pr-upp

Poster Presentations

Undergraduate Poster Presentations

UPP01

FTIR Analysis of Forensic Drug Cannabinol Detection using Gold Nanoparticles

Felesia Dukes, Britnie Hanley, and Harvey J.M. Hou*

Department of Physical/Forensic Sciences, Alabama State University, Montgomery, Alabama 36104. *Email: hhou@alasu.edu

The most trafficked illegal drug is marijuana and cannabinol is one of main metabolites of marijuana (Levin 1994). Nanoparticles, especially functionalized gold nanoparticles, are promising materials in biomedical and energy research as well as in forensic science (Tiwari et al 2011, Hou 2015, Hou et al 2017). We have previously examined the structural changes of marijuana at the elevated temperature by use of UV-vis spectrometry (Phillips 2013). In this work we explored the feasibility of FTIR detection of cannabinol using gold nanoparticles. The objective of the work is to examine the possibility for detecting cannabinol using nanoparticles and to advance the understanding of the chemical binding interaction between the nanoparticles and cannabinol. As shown in Figure 1, cannabinol showed 33 IR peaks, which tentatively assigned to the benzene ring (655, 691, 736, 814, 974, 1584, and 1626 cm-1), O-H group (1362, 1427, 3389 cm-1), C-O bond (1029, 1132, 1158, 1197, 1231, 1285, and 1383 cm-1) and alkyl group (1305, 1404, 1457, 1494, 1981, 2855, 2928, 2959 cm-1). In the presence of gold nanoparticles, comparable IR signals were found, suggesting cannobinol detection is achievable. The changes of multiple IR peaks of cannobinol after adding gold nanoparticles were observed, implying the potential binding sites of nanoparticles. The data infer that the possible binding site of the gold nanoparticles are the benzene ring and the tail of the alkyl chain (see the insert of Figure 1). The information may be used for designing new functionalized nanoparticles in cannabinol detection.

Figure 1. FTIR spectrum of cannabinol in the absence of nanoparticles. The insert is a tentative interaction model for cannobinol and gold nanoparticles

Hou HJM (2015) CSI at ASU: Probing forensic evidences using nanotechnology, Science @ ASU, pp. 16-17

Hou HJM, Najafpour MM, Moore GF, and Allakhverdiev SI, "Photosynthesis: Structures, Mechanisms, and Applications," Springer, 2017

Levine B (2014) Principles of Forensic Toxicology, 4th Ed., AACC Press

LaFaith Phillips (2013) Structural changes of marijuana and cannabinol under diverse conditions, M.S. Thesis, Alabama State University

Tiwari PM, Vig K, Dennis VA and Singh SR (2011), “Functionalized gold nanoparticle and their biomedical applications, Nanomaterials, 1, 31-63.

UPP02

A Possible Binding Model of Forensic Drug Cocaethylene and Gold Nanoparticles

James Ogoniba and Harvey J.M. Hou*

Department of Physical/Forensic Sciences, Alabama State University, Montgomery, Alabama 36104. *Email: hhou@alasu.edu

Cocaethylene is a main metabolite formed during the combined consumption of cocaine and ethanol and causes effects on brain systems (Horowitz et al 1999). In the last two Annual Research Symposia in 2015 and 2016, we have reported the possible interaction models of forensic drug cocaine and lorazepam with nanoparticles, respectively (Hanley et al 2015, Nance et al 2016). The FTIR data showed a red shift of benzene from 716 to 710 cm-1, suggesting that the binding site of cocaine is likely phenyl group (Hanley 2015). However, we noticed that the IR signal at 716 cm-1 of cocaine in the presence of gold nanoparticle is apparently very small. It may require further experimental evidence to verify or disapprove the hypothetical binding model of cocaine. In this work we analyzed the FTIR data of cocaethylene in the presence of gold nanoparticles. As shown in Figure 1, it is evident that the IR peak at 715.5 cm-1 has red shifted to 713.5 cm-1. The changes of IR peaks of cocaethylene caused by adding gold nanoparticles are summarized and interpreted. Based on these data we proposed the possible binding model of cocaethylene and gold nanoparticles (see the insert of Figure 1). The benzene ring of cocaethylene may bind to the nanoparticles in a similar manner as cocaine previously suggested. The future endeavors will be placed on examination of various functionalized gold nanoparticles and other types of nanomaterials for detection of forensic drugs.  

Figure 1. Red shift of IR peak at 715 cm-1 of cocaethylene after adding gold nanoparticles and possible binding model of cocaethylene and gold nanoparticles.

Hanley B and Hou HJM, "Forensic drug analysis using nanotechnology," Annual Research Symposium, Alabama State University, March 18-19, 2015

Horowitz JM and Torres G (1999) Cocaethylene: Effects on brain systems and behavior, Addiction Biol., 4, 127-140

Nance C and Hou HJM, "Chemical analysis of forensic drug lorazepam using nanoparticles," Annual Research Symposium, Alabama State University, March 16-17, 2016

UPP03

Chemical Detection of a Sunlight-Induced Degradation Product at Retention time 7.7 min of Cocaine in Ethanol

Jumaia Todd, Anthony Asonze, and Harvey J.M. Hou*

Department of Physical/Forensic Sciences, Alabama State University, Montgomery, Alabama 36104. *Email: hhou@alasu.edu

Cocaine is one of the top illicit drugs and increases risk of stroke, teeth abrasion, and heart infarction (Levine 2014, De Giorgi et al. 2012). We previously investigated the effects of elevated temperature for 8 h on cocaine in methanol and found one small GC peak at the retention time 7.43 min (Barrow et al 2013). In the 2017 Annual Research Symposium we analyzed the structural changes of cocaine when exposed to sunlight in a car for a week using GCMS and FTIR (Asonze et al 2017). The degradation product at 7.43 min was tentatively identified to be pseudoallococaine. In this work we further examined the GC-MS data of cocaine degradation reaction treated by sunlight. One GC component around the retention time 7-8 min are observed at retention time 7.72, 7.68, 7.63, and 7.61 min, varied over time of treatment (Figure 1). The identification of the GC peaks at 7.72 and 7.68 min via library search confirmed the previous conclusion of pserdoallococaine. However, the library search for the GC peaks at 7.63 and 7.61 min failed. We noticed that the MS spectra of the degradation product at 7.63 and 7.61 min showed two new MS peak at m/z 31 and m/z 45, which might be associated with the formation of alcohol and carboxylic acid, respectively. We proposed the possible degradation products of cocaine in ethanol by sunlight are ecgonine methyl ester and benzoyl ecgonine. These data may offer useful information on the cocaine detection in forensic community.

Figure 1. Effects of sunlight on forensic drug cocaine solution in ethanol for one week

Asonze A and Hou HJM, "GCMS and FTIR analysis of forensic drug ecgonine methyl ester treated by sunlight," Annual Research Symposium, Alabama State University, March 15-16, 2017

Barrow S (2013) Response mechanisms of cocaine and gunshot residues to environment, M.S. Thesis, Alabama State University

De Giorgi A, Fabbian F, Pala M, Bonetti F, Babini I, Bagnaresi I, Manfredini F, Portaluppi F, Mikhailidis DP and Manfredini R (2012) Cocaine and acute vascular diseases, Curr. Drug Abuse Rev., 5, 129-134

Levine B (2014) Principles of Forensic Toxicology, 4th Ed., AACC Press

UPP04

Synthesis of Imidazo[1,2-a]pyridine Ligands for Use in Difficult Cross-Coupling Reactions

William Swann and Larry Yet*

Department of Chemistry, University of South Alabama, Mobile, Alabama. 36608. *Email: lyet@southalabama.edu

Developing new or more efficient methods of synthesis is a main area of focus in organic chemistry. Imidazo[1,2-a]pyridine phosphorus ligands are being investigated as aids in cross-coupling reactions. Recent advancements show that, when paired with transition metals, electron-rich moieties allow for increased specificity and activation potential within previously difficult reactive mechanisms. Structural changes to the aforementioned scaffold were made with the addition of aromatic groups bearing different functional groups to the C-2 position of the heterocycle. Diphenyl-, di-tert-butyl-, and dicyclohexylphosphines were then added via halogenated intermediates to the C-3 position of imidazo[1,2-a]pyridine core creating highly varied ligands for use in future cross-coupling reaction studies.

UPP05

Optimization of Micro-LIBS for the Elemental Analysis of Glass

Christopher Ball and Cleon Barnett*

Department of Physical/Forensic Sciences, Alabama State University, Montgomery,

Alabama 36104. *Email: cmbarnett@alasu.edu

Laser-induced breakdown spectroscopy is a proven effective analytical tool for elemental analysis of a variety of materials. Although LIBS is considered a minimally destructive method, using less energy and reducing the spot size may further reduce damages caused during the analysis. This may be advantageous when working on precious materials or forensic samples. Typical Micro-LIBS produces craters that have diameters and depths that are much less than conventional LIBS. One of the smallest reported crater sizes was 3 mm in diameter. The purpose of this experiment is to implement a procedure in which less material of the sample is destroyed in the LIBS process, but can still be used to get accurate elemental data for samples with a range of transparencies. In this study, a 10X microscope objective is used to create LIBS plasmas on 5 glass standard reference materials. NIST standard reference materials 610, 612, 614, 620 and 1831 were used in this study.  The Nd:YAG laser is operated at 532 nm and experiments are performed at energies less than 1.50 mJ. In this experiment, the emission line from Strontium at 407.76 nm is used to optimize experimental conditions. We report on the influence that the lens to sample distance has on the intensity and percent relative standard deviation of the emission. 

UPP06

The Effect of Sodium Butyrate on Exosomal Production in Breast Cancer Cells

Qiana L. Matthews1, Gina Taylor2, Courtnee’ R. Bell1, and Sabita N. Saldanha2,*

1Microbiology Doctoral Program and  2Department of Biological Sciences, Alabama State University, Montgomery, Alabama 36104. *Email: ssaldanha@alasu.edu

Butyrate, a fatty acid obtained by the fermentation of dietary fiber has been shown to be beneficial to the colonic microenvironment and is considered a safe bioactive molecule. It has also been shown to induce differentiation and apoptosis in cell culture environments and is a potent histone deacetylase inhibitor, an epigenetic property that has crucial role in gene regulation. These mechanistic properties make it a suitable anti-cancer agent as well. Exosomes are vesicular structures that carry cargo (proteins or epi-factors) that can regulate cellular function both in-situ or at distant locations. Cancer proliferation and metastasis is also known to be mediated and extended by exosomes based on the cargo they carry. Triple negative breast cancer is a debilitating disease with no known absolute treatment or cure. Developing therapeutic strategies is therefore imperative. Our study is aimed at understanding the influence of sodium butyrate (NaB) on exosomal production and composition. Specifically, our study will evaluate the effects of sodium butyrate on the cell-type specific cargo packaging of exosomes as well as the epigenetic process influenced by the cargo in mediating cell differentiation. Our study showed that administration of 1 mM sodium butyrate after 48 h brought about change in the cellular morphology of MCF-7 and MDA-MB-231 cells and mediated a change in exosomal production in both MCF-7 and MDA-MB-231 breast cancer cells. Administration of concentrations of sodium butyrate at 500 _M and 1 mM, respectively, yielded a very slight increase of exosomal production from MCF-7 cells. However, after treating MDA-MB-231 cells with NaB at 500 _M and 1mM concentrations a decrease in exosome production was observed from this cell line. This preliminary data is interesting as MCF-7 is ER-positive cancer cell type and MDA-MB-231 is triple negative for the receptors. It could be that the receptor status may have a bearing on the response to exosomal production in these cell lines on treatment with butyrate. Further, we will analyze the changes in protein production and exosomal content of this induction in both the cell types.

UPP07

Endosomal Escape of Bacteriophage in MDA-MB 231

Mashunda Longmire1, Deepa Bedi2, and Komal Vig1,*

1Center for Nano Biotechnology Research, Alabama State University, Montgomery, Alabama 36104; 2Department of Biomedical Sciences, Tuskegee University, Tuskegee, Alabama 36088. *Email: komalvig@alasu.edu

Bacteriophage also known as “phage” is a virus that infects a bacteria and reproduces inside it. Phage display is important in cell targeting and therapy. Phage display targets selected proteins on the surface of phage. The purpose of this study was to select phages from the phage library which will specifically bind to MDA-MB 231 cells and can bypass endosomes. Chloroquine phosphate is the endosomal inhibitor that was used to disrupt the endosome in the MDA-MB 231 breast cancer cells. A series of phage selections on MDA-MB 231 breast cancer cells were performed on cells with the endosomal inhibitor, chloroquine. Binding assays were performed to investigate if the selected phages could bind specifically to the MDA-MB 231 cells. Three rounds of phage selection were performed. In the first round phages were sequentially selected to plastic, to serum and to MDA-MB 231 cells to acquire cancer cells binding specific phages. Cells were washed with elution buffer to wash unbound phages followed by lysis buffer to break the cells to release selected phages. Phage tittering was performed on all washes, input, eluate and lysate using E.coli and plaque colonies were counted. The selected phages in lysate and eluate were amplified using E. coli. Amplified phages were centrifuged and supernatant containing phages was precipitated using PEG NaCl. These steps were repeated in second and third round of phage selection in MDA-MB 231 cells. Binding of the phage to the target MDA-MB231 cells in the third round were tested by phage ELISA using phage specific M13 antibody. ELISA results show 3.8 times higher binding to the MDA-MB231 cells compared to untreated cells. Phages will be further tested for their ability to escape endosomes. This work was supported by US Dept. of Education, The Minority Science and Engineering Improvement Program (MSEIP) (P120A150008) to Dr. Komal Vig (PD) and by NSF-CREST (HRD-1241701) to Dr. Shree R. Singh (PI). 

UPP08

The In Vitro Effects of Alcohol on Exosome Biology

Leandra B. Jones1, Aliyah J. Curry2, Sparkle D. Williams3, Alexandre Krendelchtchikov3, Brian Sims3, and Qiana L. Matthews1,*

1Microbiology Program, Department of Biological Sciences, Alabama State University, Montgomery, Alabama 36104; 2Department of Biological Sciences, Alabama State University, Montgomery, Alabama 36104; 3Departments of Pediatrics, Neurobiology and Cell, Developmental and Integrative Biology, Division of Neonatology, University of Alabama at Birmingham, Birmingham, Alabama 35294. *Email: qmatthews@alasu.edu

Exosomes are extracellular vesicles that are active in cell-to-cell communication, transferring macromolecules between cells, biological markers used to detect disease states. Exosomes are found in extracellular space and made up mostly of lipids and proteins. Exosomes can be affected by external factors such as alcohol and nicotine.  Specifically, we will be focusing on the effect of alcohol on exosome biogenesis. Alcohol has factors that can be beneficial to the body in moderation. However, alcohol in high concentrations can negatively alter extracellular vesicles production and release. Our goal is to study the impact of alcohol exposure on kidney exosome biology.  Human kidney cells (293A) were subjected to (control), 50 mM, or 100 mM of Ethanol for 24 h, 48 h or 72 h. Cell viability was observed at 72 h post Ethanol treatment. 293A cell viability was significantly decreased with treatments of 50 mM or 100 mM of Ethanol. Exosomes were purified using an ultracentrifugation after dosing with alcohol or vehicle control.  The protein quantity and quality was determined using standard protein quantitation methods.  NanoSight technology was used to measure exosome count and characterize exosomes along with Enzyme-Linked immunosorbent assay (ELISA). We observed that when 293A cells were treated with alcohol, exosome biogenesis was impacted, we observed a decrease in exosome production over time as well as an increase in exosomes carrying HSP60 and HSP70 proteins. Overall, these results suggest alcohol has a negative affect on cells leading to the downstream impact on exosomes. This supports the claim that alcohol can have detrimental effects on the human body.  Our future research includes investigating the effects of alcohol consumption on exosome production in vivo.

UPP09

Synthesis of Water Soluble Zinc Oxide Quantum Dots for Biomedical Applications

Gregory Bivens1, Shreekumar Pillai2, Shree Singh2, and Sapna Jain1,*

1Department of Physical/Forensic Sciences, Alabama State University, Montgomery, Alabama 36104; 2Center for Nanobiotechnology Research, Alabama State University, Montgomery, Alabama 36104. *Email: sjain@alasu.edu

Zinc oxide (ZnO) quantum dots have been extensively used for various biomedical applications including imaging. They are inexpensive, biodegradable, and possess optical and semiconducting properties. The current work aims at synthesis of core-shell ZnO quantum dots with polyethylene glycol (PEG) as shell to improve its solubility in aqueous solutions and decrease toxicity. Synthesis is being conduct at room temperature and PEG 2000 molecular weight has been used for functionalization. Characterization has been done using FT-IR, SEM, UV-visible spectroscopy and zeta sizer and zeta potential analyzer. Fluorescence properties before and after PEG coating will be tested using Fluorescence microscopy.

UPP10

Modulation of Hsp70-Mediated Cytoprotection by Resveratrol and Staurosporine in Colon Cancer Cells

Sanjay Kumar1, Natalie White1,2, Karyn Scissum Gunn1, Manoj Mishra1, Upender Manne3,

and Sabita N. Saldanha1,2,*

1Cancer Biology Research and Training, Department of Biological Sciences, Alabama State University, Montgomery, 36101 Alabama 36101; 2Department of Biological Sciences, Alabama State University, Montgomery, Alabama 36101; 3Wallace Tumor Institute, University of Alabama at Birmingham, Alabama 35294. *Email: ssaldanha@alasu.edu

Colon cancer is a predominant cancer among African American (AA) men as compared to Caucasian men with a higher death rate in AA from the disease. Preventing the progression and metastasis of colon cancer by targeting molecules that contribute to cell survival and proliferation presents a suitable chemotherapeutic approach of the disease. In colon cancer, overexpression of heat shock protein (Hsp70) encourages the progression and metastasis of the disease. Over expression of hsp70 involves anti-apoptotic roles and induce the growth of colon cancer. Therefore, hsp70 has emerged as drug targets for colon cancer therapy. Our studies aimed at looking at the chemotherapeutic effects of naturally occurring molecules resveratrol (RES) and staurosporine (STS) on the expression of Hsp70, which are potent cytotoxic drugs. We hypothesize that the use of natural compounds will block the Hsp70-mediated protection in colon cancer. To accomplish this, HCT116 WT and SW480 colon cancer cells were treated individually with RES (100_M) and STS (1_M), for 24h, and cell killing, morphology, mitochondrial potential (__m), and expression of hsp70 was studied. Our preliminary results indicated that RES and STS treated cells showed altered cell morphology and enhanced cell killing. A decrease in protein expression of Hsp70 was observed in treated HCT116 WT and SW480 cells. Multi-drug resistance -1 (MDR-1) is upregulated in many cancers, including colon cancers, contributing to treatment resistance. Therefore, real time mRNA expression of MDR-1 will be performed to determine the effect of decreased Hsp70 on MDR-1 expression at the transcriptional level. Together, these findings will illustrate the importance of these agents as potential therapeutic molecules in controlling colon cancer

UPP11

Cage Isomers of B10N14: Stability versus Structure

Melanie Walker, Kelvin Jones, DaiQuan Noble, Marquavias Walker, and Douglas L. Strout*

Department of Physical Sciences, Alabama State University, Montgomery, Alabama 36104. *Email: dstrout@alasu.edu

Boron nitride is a material that is isoelectronic to carbon and adopts chemical structures similar to carbon, including three-dimensional cages analogous to the carbon fullerenes.  The 24-atom molecule B12N12 thermodynamically prefers a cage structure over other isomers, and one dominant cage isomer exists: a cage consisting entirely of squares and hexagons in which boron atoms and nitrogen atoms occupy alternating positions.  For B12N12, boron atoms only bond to nitrogen atoms and vice versa.  For B10N14, on the other hand, the situation is far less clear.  A number of cage structures exist that are thermodynamically comparable.  While these molecules observe the isolated boron atom rule (IBAR), nitrogen atoms of necessity bond to each other.  The properties of several of these structures are calculated and discussed, along with the principles governing their geometric makeup.

UPP12

Gold Nanoparticles Decrease Viability of S. pneumoniae

Shannon P. Williams1, Veolanda A. Peoples1,2, and Mamie T. Coats1,2,*

1Department of Biological Sciences and 2Center for Nanobiotechnology Research, Alabama State University, Montgomery, Alabama 36104. *Email: mcoats@alasu.edu

Streptococcus pneumoniae is the pathogenic bacterium that has been found to cause diseases including, ear and sinus infections, meningitis, and pneumonia. These infections are typically treated with antibiotics, but due to the highly adaptive nature of S. pneumoniae there is a need for new antimicrobials that can avoid resistance while effectively inhibiting the growth of the microbe. The purpose of this study was to investigate the efficacy of gold nanoparticles (AuNPs) as an inhibitor of S. pneumoniae. Pneumococcal strains D39 and MTC553 (penicillin resistant) were grown in Todd Hewitt Media supplemented with 10% yeast extract.   AuNPs (spherical, 10-20nm) were added at a final concentration of 1mg/ml. The samples were incubated at 37°C for 4 or 24 hours. Following incubation the number of live, non-adherent bacteria was measured using viable colony counts. A 0.1% crystal violet solution was used to stain the resulting biofilms and absorbance was read at OD600 nm. At the 4-hour timepoint the AuNP caused a statistically significant decrease in the viable non-adherent bacteria for D39 (P=0.0148).  The data for MTC553 trended towards significance at the same point. Both strains had a statistically significant decrease in the viable non-adherent bacteria at the 24 hours (P<0.05). Further studies are needed to investigate and quantitate the biofilm formation in detail. This work is being supported by National Science Foundation-CREST (HRD-1241701), NSF-HBCU- UP (HRD-1135863), NSF-AGEP (HRD-1432991) and National Institutes of Health-MBRS-RISE (1R25GM106995-01) grant, as well as the United States Department of Education and The Minority Science and Engineering Improvement Program (MSEIP) (P120A150008) at Alabama State University.

UPP13

Determining the Presence of Hg in Contaminated Soils Using Laser Induced Breakdown of Spectroscopy (LIBS)

Emaszhai Henry1, Boakai K. Robertson1,*, and Cleon Barnett2,*

1Department of Biological Sciences and 2Department of Physical Sciences, Alabama State University, Montgomery, Alabama 36104. *Email: cmbarnett@alasu.edu, brobertson@alasu.edu

It has been proven that high levels of methylmercury are present as a result of waste from the Y-12 National Security Complex at Oak Ridge National Laboratory in Tennessee. This extreme amount of contamination has raised concerns from the Department of Energy (DOE), Tennessee Department of Environment and Conservation (TDEC) and the Environmental Protection Agency (EPA) for a high-risk priority clean up due to the health implications for human and environmental health. An experiment was conducted using Laser Induced Breakdown of Spectroscopy (LIBS) technology to quantify the amount of mercury in soil samples, mercury sulfide(HgS) and samples collected from a control site in Oak Ridge, TN. A dilution process was used to produced five pellets with varying amounts of HgS in the soils samples. The pellets were completed at Florida Agricultural & Mechanical University physics laboratory. Strong emission lines for mercury were 253.65 nm, 435.00 nm and 546.07 nm, however iron emission and other elements may present significant spectral interference to the Hg lines. Magic Plot application was used to analyze data collected from Andor Software. The most abundant elements found were iron (Fe) and Tungsten (W), which are natural elements found in soil. Mercury (Hg) was present but at very low concentrations which may be below the detection limits of our current setup. With the use of the HgS, we will determine the limit of detection of Hg in soils samples.

UPP14

3-D Articular Cartilage Scaffolds Based on PVA/PLA Composites

Monet Wilson, Antwan Parker, and Derrick Dean*

Biomedical Engineering Program, Alabama State University, Montgomery, Alabama 36104. *Email: ddean@alasu.edu

 Articular cartilage is a smooth, transparent tissue that covers the ends of bones in joints such as the knee and elbow. Defects and loss of cartilage can be caused by traumatic injuries and degenerative joint diseases.  Unfortunately because of its lack of vasculature, cartilage has a limited capacity to undergo self-repair.  While some treatments are available to repair defective cartilage, they do not present a long term solution.  Tissue engineering presents an approach to synthetically develop replacements for articular cartilage.  The aim of this research is to fabricate 3-dimensional scaffolds that mimic the complex architecture and biochemical nature of articular cartilage. Methods:  I have fabricated scaffolds based on electro-spun nanofibers of poly lactic acid (PLA) infused with a hydrogel system based on polyvinyl alcohol (PVA) and sodium alginate (SA).  A highly porous, nonwoven mat of PLA was prepared using electrospinning.  A hydrogel coating of approximately 500 microns was applied to the surface of the PLA mat using a syringe.  The gel coating was then crosslinked by exposing it to a calcium chloride solution. The microstructure, chemical composition, mechanical properties and cell attachment and proliferation have been studied. Microscopy of the scaffolds reveals a two-phase, porous structure. A gradient in the structure and properties is also evident, with good mixing at the interface between the two phases.  The chemical composition of the gradient was characterized using infrared spectroscopy.  The data confirms the presence of the PLA and the hydrogel system.  The coated scaffold exhibited a modulus (i.e. stiffness) that was 20% higher than the uncoated scaffold.  Therefore the coating enhanced the mechanical properties of the electrospun scaffold. We have fabricated a hybrid, three dimensional scaffold that mimics the structure and of articular cartilage.  The samples exhibit and two phase system with interconnected porosity in each phase.  The sample also exhibits a well-defined, diffuse interphase region.  Future work will involve using a 3-D printer to precisely control the thick ad pore geometry of the hydrogel coating and characterize the mechanical properties. Supported by NSF/ CBET/RUI award 1510479.

Gibas I and Janik H (2010) Review: synthetic polymer hydrogels for biomedical Applications, Chem. & Chem. Technol., 4, 4

Rezwana K, Chena QZ, Blakera JJ, and Boccaccinia AR(2006) Biodegradable and bioactive porous polymer/inorganic composite scaffolds for bone tissue engineering, Biomaterials 27, 3413–3431

Hosseinzadeh H (2013) Synthesis and swelling properties of a poly (vinyl alcohol)-based super absorbing Hydrogel, Curr. Chem. Lett., 2, 153–158

Izadifar Z, Chen X, and Kulyk W (2012) Strategic Design and Fabrication of Engineered Scaffolds for Articular Cartilage Repair, J. Funct. Biomater. 3, 799-838.

UPP15

Effect of Scaffold Architecture and Structure on Properties and Degradation

Jordin Marshall and Derrick Dean*

Biomedical Engineering Program, Alabama State University, Montgomery, Alabama 36104. *Email: ddean@alasu.edu

Nanomaterial synthesized scaffolds have great potential in tissue engineering and are gaining importance in reconstructive surgery for anterior cruciate ligament (ACL). For tissue engineering biodegradable and biocompatible scaffolds are ideal. The scaffolds should possess certain surface characteristics, particularly hydrophilicity, to promote cell adhesion. Scaffolds should also have adequate mechanical properties to maintain the three-3-D structure and should grade gradually upon formation of new tissue and an extracellular matrix. Ideal scaffolds should be highly porous with large interconnected pores to facilitate cell growth and diffusion of nutrients and waste products into and out of the scaffold. Our research investigates properties and degradation behavior of a porous polyester tissue scaffold. Scaffolds were exposed to simulated body conditions for a period of 90 days and the morphology and physical properties were investigated.  Moisture uptake studies revealed a two step mass gain process, characterized by a rapid gain initially, a period of induction and a more subtle mass increase.  This was accompanied by significant increases in the sample dimensions, notably the thickness.  Compression tests were conducted to evaluate the effect of the simulated body conditions on the stiffness and it was found to decrease with length of exposure time.  The extend of degradation due to hydrolysis is being investigated.   Future studies will investigate the effect of varying the porosity on degradation.

UPP16

The Effect of Simulated Body Fluid Exposure on the Structure and Properties of 3-D Printed Polymeric Scaffolds

Alexis Scott, Elijah Nyairo, and Derrick Dean*

Biomedical Engineering Program, Alabama State University, Montgomery, Alabama 36104. *Email: ddean@alasu.edu

Additive manufacturing, also known as 3-D printing, is revolutionizing many fields, including the field of tissue engineering.  The ability to fabricate tissue scaffolds and tissues with complex structures is an appealing feature of this technique.  Polymeric scaffolds fabricated using 3-D printing are finding numerous applications for a wide range of tissues. Polymeric tissue scaffolds have shown great potential in tissue engineering and are gaining importance for a range of applications.  Ideal scaffolds should possess interconnected pores and surface characteristics (eg., hydrophilicity) to promote cell adhesion and growth.  Scaffolds should also have adequate mechanical properties to maintain the three-3-D structure as well as biodegradability.  Our research investigates the effect of a simulated body fluid (SBF) on the degradation behavior of polymeric tissue scaffolds. A number of scaffold materials and architectures and porosities are under investigation, including polylactic acid (PLA) and PVA based hydrogels.  The effect of SBF exposure on crystallinity, modulus, mass gain and mechanical strength have been investigated.  These properties were correlated with structural changes, as characterized by infrared spectroscopy.  Studies of the crystallinity show that 3D printed samples exhibit lower crystallinity compared to conventional processing methods.  This leads to slightly lower mechanical properties.  The effect of the crystallinity on properties and its impact on degradation behavior will be presented.

UPP17

Model of the Lorenz Attractor Through a Non-autonomous Linear System

Korynn Claiborne1, Andre Souza2, and Rajendran Swamidurai1,*

1Center for Nano Biotechnology Research, Alabama State University, Montgomery, Alabama 26104; 2Georgia Institute of Technology, Atlanta, Georgia 30332. *Email: rswamidurai@alasu.edu

Today, many weather prediction models uses nonlinear ordinary differential equations. Due to extreme sensitivity to initial conditions, it is difficult to use these models for long-term weather prediction. In this research we are planning to use numerical integration techniques and big data analytics to construct a long-term weather prediction model. We changed the system into a non-autonomous linear system by modeling the flow field “x” from the Lorenz equations as different functions, specifically periodic orbits, so that we could approximate the equations through time. We numerically approximate the solutions to the Lorenz equations using the Fourth Order Runge-Kutta. Different initial conditions of the Lorenz equations lead to drastically different long-term behaviors; however, it is possible to estimate the proportion of time spent at a certain location throughout a full trajectory. This leads us to study the statistics of the trajectories of the Lorenz equations. By using symmetry reductions and closure relations for the higher-order statistics, we were able to model the statistics for the periodic orbits in order to compare them to the original Lorenz statistics. Specific initial conditions for the Lorenz systems corresponding to periodic orbits were obtained. The negative side of the periodic trajectory is referred to as A, and the positive side is referred to as B. We separated 8 periodic orbits into the “A” and “B” loops, then added randomness by simulating a coin flip through computer code that would choose which segment to run through. We gathered the statistics of the model for 1,000 flips.The model was accurate in replicating the averages from the Lorenz equations, but not all the covariances. The model was able to replicate similar averages because of symmetry. The models were not able to replicate the variances and covariances because the trajectories did not spend the same amount of time in the locations similar to the Lorenz attractor.

Viswanath D (2003) Symbolic dynamics and periodic orbits of the Lorenz attractor http://www.math.lsa.umich.edu/divakar/papers/Viswanath2003-LorenzSymDyn.pdf

Edward Lorenz E (1963) Deterministic Nonperiodic Flow. J. Atmospheric Sci., 20, 130-141. _

Kraichnan R (1994) Anomalous Scaling of a Randomly Advected Passive Scalar. Phy. Rev. Lett., 7, 1016-1019. _

Allawala A and Marston JB. Statistics of the Stochastically-forced Lorenz attractor by the Fokker- Plack equation and cumulant expansions. https://arxiv.org/pdf/1604.00867.pdf.

Sauer T (2013) Computational Solution of Stochastic Dierential Equations. Wiley Periodicals, Inc, 5, 362-371. _

UPP18

Inhibition of Human Respiratory Syncytial Virus in HEp-2 Cells Using T118-Gold Nanoparticles

Michael Wallace and Komal Vig*

Center for Nano Biotechnology Research, Alabama State University, Montgomery, Alabama 36104. *Email: komalvig@alasu.edu

Human respiratory syncytial virus (RSV) is a crucial cause of respiratory tract infection and commonly affects the lungs and breathing passage. RSV can have life threating effects on elderly people, children, and people with compromised immune systems. Key components of RSV are the fusion protein (F protein) and the attachment protein (G protein), which are both responsible for the attachment, fusion, budding, and spread of the virus. With the use of gold nanoparticles and an anti-RSV fusion peptide (T118), the virus can be effectively inhibited from binding to cells. In this study, carboxyl-Poly (ethylene glycol) (PEG)-gold nanoparticles of 10 nm core size and the anti-RSV peptide were conjugated using EDC/NHS chemistry to further hinder the virus from binding to cells. The functionalized nanoparticles were characterized by UV-Vis/DLS to confirm conjugation. Prior to use, the safety of the nanoparticles was evaluated by MTT assay. The gold nanoparticles showed less than 25% decrease in viability with up to 50 µg/ml as compared to untreated control cells. They were then evaluated for their effectiveness against RSV using immunofluorescence and plaque reduction assays. For this, HEp-2 cells were infected with a mixture of RSV (103 PFU/mL) and various concentrations of nanoparticles. Our results showed that gold nanoparticles were able to reduce RSV infection with about 50% inhibition obtained with 50 µg/ml T118-functionalized nanoparticles. These results highlight the potential of gold nanoparticles as antiviral agents that could be useful to stop the infection and spread of RSV.

UPP19

The Effects of Nicotine on Exosome Composition

Jayde Price, Aliyah Curry, and Qiana Matthews*

Microbiology Program, Department of Biological Sciences, Alabama State University, Montgomery, Alabama 36104. *Email: qmatthews@alasu.edu

Exosomes are extracellular vesicles that act as biomarkers of certain diseased states. Specific characteristics of these extracellular vesicles have been found to be elevated in particular diseased states including malaria, various cancers, and autoimmune diseases. Our objective is to measure the effects of nicotine on exosome biogenesis in liver cells. We hypothesize that nicotine administration will affect exosome biogenesis. Our findings will provide additional information related to how nicotine usage affects cellular mechanisms in the human body. We treated liver cell lines with nicotine or left them untreated (control). Cell viability was measured and we found cell viability to be impacted by nicotine dosing. Exosomes were purified from cell culture media via high-speed ultracentrifugation or commercial kits. Exosome protein quantity was determined by standard quantitation method. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis in combination with Coomassie blue staining revealed major exosomal proteins. Enzyme-linked immunosorbent assays validated the presence of exosome-associated proteins and how they are effected by nicotine dosing. Findings indicate that the composition of exosomes derived from the liver cells were altered by nicotine dosing. Overall, these results strengthen the stigma that nicotine negatively affects the human body and that exosomes act as a strong precursor to particular diseased states. Future research involves analyzing the effects of nicotine on exosome biogenesis in various cell lines including lung and skin.

UPP20

Myosin Kinase Expression in Inflammatory Disease

Kiara Moore, Kateria Shinn, Drayvon Howard, and Lula Smith*

Department of Biological Sciences, Alabama State University, Montgomery, Alabama 36101. *Email: lbsmith@alasu.edu

There are many obstacles in the pursuit of treating cancer. One of these hindrances that health professionals encounter is the motility of cancer, as this is paramount in cancer cell metastasis and migration to other tissues in the body. One of the mechanisms of treatment that scientists are currently exploring is the blocking of the metastasis process. However, the precise mechanism of metastasis of cancer cells has not been completely delineated. It is known that the actomyosin cytoskeleton undergoes rearrangement in response to metastasis and tumor cell migration. But, not much is known about the details of myosin-dependent cytoskeleton contractile processes.

Myosin light chain kinase (MLCK) is a Ca2+/calmodulin dependent serine/threonine protein kinase that phosphorylates the regulatory light chain of myosin (RCL). This event regulates a number of myosin motor activities including cell division, muscle contraction and cell motility. Several studies indicate that myosin regulation is involved in the pathology of bacterial infections, viral infections, cardiovascular disease and cancer. These described pathologies all involve cell migration and/or remodeling of the actomyosin cytoskeleton. Towards this end, the present study focuses on the use of myosin light chain kinase (MLCK) as a therapeutic marker for inflammatory diseases. To initially address this hypothesis, a panel of cancer cell lines and tissues are analyzed for diverse expression of MLCK isoforms.

UPP21

Liposome encapsulated melittin for treatment of Staphylococcus aureus

Nathan Campbell, Sameer Joshi, Brandi Barlow, Komal Vig, Shree Singh, and Shreekumar Pillai*

Center for Nanobiotechnology Research, Department of Biological Sciences, Alabama State University, Montgomery, Alabama 36104. *Email: spillai@alasu.edu

Staphylococcus aureus is a bacterium which causes severe illness in humans ranging from mild cold symptoms to skin malformations to more severe pneumonia, meningitis and sepsis. At one point these infections were treated with penicillin but penicillin-resistant strains have been on the rise rendering the penicillin ineffective. Another issue in treating staphylocococcal infections is that the drugs may fail to reach the bacterial infection sites. The first step to overcome this is to find a way to target the bacteria through the surrounding tissue, deliver a drug and prevent resistance development in the bacteria. Based on prior research we decided to use the protein melittin, extracted from honey bee venom, as our drug of choice because it demonstrates antibiotic capabilities. This would be encapsulated in liposomes to ensure that it is degraded slowly and delivered to the target site. Our preliminary results show that the MIC of melittin was 3.125 µg/mL. We plan to encapsulate melittin and test the encapsulated drug for its efficacy to treat S. aureus.

 Bahar AA and Ren D (2013). Antimicrobial Peptides. Pharmaceuticals, 6(12), 1543–1575. http://doi.org/10.3390/ph6121543

Chaudhari AA, Jasper SL, Dosunmu E, Miller ME, Arnold RD, Singh SR, and Pillai S (2015). Novel pegylated silver coated carbon nanotubes kill Salmonella but they are non-toxic to eukaryotic cells. J. Nanobiotechnology, 13(1), 23.

UPP22

Creating Relevant Independent Research Experiences and Providing Insight into Health Careers for Biology Majors

Lydia Owens, Kayla King, and Diann Jordan*

Department of Biological Sciences, Alabama State University, Montgomery, Alabama 36104. *Email: djordan@alasu.edu

Contrary to popular belief, not all biology/pre-health majors want to be a doctor or dentist, many health career choices are available.  Although being a physician or dentist are the most cited fields of interests for Biology and Pre-Health majors, many of these students are not fully aware of the vast array of choices in the health-related and other STEM fields that would utilize their skills as Biology majors. This independent study project allows student researchers to explore a dual interest of science education, the field of physician’s assistant and pharmacy.  In this independent study, 3 films are critically evaluated by the student researchers and 3 films are selected by the student as suitable classroom teaching tools.  In the proposed poster presentation, the student researchers will discuss the process of researching the films and their experiences as independent researchers and how they would use them in the classroom (K-12 and college classroom).  Additionally, both students will research their particular health careers to better understand what their future career will entail.  The first student investigates her interests in physician’s assistant (PA) career by interviewing 3 PA’s currently in the field.  The second student will, in turn, interview 3 diverse pharmacists currently in the field.  Both students have will develop a questionnaire with approval of the instructor.  In this unique independent study, the student researchers will share their findings from the evaluation of the relevant films for the Biology classroom, the experience gained from a discussion with 3 physician assistants, and 3 pharmacists currently working in this profession.  Hopefully, the results will allow the student researchers to gain insight into their next career paths and assist the instructor in designing the most appropriate independent study projects for students interested in nontraditional health and STEM-related careers.

UPP23

Using Laser Induced Breakdown Spectroscopy to Detect Spherical Gold Nanoparticles in Hydrogels with HEp-2 Cells

Aamina Dandy1, Komal Vig2, and Cleon Barnett3,*

1Tuskegee University, Tuskegee, Alabama; 2Center for Nanobiotechnology Research, 3Department of Physical/Forensic Sciences, Alabama State University, Montgomery, Alabama 36104. *Email: cmbarnett@alasu.edu

Gold nanoparticles have many useful applications in the fields of chemistry, engineering, biology, and medicine. As interest in these applications expands, it is important to be aware of the nanoparticles introduced into the system. Laser-induced breakdown spectroscopy (LIBS) may prove to be a convenient alternative to other methods of nanoparticle concentration analysis. LIBS analysis does not require vacuum or extensive sample preparation as is necessary for some other techniques. However, it is well known that LIBS emission intensities can suffer from poor reproducibility. The purpose of this study was to determine the extent to which LIBS can be used to quantify the number of gold nanoparticles within hydrogels in a biological matrix. For this study, two harmonics of the Nd:YAG laser were used (532 nm and 1064 nm). Various amounts of 25 nm gold nanoparticles were mixed into 1% alginic acid and used to create hydrogels using 1% calcium chloride. HEp-2 cells were cultured on one set of the generated Hydrogels. Hydrogels with and without cells were cut and dried on a silicon wafer and analyzed for gold nanoparticles using 532 nm and 1064 nm to create the micro-plasmas. Poor reproducibility was observed in the LIBS emission and low R-squared values indicated poor correlation for calibration curves. However, significant improvements were made when the emission was normalized to 277.9 nm and 280.2 nm. In conclusion, further development of a system such of this will facilitate better sample introduction and possibly an internal standard to improve

reproducibility and a calibration curve. This work was supported by NSF-REU (DBI-1659166) to Dr. Komal Vig (PI) and by NSFCREST (HRD-1241701) to Dr. Shree S. Singh (PI).

UPP24

Phage Display Application for Identifying Chemotherapeutic Treated SW620 Colon Cancer Cells

Demetrius McAtee1, Deepa Bedi2, Komal Vig3,*

1University of Louisiana Lafayette, Lefayette, Louisiana 70503; 2Department of Biomedical Sciences, Tuskegee University, Tuskegee, Alabama 36088; 3Center for NanoBiotechnology Research, Alabama State University, Montgomery, Alabama 36104. *Email: komalvig@alasu.edu

Colon Cancer (CC) is a carcinoma which originates in the cells of the large intestine, and often metastasizes to the liver. Currently, the therapeutic treatments available are not specific to targeting only malignant cells and often are cytotoxic to healthy benign tissue. Bacteriophages (BPs), or “phage”, are nanosized viruses which specifically infect bacterium and can only proliferate inside the host cell. The same mechanism of infecting and rapidly killing the host cell is being genetically manipulated and bio-medically applied in nanomedicine to CC cells for diagnostic and therapeutic approaches by phage display technology. In the present study, 16 BPs were tested for their specificity binding to SW620 cells using an Enzyme Linked Immunosorbent Assay (ELISA). The in vitro assay was conducted using SW620 cells showing ≥90% viability. Cells were treated with camptothecin (CPT), a chemotherapeutic treatment for SW620 for 24-hours. Each phage had a control (no drug treatment) and a CPT treated cell. The diagnostic technique analyzed the M13 antibody’s binding affinity to the CPT treated SW620 antigen. Of the 16 BPs, the 3 phages L2, E2, and L47 showed the highest selectivity, identifying and binding to CPT treated SW620 cells and not bind to non-treated cells. To determine the concentration of selected phage binding to SW620 cells, a phage titration assay was conducted on L2, E2, and L47 phages. Further studies are being carried out to demonstrate that these phages can be used as diagnostic biomarkers or therapeutic antineoplastic on targeted SW620 cells. This work was supported by NSF-REU (DBI-1659166) to Dr. Komal Vig (PI) and by NSFCREST (HRD-1241701) to Dr. Shree S. Singh (PI).

UPP25

The Resistance of Nanomaterials in Streptococcus pneumoniae

Deja Lee, Veolanda Peoples, and Mamie T. Coats*

Center for Nanobiotechnology Research, Department of Biological Sciences, Alabama State University, Montgomery, Alabama 36104. *Email: mcoats@alasu.edu

Streptococcus pneumoniae is the major cause of potentially deadly diseases in humans especially affecting children under the age of five, immunocompromised individuals, and the elderly. Bacterial resistance to known antibiotics is an urgent issue. Our research described here and elsewhere has found that nanoparticles do exhibit some antimicrobial properties and can be used as a drug delivery agent. Here we sought to determine how metallic nanoparticles (copper, 50 nm and gold, 10-20 nm) inhibit S. pneumoniae. We found that 0.125_g/mL of copper or greater inhibits planktonic S. pneumoniae. Furthermore, copper nanoparticles display unique catalytic activity and generate low toxicity towards mammalian cells. Future studies include examining increasing concentrations of copper and gold to identify a minimum inhibitory dose and examine the genetic response of S. pneumoniae to these particles. This work was supported by NSF-REU (DBI-1659166) to Dr. Komal Vig (PI) and by NSFCREST (HRD-1241701) to Dr. Shree S. Singh (PI).

UPP26

Formulation and Characterization of Bioinks for 3-D printing

Parris Debrow and Derrick Dean*

Center for Nanobiotechnology Research, Biomedical Engineering Program, Alabama State University, Montgomery, Alabama 36104, *Email: ddean@alasu.edu

The regeneration, restoration, and replacement of defective tissues caused by genetic disease or injury involved complex and extensive procedures in the past. With the advancements in additive manufacturing technologies otherwise known as 3D printing has made it less complicated to fabricate tissues no matter how complex they are. While 3D printing holds much promise in this field, the need for bioinks that are appropriate for the tissue of interest and have the requisite viscosity is very critical. Thus, the objective of our research is to develop a hydrogel bioink for 3D printing bone tissue scaffold, and elecuidate the underlying structure-property relationships. Hydrogels play a large role in the accessibility and function of scaffolds to proliferate a composition similar to human cells that can be used for many applications. The most important components of a bioink include viscosity, shear-thinning, viscoelasticity, cytocompatibility and most of all biocompatibility to encourage healthy growth of living tissue. Extrusion bioprinting is the most common method to create 3D scaffolds. Polymeric hydrogels are advantageous due to their viscosity and high cell density. Another good aspect of hydrogels is that they can contain many chemical components for the best physical and mechanically stable biomaterials for successful tissue engineering. This research shows a multi component hydrogel consisting of polyvinylpyrrolidone (PVP), sodium alginate (SA) and hydroxyapatite (HA). Primarily different amounts of sodium alginate (ranging from 0.5 to 3%) were added to find a consistency comparable enough to be used in a bioprinter. The hydrogel was characterized by the use of rheology, Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC) and scanning electron microscopy (SEM). Rheology showed how the stress affected the sample based on the increase in viscosity of hydroxyapatite or sodium alginate. The DSC shows the effect of various additives to the gel on the melting behavior. Lastly FTIR was used to obtain an infrared spectrum of absorption or emission of the solid. The hydrogel solutions developed exhibited several characteristics similar to other bioinks. The 3.0% HA solution proved the best results in terms of viscosity, viscoelasticity, dehydration and rehydration traits. These gels have the potential to provide a promising solution to the need for advanced bioinks. This study was supported, in part, by a grant from NSF-REU (DBI-1358923) to Dr. Komal Vig (PI)

Maas M, Hess U, Rezwan K (2014) The contribution of rheology for designing hydroxyapatite biomaterials, Current Opinion in Colloid & Interface Science 19, 585–593.

Bose S, Vahabzadeh S, and Bandyopadhyay A (2013) Bone tissue engineering using 3D Printing, Materials Today, 16, 12.

Production of Untagged Recombinant ZIKA E Protein with Sumo System

Sydney Jackson and Alain B. Waffo*

Department of Biological Sciences, Alabama State University, Montgomery, Alabama 36104, USA. *Email: abopdawaffo@alasu.edu

Outbreaks of the infection from 2007 to 2016 have brought ZIKV to the forefront of the medical and scientific research communities as well as the general public.  Some known facts regarding the ZIKV are as follows:  A) It is a single-stranded RNA arbovirus which encodes a single polyprotein that cleaved into three structural proteins (prM, C & E) and seven non-structural proteins (1, 2A, 2B, 3, 4A, 4B & 5). B) The E protein is conserve among ZIKV lineages and is projected to mediate viral assembly, attachment, entry and fusion. Although several predictions have been postulated regarding cellular protein binding partners of the E protein, there has not been any experimental data to confirm these predictions.  Our goals with this project are to identify candidate proteins that bind the envelope (E) protein of the ZIKV. To achieve this aim we have been working on producing untagged E protein. Synthetic E protein was purchased from Eurofilm in pUC vector. The pESUMO was the vector used from NEB. Both plamids were restricted with NotI and PstI. The restricted pSUMO and E protein genes were agarose gel purified and ligated. The recombinant pSUMOE was sequenced and used to transform various cells. A better expression and solubility were achieved with 43(DES) cells. The purification conditions of untagged E in SUMO with better yield are to be optimized. This recombinant protein will help for future ZIKA disease investigation and efficacious vaccine development.