Document 14570793

WELCOME

Dear Colleagues,

Welcome to the 7th Annual MS-INBRE Research Symposium. We are happy you can join us again this year at The University of Southern Mississippi in the “Hub City”. The Symposium continues to grow – as you look around during the day, you will see many undergraduate students presenting some of their research. This, I think, is one of the most exciting aspects about the MS-INBRE Symposium - we see the future of Mississippi in action – the students of today. I am always impressed with the hands-on research and creativity we see in the undergraduate posters. As I say each year – the future is in good hands.

The opportunity to interact with students and faculty across the State is essential in all our efforts to enhance biomedical research and training in the Magnolia State. A lot of exciting research is ongoing in our State and this is an excellent opportunity to hear about some of it and chat with colleagues. Each year a number of research collaborations develop from these chats and I’m sure we will see the same thing happen again this year.

The work of the Mississippi INBRE depends on the generous funding from the National Institutes of Health. Please remember to acknowledge this funding on all your publications and presentations (and send us the citation information) now and in the future: "This work was supported by the Mississippi INBRE funded by an IDeA award from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20 GM103476.”

We hope you enjoy the day’s events and take opportunities to chat with your colleagues from across the State – who knows what amazing research collaborations may arise!

Best regards,

Glen Shearer

Glen Shearer

Director, Mississippi INBRE

Professor and Chair, Biological Sciences

The University of Southern Mississippi

MS-INBRE 2014 ANNUAL RESEARCH SYMPOSIUM

Saturday, February 15, 2014

Venue Trent Lott Center, The University of Southern Mississippi

8:00 - 9:00

Registration and Poster set up

Trent Lott Center Lobby

9:00 - 9:45

Oral Presentations Session 1 : Room 102

9: 45 - 10:00

Coffee Break

10:00 - 10:45


11:00 - 12:30

Poster Session 1 : Room 103

12:30 - 1:30

Lunch

1:30 - 2:15


2:15 - 2:30

Coffee Break

2:30 - 4:00

Poster Session 2 : Room 103

4:00 - 4:30

External Advisory Committee and Administrative Core Meeting : Conference Room

MS-INBRE 2014 ANNUAL RESEARCH SYMPOSIUM

Oral Session 1 9:00 - 9:45

Sabrice Guerrier, Millsaps College “The regulation of membrane curvature during cell-cell fusion in Tetrahymena thermophila” 9:00 - 9:15

Scharri Walker, Tougaloo College “The Evaluation of Natural Products as

Anticancer Therapies in Pediatric Cancers” 9:15 - 9:30

Manliang Feng, Tougaloo College “Design Of A Multifunctional Polypeptide

For Application In Cancer Diagnosis And Treatment” 9:15 - 9:30

Oral Session 2 10:00 - 10:45

Davida Crossley, Alcorn State University “Regulation Of The M46 Gene In

Histoplasma Capsulatum” 10:00 - 10:15

Dale Rosado, Mississippi College “Antimycobacterial Properties Of Novel

Ethambutol Analogues” 10:15 - 10:30

Bidisha Sengupta, Tougaloo College “Prospect Of Plant Flavonoids As Agents

For Reducing Oxidative Stress” 10:30 - 10:45

Poster Session 1 11:00 - 12:30

Oral Session 3 1:30 - 2:15

Clement Yedjou, Jackson State University “Technology-Enhanced Teaching and Learning of Exposure Science” 1:30 - 1:45

Jon Moreno, Alcorn State University “Role Of RNA-Binding Proteins In

Mitoribosome Biogenesis” 1:45 - 2:00

Jerry Reagan, Mississippi College “Lysosomal Cholesterol Accumulation

Regulates Apoptosis” 2:00 - 2:15

Poster Session 2 2:30 - 4:00

O1 The regulation of membrane curvature during cell-cell fusion in

Tetrahymena thermophila .

Samuel Rosenberg 1 , John Cannon 1 , Sabrice Guerrier 1,2

1 Biology Department, Carleton College, Northfield, MN

2 Biology Department, Millsaps College, Jackson, MS

The overall goal of our lab is to understand the molecular mechanisms that underlie the process of cell-cell fusion. Cell-cell fusion is important for the development of complex tissue including muscle and bone and has recently emerged was a method for the progression of various cancers. More specifically, several cancers have been shown to become metastatic by fusing with normal cells that have migratory abilities. As a result, there is much interest in the identification of genes responsible for facilitating cell fusion. To this end, our lab uses

Tetrahymena thermophila , a fresh water ciliate, that undergoes cell-cell fusion quite readily in the laboratory, to identify novel regulators of cell-cell fusion. Moreover, the availability of the Tetrahymena Gene

Expression Database (TGED) allows one to identify genes whose temporal pattern of expression is consistent with a role in fusion.

Using the TGED, molecular genetic and cell biological approaches we aim to identify novel proteins that regulate cell-cell fusion by regulating membrane curvature at two distinct stages: 1. During fusion initiation and 2. During fusion pore expansion. We have identified several candidate proteins at each phase and here we report the localization of PX-BAR protein. Importantly, BAR proteins have been shown to regulate membrane curvature in a variety of cell types.

Our results show that PX-BAR appears to localize to the site of cell fusion in mating Tetrahymena. Moreover, analysis of TGED reveals the potential for co-regulation as 3 additional genes show very similar expression profiles, suggesting potential interactions. One such protein, Reticulon-4 has also shown to regulate membrane curvature suggesting the existence of a potential complex.

MS-INBRE 2014 ANNUAL RESEARCH SYMPOSIUM 1

O2 The Evaluation Of Natural Products As Anticancer Therapies In

Pediatric Cancer

Scharri Ezell Walker

Department of Biology, Tougaloo College, Tougaloo, MS

Brain tumors are the most common solid tumors and exhibit relatively high recurrence and mortality rates in the pediatric patient population.

Furthermore, resistance to current therapies may develop over time, giving rise to the development of advanced and drug-resistant cancers.

Therefore, there is an urgent and unmet need to find new anti-cancer treatments in the pediatric indication. The objective of this research is to investigate the anti-cancer potential of natural products and their pharmacological properties in pediatric cancer. Previous research studies have established natural products as an abundant source of active therapeutic agents against cancer. To accomplish this objective, three hypothesis driven aims will be completed: 1) Investigate the anti-cancer effects of cabazitaxel against murine and human brain/CNS cancer cell lines and the underlying molecular mechanism(s) of action responsible for these effects; 2) Investigate the anti-cancer effects of cabazitaxel against human brain tumor tissue and the underlying molecular mechanism(s) of action responsible for these effects; and 3) Determine the pre-clinical pharmacological properties of cabazitaxel.


O3 Design Of A Multifunctional Polypeptide For Application In

Cancer Diagnosis And Treatment

Manliang Feng 1 , Jasmine Jennings 1 , Ming-Shen Wu 1 , Drazen Raucher 2 ,

Jinghe Mao 1

1 Natural Science Divison, Tougaloo College, Tougaloo, MS

2 Department of Biochemistry, University of Mississippi Medical Center,

Jackson, MS

It is desirable in cancer therapy that anticancer agent can be traced invivo during the treatment providing information not only for the anticancer drug delivery efficiency but also the effectiveness of the therapeutic treatment. In this research, we designed a multifuntional polypeptide containing a cell penetration segment, a targeted delivery segment and a tracing segment. The cell penetration segment contains a SynB1 peptide with an amino acid sequence of

RGGRLSYSRRRFSTSTGRA. The target delivery segment contains a elestin-like polypeptide (ELP) consist of repeated sequence of pentapeptides (VPGXG), where the guest residue X can be any amino acid except Proline. ELP possesses a unique inverse temperaturephase transition feature. It is employed in the designed polypeptide for thermal-targeted delivery. The last segment of the polypeptide contains a amino acid sequence that can binds rare earth metal ions.

This segment of peptide when bind to rare earth metal ions such as

Eu 3+ and Gd 3+ can produce fluorescent and magnetic feature that can be used to trace the designed polypeptide both in-vitro and in-vivo.

The designed polypeptide was expressed and purified from E. Coli .

The metal binding properties and phase transition properties of this newly designed polypeptide were characterized. The preliminary results from a T

1

-weighed scan shows that the designed peptide enhances the MR signal.


O4 Characterizing The Mold Specific M46 Gene, Of The Pathogenic

Dimorphic Fungus Histoplasma capsulatum

Davida Crossley 1 , Glenmore Shearer, Jr.

2

1 Department of Biological Sciences, Alcorn State University, Lorman,

MS

2 Department of Biological Sciences, The University of Southern

Mississippi, Hattiesburg, MS.

Histoplasma capsulatum (Hc) is a dimorphic fungus, that causes the respiratory infection histoplasmosis.The fungus exists in the environment as a saprophytic mold and converts to a parasitic yeast in the mammalian host. It is the yeast which is pathogenic, and is the morphotype that is highly studied. Therefore, studies on the mold morphotype is highly overlooked. This research focuses on characterizing the mold specific M46 gene. M46 is a single copy gene.

Northern blot analysis with four commonly used Hc strains has shown that M46 is upregulated in strains G186AS and Downs but is down regulated in strains G184AS and G217B. The reason for the lack of expression in the latter strains is unknown. Currently we are investigating via promoter analysis and electrophobility assays to determine if the reason for lack of expression is due to a cis or transacting element(s). Promoter deletion studies are also ongoing to determine the essential regions of the M46 promoter that is required for expression. According to NCBI Genbank, M46 does not have a homolog, and therefore the function is unknown. An M46 knockout revealed that M46 may not be involved in dimorphism, maintaining normal hyphal formation, and cell growth rate. RNA sequencing suggests that M46 may be involved in drug resistance. Currently we are conducting drug susceptibility assays to test this hypothesis. Other current M46 characterization studies include; confirming the predicted protein 8.5kDA protein size of M46 via western blot analysis, and determine the location of M46 in the cell via immunofluorescence.


O5 Progress Toward The Synthesis And Anti-Mycobacterial

Properties Of Ethambutol Analogues Containing 3-

Aminoalcohols

Dale Rosado 1 , Erin Norcross 2 , Benjamin Lewing 1 , Wanqing Lu 1 , Amy

Wilcosky 1 , Roshetta Williams 1 , Pradeep Bandi 1 , Nancy Espinal 1 , Joshua

Irby 2 , Hallie Hodge 2

1 Department of Chemistry and Biochemistry, Mississippi College,

Clinton, MS

2 Department of Biological Sciences, Mississippi College,

Clinton, MS

Amino alcohols are found in a wide variety of biologically active compounds and have been used as chiral ligands for transition metal catalysts and to induce chirality in asymmetric synthesis. Ethambutol

(EMB), an anti-tuberculosis drug, contains two ( S )-2-amino-1butanol moieties. Replacement of the 2-aminoalcohols of EMB with

3-aminoalcohols may yield EMB analogues with increased activity. In order to investigate the properties EMB analogues containing 3aminoalcohols, an efficient synthetic route to both enantiomers has been developed. Reduction of diethyl 2-ethylmalonate with LiAlH

4 yields a 1,3-diol. The diol is quantitatively converted to a diacetate by treatment with acetic anhydride. Hydrolysis of the diacetate with

Pseudomonas fluorescens Lipase in phosphate buffer yields an acyl alcohols that can be treated with diphenyl phosphoroazide yields an acyl azide. ( S )-2-(aminomethyl)-1-butanol is obtained after reduction of the acyl azide with LiAlH

4

. The enantiomer of the 2-

(aminomethyl)-1-butanol was synthesized by manipulation of the protecting groups and repetition of the azidation and reduction chemistry. The synthetic route outlined above using mild conditions, produces good yields in each step, and results in synthesis of both enantiomers of 2-(aminomethyl)-1-butanol from a common acyl alcohol intermediate. Minimum inhibitory concentrations (MICs) are currently being conducted for the 3-aminoalchols and all synthetic intermediates against Mycobacterium smegmatis . All EMB analogues will be evaluated for activity once synthesis is complete. Any compounds that show MICs lower than that of EMB will be tested against M. tuberculosis H37Ra.


O6 Prospect Of Plant Flavonoids As Agents For Reducing Oxidative

Stress: Cellular And Molecular Studies

Bidisha Sengupta 1 , Donald Davis 1 , Ming Shenwu 2 , Falicia Edwards 2 ,

Kisa Harris 1 , Denise Ward 1 , D’Asia Gholar 1

1 Department of Chemistry, Tougaloo College, Tougaloo, MS

2 Department of Biology, Tougaloo College, Tougaloo, MS

In 1936, Rusznyák and Szent Györgyi first drew attention to the therapeutically beneficial role of dietary flavonoids, which are the most common group of polyphenols ubiquitously present in plant based food and beverages. Recent years have witnessed a renascence of interest on these nutraceuticals, which, because of their high potency and low systemic toxicity, are gradually emerging as promising alternatives to conventional therapeutic drugs. There is a mounting evidence that various proteins frequently serve as targets for therapeutically important flavonoids. In this preliminary study, we evaluated the effects of flavonol fisetin, flavone luteolin and an anthocyanin malvidin chloride against stress induced by a well known pesticide methyl parathion (MP) in HEPG2 cell lines. The flavonoids are present in common vegetable and fruits namely onion and bilberries respectively. For cellular investigations we used MTT cell viability assay while for molecular studies we relied on UV/Vis absorption, fluorescence and melting as well as modeling studies.

There is a significant decrease in MP induced cell death in presence of flavonoids. Molecular studies are carried out on protein extracted from the cells which are exposed to MP in absence and presence of the flavonoids. The beneficial effects of the flavonoids in the prevention of stress induced by MP may be related to their abilities of scavenging free radicals produced by oxidative stress. It is very pertinent to establish the structure-activity correlations for the functionalities of the flavonoids in this case.


O7 Technology-Enhanced Teaching And Learning Of Exposure

Science

Clement G. Yedjou, Raphael Isokpehi

Department of Biology, College of Science, Engineering and

Technology, Jackson State University, Jackson, MS

Exposure science is the study of human contact with chemical, physical, or biological agents occurring in their environments, and advances knowledge of the mechanisms and dynamics of events either causing or preventing adverse health outcomes. We know that many diseases are primarily due to environmental factors while genetic factors are rare with few exceptions. Therefore, we better figure out how to best characterize the environment to which we are exposed.

That is where exposure science comes in. There are volumes of data emerging from research on the exposome, which is the totality of environmental exposures (including such factors as diet, drug use, and infection) throughout a person’s life. However, a large number of people are still unaware about the potential impacts of environmental exposure. Therefore, the central goal of the present research is to enhance biology course resource and incorporate the use of interactive visual interfaces and software that facilitate visual analytics tasks, and to offer the learners the acquire knowledge. To achieve our goal, we design and implement innovative curriculum resources on exposure science in an online Marine and Environmental Science course.

Blackboard and visual analytics technologies are used in this project to facilitate evidence-based instruction and discovery-based learning.

Our preliminary data demonstrated students perform better when teaching with technology compared to face-to-face classroom environment. Taken together, both the used of blackboard technologies and visual analytics enhanced course resources provides a new comprehensive approach to improve student productivity and outcomes of undergraduate programs.


O8 Role Of Pentatricopeptide Repeat RNA-Binding Proteins In

Mitoribosome Biogenesis

J. Ignacio Moreno, Marta A. Piva

Department of Biological Sciences, Alcorn State University, Alcorn

State, MS

Breast cancer is a particularly important issue in the state of

Mississippi where a substantial percentage of the population is African

American. Women of this race have the highest death rates and often have cancers that grow faster and are harder to treat. A direct link between tumorigenesis and augmented mitochondrial activity and biogenesis has been extensively demonstrated. Tumor epithelial cells have their increased energy requirements met by associated fibroblasts in the form of lactate. This metabolic partnership allows them to over-proliferate creating the invasive malignant tissue. Mitochondria harbor their own genome that codes few but essential proteins that belong to the respiratory chain (OXPHOS). Thus, the entire transcriptional, translational and folding machineries that produce the active proteins are compartmentalized inside the organelle. Expression of about forty nuclear-encoded mitochondrial proteins (most of them mitoribosomal) is increased in human breast cancer cells. Therefore, mitochondrial ribosomes (mitoribosome) biosynthesis and tumor growth are strongly related and the mutual dependence has been established. In spite of the intrinsic relevance of this issue, many aspects of the mitoribosome biogenesis are still unknown. Our laboratory elucidated the moonlighting nature of Ccm1p ( COB and

COX1 mRNAs maturation) a pentatricopeptide repeat (PPR) protein that stabilizes the rRNA of the mitoribosomal minor subunit in

Saccharomyces cerevisiae (15S rRNA). The human counterparts are

PTCD3 / 12S rRNA. Preliminary experiments in our laboratory suggested that Ccm1p has an additional third function related to the

OXPHOS assembly. Currently we aim to elucidate the nature of the in vivo interaction between 15S rRNA and Ccm1p. Specifically we are determining whether Ccm1p acts as a 15S rRNA chaperone that guides the RNA in acquiring an active conformation.

Characterization of other factors that interact with Ccm1p and15S rRNA is also in progress. Data obtained from these main issues will


define, for the first time, the nuclear-encoded initiator of the chain of events that leads to the mitochondrial ribosome genesis. The elucidation of mitoribosome synthesis pathways might contribute to the design of new drugs that diminish the up-regulation of the mitoribosome genesis in breast tumor cells.


O9 Effect Of Cholesterol-Mediated Regulation Of Acid

Sphingomyelinase On Sensitivity Of Human Breast Cancer Cells

(Mcf-7) To Apoptosis-Inducing Stimuli

Sucheta S. Naidu, Gary T. Hodge, II, Benjamin J. Melancon, Peyton S.

Pollard, Erin W. Norcross , , Jerry W. Reagan, Jr.

Department of Biological Sciences, Mississippi College, Clinton, MS

A number of studies indicate that functional acid sphingomyelinase

(aSMase) is required for induction of apoptosis following exposure of cultured cells to various stimuli including TNFα and UV light. Since aSMase activity is sensitive to lysosomal cholesterol concentrations, we hypothesize that intralysosomal cholesterol can modulate the apoptotic threshold indirectly by regulating aSMase activity. To test this hypothesis, CHO-K1 and MCF-7 cells were incubated under conditions that down-regulate aSMase activity. The cells were then evaluated for their response to apoptosis-inducing stimuli by assessing several parameters that are indicative of programmed cell death.

These studies indicate that free cholesterol enrichment of the lysosomal compartment alters sensitivity of cells to death-inducing stimuli. However, whether the response is directly related to a reduction in aSMase activity or secondary to other cholesterol mediated-changes in lysosomal function (e.g. increased lysosomal membrane permeability) remains unclear. Differentiating between these possibilities will require the alteration of lysosomal cholesterol mass and aSMase activity independent of each other. Regardless of the outcome of additional studies, the results presented here indicate that the response of MCF-7 cells to apoptosis-inducing stimuli is cholesterol-dependent. Thus, internalization of low density lipoprotein and/or oxidized low density lipoprotein found in the plasma of obese individuals has the potential to regulate apoptosis.

Modulation of lysosomal cholesterol, therefore, appears to be a major contributing factor to the overall cellular mechanism(s) that initiates/propagates cell death pathways or allows the cell to engage in the uncontrolled proliferation that is characteristic of cancer cells.


P1 Role Of Hepatoma Up-Regulated Protein (Hurp) In Prostate

Cancer Resistance To Chemotherapeutic Drugs

James E. Kent Jr.

1 , Abdelouahid El-Khattouti 2 , Tangeng Ma 2 , Mohamed

Hassan 2 , Ingrid Espinoza 2 , Christian R. Gomez 2

1 Mississippi University for Women, Columbus, MS

2 Cancer Institute, University of Mississippi Medical Center, Jackson, MS

The goal of this study is to investigate the tumorigenic mechanisms of

Hepatoma Up-Regulated Protein (HURP) in prostate cancer (PCa).

HURP is a cell-cycle-regulated and microtubule-associated protein. It functions as a Ran GTPase effector and is involved in the stabilization of the mitotic kinetochore fibers. We found that HURP expression in

PCa is an independent predictor of aggressive disease. To study the association between HURP levels and the efficiency of killing PCa cells using chemotherapeutic drugs, LNCaP cells were transfected with a Lenti viral Tet-On expression system for tightly regulated expression of HURP. Empty vector- (3G) and HURP-overexpressing

(HURP) LNCaP cells were treated with docetaxel and bortezomib.

Cell survival was determined by standard MTT assay and the fraction of surviving cells vs. drug concentration was expressed as IC

50 concentration using GraphPad Prism. When treated with docetaxel and bortezomib, the empty vector transfected cells showed IC

50

’s of

2.12nM and 18.59nM respectively. HURP overexpressing cells were nearly three-fold higher with IC

50

’s of 6.11nM and 49.45nM respectively. These results demonstrate that HURP overexpression increases chemoresistance in PCa cells. Additionally, our findings set the benchmarks to propose HURP as a predictor of the therapeutic response and as a potential target for PCa treatment. Support: DOD

PC094680 (CRG), PCF Creativity Award (CRG), MS INBRE research scholar award (JEK).


P2 Sex Differences In The Progression Of Renal Injury Leptin

Receptor Knockout Dahl Salt-Sensitive (SS) Rats

Devin Guillory 1,2,3 , Kasi McPherson 2 , Jan Williams 2

1 Jackson State University, Jackson, MS

2 Department of Pharmacology, University of Mississippi Medical

Center, Jackson, MS

3 The University of Southern Mississippi, Hattiesburg, MS

Obesity is a major risk factor for the development of chronic kidney disease (CKD). In the current study, we used Zinc-finger nucleases to create a 16 base pair frame-shift deletion in exon 11 of the leptin receptor gene within the genetic background of the Dahl SS rat

(SS

Lepr

-/- strain) to study the effect of obesity on the progression of kidney disease in SS rats. We observed an increase in body weight in both female and male SS

Lepr

-/- rats when compared to SS rats at 6 weeks of age. At the end of the study body weight increased in female and male SS

Lepr

-/- rats. The SS

Lepr

-/- strain exhibited impaired glucose tolerance and increased plasma insulin levels as early as 6 weeks of age suggesting insulin resistance while SS rats did not.

However, glucose levels remained in the normal range throughout the course of the study. Both female and male SS

Lepr

-/- rats developed severe systolic hypertension when compared to the values seen in SS rats. The kidneys from the SS

Lepr

-/- strain displayed increased interstitial fibrosis when compared to SS rats regardless of sex.

Interestingly, the major finding from the current study was that the survival rate in female SS

Lepr

-/- rats was markedly reduced versus male

SS

Lepr

-/- rats. Overall, these data indicate that the SS

Lepr

-/- strain may be a useful model to study signaling pathways and/or sex differences that play a role in CKD associated with obesity.


P3 Measurement Of Particle Size And Charge Distributions Of

Asthma Drug Dry Powder Inhaler Aerosols

Marina Ali 1 , Mohammed Ali 2

1 St. Andrew’s Episcopal School, Ridgeland, MS

2 Department of Technology, Jackson State University, Jackson, MS

An experimental method for analyzing electromechanical properties

(e.g., size, electrostatic charge, polarity) of therapeutic aerosols produced by three different commercially available dry powder inhalers (DPIs), including Advair Diskus™, Spiriva™, and,

Pulmicort™ is presented. In this method we have used an Electronic

Single Particle Aerodynamic Relaxation Time (ESPART) Analyzer.

The ESPART employs the principles of laser doppler velocimetry for simultaneously measuring the aerodynamic diameter and electrostatic charge of each single particle in real time. Prior to each run, the aerosol sampling chamber (ASC) was cleaned and evacuated (36 cm of Hg) so that a bolus of 4L of aerosol could be drawn from the test

DPI upon opening of the inlet valve at 30L/min for 8 seconds. The chamber’s inside walls were lined with a grounded wire mesh. Once the ASC was filled, the port to the ESPART analyzer was opened for characterizing sampled aerosols for 5 minutes. Aerosol particles from all drug delivery devices were found to not only have different size distributions but also varied charge distributions. The drug aerosols inhaled from Advair, Spiriva and Pulmicort were determined to be electropositive. Count and mass distributions were reproducible for all

DPIs. These findings can be explained by triboelectrification charging between excipient and active pharmaceutical ingredient, delivery device geometries, and drug/carrier homogeneities. In conclusion, the

ESPART provided more detailed charge information about the DPI aerosol particles.


P4 Photochemistry Of N-Alkoxy Quinolinium, Isoquinolinium, And

Phenanthridinium Tetrafluoroborates: Analysis Of The Reaction

Pathways By Ph Monitoring

Priya P. Patel, Muzamil A. Khawaja, Katie L. Odom, Brooke K. Lassiter,

GeNita N. Finley, Wolfgang H. Kramer

Department of Chemistry and Biochemistry, Millsaps College, Jackson,

MS

N-methoxy substituted aromatic heterocycles undergo a photoinduced homolytical N-O-bond cleavage. The reaction produces a methoxy radical and a heteroaromatic radical cation. The quantum yields of ion/radical formation have been determined by laser flash photolysis/quenching for the title compounds. Each transient species was produced with a yield of about 0.6 (±0.05). The energy wasting step appears to be a radical recombination reaction, which also produces a proton.

At Millsaps, we attempted to analyze the homolytic N-O bond cleaving reaction by focussing on the proton. This would give us a versatile tool for the analysis of the energy wasting side reaction and thus also the efficiency of the overall reaction. The methods we employed were titration with p-nitrophenolate and pH monitoring with buffer systems. It was important to keep the pH above about 6 to avoid acid base equilibria with other photoproducts.

Product distribution was determined by GC/MS after basic extraction. Interestingly, only four major isomers were formed.

Attempts to correlate the isomeric distribution with the solvent polarity was unsuccessful so far.


P5 Phenolic And Anthocyanin Compound Concentrations In

Blueberry (Vaccinium Corymbosum) Cultivars

Donna Marshall 1 , Ashten Rea 2

1 USDA-ARS, Poplarville, MS

2 William Carey University, Hattiesburg, MS

Phenolic and anthocyanin overall compound concentrations were analyzed in 18 blueberry cultivars and 1 blueberry selection from

Poplarville, MS. The selections were to compare with phenolic and anthocyanin concentrations of 1 corresponding blueberry selection from Crystal Springs, MS, in 2011, as well as serve for continuation of the blueberry analyses from the previous year, for comparison with

Crystal Springs and Poplarville 2008 and 2010 selections. Analysis was carried out on blueberry juice partitions extracted from the blueberry fruit skin, seeds and pulp. The major phenolics in the extracted juice were identified by their characteristic spectra.

Quantification was made by calibration curves of external standards for both the phenolics and anthocyanin. Total anthocyanins and phenolics were observed in all varieties. The highest levels of phenolics were observed in the Onslow at 251.8 while the lowest levels were observed in Baldwin at 81.2. For anthocyanins the highest value included Maru at 30.7 and the lowest with Baldwin at 0.42. The

Blueberries have special interest due to their high antioxidant capacity.

Research shows that phenolic components within blueberries may have multiple health benefits, while the anthocyanins may have health benefits that are independent of or in addition to the blueberry's antioxidant effects.


P6 The Drosophila T-Box Transcription Factor Midline Functions

Within The Notch-Delta Signaling Pathway To Specify Sensory

Organ Precursor Cell Fates And Regulates Cell Survival Within

The Eye Imaginal Disc

Sudeshna Das 1 , Q. Brent Chen 1 , Joseph D. Saucier 1 , Brandon Drescher 1 ,

Yan Zong 2 , Sarah Morgan 2 , John Forstall 1 , Andrew Meriwether 1 , Randy

Toranzo 1 , Sandra M. Leal 1


Mississippi, Hattiesburg, MS

2 Department of Polymer and High Performance Materials, The

University of Southern Mississippi, Hattiesburg, MS

We report that the T-box transcription factor Midline (Mid), an evolutionary conserved homolog of the vertebrate Tbx20 protein, functions within the Notch-Delta signaling pathway essential for specifying the fates of sensory organ precursor cells. This complements an established history of research showing that Mid regulates the cell-fate specification of diverse cell types within the developing heart, epidermis and central nervous system. Tbx20 has been detected in diverse neuronal and epithelial cells of embryonic eye tissues in both mice and humans. However, the mechanisms by which either Mid or Tbx20 function to regulate cell-fate specification or other critical aspects of eye development including cell survival have not yet been elucidated. We have also gathered preliminary evidence suggesting that Mid may play an indirect, but vital role in selecting sensory organ precursor cells within the third-instar larval eye disc by regulating the expression of the proneural gene atonal . During subsequent pupal stages, Mid specifies SOP cell fates as a member of the Notch-Delta signaling hierarchy and is essential for maintaining cell viability by inhibiting apoptotic pathways. We present several new hypotheses that seek to understand the role of Mid in regulating developmental processes downstream of the Notch receptor that are critical for specifying unique cell fates, patterning the adult eye and maintaining cellular homeostasis during eye disc morphogenesis.


P7 Investigation Of The Water Quality Of Lake Grenada, MS

Tasha Norwood 1 , Padmanava Dash 2 , Julius Ikenga 3 , James Pinckney 4

1 Department of Biology, Jackson State University, Jackson, MS

2 Department of Geosciences, Mississippi State University, Mississippi

State, MS

3 Department of Natural Sciences and Environmental Health,

Mississippi Valley State University, Itta Bena, MS

4 Marine Science Program and Department of Biological Sciences,

University of South Carolina, Columbia, SC

Lake Grenada (33.8196, -89.7737), located near the city of Grenada,

MS, is one of the largest lakes in Mississippi. The main objective of this research was to investigate the water quality of Lake Grenada.

First, a time-series of true color satellite images were generated and examined for the occurrence of algal blooms in the past. The lake was sampled on June 2, 2013. Clean Nalgene bottles were used to sample water at the twelve selected sites. A portable Hanna Instrument

(HI769828) was used to measure temperature, salinity, dissolved oxygen and pH at each site. High Performance Liquid

Chromatography (HPLC) analysis was performed to obtain algal pigment concentrations. The collected samples were also analyzed for cyanobacteria specific pigment phycocyanin (PC), Suspended

Particulate Matter (SPM), phycotoxin, toxic metal and microscopy.

The organic SPM was higher at seven sites out of the total twelve sites. The relative abundances of major algal groups revealed the dominance of euglenophytes in the lake with the presence of cryptophytes and cyanobacteria. Microcystin a hepatotoxic cyanobacterial toxin was found in the lake with concentration ranging from 0.18-0.43 µg/L. Among the four types of bacteria counted, total coliforms and heterotrophic bacteria were present at all the sites in high concentrations, but E. Coli and Enterococii bacteria were also present at few sites. Several toxic metals were also found. These findings suggest the need of continuous monitoring of the water quality of Lake Grenada.


P8 The UMMC Molecular And Genomics Facility: A Resource For

Mississippi’s Genomic Needs

Michael R. Garrett 1,2 , Zibiao Guo 1

1 Department of Pharmacology and Toxicology, University of Mississippi

Medical Center, Jackson, MS

2 UMMC Molecular and Genomics Core, University of Mississippi Medical


The University of Mississippi Medical Center (UMMC) Molecular and Genomics Core Facility (MGCF) provides centralized access to molecular and genomics expertise and services for the University and external researchers. In recent years, it has become evident that a scientific approach using high-throughput genomic technologies, such as microarray and next-generation sequencing (NGS) can provide an enormous capacity to understand the complex interaction of biological systems associated with living organisms, human health, and disease.

Mission: The mission of the MGCF is to serve as a nucleus to develop research and educational programs to increase the competitiveness and enhance biomedical discovery of researchers.

Equipment: The MGCF is equipped with several genomics platforms, including Affymetrix 3000 7G and GeneAtlas Instruments for whole genome expression profiling and an Illumina MiSeq for small RNA sequencing (miRNA), ChIP-Seq, and amplicon sequencing. Mammalian level RNA-Seq services are offered by inhouse preparation of libraries, quality control on Illumina MiSeq and deep-sequencing (Illumina HiSeq) using an outside service provider.

Services: The MGCF provides the following services: (1) sample preparation, quality control, and storage; (2) sequencing and genotyping; (3) microarray, NGS, and validation via quantitative realtime PCR; and (4) preliminary bioinformatics analysis. In summary, the MGCF provides cutting-edge genomic technologies and genomics expertise to academic institutions throughout the State of

Mississippi. Supported by P20 GM103476 [MS-INBRE-(Shearer)];

P30 GM103328 [CPN-COBRE (Stockmeier)]; and P20 GM104357

[Cardio-Renal (Hall)].


P9 Tracking Plastid (And Mitochondrial) Gene Migration In K.

Brevis

Kelly E. Scott, Timothy McLean

Department of Biological Sciences, The University of Southern


Karenia brevis is a marine dinoflagellate responsible for the harmful algal blooms (also known as red tides) in the Gulf of Mexico. K. brevis expresses antisense (AS) RNAs, each of which has a complementary region to the messenger RNA (mRNA) of a variety of genes. In dinoflagellates, many plastid (and mitochondrial) genes have migrated to the nuclear genome. It is unknown whether chloroplast genes, such as photosystem – D2, have migrated in K. brevis. It is also unknown where the gene that expresses the AS RNA for photosystem

D2 resides. The protein-coding gene and the AS RNA-expressing gene could both be in the chloroplast, both in the nucleus, or some split combination between the two genomes. Primers designed from photosystem D2 ESTs were used in a series of RACE reactions to capture the unique regions of both photosystem – D2 AS RNA and mRNA. Gel imaging showed a distinct band for the unique 5’ end of the mRNA. Sequencing of this band will allow for the design of a probe to determine which genome houses the photosystem – D2 mRNA. This work can be furthered to compile known locations for both the mRNA and AS RNA of both chloroplast and mitochondrial genes of K. brevis.


P10 The Effects Of High Energy Radiation On The Vestibular And

Immune Function In Rats

Ansley Scott 1 , Ming Shenwu 1 , Xuehui Tang 2 , Jun Huang 2 , Chunli Yang 5 ,

Wu Zhou 2,3,4 , Hong Zhu 2,3 , Jinghe Mao 1,2

1 Tougaloo College, Tougaloo, MS

2 Departments of Otolaryngology &Communicative Sciences, University of Mississippi Medical Center, Jackson, MS

3 Neurobiology & Anatomical Sciences, University of Mississippi Medical


4 Neurology, University of Mississippi Medical Center, Jackson, MS

5 Radiation Oncology, University of Mississippi Medical Center, Jackson,

MS

The objective of the project is to study radiation effects on the vestibular and immune systems in rats. It is proposed that high energy radiation will compromise vestibular/balance system and immune function since nasopharyngeal cancer patients experience post-irradiation vertigo and whole body radiation therapy causes immune suppression. However, the underlying mechanisms are not fully understood.

Sprague-Dawley rats were exposed to different doses of either unilateral cranial using 6 MeV electron beam (0 to 60Gy) or whole body using 18

MeV electron beam (0 to 10 Gy). For vestibular study, seven days post irradiation, single unit recording of vestibular afferent activity was performed. A total of 309 horizontal semicircular canal afferents’ responses to sinusoidal earth-horizontal rotations (~1Hz) were analyzed.

For immune function study, total RNAs were extracted from immune organs such as thymus and spleen. The genes involved in Stress and

Toxicity PathwayFinder were examined using RT 2 Profiler TM PCR

Array.

Preliminary analysis shows that electron irradiation caused significant decreases in the gains of the irregular horizontal semicircular canal afferents. The gene expression profiles indicated significant changes in oxidative stress, DNA damage and repair as well as apoptosis related genes.

In conclusion, these results indicate that radiation exposure induces specific functional deficiency of the vestibular and immune system in rats.


P11 Study Of The Role Of The Mold-Specific Ms95 Gene In Dna

Repair In The Pathogenic,

Dimorphic Fungus Histoplasma Capsulatum

Erin M. Smith, Logan Blancett, Davida Crossley, Glen Shearer



Histoplasma capsulatum (Hc) is a dimorphic fungus that is the etiologic agent of the respiratory infection Histoplasmosis. In the environment, Hc exists as a multicellular, differentiated mold. The organism undergoes a shift to a unicellular, undifferentiated yeast once inhaled into the host’s lungs. This transition from mold to yeast is essential in the pathogenicity of Hc . Genes have been identified that are specific to the mold or yeast phase in an attempt to understand the molecular biology of this dimorphic shift. In our lab, MS95 was identified in a subtractive cDNA library that was enriched for genes upregulated in the mold morphotype. The gene was characterized via northern and southern blot analysis of four Hc strains , confirming that it is a mold-specific gene. Using NCBI genbank and BLAST analysis, it was found that MS95 is homologous to the DNA damageresponsive gene DDR48, which functions in DNA repair in

Saccharomyces cerevisiae. Because of the homology, it is hypothesized that MS95 may be involved in DNA repair in Hc . In order to elucidate the function of MS95 , a loss of function mutant was created via allelic replacement. In order to determine if MS95 is involved in

DNA repair, wild-type as well as MS95 knockout mutant strains were grown on Histoplasma macrophage media (HMM) supplemented with varying concentrations of 4-nitroquinoline 1-oxide (4-NQO) or paraquat dichloride. These chemicals are known to cause oxidative stress, inducing DNA damage. Growth of both strains was monitored for any observable changes between the two strains indicating that MS95 plays a role in DNA repair. Future experiments include creating a complement of MS95 and over-expressing it in the yeast phase to further determine its role in Hc.


P12 Cooperative Interaction Of Human Granulin-A (Grna) With

Amyloidβ (A β ) Peptide: Implications Of Inflammatory

Processes On Alzheimer’s Disease

Gaurav Ghag, Vijay Rangachari

Department of Chemistry and Biochemistry, The University of Southern


Granulins (Grns) are small (~ 6 kDa) cysteine rich proteins that are generated upon proteolytic cleavage of a precursor protein called progranulin (PGrn) during an inflammatory response. PGrn contains

7.5 tandem repeats of Grn motifs (GrnA-G). PGrns are known play role in diverse processes like wound healing and repair as well as in tumorigenesis. While PGrns are anti-inflammatory, Grns are proinflammatory. Mutations in the PGRN gene resulting in loss of function of PGrn have been implicated in the familial form of frontotemporal lobar degeneration (FTLD), a neurodegenerative disorder characterized by behavioral and personality changes. Several circumstantial evidence indicate the involvement of both PGrn in

AD: Clinical diagnoses of Alzheimer’s (AD) and Parkinson’s diseases

(PD) have been found to be associated with certain PGrn mutations, and PGrn is observed to be upregulated in activated microglia surrounding A β plaques that interfere with neurotransmission in the brain leading to the symptoms of AD. Furthermore, the increased A β aggregates observed in patients suffered from a traumatic brain injury

(TBI) suggests a correlation between the increased PGrn production, also observed after a traumatic brain insult, and A β aggregation. Based on the evidence as well as the structural similarity between N-terminal domain of human GrnA and A β fibrils, we hypothesize a potential interaction between the two proteins. To test our hypothesis, we have characterized the molecular interactions between GrnA and A β 42 in vitro . We observed that GrnA displays a monomer  dimer equilibrium with a K

D

of ~ 150 nM. Interestingly, the protein adopts a

PP-II helical conformation as a dimer and a partial α -helical structure as a monomer in solution. We think that the interaction between

GrnA and A β depend on whether GrnA is monomeric or dimeric.

GrnA also seemed to preferentially interact with different aggregate forms of A β indicating a highly cooperative nature of the overall


interactions between the two proteins. This cooperativity may have far-reaching consequences in the inflammatory responses in AD. The data is presented and discussed.


P13 The Drosophila T-box Transcription Factor Midline Functions within Stress-Reactive Signaling Pathways to Regulate Cellular

Homeostasis

Q. Brent Chen 1 , Sudeshna Das 1 , Robert S. Smith 1 , Wisam Buti 1 , Yan

Zong 2 , Sandra M. Leal 1



2 Department of Polymer and High Performance Materials, The

University of Southern Mississippi, Hattiesburg, MS

We recently reported that the T-box transcription factor midline ( mid ) functions within the Notch-Delta signaling pathway to specify sensory organ precursor (SOP) cell fates in early-staged pupal eye imaginal discs (Das et al., 2013). Targeted reduction of mid expression in cells within and posterior of the morphogenetic furrow (MF) during third-instar larval (3 o L) and early pupal developmental stages using the UAS-Gal4 binary expression system and RNAi methods not only resulted in decreased numbers of interommatidial bristles (IOBs) generated from SOP cells, but also lead to widespread apoptosis in the adult eye (Das et al., 2013). We have now identified Drosophila FOXO

( dFOXO ) and target of rapamycin ( dTOR ), both integral members of the Insulin Receptor (InR) signaling pathway, as midinteracting genes that antagonize mid function in developing eye tissues. Carrying out an allelic modifier screen, we placed mid within the genetic hierarchy of Insulin Receptor (InR) signaling that is responsive to the nutritional needs of the organism. Consequently, inducing metabolic stress enhanced the mid mutant phenotype typified by a greater loss of

IOB complexes compared to unstressed mid mutant flies. Subjecting mid mutants throughout their life cycle to oxidative stress using paraquat also resulted in an enhancement of the mid mutant phenotype in adult flies. These comprehensive results suggest that mid is a critical gene functioning within signaling pathways that engage in crosstalk under conditions of either metabolic or oxidative stress. To complement these studies, we are currently examining whether mid regulates gene expression within the InR important for maintaining cellular homeostasis.


P14 The Effects Of Genistein And 5-Fu On Caov3 Cells

Chakiras S. Alexander, Gerri A. Wilson, Michelle A. Tucci, Hamed A.

Benghuzzi

School of Health Related Professions, University of Mississippi Medical


CAOV3 is a specific type of malignant tumor cell and is among the many types of ovarian cancer. Much research has indicated that

CAOV3 is an epithelial cell on th e surface of ovaries. Several treatments have been introduced; however, none has been successfully conclusive. In this experiment, genistein, 5 Fluorouracil (5FU), and the combination (Combo) of these two agents have been used to determine their effect o n CAOV3.

Cellular protein, total reduced cellular glutathione levels, and cytomorphological evaluations were used to determine the effects of the compounds on cellular viability. CAOV3 cells were treated with genistein, 5 FU, and the combo for periods of 24, 48, and 72 hours.

The cells and supernatants were collected and stained. Pictures were taken to observe the cell morphology.

The objective of this experiment has been achieved. The results for the assays have indicated that genistein, 5 FUl, and t he Combo have reduced CAOV3 cell population, with the combination of the two drugs showing significant changes in cell morphology.

Although the effects of genistein, 5FU, and the combo have been positive, the results also indicated that more tests are needed to determine if CAOV3 can be eliminated entirely.


P15 The Effects Of Genistein And 5fu On Ovarian Cancer Cells

Stacey Bodkins, Gerri Wilson, Michelle A. Tucci, Hamed Benghuzzi

School of Health Related Professions, University of Mississippi Medical


Ovarian cancer deaths account for 50% of cancer in women over 63.

Treatments include surgery, chemotherapy, radiation therapy, immunotherapy, and vaccine therapy. Genistein is a natural compound that is found in soybean and has shown potential to be effective in suppressing cancer cell growth. Our goal was to compare genistein with a known chemotherapeutic agent, 5FU for suppression of adenocarcinoma of the ovary using SKOV 3 ovarian cells. We evaluated cellular growth by asse ssing total cellular protein concentration, and cellular toxicity by evaluation of total glutathione levels along with cytomorphologial evaluation. Our results indicate genistein alone was not effective in reducing cell protein or inducing changes in cell ular glutathione for the duration of study. Cells treated with 5FU also did not show significant changes in total cellular protein levels, but did show evidence of cellular toxicity as indicated by the reduced level of glutathione and changes in cellular morphology. Administering both compound together resulted in a significant decline in cellular protein and glutathione compared to control and to the addition of individual compounds for the duration of study. The morphological evaluation showed changes in the combo group as early as 24 hours that remained even after 72 hours of treatment. The results show the potential to add a combination of cancer drugs that work in different parts of cell cycle that when added individually do not effectively reduced cancer cell load. Combining drugs that target different phases of the cell cycle may be more effective in reducing the cancer cell load and may potentially aid in reducing cellular resistance to drugs.


P16 Investigating The Mold- Specific M46 Gene Involvement In Drug

Resistance In The Pathogenic Dimorphic Fungus Histoplasma

Capsulatum

Alicia Blanton, Davida Crossley

Department of Biological Sciences, Alcorn State University, Lorman, MS

Histoplasma capsulatum is the etiologic agent for histoplasmosis. The fungus is dimorphic and can exist as a multi-cellular saprophytic mold in contaminated soils of birds and bat excreta. The fungus shifts to the unicellular parasitic yeast in the lungs of humans and thus cause infection. In order for pathogenesis to occur, there must be conversion of mold to the yeast morphotype. Several previous studies are focused on host parasitic interactions, yeast phase specific genes, and virulence factors. Very few studies have been on mold phase specific genes and environmental factors. Thus the mold is often over looked. This study focuses on the mold specific M46 gene. Very little is known about

M46 . It is a single copy gene that contains two exons and one intron, and encodes for a very small protein of 8.5 kDa. According to NCBI,

M46 currently does not have a homolog, and therefore its function is unknown. Previously, an M46 knockout was created to determine the function of M46 . The knockout has shown that M46 is not involved in dimorphism, maintaining the normal formation of the mold, and maintaining the normal growth rate of cells. RNA sequencing data have shown that 6 six, MFS (Major Facilitated Superfamily) transporters that are involved in a drug efflux system are up regulated in the knockout strain and down regulated in the wild type strain.

There is one, ABC (ATP Binding Cassette) transporter and one, drug resistant protein that is up regulated in the wild type and down regulated in the knockout strain. These results suggest that M46 may be involved in drug resistance. We have now currently identified the transporters substrates by using NCBI tblastn. The search suggests that the substrates include cycloheximide, florphenicol, leptomycin, and oligomycin. A drug susceptibility assay with several of the antifungal agents with various concentrations is underway to test this hypothesis.


P17 Chemo-Preventive Of Garlic In Human Leukemia Cells:

Involvement Of Oxidative Stress, Apoptosis, And Cell Cycle

Arrest

Clement G. Yedjou, Paul B. Tchounwou

Cellomics and Toxicogenomics Research Laboratory, NIH RCMI-Center for Environmental Health, College of Science, Engineering and

Technology, Jackson State University, Jackson, MS

Garlic supplementation in diet has been shown to be beneficial to cancer patients. Recently, its pharmacological role in the prevention and treatment of cancer has received increasing attention. However, the mechanisms by which garlic extract (GE) induces cytotoxicity, oxidative stress, apoptosis, and cell cycle arrest in cancer cells remain largely unknown. The present study was designed to use HL-60 cells as a test model to evaluate whether or not GE-induced cytotoxicity, apoptosis, and cell cycle arrest in human leukemia (HL-60) cells is mediated through oxidative stress. Human leukemia (HL-60) cells were treated with different concentrations of GE for 12 hr. Cell survival was determined by MTT assay. The extent of oxidative cell/tissue damage was determined by measuring malondialdehyde

(lipid peroxidation biomarker) concentrations by spectrophotometry.

Cell apoptosis was measured by flow cytometry assessment (Annexin-

V and caspase-3 assays) and agarose gel electrophoresis (DNA laddering assay). Cell cycle distribution was detected by the cellometer vision. Data obtained from the MTT assay indicated that GE significantly ( p < 0.05

) reduced the viability of HL-60 cells in a concentration-dependent manner. We detected a significant ( p < 0.05) increase in malondialdehyde (MDA) concentrations in GE-treated

HL-60 cells compared to the control. Flow cytometry data showed a strong concentration-response relationship between GE exposure and

Annexin-V positive HL-60 cells. Similarly, a statistically significant and concentration-dependent increase ( p < 0.05) were recorded with regard to caspase-3 activity in HL-60 cells undergoing late apoptosis.

Annexin V and Caspase-3 data were confirmed by the result of DNA laddering assay showing clear evidence of nucleosomal DNA fragmentation in GE-treated cells. GE treatment induced G

0

/G

1

cell cycle arrest in in GE-treated HL-60 cells. Together, our findings


indicated that garlic significantly inhibited the proliferation of human leukemia (HL-60) cells via G

0

/G

1

cell cycle arrest, induction of apoptosis, and the induction of apoptosis was associated with oxidative stress. This old drug may be valuable for both the prevention and treatment of acute promyelocytic leukemia.


P18 Toxicity And Gene Expression Assessments In Vernonia

Amygdalina-Treated Breast Cancer Cells

Lecia Robinson 2 , Clement G. Yedjou 1,2 , Ernest Izevbigie 2 , Paul B.

Tchounwou 1,3

1 Cellomics and Toxicogenomics Research Laboratory, NIH-Center for

Environmental Health, Jackson State University, Jackson, MS

2 Cellular Signaling, Phytoceuticals, and Cancer Prevention and

Therapies, Jackson State University, Jackson, MS

3 Environmental Toxicology Research Laboratory College of Science,

Engineering and Technology, Jackson State University, Jackson, MS

Cancer of the breast is the most commonly diagnosed non-skin cancer and second leading cause of cancer-related deaths in women in the

United States. Breast cancer represents 29% of new cases of all cancers. An estimated 226,870 women will be diagnosed with invasive breast cancer and 39,510 women will die from the disease this year in the United States. There is an urgent need for the discovery and development of agent(s) efficacious against breast cancer to decrease breast cancer mortality and morbidity. National surveys on the use of

Complementary and Alternative Medicine (CAM) among patients show more than eighty percent of cancer patients, representing a spectrum of malignancies and disease stages acknowledged the use of

CAM. The growing popularity of CAM usage has led to the discovery of aqueous leaf extracts of Vernonia amygdalina ( V. amygdalina ), a Nigerian edible plant as a very strong candidate.

Previous studies have shown V. amygdalina to inhibit the proliferation of estrogen receptor positive (ER+) and estrogen receptor negative

(ER-) human breast carcinoma cells in vitro. V. amygdalina may be used alone or in combination (adjuvant) with known breast cancer drugs. Therefore, the central goal of this research was to determine the therapeutic mechanisms of V. amygdalina leaf extracts in breast cancer cells. To achieve this goal, cell viability, live and death cells were determined by the means of the MTT and propidium iodine assays, respectively. Western blot analysis was performed to assess the expression of p53 tumor suppressor. Data obtained from the MTT assay indicated that V. amygdalina significantly reduced the viability of

MCF-7 cells a dose-dependent response. On one hand, the Trypan


blue dye exclusion test demonstrated the integrity of the membrane of untreated cells in culture. On the other hand, the trypan blue dye exclusion test demonstrated a loss of viability in V. amygdalina -treated cells due to membrane damage. A statistically significant was recorded with propidium iodine data, showing an increase of necrotic cell death in V. amygdalina -treated cells, indicative of membrane rupture by V. amygdalina . No statistically significant differences ( p >0.05) in p53 expression was found between V. amygdalina -treated cells and the control, suggesting that there was that was not arrested at G

1

/G

0

upon

24 hours of treatment. Taken together, our research demonstrated that V. amygdalina treatment induced cytotoxic effects and expression of p53 tumor suppressor gene in cancer cells.


P19 msaABCR Regulates Autolysis Independent Of sarA In

Staphylococcus aureus

Justin Batte, Gyan Sahukhal, Mohamed O. Elasri



Staphylococcus aureus has many different virulent factors that allow it to be successful in the colonization and infection processes within the host. It has developed a complex regulatory network that is responsible for the regulation virulence. We have identified and defined the msaABCR operon that plays a role in this complex regulation process. Deletion of msaABCR reduces the expression of the global regulator sarA, and reduce biofilm formation. In this study we show that proteases and autolysis are regulated by both the msaABCR operon and sarA in an independent fashion.

To define the regulatory relationship between msaABCR and sarA , we constructed an msaABCR/sarA double mutant in the USA300 LAC strain. We also cloned and fused the sarA ORF to a cadmiuminducible promoter for trans-complementation of sarA . We tested the effect of the double mutation in the presence and absence of sarA on various phenotypes that included protease production, autolysis, and biofilm formation.

Previously, we showed that msaABCR essential for full expression of sarA . In this study, we set out to define the regulatory relationship between sarA and msaABCR relative to autolysis. The msaABCR/sarA double mutant and the sarA mutant showed an increased rate of autolysis relative to wild type. The msaABCR deletion mutant , however , showed an intermediate rate of relative to wild type. When we restore the level of sarA expression in the double mutant using an inducible promoter, autolysis rate are reduced but they do not reach wild type level or that of msaABCR deletion mutant. This suggests that sarA is the main regulator of autolysis but msaABCR remains necessary for regulation of autolysis.

This study shows that the msaABCR operon plays a key role in autolysis and that this regulation is not mediated by sarA . This is interesting because msaABCR positively regulates the expression of sarA , however, restoration of sarA expression in the msaABCR mutant


is not sufficient to restore the autolysis defect to wild type. This indicates that msaABCR and sarA are both required to regulate autolysis. Currently, we are investigating how these regulators are responsible for the regulation observed individually as well as how they may be linked together in one system.


P20 Molecular Detection Of Spotted Fever Group Rickettsia In Ixodid

Ticks From Pakistan

Kelsey Carter, Jaclyn Williams, Asma Kausar, Nabanita Mukherjee,

Shahid Karim



Ticks are obligate hematophagous ectoparasitic members of the class

Arachnida and order Ixodida that affect every class of vertebrate. In

Pakistan, ticks transmit more diseases, to livestock and humans than any other arthropod. Among these diseases is rickettsiosis: an illness caused by spotted fever group Rickettsia (SFGR). Rickettsia spp. are obligate intracellular pathogens responsible for economic and livestock loss in Pakistan. The aim of this project was to detect spotted fever group Rickettsia (SFGR) in Ixodid tick species from the Kashmir,

Punjab, and Sindh regions of Pakistan using an outer membrane protein A (ompA) PCR assay. The data reveals the Kashmir region of

Pakistan has the highest number of Ixodid tick species infected with spotted fever group Rickettsia . This molecular epidemiology survey for

SFGR in Ixodid ticks from Kashmir also reveals that Haemaphysalis spp. of ticks is more commonly infected with Rickettsia spp. than either the Hyalomma or Rhipicephalus spp. The Punjab and Sindh regions show that Rhipicephalus spp. of ticks is more likely than

Hylomma spp. to be infected with SFGR.


P21 Expression Of Stress-Related Genes In The Red Blood Cells

Of The Atlantic Stingray ( Dasyatis Sabina)

Ethan J. Semrad 1 , Andrew N. Evans 2

1 Belhaven University, Jackson, MS

2 Department of Coastal Sciences, The University of Southern

Mississippi, Ocean Springs, MS

Unlike the red blood cells (RBCs) of many vertebrate taxa, elasmobranch RBCs are nucleated. Because of this, they may play a pivotal role in the first response to stress. The first step to determine if RBCs are part of the elasmobranch stress response is to determine what stress related genes they express. We took blood samples from

Atlantic stingray ( Dasyatis sabina ) and perf ormed PCR for the following physiological and osmotic stress related genes: Heat Shock

Protein 70 ( HSP70 ), Serum and Glucocorticoid Inducible Kinase

( SGK ), Mineralocorticoid Receptor ( MR ), Glucocorticoid Receptor

( GR ), Angiotensin Receptor ( AT ), and Natriuretic Peptide Receptor

( NPR-B ). Glyceraldehyde 3 Phosphate Dehydrogenase ( GAPDH ) is the housekeeping gene to which the expression of the other genes was compared. We saw that the RBCs expressed several of the genes involved in the stress response a nd thus may play a part in the first response to stress. There was strong expression of HSP70 , MR , and

GR , which implies that the RBCs may respond to cellular stress and corticosteroids. NPR-B was expressed in all groups but its expression was not as str ong as HSP70 , MR , and GR . SGK was not expressed under basal conditions and AT was only expressed by one of the samples so future studies are needed to determine if RBC SGK and

AT gene expression is inducible by stress or other factors.


P22 Biochemical Analysis Of Two Suppressors Of Cell Cycle

Mutations In Aspergillus Nidulans

Casey A. Spell, Sarah Lea Anglin

Biology Department, Millsaps College, Jackson, MS

The filamentous fungus Aspergillus nidulans is a model organism for the study of the eukaryotic cell cycle. In this work, two specific genes that interact with known cell cycle proteins have been studied. The snoA gene functions during S phase, and mutations in snoA suppress the effects of a mutation in nimO dbf4 , which is a DNA licensing factor required for DNA synthesis. The snxA gene functions during G1 and interacts with nimX cdc2 , the master cell cycle control protein in all eukaryotes. The goal of our work with snoA was to use single-step affinity purification using an S-tagged snoA construct, followed by mass spectrometry, to identify SNOA and co-purifying proteins. Our results showed that SNOA protein was detectable by western blot at the predicted size (~250 kD), along with several smaller bands that are likely to be proteolytic degradation products. The goal of our work with snxA was to test the hypothesis that two hypomorphic mutations, snxA 1 and snxA 2, have reduced expression of sxnA mRNA compared to wild-type cells. To test this hypothesis, isogenic strains of snxA +, snxA 1, and snxA 2 were grown and qRT-PCR was performed to quantitate levels of snxA mRNA. The levels of mRNA in the wildtype ( snxA +) strain were found to be approximately 5-fold increased compared to both mutant strains ( snxA 1 and snxA 2). These data suggest that the hypomorphic phenotype is the result of lowered snxA expression. Future experiments are aimed at understanding the mechanism leading to lowered snxA expression in mutant strains.


P23 The Effect Of Low Level Laser Therapy On LPS Stimulated

Macrophage Cells

Courtland Brown 1 , Felix Adah 2 , Michelle A. Tucci 2 , Hamed Benghuzzi 2

1 Murrah High School, Jackson, MS

2 School of Health Related Professions, University of Mississippi Medical


Low level laser therapy (LLLT) has been shown to have positive effects on pathological conditions such as tendinopathies, osteoarthritis, and wound healing. In addition, it has been shown to reduce markers of inflammation. Although it can be used clinically, there is controversy surrounding the effectiveness of the LLLT because the biochemical mechanisms, timing, applied light intensity, and effective wavelengths are not fully understood. Our goal is to determine the effects of LLLT on acute inflammation using LPS stimulated macrophages as a model. Macrophage like cells were plated at a density of 1 x 106 cells per well onto 6 well plates and divided into 4 groups (cells only, cell + LLLT, cells + LPS, and cell +

LPS + LLLT). Cells in each treatment group were evaluated at 24 and 72 hours after a single LLLT treatment. Cellular protein concentrations were similar after 24 hours, but at 72 hours the cellular protein concentrations in both laser treatments groups were significantly less than cells receiving no treatment or cells stimulated with LPS. Nitric oxide production at 24 hours was significantly higher in both treatment groups receiving LPS. At 72 hours the cells which received LLLT treatment with or without LPS stimulation were significantly elevated when compared to control cells. Cellular morphological changes were also seen in LPS stimulated and LLLT treated cells. Macrophage cells are affected by LLLT and further studies are needed to determine the effects of LLLT on inflammatory markers.


P24 Mitochondrial Ribosomes And Cancer

Marta A. Piva, J. Ignacio Moreno


State, MS

African American women have the highest death rates, being 40% more likely to die of breast cancer than white women. Moreover, black women often have cancers that grow faster and are harder to treat. Interestingly, a new paradigm has recently emerged regarding the direct relationship between breast cancer aggressiveness and mitochondrial biogenesis. According to this model, human tumors and in particular breast cancer tumors consist of two distinct metabolic compartments: cancer adjacent-associated fibroblasts that produce large quantities of lactate by fermentation and epithelial cancer cells located nearby which uptake the lactate and use it to fuel oxidative mitochondrial metabolism. The direct link between mitochondrial biogenesis and tumorigenesis has been extensively demonstrated: overexpression of molecules that increase the abundance of oxidative phosphorylation (OXPHOS) complexes in human breast cancer cell lines such as the mitochondrial RNA polymerase-DNA dependent (POLRMT) also promote tumor growth. Moreover, high expression of POLRMT, the enzyme that catalyzes the synthesis of both rRNAs and mRNAs in mitochondria, predicts poor overall patient survival. RNA chaperones are also important factors whose role in ribosome biogenesis has recently emerged. These proteins, which are loosely associated with mitoribosomes, are required for rRNAs folding and thus their stability to maintain the integrity of ribosome subunits. It has been recently proposed that aggressive epithelial cancer cells behave much like infectious metabolic “parasites”. Therefore, novel anticancer agents should be akin to antibiotics which eradicate mitochondrial biogenesis in epithelial cancer cells. This principle is even illustrated in the treatment of other type of cancers such as acute myeloid leukemia: pharmacological concentrations of the antimicrobial tigecycline, which specifically targets the mitoribosome large subunit, selectively kill leukemia stem and progenitor cells. Baker’s yeast constitutes an ideal model to study mitochondrial biogenesis because it can live


without this organelle. The similarities between human and yeast mitoribosomes have validated its use as a model to elucidate the mechanism of action of anticancer drugs. Based on these premises, unraveling the mechanisms of mitochondrial biogenesis has become essential to understand and perhaps remedy this most devastating disease.


P25 Key Role Of A 35-Residue Motif Of Ccm1p Full-Length On The

Steady-State Levels Of 15s Rrna In An Intronless Mitochondria

Variant Of Saccharomyces Cerevisiae

K. Allyne Echols, Cherrelle T. S. Gee, Marta A. Piva, J. Ignacio Moreno


State, MS

Mitochondria are required to synthesize several components of the electron transport chain, without which, the generation of energy by aerobic respiration is not possible. While in neurons it causes cell death, Saccharomyces cerevisiae (yeast) can perfectly survive without mitochondrial dysfunction via fermentation, thus making yeast a perfect tool to study mitochondria functionality. Under this condition, the microbe is unable to use non-fermentable substrates as sole source of carbon and energy. Consequently, such phenotype is a rapid and easy way to evaluate the overall activity of this organelle. Ccm1p, a protein that contains pentatricopeptide repeat (PPR) domains, is essential for stabilizing 15S rRNA, a key component of the mitochondrial ribosome small subunit. The function of Ccm1p and the importance of its PPRs motifs on stabilizing 15S rRNA were elucidated in our laboratory using intron-containing mitochondria.

15S rRNA decay in deltaccm1 mutants leads to a translation failure causing irreversible dysfunction due to total mitochondrial DNA loss.

To understand the mechanism by which PPRs in Ccm1p protects 15S rRNA, we deleted the most canonical PPR domain (PPR2) from the

CCM1 open reading frame (ORF). The mutated CCM1 ORF (delta-

PPR2) did not complement the phenotype of deltaccm1 mutants which, therefore, were unable to grow on medium with nonfermentable substrate. Reverse-transcription quantitative PCR studies reveled that cells harboring the delta-PPR2 vector which expressed the mutated protein, contained ten times less 15S rRNA than their wild-type counterparts (P =0.005; n = 5), but expressed at least six times more its corresponding CCM1 mRNA (P = 0.003; n = 5).

Immunoblot studies on enriched mitochondrial fractions isolated from cells that harbor delta-PPR2 and wild-type ORFs showed that this deletion did not affect the Ccm1p import process into mitochondria and thus validated PPR2 as a crucial domain that might participate in


15S rRNA stabilization per se. These results suggest that Ccm1p may act as a RNA chaperone via PPR-mediated protein-RNA interaction most probably in early stages of mitoribosome assembly.


P26 Finding The Molecular Partners Of Ccm1p, A Saccharomyces

Cerevisiae PPR Protein Involved In Mitoribosome Biogenesis

Jasmine L. Moore, Jacquais D. D. Dukes, J. Ignacio Moreno, Marta A.

Piva


State, MS

A new paradigm has recently emerged regarding the direct relationship between breast cancer aggressiveness and mitochondrial biogenesis. Notably, transcripts encoding 39 mitochondrial ribosomal proteins (MRPs) are specifically up-regulated in human epithelial breast cancer cells. Among these biomarkers, two MRP of the small subunit (Mrps15p and Mrps7p) are of particular interest, because they are primary binders which associate directly and stably with 12S rRNA, the RNA component of the subunit. Likewise, Mrps28p and

Rsm7p, the S. cerevisiae counterparts of Mrps15p and Mrps7p, act along with Mrps17p, early in mitoribosome assembly, by facilitating the incorporation of other proteins. Data from our laboratory have recently shown that one of the PPR domains of Ccm1p (PPR2) is essential to accumulate 15S rRNA, the S. cerevisiae equivalent of 12S rRNA. Since Ccmp1 lacking PPR2 (DeltaPPR2) is imported into mitochondria, the accumulation defect could be ascribed to this protein inability to interact with partners. The present study has two objectives: (1) Assessment of CCM1 , MRSP28 , RSM7 , MRSP17 , and

15S R_RNA gene expression in conditions of mitoribosome biogenesis. (2) Identification of molecular partners that physically interact with Ccm1p, but not with DeltaPPR2. RT-qPCR experiments showed that protein-encoding gene mRNAs and 15S rRNA levels increased by 50% and 500%, respectively, after switching the cultures from dextrose (fermentation) to glycerol (aerobic respiration). For the second objective, yeast mutant cells lacking the

CCM1 gene (delta ccm1 ) were transformed with single copy plasmids to express either Ccm1p-ZZ or DeltaPPR2-ZZ. The ZZ tag binds to IgG, which allows the affinity purification of the proteins along with any molecule(s) associated to them. Our preliminary results show that 15S rRNA specifically co-purifies with Ccm1p-ZZ indicating a direct protein-RNA interaction in vivo . Surprisingly, DeltaPPR2


binds 15S rRNA in vitro , even though cells expressing the mutant protein virtually do not accumulate this RNA. Therefore, PPR2 appears to participate only in the accumulation phase of 15S rRNA, probably mediating the interaction between Ccm1p and the primary binding ribosomal proteins.


P27 Elucidating The Binding Nature Of Ccm1p To Mitochondrial 15S

Rrna, Required For The RNA Stabilization During Mitoribosome

Assembly

Jacquais D.D. Dukes, Jasmine L. Moore, Marta A. Piva, J. Ignacio

Moreno


State, MS

Mitochondria biogenesis is significantly increased in breast cancer cells. It is correlated with an enhancement in mitochondrial ribosome

(mitoribosome) assembly rate. The regulation of this later process is still unclear. Thus, the elucidation of the mitoribosome biosynthesis pathway and how the cell controls it would open new possibilities for designing new drugs that differentially target such deregulation in tumor cells.

Saccharomyces cerevisiae was and is an excellent model to study molecular aspects of mitochondria proliferation. Reports from our laboratory demonstrated the key role of Ccm1p in maintaining the steady state of the rRNA in the minor subunit of the mitoribosome in

S. cerevisiae (15S rRNA). In humans, PTCD3 binds the rRNA of the minor mitoribosomal subunit (12S rRNA) before it is incorporated into the subunit. Interestingly, both proteins contain pentatricopeptide repeats motifs (PPR). These facts make PTCD3 a potential human counterpart of Ccm1p. Nevertheless, it is not clear the role of both PTCD3 and Ccm1p factors. The present work aims to elucidate how Ccm1p participates with 15S rRNA in the mitoribosome genesis scenario.

We created a S. cerevisiae system in which the supply of Ccm1p can be down and up regulated to cause transient and reversible mitochondrial dysfunction. The levels of Ccm1p and 15S rRNA were correlated as well as the mitochondrial functionality (growth in glycerol).

Specifically, when Ccm1p levels were up-regulated (after being silenced), 15S rRNA resumed its accumulation reaching mitochondrial function recovery status. We postulate that during this mitochondrial recuperation, Ccm1p would be saturated in the

Ccm1p-15S rRNA interaction. Currently we are investigating the

Ccm1p-15S rRNA molar ratio during this time frame. The final data


will contribute to postulate a model of how Ccm1p participate in the ribosomal minor subunit genesis in vivo .


P28 Sudden Mitochondria Deprivation Of Ccm1p: An Approach To

Elucidate The Synthesis Pathway Of The Mitochondrial

Translational Machinery In Saccharomyces Cerevisiae

Cherrelle T. S. Gee, K. Allyne Echols, J. Ignacio Moreno, Marta A. Piva


State, MS

The biogenesis of mitochondrial ribosomes has been recently associated with malignant transformation. However, in spite of its importance, the temporal sequence of events that leads to the assembly of mitoribosomes is still unknown. We have reported the function of CCM1 gene product as a crucial stabilizing factor of 15S rRNA in intron-containing mitochondria. Our preliminary experiments strongly suggested that about 90 % of the 15S rRNA produced inside the mitochondria is in transit, with Ccm1p acting as a stabilizer through a putative transient protein-RNA interaction. To study Ccm1p role, a sudden mitochondrial deprivation of Ccm1p (SMDC) was approached by (a) fusing two SPA domains (ZZ) at the C-termini of Ccmp1, thus causing an “in route” import delay, (b) down-regulating the fusion protein expression

(controlled by GAL1 promoter) by switching from galactose to glucose, and (c) expressing Ccm1p-ZZ in a low-copy vector. SMDC caused loss of 15S rRNA with the concomitant phenotype of inability to utilize non-fermentable substrates, a direct outcome of mitochondrial dysfunction. Western, Northern, and Southern blot analyses showed that after 48 hours of SMDC, Ccm1p was undetectable and 15S rRNA levels were significantly reduced in the mutant yeast cells (P = 0.004; n =3).

However, 21S rRNA and mtDNA levels were similar to those of the wild-type strain, suggesting that mitoribosome biogenesis could resume if Ccm1p levels were restored. Consequently, when Ccm1p expression was switched “on”, the ability to grow on non-fermentable substrates was recovered and 15S rRNA levels significantly increased (P = 0.006; n =3) to pre-SMDC levels. Therefore, we conclude that at “mid-log” phase of recovery, transient Ccm1p complexes support competently folded 15S rRNA to which early ribosomal binding proteins are expected to associate. The transient status is ideal to isolate intermediates whose identification is essential to elucidate the temporal pathway of mitoribosome assembly.


P29 Chemoenzymatic Synthesis Of Chiral 4-Aminoalcohols

Amy Wilcosky 1 , Benjamin Lewing 1 , Erin Norcross 2 , Dale A. Rosado 1


Clinton, MS


Clinton, MS

Ethambutol is an anti-bacterial / bacteriostatic drug that is composed of two 2-alkyl-2-aminoalcohols connected by an ethylene linker. Our group is interested in synthesizing and evaluating the antimycobacterial properties of ethambutol analogues containing 2-Alkyl-

4-aminoalcohols. Synthetic methodology that will allow for generation of both enantiomers of 2-alkyl-4-aminoalcohols is presented herein. Reduction of diethyl 2-benzylmalonate with LiAlH

4 yields a 2-benzyl-1,3-propanediol in good yields. To introduce the needed chirality, Burkholderia cepacia lipase was used to selectively acylate 2-benzyl-1,3-propanediol. The resulting acyl alcohol was used as a chiral intermediate from which both enantiomers of a 2-benzyl-

4-amino-1-butanol can be synthesized. Synthesis of the ( R) enantiomer is accomplished through oxidation of the alcohol moiety to an aldehyde, a Wittig reaction, hydrogenolysis and a Curtius reaction. Synthesis of the ( S) -enantiomer requires modification of the protecting group strategy. The free alcohol is first protected as a

TBDMS ether and the acyl protecting group is removed by treatment with K

2

CO

3

. The alcohol can then be subjected to the same oxidation/Wittig/Curtius sequence as those used to make the Renantiomer. The synthetic route outlined above produces good yields in each step and results in synthesis of both enantiomers of 2-benzyl-

4-amino-1-butanol from a common acyl alcohol intermediate.


P30 Betulinic Acid Does Not Affect Rates Of In Vivo Spontaneous

Formation Of The [URE3] Prion In Saccharomyces Cerevisiae

Samantha Humphrey, Ross Whitwam

Department of Sciences and Mathematics, Mississippi University for

Women, Columbus, MS

Prions are misfolded cellular proteins that are infectious because they can induce other copies of the same protein to misfold and aggregate into amyloids. In mammals, prions are associated with neurodegenerative diseases. In humans, misfolded, amyloid-forming (but non-infectious) proteins are associated with diseases like Alzheimer’s disease and

Parkinson’s disease. Planchard et al. (2012. ACS Chem Neurosci 3: 900-

908) showed that betulinic acid accelerates the in vitro amyloid formation from amyloidβ peptides. In mammalian prion diseases, it is believed that the amyloid less harmful than oligomers of the prion proteins. Anything that accelerates amyloid formation may be protective. We wanted to investigate whether betulinic acid accelerates amyloid formation from prion proteins in vivo. Our hypothesis was that exposure of cells of

Saccharomyces cerevisiae to betulinic acid would accelerate the spontaneous in vivo formation of the [URE3] prion by accelerating amyloid formation of misfolded [URE3] prions. However, incubation of prion-free yeast cells with 100 µ M betulinic acid did not accelerate rates of spontaneous in vivo prion formation. We then hypothesized that the betulinic acid might not be entering the cell, so we exposed prion-free yeast cells to 100

µ M betulinic acid in the presence of 0.2% saponin, a detergent known to increase cell permeability. Even in the presence of saponin, betulinic acid did not alter the rate of spontaneous in vivo prion formation in the yeast.

This suggests that either betulinic acid does not affect the rates of in vivo

[URE3] amyloid formation in vivo at the concentrations used.


P31 Subcellular Studies Of A Novel Engineered Protein Required To

Assess Non-Specific Molecular Interactions In Intronless

Mitochondria Of Saccharomyces Cerevisiae

Ineshia Coleman 1 , Marta A. Piva 2 , J. Ignacio Moreno 2

1 Division of Natural Science, Tougaloo College, Tougaloo, MS

2 Department of Biological Science, , Alcorn State University, Alcorn State,

MS

The yeast Saccharomyces cerevisiae is an excellent model to study mitochondrial biogenesis due its capacity to live without this organelle. the yeast can obtain energy via fermentation. Our target mitochondrial protein is CCM1 (Ccm1p). The first function of

Ccm1p was elucidated in our laboratory, as a vital splicing enabler factor. A second function as stabilizer of 15S ribosome RNA (15S rRNA) was also described. We also defined both activities as independent from one another. Several bioinformatics programs revealed the presence of at least two “pentatricopeptide repeats” (PPR) in the Ccm1p sequence. The predicted second PPR motif (PPR2) was proved to be essential for both functions. Therefore, a novel genetic construct consisting of a deleted PPR2 and a fusion of ZZ affinity tag at the 3’ end (Ccm1pD2ZZ protein) should be an excellent molecular tool to evaluate non-specific molecular interactions inside mitochondria. Phenotypic studies of Dccm1 segregants expressing

Ccm1pD2ZZ consisted in cell morphology assessment by light microscopy and complementation (whether Ccm1pD2ZZ enables the cell to utilize non-fermentable substrates). Molecular studies used

RT-qPCR to determine Ccm1pD2ZZ capability to maintain 15S rRNA steady state. While morphology of D ccm1 segregants expressing

Ccm1pD2ZZ of cells displayed no changes, they failed to grow with non-fermentable substrates as source of carbon and energy. Even though Ccm1pD2ZZ was imported into mitochondria, it was not able to maintain 15S rRNA steady state levels. These results demonstrated that Ccm1pD2ZZ is an excellent tool to evaluate nonfunctional molecular interactions in the characterization of Ccm1p role during mitoribosome biogenesis.


P32 Biosynthesis Of Phenazine Metabolites In Burkholderia spp.

Olga V. Mavrodi, Dmitri V. Mavrodi



Phenazines are bacterial secondary metabolites that contribute to the ecological fitness and pathogenicity of the producing strains. Our understanding of phenazine biology is largely based on the pyocyaninproducing opportunistic pathogen Pseudomonas aeruginosa . Similar knowledge for other phenazine-producing bacterial species is severely lagging. Here, we focus on phenazines synthesized by Burkholderia, a ubiquitous opportunistic bacterial pathogen that can infect plants, animals and humans. We screened a collection of Burkholderia strains of various geographic, environmental and clinical origins and demonstrated that most phenazine-producing (Phz + ) strains belonged to the B. cepacia complex, B. lata and B. glumae . We further revealed that in B. lata and B. glumae phz genes form compact operons encoding putative phenazine core biosynthesis and modifying enzymes and reside on unique IS composite transposons. Our preliminary functional analysis demonstrated that B. lata produces several different phenazines including 4,9-dihydroxyphenazine-1,6dicarboxylic acid dimethylester. Our current work is focused on the molecular signaling role that phenazines play in Burkholderia biofilms, and in mixed biofilm communities formed by Burkholderia and P. aeruginosa . At the conclusion of these experiments, we expect to unravel the organization of phz pathways in Burkholderia and prioritize gene targets for the subsequent mutational inactivation and testing in different models of infection. In the long run, the expected results will help to better understand the role of phenazine compounds in an important group of bacterial pathogens, and will aid in the development of strategies to inhibit the production of phenazines and to neutralize their toxicity.


P33 Comprehensive View Of Water Quality Of Ross Barnett

Reservoir

LaTia Peavy 1 , Padmanava Dash 2 , Julius O. Ikenga 3 , James L. Pinckney 4


2 Department of Geosciences, Mississippi State University, Mississippi

State, MS

3 Department of Natural Sciences and Environmental Health,

Mississippi Valley State University, Itta Bena, MS


University of South Carolina, Columbia, SC

Ross Barnett Reservoir, a 32.31 square miles (83.67 km ² ) freshwater body, serves as the source of drinking water for the greater Jackson area and a major recreational destination in the state of Mississippi.

The main objective of this research was to take a comprehensive view at the water quality of the Ross Barnett Reservoir. Two sampling trips were undertaken, on May 5 and July 10, 2013, to Ross Barnett

Reservoir to collect water samples from twelve sites. In addition, a

Hanna instrument was used to measure temperature, salinity, dissolved oxygen, and pH at each site. After completion of the trips, the water samples were vacuum filtered and the filter papers were processed for high performance liquid chromatography (HPLC) photopigments, cyanobacteria specific pigment phycocyanin (PC), suspended particulate matter (SPM), phycotoxins, bacterial counts, toxic metals, and microscopy analyses. The SPM analyses revealed that the organic SPM concentrations were higher than the inorganic

SPM concentrations. The relative abundances of major algal groups suggested that the cyanobacterial concentrations were higher during the second in compared to the first sampling trip. The cyanotoxin, hepatotoxic microcystin, was found at all sites on both the dates.

Bacterial counts revealed high concentration of total coliforms and heterotrophic bacteria at all sites and presence of E. Coli and

Enterococii bacteria at few sites. Several toxic metals were also found.

The finding of cyanobacteria, microcystin, toxic metals and E. Coli and Enterococii bacteria calls for continuous monitoring of the Ross

Barnett Reservoir.


P34 Human Disease Micrornas And Their Counterparts In Domestic

Animals

Teresia Buza 1, 2 , Tony Arick 2 , Hui Wang 2 , Daniel G. Peterson 2

1 Department of Basic Sciences, College of Veterinary Medicine,

Mississippi State University, Mississippi State, MS

2 Institute for Genomics, Biocomputing & Biotechnology, Mississippi

State University, Mississippi State, MS

For over a decade there has been strong evidence that miRNAs serve as diagnostic biomarkers of various human diseases including cancer and cardiovascular, immunological and gastrointestinal diseases. The role of miRNAs in diseases in domestic animals has not been studied in any depth. However, it is probable that many of the animal homologs of human disease-associated miRNAs may be involved in domestic animal diseases. We have developed a highly curated database of human disease-associated miRNAs and their domestic animal homologs which we have deemed “VetBioBase-miRNAs.” In brief, we utilized published human disease miRNAs to computationally identify homologous miRNAs in cow, chicken, pig, horse, and dog. We identified 287 human disease-associated miRNAs which had at least one 100% identical animal homolog. The 287 miRNAs were associated with 359 human diseases published in 2,863

PubMed articles. Phylogenetic analysis of miRNA precursors indicated that 60% of currently published horse miRNAs are homologous to human disease miRNAs. Not surprisingly, of the animals evaluated, chicken miRNAs had the least similarity with human disease miRNAs (5%). In summary, all data collectively provided 37,651 non-redundant lines of associations. This data is a resource that the animal/veterinary research community can leverage to study miRNA-related diseases in animals while informing research efforts aimed at identifying novel disease models. Data is deposited in http://vetbiobase.igbb.msstate.edu which will be publicly available by

April 2014.


P35 Molecular And Phenotypic Characterization Of Methicillin

Resistant Staphylococcus Aureus Isolates Causing Bacteremia In

A Major Hospital In South Mississippi

Dhritiman Samanta 1 , Justin Batte 1 , Luis Marcos 2 , Mohamed O. Elasri 1



2 Forrest General Hospital, Hattiesburg, MS

Staphylococcus aureus is the predominant cause of bacteremia around the world. The situation has been worsened by the acquisition of antibiotic resistance. The health hazard burden caused by this species is severely exacerbated by worldwide dissemination of clones resistant to beta-lactam antibiotics (Methicillin Resistant Staphylococcus aureus ;

MRSA) in hospitals and communities. Methicillin resistance is typically conferred by a mecA gene located on a large chromosomal element, SCCmec . The distribution of MRSA clones is dynamic and tends to be geographically unique. The purpose of this study is to determine the molecular characteristics and antibiotic resistance properties of the MRSA isolates causing bacteremia in a major hospital in south Mississippi, USA. We collected 30 MRSA isolates from blood infections in an 8-month time period from May through

December in 2013. The isolates were subjected to antibiogram analysis and SCC mec typing by multiplex PCR. We also grouped these isolates by Pulsed Field Gel Electrophoresis and Multi-Locus

Sequence Typing (MLST) where seven housekeeping genes from each isolate were sequenced and analyzed. The analysis showed all these isolates were mecA positive and 33% of them were SCCmec type

II of which 50 % were PVL negative indicating hospital-associated infections. The other 66% of the isolates were SCCmec type IV indicating community-associated infections. As predicted, the vast majority (97%) of the isolates were resistant to oxacillin and erythromycin and 42% were resistant to clindamycin. All isolates were resistant to Augmentin and sensitive to bactrim, linezolid and vancomycin. Diverse genetic backgrounds in terms of SCCmec types of MRSA were identified as causing bacteremia in Mississippi, USA.

Clinical MRSA isolates were shown to be resistant to multiple drugs.

However, no Vancomycin intermediate or resistant isolates were


identified. This study allows us to associate specific clonal types with bacteremia isolates and detect outbreak by any particular type in south

Mississippi.


P36 Alternate Drug Designing Strategy For Breast And Prostate

Cancers Targeting Protein Dimerization Interface

Pradip K Biswas 1 , Rajendram Rajnarayanan 2 , Sandipan Chakraborty 1 ,

Nicholas Rader 1

1 Laboratory of Computational Biophysics & Bioengineering,

Department of Physics, Tougaloo College, Tougaloo MS

2 Department of Pharmacology and Toxicology,

University of Buffalo, Buffalo, NY

In tamoxifen-resistant Estrogen Receptor alpha (ER α ) positive breast cancers, the reported over-expressions of co-activator calcium binding proteins like calmodulin (CaM), and direct binding of CaM to ER α , indicate possible ligand-independent pathways to gene transcription.

Thus, in our MS-INBRE project, we proposed an alternate drug designing strategy to counter possible ligand-independent pathways to gene transcription by targeting the protein dimerization interface. As

ER α and ER β dimerize only among themselves as homo and hetero dimers, they must possess a unique dimerization recognition surface and provide a unique target to inhibit the progression of ER-positive breast tumors. Elucidating the homo dimerization interface of ER α ligand binding domain (LBD) in molecular details, we have identified three sequence motifs responsible for dimerization – DXXTD (480-484),

LQXXHQXXAQ (497-506), LSXXRHXXNK (511-520) – and used these sequences to develop peptidic and peptidomimetic inhibitors [Mol.

Div. 16 , 441 (2012)]. Two of our designer peptides (yet to publish) have exhibited strong inhibition in in-vitro testing for MCF-7 cell lines.

As Androgen Receptor (AR) mediated transcription for prostate cancer also involve dimerization of the protein, a similar dimer interface based inhibition technique should be quite effective. But, there does not exist any crystal structures for AR-LBD dimer. Following the works of

Shaffer et al [PNAS 14 , 4758 (2004)] that AR does not dimerize like

ER in head-to-tail fashion but in symmetric form like the glucocorticoid receptors, we have developed AR-LBD dimer structure using in-silico techniques. Elucidation of the dimerization recognition sequences to develop peptidic and peptido-mimetic inhibitors for AR is underway.


P37 Influence Of Flavonoids In Reducing Oxidative Stress In Human

Serum Albumin: Spectroscopic And Molecular Modeling

Studies

D’Asia Gholar, Denise Ward, Anthony Saracino, Bidisha Sengupta

Chemistry Department, Tougaloo College, Tougaloo, MS

We have explored the interactions of fisetin, quercetin and malvidin chloride with human serum albumin (HSA), using optical spectroscopy and gel electrophoresis, combined with molecular modeling approaches. Methyl parathion (MP) is a known pesticide and oxidative stress inducing agent. Quercetin and Malvidin Chloride, are ubiquitous bioactive flavonoids, belonging to flavonol and anthocyanidin families and are abundant in onion and bilberries respectively. They are known to possess antioxidant, anticarcinogenic, hypolipidemic, vasoprotective and other important therapeutic properties. Specific interactions of the flavonoids with their carrier protein HSA are confirmed from flavonoid-induced quenching which is evident from steady state fluorescence, change in absorbance as well fluorescence anisotropy. UV/Vis melting and CD studies indicate no significant change in the secondary structure of

HSA upon flavonoid binding indicating the usefulness of this flavonoid in medicinal biology. Both temperature dependent fluorescence measurements and molecular docking investigations reveal that apart from hydrogen bonding and van der Waals interactions, electrostatic interactions also play crucial role in flavonoid-HSA interactions. Our studies indicate that the flavonoids can inhibit the MP induced structural perturbations of HSA to a significant extent. Studies are underway.


P38 Soluble AmyloidΒ Prions Promote The Formation Of Soluble Α -

Synuclein Oligomers

Matthew Planchard, Vijay Rangachari

Department of Chemistry and Biochemistry, The University of Southern


An increasing amount of research focuses on the propensity of amyloidogenic proteins involved in neurodegenerative diseases to act as prions and propagate infective states of misfolding. A number of reports have found interactions between amyloidβ (A β ) and α -synuclein ( α S), proteins that are implicated in Alzheimer’s (AD) and Parkinson’s (PD) diseases, respectively, which suggests a potential interplay between the two pathologies. Both proteins are associated with neuronal death through a toxic gain of function, in which they adopt an altered conformation and aggregate into oligomers and fibrils. Up to 60% of PD patients also have some level of A β pathology, and the converse is true for AD, in which up to 50% of cases also show Lewy Body pathology.

For both diseases, the presence of the complementary pathology leads to a more aggressive disease progression. Our research aims to discover whether toxic A β oligomers generated in our lab (LFAOs), which have been shown to replicate in a prion-like fashion, are capable of crossseeding the formation of potentially toxic α S oligomers, providing a molecular link in the association between AD and PD. Western blot visualization of coincubations of α S monomer with LFAO seeds showed the formation of α S oligomers with a similar molecular weight range to

LFAOs within 72 h, which were not observed in control incubations of

α S without LFAOs. Sedimentation assays and ThT showed these oligomeric species to be soluble and nonfibrillar. We are investigating this cross-seeding phenomenon through several additional biophysical methods. In parallel, α S oligomers are being generated to seed A β oligomer formation in a reverse cross-seeding experiment. Our observation of cross-seeding between these two amyloidogenic proteins provides a molecular basis for the interaction between AD and PD while providing evidence for a prion-like mechanism in which amyloidogenic proteins contagiously propagate their pathologically misfolded states throughout the brain.


P39 Heme Oxygenase Regulation Of Trophoblast Enac Mediated

Migration

Kayla D. Coleman, Junie P. Warrington, David E. Stec, Joey P. Granger,

Heather A. Drummond

Department of Physiology & Biophysics, University of Mississippi

Medical Center, Jackson, MS

During normal pregnancy, trophoblasts from the fetal placenta migrate into the maternal spiral arteries of the uterus to replace vascular smooth muscle cells. This remodeling of the spiral arteries accommodates increased fetal blood flow. In patients with preeclampsia, spiral artery remodelling is impaired due to impaired trophoblast migration. Recent studies suggest altered Epithelial

Sodium Channel (ENaC) expression may be a contributing mechanism for impaired trophoblast migration. A recent study suggests that heme oxygenase-1 (HO-1) protects against hypertension in a rat model of placental ischemia; however, whether HO-1 regulation of ENaC contributes to the beneficial effects of HO-1 is unknown. The purpose of this study was to determine if HO-1 enhances ENaC mediated cytotrophoblast migration. We found that silencing of β ENaC and gENaC in cultured cytotrophobasts (BeWo cells), by expression of dominant-negative constructs, reduced migration to 56 ± 13 and 26 ± 3 %, respectively (p<0.05) of control.

We then showed that HO-1 induction, using cobalt protoporphyrin, stimulates cytotrophoblast β ENaC expression by 1.5 and 1.8 fold (10 and 50 µM) and migration (43 ± 5% of control, p<0.05). Importantly, the enhanced migratory response to CoPP was entirely blocked by

ENaC inhibition with amiloride (10 µM). Taken together, our results suggest that b and gENaC mediates cytotrophoblast migration and increasing bENaC expression by HO-1 induction enhances migration. These findings suggest that HO-1 regulation of cytotrophoblast bENaC and gENaC expression and migration may be a potential therapeutic target in preeclamptic patients.


P40 Promoter Analysis Of M46 , A Mold Specific Gene, Of The

Dimorphic Pathogenic Fungus Histoplasma Capsulatum

Shavonda McDaniel, Davida Crossley

Department of Biological Sciences, Alcorn State University, Lorman, MS

Histoplasma capsulatum, Hc , is a dimorphic fungus that is primarily found in the environment in soils of contaminated birds and bat excrements as a mold. Once, the soil is disturbed, spores are released, and are inhaled into the lungs, where the fungus converts to yeast.

The yeast is the cause for the respiratory infection histoplasmosis. The mold to yeast conversion is a requirement for pathogenesis. This study focuses on M46 , a gene that is expressed in the mold phase but is not expressed in the yeast phase. Northen blot analysis of M46 from four major strains of Hc , has shown that M46 is up regulated in strains

G186AS and Downs, but is down regulated in strains G184AS and

G217B. The reason for the down regulation in strains G184AS and

G217B is unknown. To see if the reason for lack of expression of the latter strains is due to a non-functional promoter, a 1.2 kb promoter fragment of M46 from all four Hc strains will be amplified via PCR and will ultimately be placed in the vector pCR346 in frame with the reporter GFP (Green Fluorescent protein). The plasmid will be electroporated into the M46 expressing strain G186AS ura- and the non-expressing strain G184AS ura-. Fluorescence of GFP will be observed by using a Olympus BX51 fluorescence microscope. The fluorescence from GFP will determine if the M46 promoters in strains

G184AS and G217B are functional and if the promoter is the reason for lack of expression in strain G184AS and G217B.


P41 The [URE3] Prion Of Saccharomyces Cerevisiae Is Detrimental

When Prion-Containing Yeast Are In Direct Competition With

Prion-Free Yeast

Ross Whitwam, Jeremy Winders, Samantha McCorkle, Katherine

Brinkman

Department of Sciences and Mathematics, Mississippi University for


Prions are infectious proteins which, in mammals are associated with neurodegenerative diseases and have many features in common with non-infectious diseases such as Alzheimer's disease, Parkinson's disease, Huntington's Disease, and others. Yeast prions such as

[URE3] serve as model systems for mammalian prions because they share many molecule features in common with them. However, unlike mammalian prions, the yeast prions are not associated with any disease states and while the [URE3] prion has been reported to slow yeast growth, the evidence is not strong. We show that, contrary to published reports, pure cultures of [URE3] yeast grow at exactly the same rates as prion-free yeast in both nutrient-rich and nutrientlimiting media. However, when prion-containing [URE3] yeast are grown in direct competition with prion-free yeast in the same culture, the prion-free yeast out-grow the prion-containing [URE3] yeast and eventually come to dominate the culture. This is the first time the

[URE3] prion has been shown to have a deleterious effect on its host and suggests that the [URE3] system may be used to model certain aspects of the disease states of mammalian prions as well as their molecular features.


P42 Preparation And Characterization Of A Synb1-Elp-Ga(Iii)

Binding Polypeptide

Natalia I Grace 1 , Jasmine Jennings 1 , Ming-Shen Wu 1 , Drazen Raucher 2 ,

Jinghe Mao 1 , Manliang Feng 1

1 Natural Science Divison, Tougaloo College, Tougaloo, MS

2 Department of Biochemistry, University of Mississippi Medical Center,

Jackson, MS

In this work, an oligonucleotide that encode a Gd(III) binding polypeptide was introduced in a pET25 vector that contains sequences of cell penetration peptides (Synb1) and elastin -like polypeptide

(ELP). The cell penetration sequence encodes a polypeptide with an amino acid sequence of RGGRLSYSRRRFSTSTGRA. The elastinlike polypeptide (ELP) consists of repeated sequence of pentapeptides

(VPGXG), where the guest residue X can be any amino acid except

Proline. The unique inverse temperature-phase transition feature of

ELP has been employed in the designed polypeptide for thermallytargeted drug delivery. The purpose of this research is to engineer a protein that can be thermally guided to target location and imaged invivo. The designed polypeptide can be used in many areas such as drug-design, clinical diagnosis and pharmaco-kinetic studies.

Towards this goal a polypeptide contains amino acids sequence for binding rare earth metal ions such as Gd(III) ion was introduced into a snb1-ELP polypeptide. The paramagnetic property of Gd ion will be used for potential in-vivo MR imaging. The target protein was expressed in E. coli . The metal binding properties and phase transition properties of this newly designed polypeptide were characterized using

UV-Vis and fluorescence spectroscopy. A preliminary MRI scan shows that the designed protein has potential as a contrast enhancing agent for MRI.


P43 Lack Of Innate Immunity In Mouse Embryonic Stem Cells

(Mescs) And Mesc-Derived Fibroblasts

Rachel Payne, Jundi Wang, Yan-Lin Guo



Embryonic stem cells (ESCs) are a promising cell source for regenerative medicine. However, recent studies indicate that ESCderived endothelial and smooth muscle cells do not have active innate immunity against various pathogens. When used in the patient, ESCderived cells would be placed in a wound site that is exposed to various pathogens and inflammatory cytokines; therefore, their viability and functionality would be significantly compromised if the cells do not have competent immunity. Our recent study demonstrated that mESCs are intrinsically deficient in expressing type I interferons

(IFN) and inflammatory cytokines ( J. Biol. Chem.

2013, 288:15926 -

15936) .

This finding explains the lack of innate immunity in ESCdifferentiated endothelial cells and smooth muscle cells, and it also indicates that the current existing in vitro differentiation methods could not promote innate immunity development. To test if the lack of innate immunity is specific to ESC-derived endothelial cells and smooth muscle cells or is common to other types of ESC-derived cells, we used mESC-differentiated fibroblasts (ESC-FBs) as a model system because FBs are a major cell type that expresses type I IFN and cytokines and modulates immune and inflammatory responses in different tissues .

We generated mESC-FBs by three differentiation methods: 1) direct differentiation in a monolayer, 2) differentiation through embryoid body formation, and 3) retinoic acid–induced differentiation. mESC-FBs generated by all three methods have typical morphology and express markers similar to naturally differentiated FBs, but they showed no or limited responses to several major immunostimulants, including inflammatory cytokines (TNFa and IL-1b), a bacterial endotoxin (lipopolysaccharides), polyIC (a viral RNA analog), or live viral infections, whereas FBs isolated from embryonic tissues showed robust responses to all agents tested.

Further analyses revealed that the two key signaling pathways that mediate immune responses, the NF-kB and the IRF pathways, are


not active in mESC-FBs. We conclude that the innate immunity is not developed in ESCs and cannot be induced by the methods commonly used for in vitro differentiation of ESCs.


P44 Antiviral Responses In Mouse Embryonic Stem Cells:

Imbalanced Development Of Cellular Mechanisms In Type I

Interferon Production And Response

Ruoxing Wang 1 , Jundi Wang 1 , Dhiraj Acharya 1 , Amber M. Paul 1 , Fengwei

Bai 1 , Faqing Huang 2 , Yan-Lin Guo 1



2 Department of Chemistry and Biochemistry, The University of

Southern Mississippi, Hattiesburg, MS

Innate immunity has been extensively studied and is presumably acquired by most types of somatic cells , but little is known about embryonic stem cells (ESCs). Recent studies suggest that human

ESCs (hESCs) and hESC-derived cells do not respond to various infectious agents, including bacterial endotoxins and viral RNA analogs.

Mouse ESCs (mESCs) and mESC-derived cells similarly do not show inflammatory responses to cytokines, lipopolysaccharides, and even live bacteria . These findings suggest that ESCs, normally residing in the sterile womb environment, may not have functional innate immunity. In line with this hypothesis, we have recently reported that mESCs are susceptible to cytopathic effects of viral infection and synthetic RNA viral analogs , but they do not have functional antiviral mechanisms as a critical part of innate immunity, in particular, they are deficient in expressing type I interferons ( IFN)

( J. Biol. Chem.

2013, 288:15926 15936) .

In this study, we further investigated whether or n ot mESCs can respond to type I IFN, our results revealed that pretreatment mESCs with either IFNb or IFNw , two members of type I IFN, can protect the cells from La Crosse virus-induced lytic cell death and repress viral replication. Both IFNb and IFNw induced IFN stimulated genes (ISGs) in mESCs, the hallmark of cellular responses to type I IFN. Although IFNb- and

IFNw-induced cellular responses in mESCs were weaker than in differentiated fibroblasts, the patterns were similar in the two cell types. Additional data showed that IFNb and IFNw have no effects on the stem cell state as determined by the analysis of pluripotency marker expression pattern and cell cycle profile of mESCs. We conclude that the cellular mechanisms for production of and response


to type I IFN are not equally developed in mESCs; they are deficient in type I interferon expression, but have functional mechanisms to respond and mediate the antiviral effects of type I IFN. Our studies revealed a unique and previously uncharacterized property of mESCs that is important for understanding innate immunity development and

ESC physiolo


P45 Enhancing The Cell Biology Curriculum

Melissa A. Adams

Copiah-Lincoln Community College, Natchez, Mississippi

The goal of this project is to enhance the Cell Biology curriculum by adding a “hands on” component to the class and the lab. Specifically, by enhancing the academic training and experience for students who are underserved. I believe creating this Cell Biology curriculum would generate interest for those students and could possibly lead to a much larger minority population choosing a career in the sciences. Many community college students transfer to four year institutions, only to return after a semester or two because the transition from community college to senior college is overwhelming. This proposed Cell Biology course would help bridge the gap between underserved students at the community college level and the sciences at four year institutions, better preparing them for the advanced classes they will have to complete to obtain a degree in the STEM curriculum. I plan to develop a course that generates interest in the biomedical sciences, and to build the lab component so that it will be capable of running the type of experiments that would typically be used in scientific research. In this context, students would learn more and also see the importance of understanding the material given in the textbook.

Further understanding of the material, as well as structuring the course to applied methods has the potential to stimulate student’s interests in the sciences, possibly prompting them to consider a career in STEM.


P46 Inhibition Of Proteasome And Lysosomal Enzymes Can

Partially Rescue CFTR Degradation

Deanna Watson, Ghanshyam D. Heda

Department of Sciences & Mathematics, Mississippi University for


DF508 is the most common mutation affecting the function of

Cystic Fibrosis Transmembrane-conductance Regulator (CFTR) protein, and is responsible for the genetic disease cystic fibrosis.

We hypothesize that the surface instability of DF508 CFTR [Heda et al, Am J Physiol, 280, C166-C174, 2001] is may be due to its selective sorting to cellular sites different than that of wild type

CFTR. In this study we aim to determine the role of proteasomes and lysosomes in CFTR degradation. Human bronchial epithelial cells (CFBE) and pig kidney epithelial cells (LLC-PK

1

) stably transfected with DF508 or wild type CFTR were pre-treated with 5 mM sodium butyrate at 27 o C for 60 h to up-regulate the plasma membrane CFTR expression. Cells were then “chased” at 37 o C in the presence of protein synthesis inhibitor (cycloheximide) and/or inhibitors of proteasomes (MG132), or lysosomal enzymes (E64).

Cell lysates were directly immunoblotted with anti-CFTR antibody or immunoprecipitated first with anti-CFTR antibody followed by immunoblotting with anti-ubiquitin antibody. MG132 and E64 partially rescued the degradation of plasma membrane DF508

CFTR in LLC-PK

1

and CFBE cell lines. Immunoprecipitated

DF508 CFTR in LLC-PK

1

cells also appeared to be more ubiquitinylated than its wide type counterpart. Partial rescue of

CFTR degradation by MG132 and increased levels of CFTR ubiquitination suggest the role of ubiquitin dependent proteolytic system in CFTR degradation. Additionally, rescue of CFTR from degradation by E64 suggests that a portion of CFTR is degraded by lysosomal enzymes.


P47 mRNA Directed Differentiation Of Mouse Embryonic Stem Cells

Based On Their Underdeveloped Antiviral Mechanisms

Yan-Lin Guo 1 , Ruoxing Wang 1 , Chengwen Teng 1 , Joseph Spangler 1 ,

Jundi Wang 1 , Faqing Huang 2





The landmark discovery of induced pluripotent stem cells (iPSCs) has led to the new concept of cell reprogramming, but the fact that viral vectors are commonly used for effective expression of reprogramming factors prevents the therapeutic use of the reprogrammed cells.

Extensive effort has led to the development of several alternatives, among which mRNA-mediated gene expression has shown great promise .

This method directly introduces synthetic mRNA into the host cell to express reprogramming factors, therefore bypassing the need of viral/DNA vectors. The successful generation of RiPSCs

(RNA-induced iPSCs) from fibroblasts has led to the belief that this strategy is the beginning of a new era of cell reprogramming. In principle, this strategy could be expanded to reprogram any type of cells as long as the genes that control the cell fate are identified.

However, a major biological challenge is that synthetic mRNA is detected as a viral RNA analog by the host cell, resulting in a series of adverse effects associated with antiviral responses. As a result, a synthetic mRNA has to be extensively modified to reduce its immunogenicity. We have recently reported that mouse embryonic stem cells (mESCs) are deficient in expressing type I interferons

( IFN) ( J. Biol. Chem.

2013, 288:15926 15936). In this study, we further demonstrated that single-stranded RNA (ssRNA) and protein-encoding mRNA can induce strong IFN expression and cytotoxicity in differentiated cells, but none of these antiviral responses are observed in mESCs. We conclude that mESCs are intrinsically deficient in antiviral responses. While this novel feature of

ESCs is in itself important for understanding innate immunity development, it makes mESCs an ideal model for developing mRNAmediated gene expression strategies. We hereby demonstrate that


ESCs can tolerate repeated transfection and can express proteins from their respective mRNA with expected biological functions, as illustrated by the expression of green fluorescent protein and by the expression of transcription factor Etv2 which promotes vascular cell differentiation. ESCs represent a promising cell source for regenerative medicine, but the lack of effective differentiation methods to obtain specific types of cells with clinical quality is a major challenge. The mRNA-directed expression of transcription factors that control specific cell fate can provide a strong internal driving force for cell specific differentiation, thereby fundamentally improving the existing ESC differentiation methods.


P48 Endogenous CFTR Expression In Human Pancreatic Duct Cells

Ariel Finch, Nadeema Appukutti, Deanna Watson, Ghanshyam D.

Heda

Department of Sciences & Mathematics, Mississippi University for


CFTR is a membrane protein present on the surface of many epithelial cells and function as a chloride channel. A defective

CFTR is responsible for causing genetic disease cystic fibrosis (CF).

CFTR is expressed in trivial amounts and its endogenous expression is difficult to detect by biochemical techniques such as immunoblotting. In this study we aimed to improve the sensitivity of CFTR-immunoblotting by altering the concentration of SDS and methanol in transfer buffer. SDS facilitates the removal of proteins from polyacrylamide gels (PAGE) and methanol allows their binding to nitrocellulose (NC) membrane. Endogenous

(CFPAC, Capan-1) and exogenous (LLC-PK

1

and CFBE) CFTR expressing cell lines were treated with 5 mM sodium butyrate at

27 o C for 60 hrs to up-regulate the surface CFTR expression. Cell lysates were prepared, electrophoresed and transferred to NC membrane with various concentrations of SDS and methanol, followed by immunoblotting with anti-CFTR antibody. The addition of SDS at 0.015% in transfer buffer increased the detection levels of CFTR expression by maximally transferring the proteins from PAGE to NC membrane. Concentration of methanol (5 to

20%), however, had little or no influence on the sensitivity of

CFTR immunoblots. This improvement in immunoblotting, however, was not sufficient to detect the endogenous CFTR expression in CFPAC and Capan-1 cell lines. Endogenous CFTR expression in these cell lines therefore was measured by immunoprecipitation with anti-CFTR antibody. SDS increased the sensitivity of CFTR immunoblotting, however, the endogenous

CFTR expression can only be detected by immunoprecipitation followed by immunoblotting.


P49 Role Of msaABCR Operon In The Regulation Of Proteases And

Biofilm Formation

Gyan S. Sahukhal, Mohamed O. Elasri



Community-acquired, methicillin-resistant Staphylococcus aureus (CA-

MRSA) strains are causing severe infections among healthy individuals with no predisposing risk factors. Regulation of virulence factors in these prevalent strains is not yet understood. Previously, we identified the msa locus as a regulator of virulence, biofilm development, and antibiotic resistance. In this study, we show that the msa locus is a four-gene operon and further characterize its mode of regulation.

In order to characterize the msa locus, we deleted each individual gene in the operon by allelic gene replacement method. We also performed

5’ & 3’ Rapid Amplification of cDNA ends (RACE) analysis and mutation analysis on the operon. We examined the phenotypes

(Protease production, Pigmentation, and Biofilm formation) for all mutants.

We defined the new msaABCR operon which includes and noncoding RNA gene ( msaC ) and a regulatory anti-sense ( msaR ) gene.

The msaR transcript is complementary to the 5’ end of the msaB gene and is expressed in a growth-phase dependent manner suggesting that it might involve in regulation of the expression of the operon. We also show that msaA gene acts as a negative regulator of the msaB genes.

These findings allow us to define the mechanism of regulation of virulence by the msaABCR operon and to investigate the stimulatory signals that the msaABCR operon responds to during pathogenesis.


P50 Role Of msa In Regulation Of Cell Morphology And Tellurite

Resistance In Staphylococcus aureus

Ahmed Alzuway, Amelsaad El barasi, Bina L Jayana, Gyan S. Sahukhal,

Mohamed O. Elasri



Emergence of community-acquired methicillin resistant

Staphylococcus aureus (CA-MRSA) strains is alarming. CA-MRSA strains cause severe infections among healthy individuals without predisposing risk factors outside the hospital setting. Their ability to quickly respond to environmental conditions by regulating virulence factors is a fundamental key behind the success of these strains. We have previously identified the msa operon as a new global regulator of virulence. Deletion of the msa operon in the CA-MRSA strain

USA300_LAC, leads to a reduction of expression (-3.48 fold) in a neighboring divergent operon. String analysis for protein-protein interaction study also showed that the msa operon interacts with this operon. In this study, we investigate the potential functional relationship between this operon and msa. Preliminary bioinformatics analysis showed that SAUSA300_1297-99 codes for three hypothetical proteins that may be involved in cell morphology and tellurite resistance. We have deleted this operon in CA-MRSA

USA300_LAC strain and compared its phenotypes to the msa mutant. This analysis includes biofilm formation, autolysis, pigmentation and extracellular protease production. This study will define the potential role of msa in tellurite resistance.


P51 Roles Of msaA In msaABCR Operon In The Regulation Of

Virulence And Biofilm Formation

Amelsaad El barasi, Ahmed Alzuway, Bina L Jayana, Gyan S. Sahukhal,

Mohamed O. Elasri



Community-acquired, methicillin-resistant Staphylococcus aureus (CA-

MRSA) strains are causing severe infections among healthy individuals with no predisposing risk factors. Yet, we do not fully understand the regulation of virulence factors that allow such strains to be so prevalent. Previously, we identified the msaC gene as a regulator of several virulence genes, biofilm development, and antibiotic resistance.

We performed 5’ & 3’ Rapid Amplification of cDNA ends (RACE) to find the starts and ends of msaABCR operon. We deleted the first gene of msaABCR operon by allelic gene replacement method, and studied the expression of global regulator, sarA . We also studied the phenotypes (Protease production, Pigmentation, and Biofilm formation).

We showed that msaC is part of a newly defined four-gene operon

( msaABCR ), in which the original locus msaC is a non-protein coding

RNA that is essential for the function of the operon. We also showed that msaA gene plays an opposing role to msaC in the regulation of sarA , extracellular protease production and biofilm formation.

Deletion of msaA led to the over-expression of sarA , decrease protease production and increase biofilm formation, whereas, deletion of msaC led to the decrease expression of sarA , increase protease production and decrease biofilm formation.

These findings allow us to define the mechanism of regulation of virulence by the msaABCR operons and to begin to investigate the stimulatory signals that the msaABCR operon responds to during pathogenesis.


P52 Characterization Of Gene msaB Gene In The msaABCR Operon

Of Staphylococcus aureus

Mounir R. Saleh, Bina L. Jayana, Gyan S. Sahukhal, Mohamed O. Elasri



Staphylococcus aureus is an important human pathogen that causes skin and soft-tissue infections, endovascular infections, pneumonia, septic arthritis, endocarditis, osteomyelitis, foreign-body infections, and sepsis. S. aureus produces a vast array of virulence factors that are controlled by global virulence regulators. Previously we identified msa as a new global virulence regulator that controls the expression of sarA and biofilm development. We have also shown that msa is a part of three gene operon which includes SAUSA300_1296,

SAUSA300_1295 and SAUSA300_1294. To define the role of the individual genes in the operon, we deleted the SAUSA300_1295 from

USA300_LAC strains and compared its phenotype with single

SAUSA300_1294 single mutant and the msa operon deletion mutant.

Our preliminary data showed that SAUSA300_1295 single deletion mutants’ phenotype is similar to single msa deletion mutant and msa operon deletion mutant. All three mutants showed decreased pigmentation, increased extracellular protease activity, increased cell death and reduced biofilm formation. SAUSA300_1295 alone does not complement the phenotype which confirms that the genes in the msa operon are co-transcribed and functionally related.

Identification of this operon and expression of the three open reading frames will allow us to define the mechanism of action of the msa operon locus.


P53 Determining The Role Of The Nitrogen Regulatory Protein Area

In The Dimorphic Fungus Histoplasma Capsulatum

Logan Blancett, Thomas Buford, Glen Shearer



Histoplasma capsulatum (Hc) is the etiological agent of histoplasmosis, a common cause of respiratory mycoses in humans. Hc is a dimorphic organism existing as a mold (M) at 25°C and once inhaled by host

(37°C) undergoes a dimorphic shift to the yeast (Y) phase. This dimorphic shift is essential for the pathogenesis of the organism within the host. It is most commonly found in the United States along the Mississippi and Ohio River Valley regions where high levels of bird and bat excrements can be found. Earlier experiments revealed that Hc is able to metabolize many complex nitrogen sources such as uric acid and allantoin. We also determined that many sole nitrogen sources are favored in only one phase of Hc respectively. AreA has been characterized as the key positive-regulator of nitrogen metabolism in closely related ascomycetes. AreA induces utilization of less-energetically favorable nitrogen sources by binding to the promoters of target genes and activating transcription. In this study, it has been found that AreA is up regulated by 3-fold when Hc is subjected to nitrogen-starvation conditions. It has also been found that downstream targets of AreA such as Gap1 and Dal5 are up regulated by 12-fold and 10 fold respectively. Current work is underway-utilizing RNA interference as well as indirect immunofluorescence to further characterize AreA .


P54 Determining The Role Of The Nitrogen Regulatory Protein Area

In The Dimorphic Fungus Histoplasma Capsulatum

Terra Parker, Logan Blancett, Thomas Buford, Glen Shearer



Histoplasma capsulatum ( Hc ) is a dimorphic fungus most commonly found in the United States in the Mississippi-Ohio River Valley flyways where high levels of bird and bat excrement can be found. Hc is the etiological agent of the disease Histoplasmosis, the most common respiratory mycosis of humans. Hc is a dimorphic organism existing as a mold (M) at 25 ⁰ C and once inhaled by a mammalian host (37 ⁰ C) undergoes a dimorphic shift to the yeast (Y) phase. This dimorphic shift is required for the disease progression of Hc . In previous experiments it has been shown that it can metabolize many complex nitrogen sources, such as allantoin and uric acid. We hypothesize that Hc is under the control of Nitrogen Catabolite

Repression (NCR). NCR is the process by which fungi selectively utilize the most energetically favorable nitrogen source available. AreA is the key positive regulator of NCR in other closely related ascomycetes. In the presence of a favorable nitrogen source AreA is down-regulated and unable to activate target genes. When subjected to a nitrogen-poor environment AreA binds to the promoter of target nitrogen metabolic genes and activates transcription. This study is focused on determining if AreA is the key positive regular of NCR in

Hc . Quantitative RT-PCR data suggests that AreA is up-regulated when subjected to a nitrogen-poor environment. Current work is underway using RNA interference as well as indirect immunofluorescence to further characterize the function of AreA .


P55 Mass Spectrometry For Mississippi Scientists

Xuan (Jo-Ann) Ding, Daniel G. Peterson

Institute for Genomics, Biocomputing & Biotechnology (IGBB),


Liquid chromatography (LC) and mass spectrometry (MS) are essential analytical techniques in biomedical research. As the proteomics core facility of MS-INBRE, Mississippi State’s Institute for Genomics, Biocomputing & Biotechnology (IGBB) is equipped with multiple LC-MS and related instruments which are utilized in proteomics, metabolomics, pharmaceutical, food safety, and other research fields. Instruments include a Waters Xevo G2-S QTof,

Thermo LTQ-Orbitrap-Velos, Thermo LTQ Velos, ABI 4700

MALDI TOF/TOF, Genomic Solutions ProPrep robot, and Covaris ultrasonicator. These instruments are capable of regular and 2dimensional HPLC and UPLC separations, protein and small molecule identification and quantification, semi-automated sample preparation and clean-up, robotic in-gel digestion, and plate handling.

They have been used for (a) global and targeted chemical proteomics studies to identify potential cancer drug targets; (b) global proteome profiling (including bacteria, fungi, and plants); (c) tissue/organismspecific proteome quantification in plants and animals; (d) theoretical and application-oriented studies of peptide fragmentation; and (e) programed deposition of proteins for surface coating. The IGBB team is here to provide services and collaboration opportunities for all

Mississippi researchers.


P56 Lysosomal Accumulation Of Free Cholesterol Regulates Acid

Sphingomyelinase In MCF-7 Cells

Gary T. Hodge II, Benjamin J. Melancon, Sucheta S. Naidu, Peyton S.

Pollard, Angad Bedi, Jerry W. Reagan, Jr.


Maintenance of cellular cholesterol homeostasis requires the transfer of lipoprotein-derived cholesterol from lysosomes to the plasma membrane or the endoplasmic reticulum where it is converted into cytoplasmic cholesterol ester droplets. Exposure to hydrophobic amines (e.g. desipramine) and steroids such as progesterone or mutations that inactivate Niemann-Pick proteins 1 and 2 induces a block in the translocation of lysosomal cholesterol and lead to excessive accumulation of intralyosomal free cholesterol. The increased cholesterol concentration down regulates acid sphingomyelinase (aSMase) in human skin fibroblasts and CHO cells

(Reagan et al, J. Biol. Chem ., 2000, 274:38104-10) by an unknown mechanism. Since this enzyme plays a crucial role in the generation of ceramide, a second messenger that induces apoptosis, we sought to determine the effect of lysosomal cholesterol accumulation on aSMase activity in MCF-7 cells, a breast adenocarcinoma cell line. Following incubation with human LDL, in the presence and absence of progesterone, cholesterol mass and subcellular localization were determined. In addition, the activity and proteolytic processing of aSMase was assessed by Western blotting and indirect immunofluorescence. The data demonstrate that lysosomal cholesterol accumulation in MCF-7 cells regulates aSMase at the posttranslational level by a mechanism that does not appear to involve altered intracellular trafficking. Such regulation could play an important role in the ability of MCF-7 cells to evade apoptosis.

Moreover, these studies lead to a greater understanding of the fundamental role that cholesterol plays in the molecular mechanisms that regulate cell death.


P57 A Highly-Sensitive, SERS-Based Assay For Dengue Virus

Detection

Amber M. Paul 1 , Zhen Fan 2 , Dhiraj Acharya 1 , Paresh C. Ray 2 , Fengwei

Bai 1



2 Department of Chemistry, Jackson State University, Jackson, MS

Dengue virus (DENV) is a mosquito-transmitted RNA virus that can cause human hemorrhagic fever, shock and even death. DENV has also been recently identified as a high-priority emerging infectious agent with the potential risk of blood transfusion-transmission by the

American Association of Blood Banks (AABBs). There is currently, no blood screening measure for DENV. Herein, we developed a gold nanoparticle (AuNP)-based, highly-sensitive Surface Enhanced

Raman Spectroscopy (SERS) for a DENV detection. Monoclonal anti-DENV antibody (4G2) was conjugated with AuNPs. These

AuNP:4G2 constructs can selectively bind to DENV particles, which was confirmed by transmission electron microscopy. The AuNP:4G2 constructs were then subject to serially diluted DENV serotype-2 in

PBS for detection limit determination. We found that AuNP:4G2 were able to detect, as few as, 10 viral particles/ml in a SERS-based assay. In brief, SERS-based detection of DENV-2 is rapid, sensitive, and cost-efficient and it can be adapted to improve DENV blood screening methods and prevent possible DENV blood transfusiontransmission from occurring.


P58 Effects Of Ethanol Treatment On Host Immune Response In A

Binge Drinking Model

Maria D.S. Basco, M Bhatty, T Wei, Stephen Pruett, Bindu Nanduri


Sepsis has been a growing concern in the health care sector. Chronic alcoholism has exacerbated this burden as it has been reported to increase the susceptibility to infections and infection related mortality.

In this study, we investigated the mechanism of alcohol-induced susceptibility to infection with E.coli

using a binge-drinking murine model. Survival studies have shown late ethanol treatment (18 hr) after E. coli infection to drastically decrease the survival of the mice to less than 20 hr compared to those that survived till 72 hr when ethanol was administered 30 min prior or 2 hr post infection. This late administration of ethanol led to an increase in the viability of E. coli in the peritoneal fluid. Some observed effects of late ethanol administration were altered host immune response via decreased phagocytic clearance and altered cytokine and chemokine responses.

This finding suggests that the increased prevalence of the pathogen observed in the host could be due to the alcohol-induced suppression of the immune response. In addition, the late ethanol treatment induced hypothermia that could contribute to the pathogenesis of the infection. Mice placed on warming pads survived the infection for 72 hrs compared to the mice without heating that survived only 36 hours.

Based on these results, the timing of ethanol administration post infection influences the outcome of an infection by altering the host immune response, immunological clearance, and hypothermiainduced susceptibility to infection.


P59 Role Of Interleukin-17 During West Nile Virus Infection

Dhiraj Acharya, Amber M Paul, Jordan E Lowery, Fengwei Bai



Interleukin-17 (IL-17) has been implicated in diverse immune functions including inflammation and neuroinflammatory conditions, such as experimental autoimmune encephalitis. West Nile virus

(WNV) can cause severe encephalitis in human for which specific antiviral drug or vaccine is currently unavailable. The pathogenesis of

WNV encephalitis and the role of host immune system in recovery are still not adequately investigated. Whether IL-17 play role during

WNV infection has not been studied before. Here, we found that IL-

17-/- mice developed higher viral load in blood and brain compared to wild-type (WT) control mice and were susceptible to severe WNV infection (IL-17-/- 20% survival; WT 60% survival). Interestingly, we found that WNV specific effector CD8+ T cells isolated from IL-17-

/- mice showed reduced cytotoxicity when compared to those from

WT mice. We also detected lower expression of cytotoxicity marker

(perforin and fas L) in CD8 T cell isolated from IL17-/- mice after

WNV infection. These results suggested that IL-17 is involved in

CD8 T cell mediated clearance of WNV infection. Further studies are warranted to elucidate this novel mechanism by which IL-17 signaling regulates CD8+T cell responses during WNV infection.


P60 The Photochemistry Of N-Substituted Heteroaromatic Salts

Wolfgang H. Kramer, Lauren M. Hoth, Priya P. Patel, Courtney B.

Mullins, GeNita N. Finley, Muzamil A. Khawaja, Irene S. Corrao, Emily H.

Stewart, B. Woods Curry


MS

Photoinduced cell death (Photodynamic Therapy, PDT) is becoming increasingly popular as a non-invasive cancer treatment. Current

FDA-approved Photodynamic Therapy agents are singlet-oxygen sensitizers and the photoinduced toxicity consequently depends on oxygen availability. Since oxygen levels are usually low in tumors, our approach for a PDT reagent uses N-substituted heteroaromatic salts which do not require an external oxygen source.

N-substituted heteroaromatic salts contain a fragmentable nitrogenoxygen bond that yield a heteroaromatic radical cation and an alkoxy radical upon photoinduced homolytic cleavage. Both transient species are known DNA cleavers and have been individually used for DNA cleaving applications. Despite the efficient production of DNA cleaving species, DNA cleaving efficiency with the simple salts based on quinoline, isoquinoline, phenathridine is extremely low. This is likely caused by weak ground-state association which causes the transient species to decay without interacting with DNA.

To increase DNA binding an aromatic imide (1,8-naphthalimide, a known DNA intercalator) was synthetically attached with a flexible linker. A simple condensation reaction, followed by N-oxidation and alkylation yielded the photochemical starting material.

DNA binding is determined by UV/VIS and fluorescence titration with the isoelectronic, photostable N-ethyl derivative. CD spectroscopy is used to observe changes in the three-dimensional structure of DNA.

DNA-cleaving is determined by gel electrophoresis and CD spectroscopy. Comparison with restriction endonucleases indicated the N-substituted heteroaromatic salts to be double-strand cleavers.


P61 Analysis Of The DNA-Cleaving Efficiencies Of Bifunctional

Heteroaromatic Salts

Courtney B. Mullins, Gurjit Kaur, Lauren M. Hoth, B. Woods Curry, Emily

H. Stewart, Jonathan P. Giurintano, Wolfgang H. Kramer


MS

Current Photodynamic therapy uses sensitizers to generate singlet oxygen which causes cell death. The hypoxic environment of most cancer tissues makes oxygen a limiting reagent for this approach and several methods have recently been developed to circumvent this problem.

The photoinduced homolytic N-O bond cleavage of N-

Heteroaromatic compounds with an N-alkoxy substituent (onium salts) leads to the formation of a heteroaromatic radical cation and an alkoxy radical. Both of these species have been shown to induce DNA cleavage, each with a different mechanism. The synthesis of the nitrogen onium salts includes the oxidation of the heteroaromatic nitrogen and subsequent O-alkylation. To increase the DNA cleaving effiency by enhancing ground-state association we synthetically attached a known DNA-binder, 1,8-naphthalimide.

Several binfunctional compounds have been synthesized and their photochemistry has been investigated. Here we present the DNA cleaving efficiency of a series of bifunctional DNA-cleavers which has been analyzed by gel electrophoresis and CD spectroscopy.

Comparison with restriction endonucleases indicated the Nsubstituted heteroaromatic salts to be double-strand cleavers.


P62 Photochemistry Of N-Alkoxy Quinolinium, Isoquinolinium, And

Phenanthridinium Tetrafluoroborates: Analysis Of The Reaction

Pathways By pH Monitoring

Priya P. Patel, Muzamil A. Khawaja, Katie L. Odom, Brooke K. Lassiter,

GeNita N. Finley, Wolfgang H. Kramer


MS

N-methoxy substituted aromatic heterocycles undergo a photoinduced homolytical N-O-bond cleavage. The reaction produces a methoxy radical and a heteroaromatic radical cation. The quantum yields of ion/radical formation have been determined by laser flash photolysis/quenching for the title compounds. Each transient species was produced with a yield of about 0.6 (±0.05). The energy wasting step appears to be a radical recombination reaction, which also produces a proton.

At Millsaps, we attempted to analyze the homolytic N-O bond cleaving reaction by focussing on the proton. This would give us a versatile tool for the analysis of the energy wasting side reaction and thus also the efficiency of the overall reaction. The methods we employed were titration with p-nitrophenolate and pH monitoring with buffer systems. It was important to keep the pH above about 6 to avoid acid base equilibria with other photoproducts.

Product distribution was determined by GC/MS after basic extraction. Interestingly, only four major isomers were formed.

Attempts to correlate the isomeric distribution with the solvent polarity was unsuccessful so far.


P63 Design Of Solar Photoreactors: Green Chemistry Applications

At Millsaps College

Greyson J. Smothers, Wolfgang H. Kramer


MS

In 1912, at the International Congress of Applied Chemistry in New

York, Giacomo Ciamician, the father of organic photochemistry, presented his vision of ‘the Photochemistry of the Future’:

“On the arid lands there will spring up industrial colonies without smoke and without smokestacks; forests of glass tubes will extend over the plains, and glass buildings will rise everywhere; inside of these will take place the photochemical processes that hitherto have been the guarded secret of the plants, but that will have been mastered by human industry which will know how to make them even more abundant fruit than nature, for nature is not in a hurry and mankind is.” (Science 1912, 36, 385-394)

The requirement of specialized equipment and reaction conditions as well as high costs, however, has been used as an excuse by the chemical industry to widely ignore chemical transformations with light. More than 100 years later Ciamician’s vision thus awaits its realization today.

We have built a cost effective solar photoreactors for a variety of applications. It is currently tested for photochemical transformations.

For our purpose, we hope to optimize synthetic pathways we employ in our lab to synthesize new compounds.

The concept of Green Chemistry is discussed and several solar photoreactors are shown to evaluate different design features. The focus is on optimizing the solar photoreactors for use in Mississippi.

Solar radiation is impaired by cloudiness compared to states such as

California or Arizona.


P64 Binfunctional 1,8-Naphthalimides as Fluorescence Sensors

Melinda K. Solomon, Anna K. Allred, Irene S. Corrao, Resham Rahat,

James P. McVaugh, Emily H. Stewart, B. Woods Curry, Wolfgang H.

Kramer


MS

1,8-Naphthalimides exhibit a moderate fluorescence that can be used as a sensor for oxidative power and pH of a solution if connected to an appropriate N-substituent. We have used pyridine derivatives as substituents and the protonation as well as the oxidation state of the pyridine nitrogen determines the quenching efficiency. The electron rich pyridine N-oxides diminish the fluorescence tremendously, and an interesting dependency on substitution pattern is observed.

Protonated pyridines do not quench naphthalimide fluorescence and thus the pH of the solution directly influences luminescence. The Nalkylated pyridine substitutents can also be used for this purpose, they mimic the protonated pyridines and can be used as standards. Stern-

Vollmer analysis suggests different modes of quenching for the Noxides and parent heterocycles. Solvent polarity strongly influences fluorescence behavior by changing the energy levels of the excited states.

To obtain more information about the quenching process, intermolecular quenching experiments are employed. Quenching of

N-methyl 1,8-naphthalimide with various pyridine derivatives with electon donating and withdrawing substitutents shows that certain substitution positions are more efficienctly quenching than others.

Cellular uptake is modeled by reverse micelles. Varying ratios of water to surfactant gives an accurate description of the interfacial water layer which might interact with our compounds.


P65 Trichomonas Vaginalis Virus In Trichomoniasis

Karen E. Ezelle 1 , Lizhuo Ai 1 , Sara M. Barker 1 , Erika M. Coleman 1 , Kellen K.

Dawson 1 , Giancarlo P. Fernandez 1 , Amanda R. Kaminski 1 , Hirni Y.

Patel 1 , Payal B. Patel 1 , Brad S. Williams 1 , S. Skye Williams 1 , C. Austin

Zamarippa 1 , J. Chris Meade 2 , Cory G. Toyota 1

1 Department of Chemistry and Biochemistry, Millsaps College, Jackson,

MS

2 Department of Microbiology, University of Mississippi Medical Center,

Jackson, MS

Trichomoniasis, caused by the protozoan Trichomonas vaginalis , is the world’s most common non-viral sexually transmitted disease. It responds well to metronidazole treatment, but is linked to increased susceptibility to HIV infection, pregnancy complications, and demonstrates a pronounced racial disparity, infecting ten times more

African American women than Caucasian women. T. vaginalis isolates are also infected with up to four distinct double stranded

RNA viruses (TVV1-4) which have been proposed to enhance pathogenesis of trichomoniasis by provoking inflammation. We are working to determine the presence of the four strains of TVV from two subtypes of T. vaginalis and assess their relationship with the clinical manifestations of trichomoniasis. RNA is purified from clinical isolates of the protist, cDNA produced by RT, strain-specific

PCR products are amplified, and results are analysed by agarose gel electrophoresis.


P66 Oxalate Oxidase Activity in E. coli

Maryam N. Qureshi, Cory G. Toyota


MS

The Escherichia coli enzymes YfdW and YfdU are an oxalate:formyl-

CoA transferase and an oxalyl-CoA decarboxylase, respectively, and are homologues of the two Oxalobacter formigenes enzymes FRC and

OXC. Coupled, these enzymes catalyze the decarboxylation of oxalate to yield formate. We have shown that YfdW and YfdU are indeed necessary for an oxalate-dependent acid tolerance system in E. coli . Membrane inlet mass spectrometry (MIMS) has been used to effectively measure concentrations of small molecules, like NO and

CO

2

, in aqueous solution and to determine the Michaelis-Menten kinetic constants for enzymes like oxalate decarboxylase. In MIMS experiments, we detect [ 13 C]-CO

2 from [ 13 C]-oxalate in whole cell extracts, however, these are in low concentration and present even in

Δ yfdW and Δ yfdU deletion strains of E. coli . Concomittant O

2 consumption suggests the presence of a so-far unidentified oxalate oxidase.


P67 Chemoenzymatic Synthesis Of Both Enantiomers Of 3-

Aminoalcohols From A Common Intermediate

Pradeep Bandi 1 , Roshetta Williams 1 , Benjamin Lewing 1 , Wanqing Lu 1 ,

Erin Norcross, Dale Rosado 1


Clinton, MS


Clinton, MS

Amino alcohols are found in a wide variety of biologically active compounds and have been used as chiral ligands for transition metal catalysts and to induce chirality in asymmetric synthesis. Ethambutol, an anti-tuberculosis drug, contains two (S)-2-amino-1-butanol moieties. Replacement of the 2-aminoalcohols of ethambutol with 3aminoalcohols may yield ethambutol analogues with increased activity. In order to investigate the properties ethambutol analogues containing 3-aminoalcohols, an efficient synthetic route to both enantiomers has been developed. Reduction of diethyl 2ethylmalonate with LiAlH

4

yields a 1,3-diol. The diol is quantitatively converted to a diacetate by treatment with acetic anhydride.

Hydrolysis of the diacetate with Pseudomonas fluorescens lipase in phosphate buffer yields an acyl alcohols that can be treated with diphenyl phosphoroazide yields an acyl azide. (S)-2-(aminomethyl)-1butanol is obtained after reduction of the acyl azide with LiAlH4.

The enantiomer of the 2-(aminomethyl)-1-butanol was synthesized by manipulation of the protecting groups and repetition of the azidation and reduction chemistry. The synthetic route outlined above using mild conditions, produces good yields in each step, and results in synthesis of both enantiomers of 2-(aminomethyl)-1-butanol from a common acyl alcohol intermediate.


P68 Computational Analysis Of The Gold Clusters Interaction With

DNA Bases

Jeanne A. Ishimwe

Department of Chemistry, Belhaven University, Jackson, MS

Cancer is the number two killer in America. Cells become cancerous because DNA damage thus the study of DNA structure and its interactions is important .The computational study of the interaction of Adenine and Thymine (AT) Watson Crick base pairs with gold dimers was studied using density functional theory (DFT). Structures of Adenine, Thymine, H

2

0 and Au:Au are drawn using GaussView5 software and so are the structures of the Adenine-Thymine complexes. The structures of the complex differ only in that the position of H20 and Au:Au varies from one complex to another. The

6-31+G** is used to optimize the structures except for Au:Au whose atomic number is more than 6-31+G** can optimize. The basis set

Los Alamos effective core potential LanL2DZ is used for gold

(Au:Au). The optimized structure, gold dimer binds to one of the nitrogen atoms of a DNA base. Water molecule forms a hydrogen bond with oxygen and Nitrogen of either Adenine or Thymine.

Adiabatic electron affinity (AEA) calculated values show that both water and gold dimer placed on Adenine molecule give higher results than their position on Thymine. The difference in the AEAs is thus a result of Adenine’s high affinity for gold clusters.


P69 HMGB1b Readily Recognizes Hybrid DNA-PNA Four-Way

Junctions.

Anthony J. Bell Jr 1 , Filbert Totsingan 2



2 Department of Chemistry, New York University, New York, NY

The objective of this study was to evaluate hybrid four-way junctions

(4WJs) as potential therapeutic inhibitors against HMGB1. The premise for using a nucleic acid strategy is based upon in vitro studies that show HMG proteins bind 4WJs with very high affinity 1-6 . To improve the in vivo stability of 4WJs, hybrids have been constructed of DNA and PNA (peptide nucleic acid). PNA possesses a polyamide backbone that permits stronger Watson-Crick base-pairing between

PNA and DNA strands ( vs.

classical DNA duplexes) 7 . Moreover, the

PNA backbone is resistant to endogenous nucleases 8 . Hence, PNAs are being explored as novel therapeutics 9,10 . In this study, two hybrid

4WJs (4WJ-PNA

1

and 4WJ-PNA

3

) were investigated extensively.

Individual PNA strands were inserted into the immobilized 4WJ (J1) to evaluate the influence on: i) nucleic acid secondary structure and ii) protein recognition. Circular dichroism (CD) analysis clearly showed that the structural properties of hybrid 4WJs were similar to native

(DNA) 4WJs. 4WJ-PNA

3

, like the control 4WJ (J1), shifts readily from an open to stacked conformation in the presence of Mg +2 , while

4WJ-PNA

1

did not. Results that indicate 4WJ-PNA

1

possessed a more amorphous secondary structure. The protein recognition properties were evaluated with HMGB1b and Histone H1. CD studies indicate that HMGB1b recognizes hybrid junctions in an analogous manner. H1, however, displays a preference toward J1 vs.

hybrids. More stringent binding analyses (EMSAs) reveal that

HMGB1b binds J1 and 4WJ-PNA

3

with identical affinity (K

D s) and

4WJ-PNA

1

with two-fold lower affinity. Our data clearly shows that the sequence/location of the PNA strand strongly influences the structural and protein recognition properties of 4WJs. In the case of

4WJ-PNA

3

, its structure-function properties mimic J1 and may prove to be a viable inhibitor of HMGB1. Our long-term goal is to


introduce hybrid 4WJs, such as 4WJ-PNA

3

, into the extracellular matrix to shut off deleterious HMGB1-mediated signaling reactions.

References

1. Pohler, J. R. G. HMG box proteins bind to four-way DNA junctions in

2.

3. their open conformation. The EMBO Journal 17 , 817–826 (1998).

Taudte, S., Xin, H. & Kallenbach, N. R. Alanine mutagenesis of highmobility-group-protein-1 box B (HMG1-B). Biochem. J.

347 Pt 3 , 807–

814 (2000).

Xin, H., Taudte, S., Kallenbach, N. R., Limbach, M. P. & Zitomer, R. S.

4.

5.

6.

DNA binding by single HMG box model proteins. Nucleic Acids Res.

28 ,

4044–4050 (2000).

Varga-Weisz, P., Zlatanova, J., Leuba, S. H., Schroth, G. P. & van

Holde, K. Binding of histones H1 and H5 and their globular domains to four-way junction DNA. Proc. Natl. Acad. Sci. U.S.A.

91 , 3525–3529

(1994).

Hill, D. A. & Reeves, R. Competition between HMG-I(Y), HMG-1 and histone H1 on four-way junction DNA. (1997).

Hill, D. A., Pedulla, M. L. & Reeves, R. Directional binding of HMG-I

(Y) on four-way junction DNA and the molecular basis for competitive

7.

8.

9. binding with HMG-1 and histone H1. Nucleic Acids Res.

27 , 2135–2144

(1999).

Egholm, M., Buchardt, O., Nielsen, P. E. & Berg, R. H. Peptide nucleic acids (PNA). Oligonucleotide analogs with an achiral peptide backbone. J.

Am. Chem. Soc.

114 , 1895–1897 (1992).

Demidov, V. V. et al.

Stability of peptide nucleic acids in human serum and cellular extracts. Biochemical Pharmacology 48 , 1310–1313 (1994).

Hyrup, B. & Nielsen, P. E. Peptide nucleic acids (PNA): synthesis, properties and potential applications. Bioorganic & medicinal chemistry 4 ,

5–23 (1996).

10. Nielsen, P. E. Peptide nucleic acids (PNA) in chemical biology and drug discovery. Chem. Biodivers.

7 , 786–804 (2010).


P70 The Effects Of Inflammatory Cytokines On Melanoma

Proliferation And Migration

Elizabeth Brandon, Maxwell Schwam, Mawussi Kamassa, Omama

Ahmad, Yassmin Hegazy, Zac Burns, Alexandria Niswonger


Experimental and clinical data indicate that obesity increases the risk for colorectal, esophageal, pancreatic, endometrial, renal cell carcinoma, and breast cancer (post-menopausal). Several metabolic, endocrine, and immune system factors are positively correlated with a poor prognosis and shorter disease-free survival times in obesity associated cancers. The paracrine signaling between tumor cells and stromal cells is a hallmark of cancer that has only recently become appreciated as a catalyst for mutagenesis, immune evasion, and metastasis. Although some mechanistic knowledge has been obtained from a “one growth factor, one effect” approach, this must be paired with a systems analysis to gain an understanding of the interactions between stromal cells and melanoma cells. Our data reveals that the inflammatory cytokines, resistin and IL-6, promote melanoma proliferation (6.1x10

5 cells/day and 1.0x10

6 cells/day v. 5.5x10

5 cells/day for controls, p=0.05). We also found that leptin protects cells from necrotic and apototic cell death. There is still a large gap in our knowledge of how obesity promotes melanoma metastasis. Data from migration experiments in which cells were treated with cytokines individually have shown no difference in cell migration between experimental and control cells. Therefore, our future research will test the hypothesis that melanoma invasion and metastasis are promoted through paracrine signaling from macrophages and other stromal cells. Our initial approach will consist of co-culture experiments with macrophages to characterize these crucial interactions.


P71 Interleukin-6 And Resistin Promote Melanoma Proliferation

Mawusi Kamassa, Courtney Skaggs, Omama Ahmad, Yassmin Hegazy,

Zac Burns, Alexandria Niswonger, Elizabeth Brandon


Obesity adversely affects the prognosis of many cancers in part, by altering the levels of several circulating hormones, inflammatory cytokines, and adipokines. Melanoma tumors grow more rapidly in obese mice than in lean mice, though the mechanism is poorly understood. To better understand the role these cytokines play in melanoma growth, we developed a cocktail simulating the concentrations of some of these cytokines in obese animals and monitored its effect on cell growth rates and migration. The cocktail was comprised of regular growth medium supplemented with leptin, insulin, insulin-like growth factor 1 (IGF-1), interleukin-4 and 6 (IL-

4, IL-6), and tumor necrosis factor alpha (TNFα ). We also examined how the individual cytokines affected proliferation of B16F10 mouse melanoma cells. Relative to controls, the obesity cocktail and high concentrations of TNFα or resistin (100 ng/mL) significantly decreased cell proliferation. IL-6 and resistin at 10 ng/mL significantly increased cell proliferation. Preliminary data indicates that resistin increases the expression of the survival protein, Bcl-X

L

.

We also tested the hypothesis that inflammatory cytokines might affect melanoma migration. Our previous research showed that medium from mouse macrophages stimulated melanoma cell migration slightly, but had no effect on proliferation. To identify the unique migratory effect of some inflammatory factors, we cultured cells in IL-4, IL-6, and resistin. Cells were plated in modified Boyden chambers and then treated with each individual cytokine for 24 hours.

Inserts were removed and the cells fixed and stained with Dif-Quik™.

Cells on the proximal side of the membrane were removed and those on the distal side were counted. There was no difference in the migration of the treated cells compared to control cells. Greater insight into the mechanisms that stimulate melanoma cell migration may be gained from future studies employing co-culture systems with macrophages and adipocytes.


P72 Curly Kale Brassica oleracea var. sabellica Induces Apoptosis in

Cultured Mouse Melanoma Cells

Bilal Qizilbash, Matthew Bear, Jacob Morgan, Alexandria Niswonger,

Maxwell Schwam, Matthew Taitano, Elizabeth Brandon


The challenge with many cancers is not just killing the malignant cancer cells, but doing so in a non-toxic manner. Numerous studies have been conducted on curly kale, B. oleracea sabellica, to identify some of the compounds responsible for the health benefits of consuming the plant, in its raw or juiced form. Much of this research focuses on sulforaphane, an isothiocyanate that’s also found in foods such as broccoli, brussel sprouts, and cauliflower. Sulforaphanes have been shown to decrease cell proliferation, reduce inflammation, and induce protective autophagy in vitro . Our idea is that the natural context in which sulforaphanes and other bioactive compounds exist in curly kale creates a concert of both agonist and antagonistic effects that could have significant anticancer effects. To test this hypothesis, we prepared kale juice in two forms: unfiltered and sonicated juice that was subsequently filter sterilized. Serial dilutions were tested on melanoma cells to determine the optimum dosage for a four day treatment. We found a dose-dependent decrease in cell growth and chose the lowest concentration of juice for our experiment. Melanoma cells treated with unfiltered juice were killed by day four. Cells treated with the sonicated filtered juice had significantly reduced growth

(8.63e

5 cells/day ± 52,700 cells for control v. 1.05e

5 cells/day ± 8,660 for kale juice, p=0.01). Western blot analysis with a poly-ADP ribose polymerase (PARP) antibody showed two PARP fragments in the lysates from cells treated with kale juice. This suggests that the sonicated filtered juice induces apoptosis. These experiments will be repeated in a normal mouse epithelial cell line to determine the toxicity in non-cancerous cells. If the sonicated filtered juice is significantly less toxic to normal cells, then we will work toward examining the safety and efficacy of kale juice for treating or augmenting the treatment of melanoma in vivo.


P73 Comprehensive View Of Water Quality Of Lake Enid

Michael Collins 3 , Padmanava Dash 1 , Julius O. Ikenga 3 , Wellington

Ayensu 1 , James L. Pinckney 2 , Daniel Kibet 3



University of South Carolina, Columbia, South Carolina

3 Department of Natural Science and Environmental Health, Mississippi

Valley State University, Itta Bena, MS

Lake Enid represents one of the best recreational lakes in the state of

Mississippi. The main objective of this research work was to take a comprehensive approach to investigate the current state of water quality of Lake Enid. A sampling trip was organized to Lake Enid on

June 11, 2013. Prior to the sampling trip, a time-series of satellite data were processed and true color images were generated to examine the past occurrence of algal blooms in the lake. On the sampling trip, water samples were collected in clean Nalgene bottles and physical parameters of water (temperature, salinity, dissolved oxygen and pH) were measured using a Hanna instrument. The collected water samples were analyzed for photo-pigments using high performance liquid chromatography (HPLC), for cyanobacteria-specific pigment phycocyanin (PC), suspended particulate matter (SPM), phycotoxins, bacteria and toxic metals. The “greenness” in the time-series of true color images demonstrated the occurrence of algal blooms in lake in the past. The higher organic SPM than the inorganic counterpart suggested the presence of algal blooms in the lake. The HPLC pigment derived relative abundances of major algal groups suggested the dominance of cyanobacteria. Microcystin, a hepatotoxic cyanotoxin ranged between 0.18-0.39 µg/L. The bacterial counts revealed the dominance of heterotrophic bacteria and total coliform bacteria, but E. Coli and Enterococii bacteria were also present at many of the sites. Toxic metals were also found at all sites. All these findings suggest that the water quality of Lake Enid needs to closely monitored.


P74 Mapping And Identification Of A Genetic Mutation That Causes

Cataracts

Zaliya Morris 1 , Ashley Johnson 1 , Jonathan Lee 1 , Ashlyn Harmon 1 ,

Xuexiang Wang 1 , Elise Gomez-Sanchez 1,2,3 , Michael R. Garrett 1,2

1 Pharmacology, University of Mississippi Medical Center, Jackson, MS

2 Medicine, University of Mississippi Medical Center, Jackson, MS

3 GV(sonny) Montgomery VAMC, University of Mississippi Medical


Cataracts are a major cause of blindness. The most common forms of cataracts are age and UV- related and develops mostly in the elderly, while congenital cataracts appear at birth or in early childhood. The

Dahl salt-sensitive (SS/Jr) rat is an extensively used model of saltsensitive hypertension. In the mid 1980’s, cataracts appeared in a few animals in the Dahl S colony, presumably the result of a spontaneous mutation. The mutation was fixed and bred to establish the SS/Jrcat substrain. The SS/Jrcat substrain has been exclusively used by a single investigator to study the role of steroids and hypertension. Using a classical genetic analysis approach, we localized the cataract gene with high-resolution to a less than 1 Mbp region on chromosome 9 using an F

1

[SS/Jrcat X Spontaneous hypertensive rat (SHR)] X SHR] segregating population. The 1 Mbp region was found to contain only

13 genes, including 4 genes from the γ -crystallin ( Cryg-b,-c,-d,-e) gene family. Mutations in the many of the γ -crystallins are known to play a role in cataract formation in both humans and rodent models.

All of the γ -crystallins were sequenced and a novel point mutation in the start codon (ATGàGTG) of the Crygd gene was identified which led to the complete absence of CRYGD protein and the likely cause of cataracts in the SS/Jrcat strain. In summary, the identification of the genetic cause in this novel cataract model may provide an opportunity to better under the development of cataracts, particularly in the context of hypertension.


P75 Toxicity Of Vibrio Cholera In Mississippi Coastal Waters

Robert Buntyn , Reid Warren , Shuo Shen , D. Jay Grimes

Department of Coastal Sciences, The University of Southern Mississippi,

Long Beach, MS

Less than one percent of Vibrio cholerae strains in the U.S. coastal waters possess the cholera toxin. Cholera toxin is expressed in the ctxA gene and is responsible for the illnesses caused with V. cholerae .

Our hypothesis was that Mississippi coastal waters would reflect similar statistics and to investigate this hypothesis water samples were collected from three different locations (2 estuaries and 1 bathing beach) in Ocean Spring, MS. Salinity, water temperature, time, and

GPS coordinates were recorded for each sample. Water samples were placed in alkaline peptone water, incubated overnight at 40 o C, and the surface pellicle was plated onto thiosulfate-citrate-bile salts-sucrose agar and incubated at 40 o C. Sucrose positive colonies were purified by re-streaking onto Marine Agar 2216 (MA) and cultures were then maintained in slants. Each isolate was then plated onto MA and

DNA was extracted from this growth. A DNA primer for outer membrane ompW was then used in a polymerase chain reaction (PCR) to verify the putative V. cholerae isolates. All 37 isolates were ompW positive and these strains are now being tested for the presence of ctx

A. Thus far, five strains have tested positive for ctx A, which is an unusually high percentage.


P76 Are Peptide Nucleic Acids (Pnas) Capable Of Transferring

Genetic Information Via Aminoacyl-Trna Synthetases?

Crystal Cox Serrano 1 , Filbert Totsingan 2 , Anthony Bell Jr.

1



2 Department of Chemistry, New York University, New York, New York

The RNA (Ribonucleic Acid) World Hypothesis is widely accepted throughout Biochemistry. However, flaws exist in this theory such as the instability of ribose. Biochemical reactions that could have occurred prebiotically have not been show to produce nucleosides or nucleotides. Also, polymerization of macromolecules is universally difficult without enzymatic activity.

Over the past two decades, there has been an interest in PNA, a synthetic molecule because of experimental evidence showing that

PNAs can Watson-Crick base pair with Deoxyribonucleic Acid

(DNA). PNAs consist of a polypeptide, achiral, neutral backbone typically composed of N-(2-aminoethyl) glycine (AEG) units with adenine, cytosine, guanine and uracil-N-acetic acid substituents.

These components have been synthesized under prebiotic conditions

(Spark discharge reactions) and AEG polymerizes at 100 ° Celsius.

Moreover, PNAs have been shown to form double and triple helix forms with DNA, causing an increase in stability of a DNA/PNA duplex over DNA/DNA duplexes.

Our goal is to provide supporting evidence of PNA as a primordial molecule capable of processes now conducted by RNA or DNA, including the transfer of genetic information. To do so, we anneal a nonameric PNA and tridecameric RNA to form a RNA/PNA duplex, mimicking the acceptor stem of Alanyl-transfer ribonucleic acid

(tRNA Ala ). Aminoacylation assays will be performed to aminoacylate alanine onto the duplex using Alanyl-tRNA Synthetase (AlaRS) enzyme .


P77 Increased Expression Of SNAT2 And GLUT Transporters

Enhances Placental Efficiency In A Low Birth Weight Rat Model

Symone Booth 1 , Nicholas Porter 1 , Lydia Ayanwale 1 , Danielle T.

Williamson 1 , Norma B. Ojeda 2,4 , Barbara T. Alexander 3,4 , Bettye Sue

Hennington 1


2 Departments of Pediatrics, University of Mississippi Medical Center,

Jackson, MS

3 Physiology, University of Mississippi Medical Center, Jackson, MS

4 Women’s Health Research Center, University of Mississippi Medical


Low birth weight (LBW) has been linked to a higher risk of development of adult diseases including hypertension, type II diabetes, obesity, and renal disease. There have been no definitive studies that elucidate the metabolic pathways affected during gestation when fetal nutrition is below optimal levels. We utilized a reduced uterine perfusion programming (RUPP) rat model of LBW. For the current study, our hypothesis was that RUPP during late gestation can result in decreased fetal and placental weight; however, placental efficiency will increase. Time pregnant rats underwent

RUPP or sham surgery at 14-days of gestation. Rat pups, tissues and placentas were harvested at 19 day gestation (E19). Pups and placentas were weighed and calculations for placental efficiency (PE) were performed. RUPP offspring weight and placental weight was significantly decreased (P ≤ 0.0004; P ≤ 0.0001). PE was significantly increased in the offspring from RUPP Dams (P ≤ 0.0002). Western blot analysis using placenta homogenates revealed a significant increase in SNAT2, a neutral amino acid transport protein, and a significant increase in GLUT2 and

GLUT3 transport proteins in the RUPP placentas (P ≤ 0.02; P ≤ 0.001;

P ≤ 0.02, respectively). qPCR analysis indicated a greater than 2-fold increase in SNAT2 message in the RUPP placentas compared to the control group In conclusion, significantly increased placental efficiency in the RUPP offspring was supported by the upregulation of amino acid transporter SNAT2 and glucose transporters GLUT2 and GLUT3. The upregulation of nutrient transporters suggests a compensatory mechanism occurred upon decreased available fetal nutrition even though offspring were still born at a low birth weight.


P78 Bisulfite Sequencing Of Estrogen Receptor-Alpha Reveals

Changes In Methylation In Exon 1

Apiffani Binion 1 , Symone Booth 1 , Lydia Ayanwale 1 , Danielle T.

Williamson 1 , Suttira Intapad 3,4 , Norma B. Ojeda 2,4 , Barbara T.

Alexander 3,4 , Bettye Sue Hennington 1



Jackson, MS




The term “epigenetics” is defined literally as “in addition to genetics” but in reality refers to changes in the DNA or surrounding chromatin that influence gene expression but do not change genetic composition. DNA methylation is generally repressive of gene expression, but exceptions are beginning to emerge. There are two identified ways in which meaningful epigenetic changes can occur: (1) the addition of a methyl group to a cytosine that sits just upstream of a guanine, and is referred to as a CpG island, and (2) changes to the histones that form the core of nucleosomes around which DNA is tightly packed. Evidence for DNA methylation continuing throughout life is beginning to emerge. Studies show DNA methylation of Estrogen

Receptor-a (ER-a) was altered in a gender-specific manner during maternal grooming. Altered ER-a promoter methylation has been found in colon cancer and breast cancers. Evidence of hypermethylation and hypomethylation have been reported in various tissues in studies on artherosclerosis. Little is known about epigenetic changes caused by aging on ER-a. While studying the state of cardiovascular health of aging female Sprague Dawley rats born at low birth weight (LBW), it was found that ER-a protein in the pancreas was decreased compared to the age-matched control group. ER-a has been reported to protect against atherosclerosis and aging in the cardiovascular system. Further, it has been reported that DNA methylation in the promoter region of the ER-a gene can reduce transcription of ER-a leading to a higher risk for several


cardiovascular diseases. Our lab utilizes a reduced uterine perfusion model of intrauterine growth restriction (IUGR) that results in low birth weight offspring. Epidemiological studies have shown adults born a low birth weight are at greater risk for cardiovascular disease.

Studying LBW Sprague-Dawley females that have aged to 12 months, we found that the mean arterial pressure of the IUGR female was significantly higher (P ≤ 0.05). Preliminary data also indicated that estradiol levels in IUGR one-year-old females were not different from the age-matched control group. Our hypothesis was that there is increased methylation in the ER-a gene of the LBW females leading to decreased transcription of the ER-a gene.

The gene for ER-alpha, the dominant ER isoform, is unique in that part of the promoter region is in exon 1. To determine the degree of methylation, bisulfite sequencing of DNA was performed. DNA was isolated from pancreatic tissues, liver tissues, and kidney tissues. Sites chosen included 12 CpG sites upstream of the start site and 7 CpG sites in exon 1 of the ER-a gene. There was no significant difference in overall DNA methylation in these combined sites in the liver, pancreas or kidney. Using ANOVA analysis, there was a significant increase in DNA methylation in the liver compared to the pancreas and kidney (P ≤ 0.0001) with the least methylation occuring in the pancreas. There was a significant increase in methylation in the

IUGR kidney compared to the IUGR pancreas (P ≤ 0.0005) and significant increase in methylation in the IUGR liver compared to the

IUGR kidney (P ≤ 0.03). These data may indicate a silencing of ER-a expression in these tissues with hypermethylation. More importantly, the sequencing evidence indicates significantly increased methylation in exon 1 of the pancreatic ER-a compared to the kidney and the liver

(P ≤ 0.05). In conclusion, the evidence for no change in estradiol levels between one-year-old female rats should not contribute to elevated

MAP; however, if ER-a is not available, then estradiol-ER-a complex would be decreased in the IUGR older females.


P79 Changes In PPAR Isoform Expression And PEPCK Expression In

LBW Aging Female Rats Indicates Altered Glucose Metabolism

Bettye Sue Hennington 1 , Lydia Ayanwale 1 , Symone Booth 1 , Apiffani

Binion 1 , Danielle T. Williamson 1 , Suttira Intapad 3,4 , Norma B. Ojeda 2,4 ,

Barbara T. Alexander 3,4



Jackson, MS




Dietary fatty acids are natural activators of peroxisome proliferatoractivated receptors (PPARs) which strongly suggest that lipoproteins serve as ligand carriers for PPARs; transcription factors, which, in turn, modulate lipid homeostasis of the body. PPAR-alpha reduces triglyceride levels and is involved in regulation of energy homeostasis.

It is activated under conditions of energy deprivation. Activation of

PPAR-gamma causes insulin sensitization and enhances glucose metabolism, whereas activation of PPAR-delta enhances fatty acids metabolism. Studies have shown that estrogen inhibits the actions of

PPAR-alpha on obesity and lipid metabolism through its effects on

PPAR-alpha-dependent regulation of target genes such as carnitine palmitoyltransferase 1 ( CPT-1). CPT-1 is activated by PPAR-alpha when increased fatty acid beta-oxidation is required to yield molecules that can be used for energy production. Little is known about the actions of PPAR isoforms when estrogen levels are at menopausal levels. In parallel, our previous studies indicate low birth weight

(LBW), at full term, is a risk factor for the development of adult disease including those diseases found in metabolic syndrome. To determine the effects of fetal undernutrition on energy transformation and metabolic disorders in the aging female, we utilized a reduced uterine perfusion programming model (RUPP) that results in intrauterine growth restriction (IUGR). Previous data from our lab indicates no significant difference in levels of estradiol between the

IUGR one-year-old females and the age-matched control females.

Further, mean arterial pressure (MAP) was significantly higher in the


IUGR one-year-old females compared to the age-matched controls.

Using liver tissue from the one-year old females and qPCR, expression studies indicate 2-fold increase in PPAR-alpha in IUGR compared to control (P ≤ 0.05) along with a 3-fold increase in CPT-1 in IUGR compared to control (P ≤ 0.02). Generally expressed in adipocytes, PPAR-delta expression in liver tissue of IUGR aging females increased 4-fold compared to control (P ≤ 0.009). Our Western blot data indicates a significant increase in PEPCK, phosphoenolpyruvate carboxykinase, in IUGR whole liver homogenate compared to control. Finally, we found an elevated increase in PPAR-gamma in IUGR liver tissue compared to control

(P ≤ 0.04). Increases in PEPCK and PPAR-gamma may indicate a decrease in available glucose and an increase in gluconeogenesis. The significantly elevated CPT-1 expression in the IUGR liver indicates an increase in beta-oxidation of fatty acids. Significantly increased

PPAR-delta expression generally is needed when FA concentrations are too high. Taken together, significantly increased PPAR-alpha and CPT-1, significantly increased PEPCK and PPAR-gamma and

PPAR-delta may occur when there is an impairment in the IUGR aging females in control of molecules needed for energy homeostasis.

Future experiments will determine levels of triglycerides, cholesterol and fatty acids in order to gain a better understanding of the cellular conditions for energy homeostasis in the aging female liver.


P80 Differential Expressions Of Lysophospholipid Receptors (Lprs)

In Benign And Malignant Tissues

Jinghe Mao 1 , Chunyi Wang 2 , Samantha Redfield 2 , Yinyuan Mo 3 ,

Xinchun Zhou 2, 3


2 Department of Pathology, University of Mississippi Medical Center, MS

3 Cancer Institute, University of Mississippi Medical Center, MS

Lysophospholipid receptors (LPRs), including Lysophosphatidic acid

(LPA) receptors and sphingosine 1-phosphate (S1P) receptors, are a group of G protein-coupled receptors. LPRs are involved in a wide range of biological processes, such as embryogenesis, systemic development, immune cell trafficking, and inflammatory reactions.

The emerging evidence has suggested that the activities of LPRs may be associated with pathogenesis and metastasis of several human cancers. In this study, the expression levels of several LPRs were determined by immunohistochemistry (IHC) in various benign and malignant tissues from various human system/organs, in order to correlate the expression levels of these LPRs with different malignancies. IHC for EDG1 (S1Preceptor 1, S1P1, C- and Nterminals), EDG8 (S1P receptor 5, S1P5), EDG2 (LPA receptor 1,

LPA1) and EDG4 (LPA receptor 2, LPA2) was performed on tissue microarray slides containing 384 formalin-fixed paraffin-embedded benign and malignant tissues from 33 human system/organs. The expression level of each LPR was reported as a final IHC score calculated as staining extent score (0-3) multiplied by intensity score

(0-3) with a maximal score of 9. Student t-test was employed to analyze the differences in IHC score of each studied LPR between benign and malignant tissuesin overall and in individual system/organ.

The IHC signals for all studied LPRs were seen in both cytoplasm and nuclei of most tissues. In the level of overall benign and malignant tissues, the expression pattern was similar for all studied LPRs: the


cytoplasmic IHC signals were decreased, but the nuclear IHC signals were increased in malignant tissues as compared with benign tissues.

Among them, the expression level of cytoplasmic S1P1 (C-terminal) was significantly decreased, andexpression levels of nuclear S1P1 (Nterminal) and LPA1 were significantly increased in overall malignant tissues as compared with overall benign tissues. At the organ level, a significant increase in nuclear expression was observed in liver malignancies for all studied LPRs, whereas a decrease innuclear expressionwas observed in lung malignancies for all studied LPRs. In colon malignancies, the expression levels of S1P1 (both N- and Cterminals) were significantly increased, but the expression level of

LPA1 was significantly increased as compared with benign colon tissues.

The constitutive expression of cytoplasmic LPRs may be essential for physiological processes in all tissues. However alterations in nuclear expression of studied LPRs may differentially correlate with the tumorogenesis in different system/organs.


P81 Regulation Of Catechol Production In Response To Transferrin

Is Temperature Dependent In Bacillus cereus

Bianca L. Garner 1 , Baraka Williams 1,2 , Claressa Youngblood 1

1 Biology Department, Tougaloo College, Tougaloo, MS

2 Department of Environmental Science, Jackson State University,

Jackson, MS

Bacillus cereus ATCC 14579 excretes the siderophores bacillibactin and petrobactin, as well as the simple catechols

2,3-dihydroxybenzoic acid and 3,4-dihydroxybenzoic acid.

Independently, temperature and iron concentration regulate catechol expression in Bacillus species. Catechols were detected in culture filtrates in the presence of low levels of hemoglobin and transferrin at both 30 ° C and 37 ° C. Regardless of the growth temperature, the addition of ferric ammonium citrate as an iron source resulted in low catechol levels. Catechol production was greatest when cells were grown at

37 ° C.Catechol production was maximum when B. cereus was cultured at 37 ° C with 10 micrograms of transferrin, after which catechol production decreased to levels below that observed for no added iron sources. A similar trend was observed when B. cereus was cultured at 30 ° C, however, the maximum catechol production did not reach levels observed at

37 ° C. While temperature may impact siderophore mediate iron acquisition, the mechanism governing this process may be different from that associated with the ferric ammonium citrate systems.


P82 Morphological Effects Of Genistein, Thymoquinone,

Fluorouracil, And Laser Therapy On Laryngeal Carcinoma Cells

Osasu Adah, Michelle Tucci, Gerri Wilson, Felix Adah, Hamed Benghuzzi

University of Mississippi Medical Center, Jackson, MS

Low level laser therapy (LLLT) involves exposing cells or tissue to low levels of red and near infrared (NIR) light, and is referred to as

“low level” because of its use of light at energy densities that are low compared to other forms of laser therapy that are used for ablation, cutting, and thermally coagulating tissue. The precise biochemical mechanisms underlying the therapeutic effects of LLLT are not yet well-established. From observation, it appears that LLLT has a wide range of effects at the molecular, cellular, and tissue levels. In addition, its specific modes of action may vary among different applications. Within the cell, there is strong evidence to suggest that

LLLT acts on the mitochondria to increase adenosine triphosphate

(ATP) production, modulation of reactive oxygen species (ROS), and induction of transcription factors. LLT has shown promise for down regulating inflammation by reducing the presence of reactive oxygen species (ROS). In normal cells, high levels of ROS are damaging to the cells and the cells have the ability to squelch the production of

ROS enzymatically. Cancer cells exhibit elevated levels of ROS and due to their accelerated metabolism needed to maintain proliferation.

The goals of this experiment were : (1) to determine the effects of laryngeal cancer cells exposed to LLLT for a period of 30 minutes to determine the effects on cell survival; and (2) to determine the effects of natural chemotherapeutic agents or a known conventional chemotherapeutic agent, 5FU along with exposure to LLLT on cancer cell growth. Cells were treated with laser, Thymoquinone

(TQ), Genistein (G), 5FU, or laser in the presence of TQ, G, or 5FU for 30 minutes followed by incubation for a period of 24 hours. The cells were harvested and cellular protein, intracellular glutathione, and morphology were evaluated. The results show a decrease in cell numbers following treatment with TQ and 5 FU for 24, 48, and 72 hours, while genistein treatment showed changes in cell number after

72 hours. Interestingly, the cells in the presence of laser were reduced within 24 hours, and treatments with Laser + 5FU, laser + G, and


laser + TQ were significantly reduced further than when given the compound alone. The results show that laser and chemotherapeutic interventions may be synergistic and beneficial treatment for laryngeal cancer. These findings are important since laryngeal cancer is difficult to resect, but laser therapy could be guided into the area to reduce the tumor size or used following resection.


Type – Number Author – Institution

Poster 44 Acharya, Dhiraj - The University of Southern Mississippi



Poster 23 Adah, Felix - University of Mississippi Medical Center

Poster 82 Adah, Felix - University of Mississippi Medical Center

Poster 82 Adah, Osasu - University of Mississippi Medical Center

Poster 45 Adams, Melissa - Copiah-Lincoln Community College

Poster 70 Ahmad, Omama - Mississippi College

Poster 71 Ahmad, Omama - Mississippi College

Poster 77 Alexander, Barbara - University of Mississippi Medical Center



Poster 14 Alexander, Chakiras - University of Mississippi Medical Center

Poster 65 Ali, Lizhuo - Millsaps College

Poster 3

Poster 3

Ali, Marina - St. Andrew's Episcopal School

Ali, Mohamed - Jackson State University

Poster 64 Allred, Anna - Millsaps College

Poster 50 Alzuway, Ahmed - The University of Southern Mississippi

Poster 51 Alzuway, Ahmed - The University of Southern Mississippi

Poster 22 Anglin, Sarah Lea - Millsaps College

Poster 48 Appukutti, Nadeema - Mississippi University for Women

Poster 34 Arick, Tony - Mississippi State University

Poster 77 Ayanwale, Lydia - Tougaloo College



Poster 73 Ayensu, Wellington - Jackson State University

Poster 44 Bai, Fengwei - The University of Southern Mississippi



Oral 5 Bandi, Pradeep - Mississippi College

Poster 67 Bandi, Pradeep - Mississippi College

Poster 65 Barker, Sara - Millsaps College

Poster 58 Basco, Maria - Mississippi State University

Poster 19 Batte, Justin - The University of Southern Mississippi

Poster 35 Batte, Justin - The University of Southern Mississippi

Poster 72 Bear, Matthew - Mississippi College

Poster 56 Bedi, Angad - Mississippi College

Poster 69 Bell Jr., Anthony - The University of Southern Mississippi

Poster 76 Bell Jr., Anthony - The University of Southern Mississippi

Poster 14 Benghuzzi, Hamed - University of Mississippi Medical Center




MS-INBRE 2014 ANNUAL RESEARCH SYMPOSIUM

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Poster 58 Bhatty, M - Mississippi State University

Poster 78 Binion, Apiffani - Tougaloo College

Poster 79 Binion, Apiffani - Tougaloo College

Poster 36 Biswas, Pradip - Tougaloo College

Poster 11 Blancett, Logan - The University of Southern Mississippi



Poster 16 Blanton, Alicia - Alcorn State University

Poster 15 Bodkins, Stacey - University of Mississippi Medical Center

Poster 77 Booth, Symone - Tougaloo College



Poster 70 Brandon, Elizabeth - Mississippi College



Poster 41 Brinkman, Katherine - Mississippi University for Women

Poster 23 Brown, Courtland - Murrah High School

Poster 53 Buford, Thomas - The University of Southern Mississippi

Poster 54 Buford, Thomas - The University of Southern Mississippi

Poster 75 Buntyn, Robert - The University of Southern Mississippi

Poster 70 Burns, Zac - Mississippi College

Poster 71 Burns, Zac - Mississippi College

Poster 13 Buti, Wisam - The University of Southern Mississippi

Poster 34 Buza, Teresia - Mississippi State University

Oral 1 Cannon, John - Carleton College

Poster 20 Carter, Kelsey - The University of Southern Mississippi

Poster 36 Chakraborty, Sandipan - Tougaloo College

Poster 13 Chen, Q. Brent - The University of Southern Mississippi

Poster 6 Chen, Q. Brent - The University of Southern Mississippi

Poster 65 Coleman, Erika - Millsaps College

Poster 31 Coleman, Ineshia - Tougaloo College

Poster 39 Coleman, Kayla - University of Mississippi Medical Center

Poster 73 Collins, Michael - Mississippi Valley State University

Poster 60 Corrao, Irene - Millsaps College

Poster 64 Corrao, Irene - Millsaps College

Oral 4 Crossley, Davida - Alcorn State University

Poster 16 Crossley, Davida - Alcorn State University

Poster 40 Crossley, Davida - Alcorn State University

Poster 11 Crossley, Davida - The University of Southern Mississippi

Poster 60 Curry, B. Woods - Millsaps College



Poster 13 Das, Sudeshna - The University of Southern Mississippi


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Poster 6 Das, Sudeshna - The University of Southern Mississippi

Poster 73 Dash, Padmanava - Jackson State University

Poster 33 Dash, Padmanava - Mississippi State University

Poster 7 Dash, Padmanava - Mississippi State University

Oral 6 Davis, Donald - Tougaloo College

Poster 65 Dawson, Kellen - Millsaps College

Poster 55 Ding, Xuan - Mississippi State University

Poster 6 Drescher, Brandon - The University of Southern Mississippi

Poster 39 Drummond, Heather - University of Mississippi Medical Center

Poster 26 Dukes, Jacquais - Alcorn State University

Poster 27 Dukes, Jacquais - Alcorn State University

Poster 25 Echols, K. Allyne - Alcorn State University

Poster 28 Echols, K. Allyne - Alcorn State University

Oral 6 Edwards, Falicia - Tougaloo College

Poster 50 El barasi, Amelsaad - The University of Southern Mississippi

Poster 51 El barasi, Amelsaad - The University of Southern Mississippi

Poster 1 El-Khattouti, Abdelouahid - University of Mississippi Medical Center

Poster 19 Elasri, Mohamed - The University of Southern Mississippi






Oral 5 Espinal, Nancy - Mississippi College

Poster 1 Espinoza, Ingrid - University of Mississippi Medical Center

Poster 21 Evans, Andrew - The University of Southern Mississippi

Poster 65 Ezelle, Karen - Millsaps College

Poster 57 Fan, Zhen - Jackson State University

Oral 3 Feng, Manliang - Tougaloo College

Poster 42 Feng, Manliang - Tougaloo College

Poster 65 Fernandez, Giancarlo - Millsaps College

Poster 48 Finch, Ariel - Mississippi University for Women

Poster 4 Finley, GeNita - Millsaps College



Poster 6 Forstall, John - The University of Southern Mississippi

Poster 81 Garner, Bianca - Tougaloo College

Poster 74 Garrett, Michael - University of Mississippi Medical Center

Poster 8 Garrett, Michael - University of Mississippi Medical Center

Poster 25 Gee, Cherrelle - Alcorn State University

Poster 28 Gee, Cherrelle - Alcorn State University

Poster 12 Ghag, Gaurav - The University of Southern Mississippi

Oral 6 Gholar, D'Asia - Tougaloo College


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Poster 37 Gholar, D'Asia - Tougaloo College

Poster 61 Giurintano, Jonathan - Millsaps College

Poster 74 Gomez-Sanchez, Elise - University of Mississippi Medical Center

Poster 1 Gomez, Christian - University of Mississippi Medical Center

Poster 42 Grace, Natalia - Tougaloo College

Poster 39 Granger, Joey - University of Mississippi Medical Center

Poster 75 Grimes, D. Jay - The University of Southern Mississippi

Oral 1 Guerrier, Sabrice - Millsaps College

Poster 2 Guillory, Devin - Jackson State University, UMC, USM

Poster 43 Guo, Yan-Lin - The University of Southern Mississippi



Poster 8 Guo, Zibiao - University of Mississippi Medical Center

Poster 74 Harmon, Ashlyn - University of Mississippi Medical Center

Oral 6

Poster 1

Harris, Kisa - Tougaloo College

Hassan, Mohamed - University of Mississippi Medical Center

Poster 46 Heda, Ghanshyam - Mississippi University for Women

Poster 48 Heda, Ghanshyam - Mississippi University for Women

Poster 70 Hegazy, Yassmin - Mississippi College

Poster 71 Hegazy, Yassmin - Mississippi College

Poster 77 Hennington, Bettye Sue - Tougaloo College



Oral 9 Hodge II, Gary - Mississippi College

Poster 56 Hodge II, Gary - Mississippi College

Oral 5 Hodge, Hallie - Mississippi College

Poster 60 Hoth, Lauren - Millsaps College

Poster 61 Hoth, Lauren - Millsaps College

Poster 44 Huang, Faqing - The University of Southern Mississippi

Poster 47 Huang, Faqing - The University of Southern Mississippi

Poster 10 Huang, Jun - University of Mississippi Medical Center

Poster 30 Humphrey, Samantha - Mississippi University for Women

Poster 33 Ikenga, Julius - Mississippi Valley State University



Poster 78 Intapad, Suttira - University of Mississippi Medical Center

Poster 79 Intapad, Suttira - University of Mississippi Medical Center

Oral 5 Irby, Joshua - Mississippi College

Poster 68 Ishimwe, Jeanne - Belhaven University

Oral 7 Isokpehi, Raphael - Jackson State University

Poster 18 Izevbigie, Ernest - Jackson State University

Oral 3 Jasmine Jennings - Tougaloo College

Poster 50 Jayana, Bina - The University of Southern Mississippi


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Poster 42 Jennings, Jasmine - Tougaloo College

Poster 74 Johnson, Ashley - University of Mississippi Medical Center

Poster 71 Kamassa, Mawusi - Mississippi College

Poster 70 Kamassa, Mawussi - Mississippi College

Poster 65 Kaminski, Amanda - Millsaps College

Poster 20 Karim, Shahid - The University of Southern Mississippi

Poster 61 Kaur, Gurjit - Millsaps College

Poster 20 Kausar, Asma - The University of Southern Mississippi

Poster 1

Poster 4

Kent Jr., James - Mississippi University for Women

Khawaja, Muzamil - Millsaps College

Poster 60 Khawaja, Muzamil - Millsaps College

Poster 62 Khawaja, Muzamil - Millsaps College

Poster 73 Kibet, Daniel - Mississippi Valley State University

Poster 4 Kramer, Wolfgang - Millsaps College






Poster 4 Lassiter, Brooke - Millsaps College

Poster 62 Lassiter, Brooke - Millsaps College

Poster 13 Leal, Sandra - The University of Southern Mississippi

Poster 6 Leal, Sandra - The University of Southern Mississippi

Poster 74 Lee, Jonathan - University of Mississippi Medical Center

Oral 5 Lewing, Benjamin - Mississippi College

Poster 29 Lewing, Benjamin - Mississippi College

Poster 67 Lewing, Benjamin - Mississippi College

Poster 59 Lowery, Jordan - The University of Southern Mississippi

Poster 67 Lu, Wanqing - Mississippi College

Oral 5 Lu, Wanquig - Mississippi College

Poster 1

Oral 3

Ma, Tangeng - University of Mississippi Medical Center

Mao, Jinghe - Tougaloo College

Poster 10 Mao, Jinghe - Tougaloo College



Poster 35 Marcos, Luis - Forrest General Hospital

Poster 5 Marshall, Donna - USDA-ARS

Poster 32 Mavrodi, Mavrodi - The University of Southern Mississippi

Poster 32 Mavrodi, Olga - The University of Southern Mississippi

Poster 41 McCorkle, Samantha - Mississippi University for Women

Poster 40 McDaniel, Shavonda - Alcorn State University


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Poster 9

Poster 2

McLean, Timothy - The University of Southern Mississippi

McPherson, Kasi - University of Mississippi Medical Center

Poster 64 McVaugh, James - Millsaps College

Poster 65 Meade, J. Chris - University of Mississippi Medical Center

Poster 56 Melacon, Benjamin - Mississippi College

Oral 9 Melancon, Benjamin - Mississippi College

Poster 6 Meriwether, Andrew - The University of Southern Mississippi

Poster 80 Mo, Yinyuan - University of Mississippi Medical Center

Poster 26 Moore, Jasmine - Alcorn State University

Poster 27 Moore, Jasmine - Alcorn State University

Oral 8 Moreno, J. Ignacio - Alcorn State University

Poster 24 Moreno, J. Ignacio - Alcorn State University






Poster 72 Morgan, Jacob - Mississippi College

Poster 6 Morgan, Sarah - The University of Southern Mississippi

Poster 74 Morris, Zaliya - University of Mississippi Medical Center

Poster 20 Mukherjee, Nabanita - The University of Southern Mississippi

Poster 60 Mullins, Courtney - Millsaps College

Poster 61 Mullins, Courtney - Millsaps College

Oral 9 Naidu, Sucheta - Mississippi College

Poster 56 Naidu, Sucheta - Mississippi College

Poster 58 Nanduri, Bindu - Mississippi State University

Poster 70 Niswonger, Alexandria - Mississippi College



Oral 5 Norcross, Erin - Mississippi College

Oral 9 Norcross, Erin - Mississippi College

Poster 29 Norcross, Erin - Mississippi College

Poster 67 Norcross, Erin - Mississippi College

Poster 7 Norwood, Tasha - Jackson State University

Poster 4 Odom, Katie - Millsaps College

Poster 62 Odom, Katie - Millsaps College

Poster 77 Ojeda, Norma - University of Mississippi Medical Center



Poster 54 Parker, Terra - The University of Southern Mississippi

Poster 65 Patel, Hirni - Millsaps College

Poster 65 Patel, Payal - Millsaps College

Poster 4 Patel, Priya - Millsaps College


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Poster 44 Paul, Amber - The University of Southern Mississippi



Poster 43 Payne, Rachel - The University of Southern Mississippi

Poster 33 Peavy, LaTia - Jackson State University

Poster 34 Peterson, Daniel - Mississippi State University

Poster 55 Peterson, Daniel - Mississippi State University

Poster 33 Pinckney, James - University of South Carolina



Oral 8 Piva, Marta - Alcorn State University

Poster 24 Piva, Marta - Alcorn State University






Poster 38 Planchard, Matthew - The University of Southern Mississippi

Oral 9 Pollard, Peyton - Mississippi College

Poster 56 Pollard, Peyton - Mississippi College

Poster 77 Porter, Nicholas - Tougaloo College

Poster 58 Pruett, Stephen - Mississippi State University

Poster 72 Qizilbash, Bilal - Mississippi College

Poster 66 Qureshi, Maryam - Millsaps College

Poster 36 Rader, Nicholas - Tougaloo College

Poster 64 Rahat, Resham - Millsaps College

Poster 36 Rajnarayanan, Rajendram - University of Buffalo

Poster 12 Rangachari, Vijay - The University of Southern Mississippi

Poster 38 Rangachari, Vijay - The University of Southern Mississippi

Oral 3 Raucher, Drazen - University of Mississippi Medical Center

Poster 42 Raucher, Drazen - University of Mississippi Medical Center

Poster 57 Ray, Paresh - Jackson State University

Poster 5

Oral 9

Rea, Ashten - William Carey University

Reagan, Jerry - Mississippi College

Poster 56 Reagan, Jerry - Mississippi College

Poster 80 Redfield, Samantha - University of Mississippi Medical Center

Poster 18 Robinson, Lecia - Jackson State University

Oral 5 Rosado, Dale - Mississippi College

Poster 29 Rosado, Dale - Mississippi College

Poster 67 Rosado, Dale - Mississippi College

Oral 1 Rosenberg, Samuel - Carleton College


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Poster 19 Sahukhal, Gyan - The University of Southern Mississippi





Poster 52 Saleh, Maunir - The University of Southern Mississippi

Poster 35 Samanta, Dhritiman - The University of Southern Mississippi

Poster 37 Saracino, Anthony - Tougaloo College

Poster 6 Saucier, Joseph - The University of Southern Mississippi

Poster 70 Schwam, Maxwell - Mississippi College

Poster 72 Schwam, Maxwell - Mississippi College

Poster 10 Scott, Ansley - Tougaloo College

Poster 9 Scott, Kelly - The University of Mississippi Medical Center

Poster 21 Semrad, Ethan - Belhaven University

Oral 6 Sengupta, Bidisha - Tougaloo College

Poster 37 Sengupta, Bidisha - Tougaloo College

Poster 76 Serrano, Crystal - The University of Southern Mississippi

Oral 4 Shearer Jr., Glenmore - The University of Southern Mississippi

Poster 11 Shearer, Glen - The University of Southern Mississippi



Poster 75 Shen, Shuo - The University of Southern Mississippi

Oral 6 Shenwu, Ming - Tougaloo College

Poster 10 Shenwu, Ming - Tougaloo College

Poster 71 Skaggs, Courtney - Mississippi College

Poster 11 Smith, Erin - The University of Southern Mississippi

Poster 13 Smith, Robert - The University of Southern Mississippi

Poster 63 Smothers, Greyson - Millsaps College

Poster 64 Solomon, Melinda - Millsaps College

Poster 47 Spangler, Joseph - The University of Southern Mississippi

Poster 22 Spell, Casey - Millsaps College

Poster 39 Stec, David - University of Mississippi Medical Center

Poster 60 Stewart, Emily - Millsaps College



Poster 72 Taitano, Matthew - Mississippi College

Poster 10 Tang, Xuehui - University of Mississippi Medical Center

Poster 17 Tchounwou, Paul - Jackson State University

Poster 18 Tchounwou, Paul - Jackson State University

Poster 47 Teng, Chengwen - The University of Southern Mississippi

Poster 6 Toranzo, Randy - The University of Southern Mississippi

Poster 69 Totsingan, Filbert - New York University

Poster 76 Totsingan, Filbert - New York University


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Poster 65 Toyota, Cory - Millsaps College

Poster 66 Toyota, Cory - Millsaps College

Poster 14 Tucci, Michelle - University of Mississippi Medical Center




Oral 2 Walker, Scharri - Tougaloo College

Poster 80 Wang, Chunyi - University of Mississippi Medical Center

Poster 34 Wang, Hui - Mississippi State University

Poster 43 Wang, Jundi - The University of Southern Mississippi



Poster 44 Wang, Ruoxing - The University of Southern Mississippi

Poster 47 Wang, Ruoxing - The University of Southern Mississippi

Poster 74 Wang, Xuexiang - University of Mississippi Medical Center

Oral 6 Ward, Denise - Tougaloo College

Poster 37 Ward, Denise - Tougaloo College

Poster 75 Warren, Reid - The University of Southern Mississippi

Poster 39 Warrington, Junie - University of Mississippi Medical Center

Poster 46 Watson, Deanna - Mississippi University for Women

Poster 48 Watson, Deanna - Mississippi University for Women

Poster 58 Wei, T - Mississippi State University

Poster 30 Whitwam, Ross - Mississippi University for Women

Poster 41 Whitwam, Ross - Mississippi University for Women

Oral 5 Wilcosky, Amy - Mississippi College

Poster 29 Wilcosky, Amy - Mississippi College

Poster 81 Williams, Baraka - Tougaloo College/Jackson State University

Poster 65 Williams, Brad - Millsaps College

Poster 20 Williams, Jaclyn - The University of Southern Mississippi

Poster 2 Williams, Jan - University of Mississippi Medical Center

Oral 5 Williams, Roshetta - Mississippi College

Poster 67 Williams, Roshetta - Mississippi College

Poster 65 Williams, S. Skye - Millsaps College

Poster 77 Williamson, Danielle - Tougaloo College



Poster 14 Wilson, Gerri - University of Mississippi Medical Center



Poster 41 Winders, Jeremy - Mississippi University for Women

Oral 3 Wu, Ming-Shen - Tougaloo College

Poster 42 Wu, Ming-Shen - Tougaloo College

Poster 10 Yang, Chunli - University of Mississippi Medical Center


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Oral 7 Yedjou, Clement - Jackson State University

Poster 17 Yedjou, Clement - Jackson State University

Poster 18 Yedjou, Clement - Jackson State University

Poster 81 Youngblood, Claressa - Tougaloo College

Poster 65 Zamarippa, C. Austin - Millsaps College

Poster 10 Zhou, Wu - University of Mississippi Medical Center

Poster 80 Zhou, Xinchun - University of Mississippi Medical Center

Poster 10 Zhu, Hong - University of Mississippi Medical Center

Poster 13 Zong, Yan - The University of Southern Mississippi

Poster 6 Zong, Yan - The University of Southern Mississippi

_____

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Document 14570793

WELCOME

Glen Shearer

MS-INBRE 2014 ANNUAL RESEARCH SYMPOSIUM

MS-INBRE 2014 ANNUAL RESEARCH SYMPOSIUM

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