS NOVEL ANTIBACTERIALS. Tao Meng and Norma Dunlap. Department of Chemistry.

SEROTYPE-DEPENDENT INDUCTION OF TUMOR NECROSIS FACTOR-ALPHA AND REPLICATION PATTERNS IN HUMAN MONOCYTES BY REOVIRUS. Chad Brooks and Anthony Farone. Department of Biology, Middle Tennessee State University, Murfreesboro, Tennessee 37132.

Airway inflammation is observed when reovirus serotype 1 Lang (T1L) or serotype 3 Dearing (T3D) is administered in a reovirus pneumonia model in the rat, however, T3D causes a more prominent inflammatory response than T1L.  Previous studies from our laboratory have shown that inflammatory cytokine gene expression was stimulated in rat alveolar macrophage cell-line by reovirus stimulation of inflammatory cytokines in the human monocyte cell-line, THP-1, was serotype-dependent. Cells were treated with T1L, T3D, or medium alone.  After 24 h, supernatants were tested for tumor necrosis factor-alpha (TNF-alpha) by ELISA.  We found that TPH-1 cells treated cells with T3D had statistically significantly increased levels of TNF-alpha expression compared to cells treated with T1L.  T1l-treated cells also exhibited increased levels of TNF-alpha compared to medium alone.  These results support the hypothesis that monocytes respond differently to T1L and T3D.  Further research has shown similarities between T1L and T3D replication patterns but viral titers indicate that T1L yields significantly greater infectious virus particles after 72 h than T3D.
 
 
 

LEGIONELLA MULTIPLICATION IN AMOEBAE SUSPENDED IN SPRING WATER WITH EXOGENOUS BACTERIA. Tamara Dodson and Anthony Newsome. Department of Biology, Middle Tennessee State University, Murfreesboro, Tennessee 37132.

Legionella pneumophila has been reported to live in surface waters.  Laboratory studies have clearly demonstrated its ability to multiply within free-living amoebae.  Typically these studies are based on a high multiplicity of infection, (MOI, bacteria to amoebae ratio) and use nutrient-rich laboratory media.  To document potential interactions that might occur in surface water, this study evaluated the ability of L. pneumophila to grow in the free-living amoebae Acanthamoebae polyphaga in nutrient-poor spring water.  Results showed that at a MOI of 0.001 bacteria/amoebae, an average of a 7000-fold increase in L. pneumophila colony forming units occurred over a 144-hour time period at 30 degrees C.  The addition of non-Legionella bacteria species, added at a MOI of 100/1, had no adverse effect on the ability of L. pneumophila multiplication with the amoebae.  These results suggest a remarkable ability of L. pneumophila to multiply in A. polyphaga at a low MOI in natural waters.  Further, these results support a specific mechanism, either by receptor and/or active penetration by the bacteria, for uptake of L. pneumophila by A. polyphaga.
 
 
 
 

IMMUNOGOLD IDENTIFICATION OF LEGIONELLA SPECIES IN BIOFILMS. Michele L. Summy and Anthony Newsome. Department of Biology, Middle Tennessee State University, Murfreesboro, Tennessee 37132.

Current methods for detection of Legionella species in environmental water samples are not designed to document its associations with other microorganisms such as free-living amoebae.  Methods capable of documenting naturally occurring relationships are needed to understand the natural history of the genus Legionella.  An antibody-mediated approach was evaluated.  It was based on the commercially available Remel Legionella Poly-ID kit, which consists of a primary antibody to 22 Legionella species and a secondary fluorescein antibody conjugate.  For this study, however, a secondary gold-labeled conjugate was used.  This allowed us to subsequently Giemsa stain and visualizes the relationship between antibody reactive bacteria and associations with other microorganisms.  Placing slides in water samples for one week formed Biofilms.  Cells were then fixed prior to Immunogold and Giemsa staining.  Dark stained immuno-reactive bacteria often occurred as isolated cells.  In addition reactive cells could also be seen in proximity to amoebae and occasionally appeared to be in contact with eukaryotic cells.  Results suggest this Immunogold technique may be useful in examining naturally occurring relationships between Legionella species and other aquatic microorganisms.
 
 
 
 

INTEGRATING VRML, JAVA, AND HTML IN A WEB-BASED TOOL FOR COMPUTER LITERACY. Warren Patterson and Judy Hankins. Department of Computer Science, Middle Tennessee State University, Murfreesboro, Tennessee 37132.

Since computer technology changes constantly, computer literacy instructors are faced with the challenge of presenting the fundamental concepts of computer science as well as the leading edge of technology.  Many concepts in computer science are new to students and are difficult to understand. When learning about computer hardware, students are faced with the difficult task of  visualizing the
placement and interaction of numerous internal components.  Combining Virtual Reality Modeling Language (VRML), Java, and Hyper Text Mark-up Language (HTML), a Web-based tool was developed to help students visualize computer hardware as well as to give students experience manipulating objects in a virtual reality world.  This tool allows students to select a hardware component for investigation.  Students may display an interactive virtual representation of the object, a hyper text information document, or a list of web links on the topic.  The virtual display is designed to provide students with a simplified view of the component which they can rotate and view from other perspectives thus providing a more concrete visualization.  Additional information provided on the
remaining displays applies directly to the virtual scene and supplements class instruction.  Virtual reality is gaining importance in many fields including medical training, elementary and secondary school education, and ecology, among others.  As this technology broadens to other fields, students will be expected to be literate in the application of virtual reality.  In addition to enjoying an enhanced visual aid
which will strengthen the understanding of computer hardware concepts traditionally taught in computer literacy courses, students can gain experience navigating scenes and manipulating objects with the tools of virtual reality.
 
 
 
 

FROM MICROPROCESSOR TO MICROCONTROLLER THE EVOLUTION OF A DIGITAL READOUT SYSTEM. Preston Baxter and Saleh Sbenaty. Industrial Studies, Middle Tennessee State University, Murfreesboro, Tennessee 37132.

I designed and built a milling machine digital readout system (DRO) for an ET-467 microprocessor design project in 1998 as an undergraduate student at MTSU.  The project was designed using a Z-80 microprocessor and it's supporting components.  The DRO reads optical encoders attached to the leadscrews and spindle of a milling machine, digitally displays the position of its three axes, and displays the mill's spindle speed in revolutions per minute.  The system was successfully constructed and tested using a Sherline Milling machine.  This year as an ETIS graduate student I am improving the DRO system by converting it from a microprocessor-based design to a microcontroller-based design.

The original Z-80 microprocessor design required external RAM, ROM, I/O, and decoding logic, bringing the total chip count to over ten integrated circuits plus a twelve digit L.E.D. display.  Backlash settings, used to match the DRO to the machine, were set using mechanical rotary switches.  These switches, as well as timing circuitry for the tachometer, took up even more board space.

A microcontroller contains many of the external components of a microprocessor-based system on a single chip.  The DRO system was re-designed using the Zilog Z-8 microcontroller and an alpha-numeric display module.  The alpha-numeric display has an added bonus of making the DRO more user friendly by providing on-screen directions for system setup.  The new design  only requires the Z-8 MCU and the display module, thus greatly reducing the unit's cost, size, and power consumption.
 
 
 
 

THE RELATIONSHIP OF RELIGIOUS SELF-DISCREPANCIES TO DEPRESSION AND ANXIETY. Clinton Craun and Tom Brinthaupt. Department of Psychology, Middle Tennessee State University, Murfreesboro, Tennessee 37132.

Previous research on self-concept discrepancies has identified a model whereby discrepancies between one’s actual self (i.e. Who I really am) and one’s ideal self (i.e. Who I ideally would be) are more closely related to depression whereas discrepancies between one’s actual self and one’s ought self (i.e. Who I should be) are more closely associated with anxiety. The present research is attempting to extend this general model to a specific domain, the religious self. Analyses will examine the generalizability of self-discrepancy theory under conditions of domain specificity (e.g., the religious self).
 
 
 
 

COMPETITIVENESS IN SAME-SEX FRIENDSHIPS. Stacia Brantley and Tom Brinthaupt. Department of Psychology, Middle Tennessee State University, Murfreesboro, Tennessee 37132.

Are close same-sex friends more competitive in their friendships on the things they feel are more important to them?  One hundred twenty undergraduate psychology students listed the ways that they are similar and dissimilar to their same-sex friends.  They also rated their friendship based upon who was more competitive in those areas, and to whom the area was more important.  I am testing the hypothesis that participants will be more competitive in those areas that they feel are more important to them.  Also, it was expected that the participants would be more competitive than their friends on the items of similarity as opposed to dissimilarity.  Gender differences were also investigated to determine if males are more competitive than females in their friendships.
 
 
 
 

METHOD DEVELOPMENT FOR ANALYZING THE AQUEOUS HYDROLYSIS OF CHLORPYRIFOS. Meredith Rogers and John DiVincenzo. Department of Chemistry, Middle Tennessee State University, Murfreesboro, Tennessee 37132.

Many insecticides are nerve agents which are not only lethal to pests but harmful to numerous other organisms in the environment.  Chlorpyrifos is an organophosphorous insecticide commonly known as Dursban? and is used both around residential houses and on golf courses.  One way of determining the fate of Dursban in nearby rivers, streams, or lakes is to look as its hydrolytic degradation rate.  A method of liquid-liquid extraction followed by gas chromatography with an electron capture detector was developed for following the hydrolysis kinetics.  Since liquid-liquid extraction is ineffective in removing some hydrolysis products, a method of solid-phase extraction followed by high pressure liquid chromatography was explored as a potential means for identifying some of these products.
 
 
 
 

TRANSITION STATE ANALYSIS OF THYMIDINE PHOSPHORYLASE. Mansoureh Rezaee and Paul Kline. Department of Chemistry, Middle Tennessee State University, Murfreesboro, Tennessee 37132.

Thymidine phophorylase is one of two pyrimidine phosphorylases that operates in the salvage pathway.  It catalyzes the irreversible phosphoralysis of uridine to yield uracil and ribose-1-phosphate.  In addition, thymidine phosphorylase has been shown to be identical to platelet-derived endothelial cell growth factor.  Recent studies have shown that platelet-derived endothelial cell growth factor (PD-ECGF) is associated with the development of new blood vessels (angiogenesis) necessary for tumor growth and metastasis in a variety of malignant tumors including breast, stomach, colon, and head and neck tumors.

A complete understanding of thymidine phosphorylase requires knowledge of the chemical mechanism by which it carries out its reaction.  A family of KIE was determined for E.coli thymidine phosphorylase using stable isotopes.  The experimentally determined kinetic isotope effects were obtained [1'-13C], [2'-13C], [1'-2H], [2'-2H], [5'-2H, and [9-15N].  The commitment to catalysis was determined by isotope trapping and used to correct the observed KIE to their intrinsic values.  A combination of intrinsic kinetic isotope effects and computational analysis using a bond energy, bond order vibrational analysis was used to model the transitional state of the enzyme.
 
 
 
 

ALDOL CONDENSATION PRODUCTS PRODUCED DURING THE STATIC DRYING OF ACETONE OVER 4A MOLECULAR SIEVE. Xiaoming Want and Martin V. Stewart. Department of Chemistry, Middle Tennessee State University, Murfreesboro, Tennessee 37132.

Static drying of acetone over 4A molecular sieve at room temperature causes contamination with aldol condensation products.  GC-MS was employed to analyze the product mixture, which consists of the dimers diacetone alcohol and mesityl oxide together with lesser amounts of the trimers triacetone alcohol and semiphorone.  Synthetic methods were developed to prepare the latter two components for use  as chromatographic standards to quantify the product distribution.  Triacetone alcohol was obtained from phorone through hydrolysis by an oxymercuration/demercuration route.  Formation of the cyclic pyronyl ether, the major product of hydrolysis conducted with aqueous sulfuric acid in THF, is negligible during oxymercuration/demercuration because carbocation formation is avoided.  Partial dehydration of triacetone alcohol to semiphorone is accomplished during vacuum distillation from a trace amount of 85% phosphoric acid.  A graduate student stipend from the College of Graduate Studies at Middle Tennessee State University for the 1998 summer semester is gratefully acknowledged.
 
 
 
 
 

SYNTHESIS OF D-ALA-D-ALA PEPTIDOMIMETICS AS NOVEL ANTIBACTERIALS. Tao Meng and Norma Dunlap. Department of Chemistry, Middle Tennessee State University, Murfreesboro, Tennessee 37132.

The research project is focused on seeking novel antibacterials due to the fast emergence of resistant strains. These types of antibacterials are peptidomimetics, which may function like penicillin. The target of these antibacterials is peptidoglycan transpeptidase (PBP), which catalyzes the combination of peptidoglycan polymers to form cell wall proteins.  We have proposed a procedure to synthesize these new peptidomimetics and we will discuss some initial steps that we took to synthesize this compound by following the procedure and using (S)-prolinol as the starting material.