Expression and Purification of Tumor Suppressor Protein TIP30 for Structural Studies by NMR Spectroscopy
Joseph Blas
ABSTRACT
TIP30 (Tat-Interacting Protein, 30 kD) is a putative metastasis suppressor that promotes apoptosis and inhibits angiogenesis. To provide structural insight into the function of TIP30, structural studies by NMR spectroscopy were pursued. A TIP30 segment (101-166) was expressed and purified by His-tagged affinity chromatography followed by FPLC purification. Successful 15N-labeling of this protein fragment by two-dimensional heteronuclear single-quantum proton-nitrogen correlated spectroscopy was demonstrated. The fragment exhibited no single favored conformation under the studied conditions.
Keywords: FPLC, HSQC, IPTG, NMR, TIP30, Tumorigenesis.
The Design and Synthesis of Pyrazine Ligands Suitable for Molecular Weaving with Octahedral Metal Ions
David L Cockriel
ABSTRACT
Pyrazine-2-carboxylicAcid(pyridine-2-ylmethyl)amide and 2,4-di(n(pyridalmethyl)formamide)pyrazine, two new ligands designed to require a weaving topology around octahedral metal ions have been synthesized in high yield and characterized. These new ligands utilize the nitrogens in pyridines and pyrazines in their structures to produce poly-dentate binding sites for coordinating to metals producing multiple crossovers with the goal of forcing over/under repeats in metal complexes. Characterization of the ligands was achieved by Mass Spectroscopy, 1H NMR, 13C NMR, mp, IR spectroscopy and Elemental analysis.
Keywords: Molecular Weaving, oligomeric pyridine, pyrazine, supramolecular chemistry.
The Effects of Organic Selenium on Rumen Volatile Fatty Acid Production in Continuous Culture
Dean Feasenhiser
ABSTRACT
Selenium is a potentially toxic trace mineral that is known to be a requirement for ruminant animals. Selenium is most easily absorbed in the form of selenomethionine, but is commonly supplied in the form of sodium selenite. The goal of this experiment was to recreate the rumen environment to determine the effects of selenomethionine at levels of 1.1 and 2.5 ppm. Rumen effects were measured by changes in volatile fatty acid (VFA) concentrations using gas chromatography. The study concluded that there were no significant effects in changing from a normal level (1.1 ppm) of selenomethionine to the higher level (2.5 ppm). However, a trend was detected that showed minor decreases in most VFA levels at the high level of selenomethionine.
Keywords: rumen, ruminant, selenomethionine, selenium, volatile fatty acid.
Influence of Lactose Utilization and Population Dynamics of Eschericia coli var. mutabilis
Jenny L. Harper
ABSTRACT
30-50 million American are lactose intolerant and suffer with intestinal gas, nausea, diarrhea or abdominal cramping when they eat foods that contain lactose. The digestive track is a microecosystem that contains numerous bacterial species, eight of which have been identified by Dr. Schaedler and his colleagues. This research was intent on investigating how one of these bacteria, Eschericia coli var. mutabilis (E. coli), utilized lactose when lactose was added into the bacteria’s environment. E. coli was cultured in a Tryptic Soy Broth (TSB) containing 0%, 10%, 20% and 40% lactose (LTSB) in relation to the total amount of dextrose used in the media. Kinetics tests were run to find an approximate growth rate for the bacteria. Serial dilutions and population counts were performed from a 24-hour culture. The greatest amount of growth after 25 hours was found in the 0% LTSB and growth continued to decrease through the experiment with the least amount of growth found in the 40% LTSB. Statistical analysis of the population counts illustrated only the 20% lactose run had a significant P-value of 0.0048, therefore proving the null hypothesis incorrect-that bacteria can grow with lactose in the environment.
Keywords: Eschericia coli var. mutabilis, Lactase, Lactose, Lactose intolerance, and Population dynamics.
Human Performance Assessment Using the DYFORMON Exercise System
Dave Hoffman
ABSTRACT
The DYFORMON (Dynamic Force Monitor) system was used to assess human exercise performance by integrating user output force over time. This result, the momentum generated, was determined to be a better measure of exercise effort than power output. One subject exercised with the DYFORMON machine twice a week for 5 weeks, varying between two different workouts to prevent muscle fatigue. Workout sessions consisted of bench press, pull-downs on the bench, and a set of 50 interval bench press reps on Mondays and pull-downs from the ground, bar curls, and military presses on Wednesdays. The bench press with a protocol of 5 sets at 10 reps per set was the only exercise evaluated. Results showed an increase in total generated momentum from week to week for the bench press. The 50-interval reps bench press protocol seemed to show better improvement with a two-week recovery period, rather than a one-week. The goals of this study were to recalibrate the DYFORMON machine and to find meaningful ways to convert raw data to better understand human performance on the DYFORMON. This was not a statistical study, but demonstrated the basis for such a study.
Keywords: Dynamic Force Monitor, momentum, eccentric, concentric, impingement, exercise
The Mutagenesis of Benzo(a)pyrene on Yeast
Danielle Lucore
ABSTRACT
Benzo(a)pyrene (BaP) is one of many polycyclic aromatic hydrocarbons (PAHs), which are pollutant by-products of incomplete combustion that have been detected in the air, soils, sediments, water, and char-grilled or smoked foods. Recurrent exposure is a cause for concern, because a number of PAHs, such as BaP, have been identified as probable human carcinogens by the EPA. The purpose of this study was to test the mutagenic effects of BaP on a model organism: Saccharomyces cerevisiae (yeast). DNA was isolated from a 37 C, BaP-treated culture, amplified by the polymerase chain reaction (PCR), digested with the Hae III and HindIII restriction enzymes, and separated by electrophoresis on a 1.5% agarose gel. The DNA was then stained and the banding patterns were compared. The presence or absence of bands would indicate a mutation. Although it was expected that BaP would mutate the yeast DNA, no alteration was detected in the DNA sequence that was amplified.
Keywords: Benzo(a)pyrene (BaP), polycyclic aromatic hydrocarbon (PAH), Saccharomyces cerevisiae, polymerase chain reaction (PCR), restriction enzyme, electrophoresis.
Synthesis and Characterization of Cross-Bridged AMD3100 Analogs
Tau Lyn Snell
ABSTRACT
In 1988 AMD3100, a xylyl-bicyclam was discovered to exhibit anti-HIV activity by preventing the HIV virion from successfully interacting with the CXCR4 coreceptor and infecting the cell. AMD3100 was thus a potentially new fusion inhibitor drug and is currently undergoing clinical trials. Further research performed on AMD3100 analogs have revealed that if it is complexed with certain metals it will increase the bonding affinity to CXCR4 by causing the cyclam rings to take on a folded cis configuration. According to these results, an AMD3100 analog was synthesized with the addition of an ethyl cross-bridge that locked it into the cis folded configuration and then copper was bound to the ligand. In this complex, a metaxylyl linker was used instead of the paraxylyl linker present in AMD3100 to probe the importance of this bridging group. The copper bicyclam has been characterized and is currently being assayed for anti-HIV properties.
Keywords: AMD3100, Anti-HIV drugs, Bi-cyclam, CXCR4, SDF-1, Xylyl-bicyclam, fusion inhibitor