11th Annual Celebration of Scholarship Symposium (April 8-9, 2013)

COMPLETED RESEARCH

Analysis of the changes in acquisition of resistance to bacteriostatic and bactericidal drugs by Escherichia coli, Besil Rodriguez, Gloria
College of Science and Mathematics
Professor: Dr. Rachel Hopp
In the last few decades, hospital-acquired infections have become a major cause of illness and death in patients, despite the ever-increasing use of antimicrobial agents. Nosocomial infections are one of the most difficult to combat because very often they are the result of antibiotic resistant bacteria. After being subjected to selective pressure from the environment, these bacteria have found a way to develop mechanisms of resistance to several antibiotics. This study will concentrate on the effects that bacteriostatic versus bactericidal drugs have on Escherichia coli. Three bacteriostatic drugs have been selected: 1. Sulfamethoxazole because it is a competitive antagonist and structural analogue of para-aminobenzoic acid (PABA), therefore inhibiting folic acid synthesis. 2.Spectinomycin, which  binds to the 30S subunit of the ribosome and inhibits peptide chain elongation by interfering with peptidyl tRNA translocation . 3.Tetracycline, because it combines with the 30S subunit of the ribosome and inhibits the binding of the amino acyl tRNA (aatRNA) to the A site. A bactericidal drug, streptomycin, has been selected as a standard to measure how close and how much more E. coli will mutate and become resistant to the bacteriostatic versus the bactericidal drug. A growth curve was constructed for the E. coli strain used in this study to determine growth rate, and generation time. Since the bacterial population is most uniform in terms of chemical and physiological properties during the exponential (log) phase, this stage of growth was used to measure the minimal inhibitory concentrations (MIC). After determination of spontaneous mutations, the cells will be subjected to selective pressure. E. coli will be grown for 5 days at constant sublethal concentrations which will allow adaptation to antibiotics. The resistance rate will be measured using the fluctuation analysis calculator (FALCOR), a web tool for the determination of mutation rate using Luria-Delbruck fluctuation analysis. Results will be presented at the symposium.
 

DNA barcoding for Ligustrum japonicum, Bhatt, Asha
College of Science and Mathematics
Professor: Dr. Brenda Whaley
DNA barcoding is a molecular tool that uses a marker within an organism’s DNA sequence to identify it with a particular species group. The main purpose of DNA barcoding is to identify an unknown sample with a known specie group. The distinct differences between the genomic regions within the plants are adequate to provide specific specie information. However these genomic regions are still conserved to the point that generic primers can be used to amplify DNA using the polymerase chain reaction method. The specific DNA barcodes for our chosen plant contains the sequence for the gene that codes for ribulose-1,5-bisphoshpate carboxylase/oxygenase (RuBisCO), which is found on the chloroplast DNA. This gene is called rbcL. DNA barcoding was performed on a leaf tissue obtained from a plant garden at the following address, 4107 Vaughn Creek Court, Sugar Land, Texas. A small amount of the plant tissue was obtained and placed in nuclei lysis solution to dissolve the membrane bound organelles. The samples were then incubated and placed in RNase solution, protein precipitation solution, and isopropanol. PCR was then performed to amplify about 600 bases from the 5’ end of the rbcL gene. The primers used for rbcL were:
5’-TGTAAAACGGCCAGTATGTCACCACAAACAGAGACTAAAGC-3’
(forward primer – rbcLaf-M13)
5’-CAGGAAACAGCTATGACGTAAAATCAAGTCCACCTCG-3’
(reverse primer – rbcLa-revM13)
After PCR, gel electrophoresis was done to analyze the PCR products. Gel electrophoresis analyzes the PCR products for correct length and purity. The gels were then stained and photographed. The stained pictures and 10 µL of the PCR product was sent to GeneWiz Inc2 to be further analyzed and sequenced. A BLAST search was then used to determine the specie with the closest match to the obtained rbcL sequence. Based on the BLAST results the plant of interest was identified as Ligustrum japonicum.
 

The role of dpy-11 within C. elegans in maintaining organ morphogenesis and observable reversion in dpy-11 mutants, Bhatt, Asha; Foster, Andrea
College of Science and Mathematics
Professor: Dr. Rachel Hopp
Dpy-11, a gene found in Caenorhabditis elegans, encodes a membrane associated thioredoxin- like protein (TRX) that influences body shape and morphogenesis. The dpy-11 protein is solely expressed in the hypodermis and is essential in producing a normal body form in C. elegans. The TRX protein produced by C. elegans is homologous to the human TMX4 protein which is up-regulated in melanoma cells. The knockdown of the human TMX4 protein can be used as a cancer therapy to down-regulate TMX4 in melanoma cells. When the dpy-11 is knocked-down, these nematodes appear to have a dumpy phenotype which is observed by their short and plump characteristics. RNAi was used to induce the knockdown of the dpy-11 gene. A vector containing the RNAi insert targeting the dpy-11 gene as well as the ampicillin resistance gene was obtained from the DNA Dolan Learning Center at Cold Spring Harbor Laboratories. Escherichia coli were transformed with this vector, and then fed to nematodes to induce the knockdown of the dpy-11 gene. The resulting dumpy phenotype, specifically the area of the worm, was measured and compared to wild-type C. elegans through a computer program, Image J. Furthermore the concentrations of IPTG were manipulated to determine its effect on the dumpy phenotype. IPTG induces the promoter and therefore we expect that at higher concentrations of IPTG the worms will be more dumpy. Taurine, a dietary supplement, reduces ER stress and was used to assess reversion of the dpy-11 mutants. Nitric oxide promotes longevity of C. elegans and was therefore used to evaluate survival rate of mutants compared to wild-type C. elegans. Both taurine and nitric oxide were used by dissolving them in the NGM agar and using the RNAi methods mentioned above. We expect the dpy-11 mutants to revert back to their wild-type state with taurine in the NGM agar. We also expect the survival rate of the mutants to increase with the addition of nitric oxide to the NGM agar. Results will be presented at the symposium.
 

DNA Barcoding, Cantu, Eddie
College of Science and Mathematics
Professor: Dr. Brenda Whaley
DNA barcoding is a tool utilized for species identification. It involves the sequencing of a standard region of DNA from the genome of individual species, this sequences are then stored in a database called GenBank. Once in the sequences are stored in GenBank they can be retrieved for comparison and identification of genus and species. In order to identify a land plant species the DNA barcode must be retrievable with a single pair of primers and provide maximal discrimination among species. The perfect candidate for this in land plants was the long chain of the enzyme ribulose-1,5-bisphosphate carboxylase / oxygenase (RuBisCO) which is involved in photosynthesis with the gene found in the chloroplasts DNA with the name of rbcL.
Since the gene that has been chosen is present in the chloroplast DNA then this technique can be performed on a single leaf from a land plant. One leaf from a land plant was collected in the morning and taken into lab for experimentation the same day. The leaf was collected at this coordinates N29.692549, W95.53997. DNA kit was used in order to isolate rbcl for DNA barcoding. 10-20 mg (1/4 inch) of leaf tissue was collected and treated with lysis solution for extraction of DNA (rbcL). For purifying of the solution a proteins precipitation solution was utilized. After the DNA was extracted polymerase chain reaction (PCR) was performed with the addition of a set of primers specific for rbcL. Following PCR a confirmation was done via Gel Electrophoresis and UV blue light observation.
The identification protocol yielded the genus and species of the land plant to be Gelsemium elegans (yellow jasmine).

 
Use of DNA Sequencing for the Identification of Epipremnum aureum, Contreras, Loren
College of Science and Mathematics
Professor: Dr. Brenda Whaley
DNA barcoding is an important method for the identification of species within a genus taxonomic group.   The sequence of a standard sequence of the genome is obtained and then stored in database such as GenBank. The sequenced region is highly conserved; therefore common primers can be used to amplify this sequence by polymerase chain reaction. For plants, a particular gene for a protein involved in the chemical conversion of light to chemical energy necessary for plants, enzyme ribulose-1,5-biphosphate carboxylase/oxygenase (RuBisCo) the product of the gene rbcl was used. For this experiment a leaf was removed from my chosen plant for the isolation of its DNA followed by amplification of it through Polymerase Chain Reaction (PCR). PCR is a technology in molecular biology used to produce millions of copies of a single DNA sequence and this technique is commonly used in applications such as diagnosis of hereditary diseases, identification of genetic fingerprints and DNA-based phylogeny.. The PCR product of amplified DNA was visible after gel electrophoresis was performed and indicated fragments of appropriate size and purity. DNA sequencing of the PCR product was done by GeneWiz. DNA sequences were obtained and using a BLAST search and it identified the closest genus and species for the rbcl sequence of the leaf used in the experiment. My organism is Epipremnum aureum; its common name is Golden Pothos. The latitude and longitude for my plant was found using itouchmap at 29.693999 and -95.516623.

 
Analysis of Chloride and TDS Levels along the Buffalo Bayou, Jacob, Marian; Eapen, Jeffery
College of Science and Mathematics
Professor: Dr. Betty Thompson
The chloride measurement is a measure of the amount of chloride deposit found in water. This value can be indicative of pollution in the water. The data indicates Chloride levels to be lower at a certain undisturbed location at Gayhills with a mean value of 0.8. At the other undisturbed location, Mayde Creek, the Chloride levels were off the charts at a mean value of 173.5. Further investigation should be conducted to understand the reason for the high values at Mayde Creek. Further down the buffalo bayou the levels begin to increase in the Ship channel and substantially at Galveston Bay but remain stable in Arboretum water sources. The ranges of the values vary from close to far. The range for the entire Buffalo Bayou was 4006.7 indicating a wide distribution of chloride values throughout the bayou.
TDS is a measurement of the total dissolved solids in a sample of water. A high concentration of ions indicates that a body of water is polluted or heavily contaminated. The Environmental Protection Agency has set certain TDS parameters for bodies of water to comply under. The objective of this research is to measure and analyze water samples along the Buffalo Bayou and test whether or not the TDS levels increase as one goes down the bayou, starting from Mayde Creek to the Galveston Bay. According to our results, it appears that as one goes along the bayou, the total dissolved solids concentration increases. The Galveston Bay had the highest level of TDS while Mayde Creek had the lowest level of TDS.
 

DNA Barcoding, Mathew, Ebby
College of Science and Mathematics
Professor: Dr. Brenda Whaley
DNA barcoding is a tool that uses a region of DNA for species identification.  The region that is used in plants is ribulose-1,5-bisphosphate carboxylase / oxygenase (rbcL) which is  found in chloroplast DNA and is involved in photosynthesis. rbcL is chosen because it is easily retrievable, well suited for high-quality bidirectional sequences, and is the best option for species discrimination from previous studies. In this study, a leaf was removed from a plant at HBU (29.69531 °N, -95.51533 °W). The DNA sequence was extracted from the rbcL gene, amplified (Polymerase chain reaction), underwent thermal cycling and gel electrophoresis, and was sent to GeneWiz Inc for DNA sequencing. Based on the results from sequencing, a BLAST search matched the plant the rbcL sequence to one that was the most similar, or has highest identity. The BLAST search suggested that leaf extracted at 29.69531 °N, -95.51533 °W belongs to the Alpinia zerumbet plastid, also known as Shell ginger, and had a 99% identity.
 

Analysis of unc-22 mutant phenotype in Caenorhabditis elegans, Odey, Ochuko; Villanueva, Grace
College of Science and Mathematics
Professor: Dr. Rachel Hopp
Caenorhabditis elegan’s unc-22 codes for UNC-22 protein (twitchin), a homolog of proteins MyBP-C (cardiac myosin binding protein) and titin in humans. The unc-22 gene was identified as the founding member of a family of very large cytoplasmic proteins, on which muscle structure and function depends. MyBP-C is found to be frequently mutated in cardiomyopathy patients, while mutations in titin are associated with loss of muscle function which is one of the characteristics of muscular dystrophy. The knockdown of unc-22 was initiated by RNA interference (RNAi). C. elegans are the ideal specimens when working with RNAi due to the many effective techniques in which they can be introduced to specific double stranded RNA by feeding them bacteria. In this experiment, we fed the nematodes bacteria expressing dsRNA of unc-22 gene to trigger the silencing of the protein (twitchin). Knockdown of this gene is expected to produce a twitching muscular movement phenotype. In this case, we used IPTG and Carbenicillin in nematode growth agar (NGM) to induce silencing of the unc-22 gene and select for antibiotic resistant E.coli respectively. Varying the concentration of IPTG we will observe the number of twitches, and the distance traveled per/min of mutant worms growing in the different concentrations. We will then synchronize the worms by bleaching in order to place them in the same stage of development, and allow us to plate the eggs on HT115/NC and HT115/NIC in order to compare their hatching rates.  In a follow up experiment, we will also place eggs from L2 stage worms on new growth media plates containing HT115 E. coli/NC and observe if gene silencing will continue to the F1 generation. The number of twitches/min in the F1 generation will be recorded. Results will be presented at the symposium.
 

Analysis of Buffalo Bayou Waterway, Odey, Ochuko; Tadros, Meena; To, Alyce
College of Science and Mathematics
Professor: Dr. Betty Thompson
Fertilizers, high amounts of sewage wastes and vehicles in the cities contribute to elevated levels of nitrates, TDS, and chloride in waterways.  Due to these exceedingly high levels of nutrients in the water (eutrophication), water organisms are deprived of light and dissolved oxygen leading to death. In this experiment we tested nitrate, TDS, and chloride levels of various sampling sites on the Buffalo Bayou Waterway. We observed the upstream sites contained lower nitrate, TDS, and chloride levels and downstream through the urban parts of the city, nitrate and TDS levels greatly increased.  Chloride had increased levels at Mayde Creek and Galveston, but low levels in the other sites.  Gay Hills, is far from polluted city life and the nitrate level of 1.1 ppm, TDS level of 98 ppm, and chloride level of 0.8 ppm reflected that.  The next two collection sites, Mayde Creek and White Oak, had similar nitrates levels (13.97 ppm and 15.2 ppm).  TDS levels for Mayde Creek and White Oak were 180.5 ppm and 566.5 ppm, respectively.  On the other hand, chloride level for Mayde Creek (179 ppm) was higher than White Oak (3.2 ppm). The Ship Channel and Galveston had highest levels of nitrates and TDS.  The Ship Channel contained nitrates at 77.82 ppm and TDS at 6827 ppm.  Galveston contained 103.6 ppm of nitrates and 32150 ppm of TDS.  Chloride level for the Ship Channel was 4 ppm.  Galveston had high chloride levels, as expected (3394.7 ppm).  These high levels were caused by large amounts of pollution from factories and fertilizers.  The Arboretum, although located midway in the collection sites, contained the lowest amounts of all three parameters.  This shows how the natural environment surrounding the Arboretum provided the site with efficient ways of keeping the waterways clean and filtering out pollution.
 
 
Growth and Characterization of PEDOT:PSS and Carbon Nanotube Composite Structures for Excitonic Solar Cells, Owens, Constance
Owens, Constancea, Hettiarachchi, Chamindab, Feliciano, Domingob, Hyde, Robertb, Witanachchi, Sarathb
aCollege of Science and Mathematics, Houston Baptist University
bDepartment of Physics, University of South Florida
Harnessing solar energy is one of the most promising ways to tackle today’s energy issues. Though solar cells are comprised of many different layers, our focus is on a single layer. The main goal of this study is to create thin film composite structures of poly(3,4-ethylenedioxythiophene) poly(styrenesulfonate) (PEDOT:PSS) and carbon nanotubes (CNT), more specifically with multiple wall carbon nanotubes (MWCNT). In this study we employ a spray method that utilizes Bernoulli’s principle. We believe that a spray method will produce a better uniform layer than other methods that are utilized for creating thin films. Uniformity within a thin film is of the upmost importance because as uniformity is improved, many properties are enhanced. PEDOT:PSS was mixed separately with both dimethylformamide (DMF) and water. By the Dektak 3030ST, a profilometer device, it was discovered that the PEDOT:PSS containing DMF dispersed better than the PEDOT:PSS mixed with water, thus creating a more uniform film. Also it is well known that MWCNTs possess many excellent properties that can make them very useful in the field of solar technology, such as they are durable, and have a broad range of electronic, thermal and structural properties. In this study we also combine MWCNTs into our thin films to see how they affect thickness, transparency and conductivity by using the Lambda 950, a UV/VIS spectrometer, and a four point probe.
 

Measuring Nitrate Levels Along Buffalo Bayou, Patail, Nimrah
College of Science and Mathematics
Professor: Dr. Betty Thompson
Buffalo Bayou is a large body of water flowing through Houston, Texas. It begins in Katy, Texas, and flows approximately 53 miles east to the Houston Ship Channel and then into Galveston Bay and the Gulf of Mexico. One purpose for conducting a study on Nitrate values across various points across Buffalo Bayou was to hopefully identify areas suggestive of point-source pollution. The data collected shows the only nitrate values that were low enough to meet the water quality standard were found the Gay Hills area with a mean value of 1.1 mg/L. Nitrate levels greatly increase at Mayde Creek, which should ideally be another undisturbed area that neighbors Gay Hills. Instead, Mayde Creek water exhibited an average Nitrate value of 7.6 (including the outlier of .4 mg/L, which makes our average seem smaller than it probably is). We continued to measure the trail of Buffalo Bayou at various points to monitor the nitrate trend as the bayou emptied out into Galveston Bay. The average nitrate level grows in an exponential manner by the time the bayou empties out into Galveston with an average ppm of 103.6. The three Arboretum ponds (which serve as a model of an undisturbed ecosystem) sustain fairly constant, low nitrate values, suggesting that the large values of Nitrates in surrounding bodies of water are the result of human activity.
 

Measuring Lysozyme Solubility as a Function of pH, Portillo, Carmen
College of Science and Mathematics
Professor: Dr. Saul Trevino
Poor protein solubility is a major problem in the development of protein pharmaceuticals. Solubility determines how concentrated drugs are in the system. When medication is injected into the system, higher concentrations are needed for effective results. Therefore, strategies for improving protein solubility are of great interest to the pharmaceutical industry. In this study, the solubility of lysozyme was measured as a function of pH. Although these proteins are not necessarily pharmaceutically relevant, the information obtained from them could hopefully be applied to pharmaceutically relevant proteins. It was previously thought that solubility would be minimal at the isoelectric point and then would continually increase with increasing net charge on both sides of the isoelectric point. Up to this point, results have remained consistent within a minimum selected range of pH values. When the range is extended, deviations in the expected pattern of increased solubility with respect to pH are observed. In the pH range of 4.0 to 5.5, predicted solubility values at 1.1 M consistently increased with pH. These values are trusted since they were predicted from calibration curves that contained a R2 values very close to 1, and were verified experimentally. We infer that with lower pH values, the solubility of lysozyme protein will continue to decrease; and thus will be verified with future experimentation.
 

Pollution along Buffalo Bayou, Quach, Jessica; Robinson, Corey; Maradiago, David
College of Science and Mathematics
Professor: Dr. Betty Thompson
The purpose of this study was to identify the potential pollution present by testing certain parameters of water, such as the levels of nitrate, ammonium, TDS (total dissolved solids), and chloride, for different bodies of water throughout Houston along the Buffalo Bayou. The different locations chosen for sampling included: Gay Hill, Mayde Creek, White Oak Bayou, the Houston Arboretum, the Port of Houston Ship Channel, and Galveston Bay. Water samples from the aforementioned sources were collected and then tested and analyzed using certain instruments such as TDS meters and probes like the Vernier Nitrate Ion-Selective Electrode (ISE). Water from the Houston Arboretum served as the control, since it is a relatively undisturbed ecosystem. It was hypothesized that the levels of all the parameters to be tested should increase from body of water to body of water as the water travels farther down the Buffalo Bayou due to the pollution accumulated.  Results showed that the amount of only certain parameters, such as nitrate levels, increased in a direct manner as predicted.
 

Muscle dystrophy in Caenorhabditis elegans, Roy, Sarath; Brandow, Christopher
College of Science and Mathematics
Professor: Dr. Rachel Hopp
Caenorhabditis elegan’s dys-1, a homolog of human (Duchenne Muscle Dystrophy) DMD dystrophin gene, is a dystrophin like gene that when mutated leads to hyperactivity and hypercontractivity of muscles in animals. DMD is an X-linked muscle disease and according to previous studies many of these mutations can be corrected at the posttranscriptional level by alternative splicing which removes the mutated exon. Research in this field can provide an indisputable prospect into correction by alternative splicing and for further gene therapy. Dystrophin is essential for assembly of the dystrophin-glycoprotein complex and provides a strong connection between the cytoskeleton and the extracellular matrix. RNA interference, RNAi, is used to induce the specific knockdown of dys-1 gene. We received a double stranded RNAi feeding strain (targeting dys-1) from the Dolan DNA Learning Center at the Cold spring Harbor Laboratories. A dys-1 knockdown in C. elegans is expected to exhibit hyperactivity and hypercontractivity of muscular functions. In this experiment, features of the dys-1 knockdown nematodes are observed by placing the worms on plates that contain a bacteria food source only around the outer edge. Worms were placed in the center of each plate and their movements and speed were observed using a dissecting microscope. Photographs and video recordings of both control and dys-1 knockdown worms were obtained via iPhone and analyzed to visualize the hyperactivity in the knockdowns as compared to the wild type. Data analysis was done as distance over time as they moved from the center to the outer edges. Since dys-1 knockdown worms are expected to be more hyperactive in their movement, the video recordings should display the movement difference when compared to the wild type in that mutated worms had a hyper body movement or what appears as an uncoordinated twitch. Results will be presented at the symposium.
 

The effects of Silencing ERM-1 gene in Caenorhabditis elegans, Sabharwal, Kiran
College of Science and Mathematics
Professor: Dr. Rachel Hopp
Caenorhabditis elegans, transparent nematodes, have been widely used as a model organism system for research in neurology and biology. When combined with the uses of RNAi, C. elegans have proven to help understand just how gene regulation is altered.  The ERM protein family consists of three closely related proteins, ezrin, radixin, and moesin, which are known to serve as linkers between membrane proteins and F-actin cytoskeleton in many organisms. Silencing of erm-1 by RNAi is expected to interfere with the apical junction (AJ), resulting in abnormalities in gut formation, leading to intestinal constriction and obstructions. This intestine-specific phenotype can then be used to gain a better understanding of gut development and its importance in nutrient absorption and waste production, not only in C. elegans, but in humans as well. Therefore, it can be hypothesized that silencing of erm-1 will result in impaired gut formation, leading to deficiencies in nutrient absorption and waste production output. Objectives of this study would be to generate a feeding vector using the pPR244 plasmid containing the dsRNA of erm-1 – this will introduce the gene into a vector that can be fed to the C. elegans, in order to induce erm-1 knockdown by RNAi. Through further research we will be able to observe how erm-1, when silenced, causes major changes and alterations in the gut of C. elegans and what this means for gut development in humans.
 

Oil-Degrading Microbes: Bioremediation of Cooking Oils, Sabharwal, Kiran; Nguyen, Trinh; Salinas, Rure
College of Science and Mathematics
Professor: Dr . Jacqueline Horn
The purpose of this experiment is to isolate, grow, and enhance the oil degradation of microbes from different environments. We isolated our microbes from a fast food sink, Friendship pond, and Hodo Residence College Fountain based on an inference that these environments would contain microbes with the ability to degrade oil because of their exposure to hydrocarbons.  Minimal nutrient agars were used to isolate and culture the microbes. Stained vegetable oil and Crisco were added to agars as the only carbon source. Therefore, the growth observed on the agars would confirm that these isolated microbes indeed have oil degrading capabilities. In order to precisely measure the oil degradation, a minimal nutrient broth was used with the stained oil and Crisco shortening added on top. Although microbial growth was observed in the tubes with Crisco, a measurement of degradation could not be obtained due to the solid properties of shortening. Different sizes of tubes were used to further enhance the rate of degradation. By increasing the surface area of the tube, microbes present were able to gain better access to the oil and show an increase in lipolytic activity. Light and dark conditions were studied to test for enhanced growth of photosynthesizing microbes. Finally, the pH of the broth was altered to find the preferred pH environment for the microbes.
 
 
Applying the Method of DNA Barcoding to Identify Ilex x attenuata, Truong, Julia
College of Science and Mathematics
Professor: Dr. Brenda Whaley
DNA barcoding is a technique that uses a standard region of an organism’s DNA to sequence and identify the corresponding genus or species.  An ideal genomic region for DNA barcoding should be able to be amplified by polymerase chain reaction using common primers, demonstrate high quality bidirectional sequencing with minimal editing, and provide maximal discriminating power among species.  The rbcL gene, which is found on the chloroplast DNA and codes for the enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCoO) long chain, offers these important qualities, and is therefore used as the DNA barcode for plants.
To begin, a single leaf was obtained from the plant of interest, which is located at 29.861011°N and -95.600770°W, and pictures were taken of both the individual leaf and the whole plant.  A small section of the leaf was used to isolate the DNA from the plant tissue.  The rbcL gene was then amplified by polymerase chain reaction (PCR) using two different primers.  Thermal cycling made copies of the rbcL gene by denaturing, annealing, and extending the DNA region.  Next, gel electrophoresis was used to analyze the amplification products for correct length and purity.  Gel staining allowed the PCR samples to be visible, which were photographed and sent along with a sample of the PCR product to a sequencing company.  After receiving the sequences of the gene, they were used to perform a BLAST search.  By using the closest match for the rbcL sequence identified by BLAST and evaluating the leaf anatomy, the unknown plant was identified as Ilex x attenuata, also known as Savannah holly.
 

Using Microbial Communities with Cellulolytic Activity to Create Biofuels, Truong, Lindy; Cerda, Vanessa; Jimenez, Steven
College of Science and Mathematics
Professor: Dr. Jacqueline Horn
The current endeavor of the biofuel industry is to use microorganisms that secrete cellulolytic enzymes to break down cellulose into simple sugars that can be fermented into ethanol and biobutanol. The focus thus far, has been largely on singular organisms in hopes of discovering the most efficient for cellulose hydrolysis. Little has been done to study the efficiency of microbial communities. In this experiment, lichen, a microbial symbiotic community between fungi and algae or photosynthetic bacteria, is used to measure for cellulolytic activity against that of individual microorganisms. Locally selected samples at Houston Baptist University are collected for fungi, algae, bacteria, and lichen, and grown in a laboratory setting on agar plates using carboxymethylcellulose as the sole carbon source. Congo Red solution, a biological stain that forms strong non-covalent bonds to cellulose, is then used to detect the presence of the enzymatic activity as the organisms degrade the cellulose present through the clear zones that form where cellulose has been broken down. The specimens are also tested on minimal media that has Filter paper as its sole carbon source to measure their efficiency with crystalline cellulose. This experiment hopes to provide support to using the synergism between certain organisms as a possible venue for the advancement in biofuel technology.
 
 
PROPOSED RESEARCH
 
Determining the binding domain of the Z-DNA binding protein for the negative regulatory element of the ADAM-12 gene, Chang, Jordan
College of Science and Mathematics
Professor: Dr. Saul Trevino
ADAM-12 is a multifunctional protein that is involved in cell communication and cell growth. This gene appears to play a vital role in many cancers and tumor formations. While expression of ADAM-12 is generally low in adult human tissues, cancer cells are seen to have remarkably high levels of expression of this protein. In an attempt to regulate ADAM-12 expression, negative regulatory elements (NRE), located upstream of the ADAM-12 gene, are suspected to inhibit the translation of the ADAM-12 gene. This is done through the interaction of a Z-DNA binding protein. While the identity of the Z-DNA binding protein is unknown as well as how this interaction occurs, we hypothesize that the Z-DNA binding protein blocks DNA polymerase from initiating the translation of the ADAM-12 gene. We propose to identify the sequence and structure of the Z-DNA binding protein for the NRE of the ADAM-12 gene. This can be done by first isolating the protein through affinity chromatography from human cancer cells and then, using X-ray crystallography or NMR, determine the sequence and structure of the protein. To identify the Z-DNA binding domain of the protein, we propose to use the sequence we determine in a multiple sequence alignment with other known Z-DNA binding proteins. This will provide a better understanding of this inhibitory mechanism and possibly present new therapies to battle against cancer and tumor growth.
 

Developing an oligobody for highly sensitive detection of FPN1 on O-2A progenitor cells, Contreras, Loren
College of Science and Mathematics
Professor: Dr. Saul Trevino
The purpose of my research is to develop an oligobody for highly sensitive detection of FPN1 on O-2A progenitor cells. An oligobody is a sequence of short, single strand of DNA or RNA molecules attached to an antibody that is routinely used for diagnostic detections in genetic tests, research and forensics. The importance of building an oligonucleotide relies on specific binding to specific-mutated proteins benefiting in higher specificity and thus allows for improved detection of antigen. FPN1 is an iron exporter protein whose expression of mRNA and protein are seen in O-2A progenitor cells. O-2A are progenitor cells in which FPN1 has a function of promoting iron release; this is important because the accumulation of iron is affected by the downregulation of FPN1 and thus lead to neurological diseases as Parkinson’s disease and other neurological diseases such as cerebral ischemia. O-2A progenitor cells were the main component of neonatal periventricular white matter, susceptible to the hypoxia-ischemia and thus became the target cells of periventricular leukomalacia (PVL). The model system I propose is based on the mutation of FPN-1 antibody and conjugate an oligonucleotide to the anti-monoclonal FPN1 antibody mutated with in Lys residue to phenylalanine (pAcF) in order to identify the level of expression and distribution of FPN1 in O2-A; in this proposal, the detection of O-2A detection serves to detect iron transport necessary for the prevention and diagnosis of neonates. If this theoretical research was to be carried in lab, we would expected higher levels of of FPN-1 detected expression through the use of the synthesized oligobody than the ones measured in previous experiments by RT-PCR and western blot. In conclusion, the detection of FPN-1 will allow researchers to make more accurate diagnosis and prevention of diseases related to iron accumulation and/or deficiency in neonates.

 
Regulation of tissue transglutaminase in Celiac Disease, Ng, Zui Keat
College of Science and Mathematics
Professor: Dr. Saul Trevino
Celiac Disease (CD) is a medical condition characterized by mal-absorption, weight loss and increased energy expenditure, and the exacerbation of the disease is usually caused by foods that contain gluten (Capristo et. al., 2005).  Gluten is a peptide that has glutamine residues in its chemical structure, and glutamine residues contain amide group in which transglutaminase (tTG) deamidates.  Patients who have CD usually have high levels of tissue (tTG), and they are sensitive to gluten due to the deamidating activity of tTG.  The specific aim of this study is to regulate tTG activity using RNAi.  After the treatment of RNAi, the level of tTG will be measured using a technique called Western blot.  A decreased level of activity of tTG in patients with CD is expected. Finally, several biochemical values related to Celiac Disease will be measured to assess the effect of the RNAi treatment to reduce tTG activity.
 
 
ORAL PRESENTATIONS
 
PCR Detection of Naegleria fowleri in Texas Lakes, Ghosn, Jean
College of Science and Mathematics
Professor: Dr. Hannah Wingate
Primary Amoebic Meningoencephalitis (PAM), a 98% fatal disease caused by the free-living amoeboflagellate Naegleria fowleri, has reached a total of 257 documented cases worldwide and 28 cases in Texas since 1983. With no available cure and rapid proliferation, insufflated N. fowleri invades the central nervous system primarily in immunocompetent children engaging in freshwater activities. Little is currently known regarding the specific distribution of N. fowleri, especially in Texas lakes. Efforts in correlating physical, chemical, or biological parameters with N. fowleri proliferation have proven inconclusive. In this project, we directly detected N. fowleri from three popular Texas lakes through PCR using species-specific 18S rRNA gene primers. Forty five two-liter samples collected from various locations from each lake were filtered, and the resulting filter papers were exposed to cell lysis solution and phenol-chloroform to extract purified DNA. N. fowleri was detected in several locations in Lake Arlington, but not in Joe Pool Lake or Lake Waxahachie. With this data, we aim to compile a database of N. fowleri-positive lakes in Texas for further PAM prevention efforts.
 
 
Development of a General Chemistry Lab Experiment, Jara, Rosario
College of Science and Mathematics
Professor: Dr. Taiya Fabre
A two day general chemistry laboratory experiment was developed to allow students to determine the solubility product (Ksp) of a sparingly soluble salt. A sparingly soluble salt in contact with water maintains equilibrium between the solid and its ions. Solubility can very according to each solution, but the solubility product must satisfy the Ksp expression. In this experiment, the Ksp of calcium sulfate dihydrate is obtained.

 
Quantum Gravitational Effects and the Generalized Uncertainty Principle, Meyers, Vincent; Owens, Constance
College of Science and Mathematics
Professor: Dr. Gardo Blado
Two physical theories share between them nearly the sum of all thought given to the field of physics: General Relativity (GR) and Quantum Mechanics (QM). GR describes the behavior of the very large, and QM describes that of the very small. There has been and continues to be a great rift between them: that they appear fundamentally to disagree in their descriptions of aggregate particle interactions. The least contentious proposition to reconcile this rift increasingly appears to be the theory of Quantum Gravity (QG). It holds implications for the whole of physics, and one of the least frequently treated of these is the Minimal Length Uncertainty (MLU). Broad conjecture holds this to be on the order of the Planck Scale, or , though no serious effort has been devoted to ascertaining its exact size. While seemingly trivial, the fact of a MLU would dramatically restructure another of the most venerable and fundamental notions in physics, that of the Heisenberg Uncertainty Principle, which says that location and momentum are acutely related and limited. We propose that the application of QG to the Heisenberg Uncertainty Principle within the context of a finite square well is the most expeditious means of gaining an understanding of the MLU. In our research, we model the dichotomous bound and scattering states of a particle in a finite square well with a QG-corrected set of equations. Indeed, the application of QG to the uncertainty principle and the Schrödinger equation have a potentially dramatic effect on the modeled behavior of a particle in this most seminal of environments.
 
 
An Investigation of Porphyrin Macrocycles With and Without Coordinated Organic Substituents in Aqueous Media by UV-Visible and Emission Spectroscopy, Portillo, Carmen; Ramirez, Kevin
College of Science and Mathematics
Professor: Dr. Eric Van Caemelbecke
The objective of this senior seminar experiment was to observe the increase fluorescence lifetime of various sample solutions of porphyrin when factors such as concentration, pH and solvent are manipulated. The Shimadzu Spectrofluorophotometer is a new instrument acquired by the Houston Baptist University Chemistry Department through Welch Foundation Funding and was primarily used during this study. After acquiring many spectrums and placing various solutions under analysis, it was determined that the properties that enhanced the yield of a solution were those external factors that were manipulated such as the pH of the solution, and the presence of quinine; and the molecular structure of the compound under study. Further examination is recommended in order to determine the factors than contribute to the increase in quantum yield of the solution of quinine and porphyrin.
 
 
Synthesis, Structure, and Electrochemical Characterization of Ru2(2,4,6-(CH3)3ap)3(O2CCH3)Cl (ap = 2-anilinopyridinate anion); A Mixed-Ligand Diruthenium(III,II) Complex with an Unusual Arrangement of the Bridging Ligands, Ramirez, Kevin
College of Science and Mathematics
Professor: Dr. Eric Van Caemelbecke
A mixed-ligand metal-metal bonded diruthenium complex having the formula Ru2(2,4,6-(CH3)3ap)3(O2CCH3)Cl where ap is the anilinopyridinate anion was synthesized from the reaction of Ru2(O2CCH3)4Cl and H(2,4,6-(CH3)3ap), after which the isolated product was structurally, spectroscopically and electrochemically characterized.  The crystal structure reveals an unusual arrangement of the bridging ligands around the dimetal unit where one ruthenium atom is coordinated to one anilino and two pyridyl nitrogen atoms while the other ruthenium atom is coordinated to one pyridyl and two anilino nitrogen atoms. To our knowledge, Ru2(2,4,6-(CH3)3ap)3(O2CCH3)Cl is the only example of a mixed-ligand diruthenium complex of the type [Ru2L3(O2CCH3)]+, where L is an unsymmetrical anionic bridging ligand that has been structurally characterized with a “(2,1)” geometric conformation of the bridging ligands, all others being “(3,0)”.   The initial Ru25+ compound in CH2Cl2 or CH3CN containing 0.1 M tetra-n-butylammonium perchlorate (TBAP) undergoes up to four one-electron redox processes involving the dimetal unit. The Ru25+/4+ and Ru25+/6+ processes were characterized under N2 using thin-layer UV-visible spectroelectrochemistry and this data is compared to UV-visible spectral changes obtained during similar electrode reactions for related diruthenium compounds having the formula Ru2L4Cl or Ru2L3(OCCH3)Cl where L is an anionic bridging ligand. Ru2(2,4,6-(CH3)3ap)3(O2CCH3)Cl was also examined by UV-visible and FTIR spectroelectrochemistry under a CO atmosphere and two singly reduced Ru24+ species, [Ru2(2,4,6-(CH3)3ap)3(O2CCH3)(CO)Cl]- and Ru2(2,4,6-(CH3)3ap)3(O2CCH3)(CO) were in-situ generated for further characterization. The CO-bound complexes could be further reduced and exhibited additional reductions to their Ru23+ and Ru22+ oxidation states.

 
Determination of Ethanol in Solutions by Gas Chromatography, Ramirez, Kevin
College of Science and Mathematics
Professor: Dr. Robert Towery
A Shimadzu Gas Chromatograph 2014 equipped with a Zebron column was used to evaluate the percent ethanol content of prepared ethanol standards as well as collected ethanol containing samples of both known and unknown ethanol content. From the prepared ethanol standards, it was determined that the equation of the line for the calibration curve was y = 1100049.7351x + 1157403.1115 with an R2 value of 0.9987. With this calibration curve and the equation of the line, a sample of known ethanol content, McCormick lemon extract, was evaluated and found to have a mean percent ethanol of 50.63622. From this value, it was then determined that the percent error present in the McCormick lemon samples was 38.99%. This gross error was then attributed to the fact that the McCormick bottle was old and left to sit on a shelf for an extended period of time, allowing ethanol to volatilize out of solution. Two other collected samples, later referred to as ED 32 and ED io, were also run on the GC to determine their percent ethanol content. However, since their true percent ethanol contents were unknown, it was decided that their values were to be expressed as a range of possible percent ethanol contents. From experimentation, it was found that ED 12 had a percent ethanol content in the range of 1.16 – 3.22 % ethanol and ED io had a percent ethanol content in the range of 8.89 – 12.81 % ethanol.