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Myers CV_2015

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Myers CV_2015

  1. 1. Amanda Myers Cell: (937) 232-8023 1861A Brattleboro Ct Kettering, OH, 45440 myers.k.amanda@gmail.com CAREER STATEMENT I intend to be strategically indispensable while developing new skills and completing meaningful research in areas such as genetics, bioinformatics, and human health. TECHNICAL EXPERTISE Specialized Skills Chemical hazard identification Substance risk assessment Technical report review Study quality determination Creation of toxicological profiles Review of chemical read-across Comprehensive chemical evaluations Office & Software Skills Client confidentiality Local and international coordination Microsoft Word Microsoft Excel Microsoft PowerPoint Microsoft Outlook OECD QSAR Toolbox Minitab (Statistical Program) Cn3D (3D protein viewing software) Vector NTI (plasmid design software) Adobe Professional (PDF software) NCBI BLAST (Protein/neucleotide comparison tool) NCBI Neucleotide (Gene sequence database) Laboratory Skills Gel electrophoresis (agarose, PAGE) DNA and RNA extraction Western blot analysis Northern blot analysis Southern blot analysis Stock solution preparation PCR Primer design Mammalian cell transfection
  2. 2. Light microscope use Site-directed mutagenesis Cell counting Primary explants Chloroform extraction EMPLOYMENT HISTORY Jul. 2012 – Present Advanced Testing Laboratory Thorough human risk assessments of product and raw material formulations Generation of complete toxicological profiles on specific compounds Quantitative Structure-Activity Relationship Modeling Local and international correspondence and coordination for issue resolution Handling of proprietary client information while maintaining confidentiality Database update On-boarding new employees EDUCATION Aug. 2008 – May 2012 Ball State University, Muncie, IN BS Biology (Genetics Concentration) BS minor, Chemistry Biotechnology Certificate GPA: 3.474/4.000 RESEARCH AND PROJECT EXPERIENCE Aug. 2014 — Present Employee On-Boarding Revamp, Advanced Testing Laboratory, Cincinnati, OH Leads: Amanda Myers and Amanda Dysert Purpose: This project seeks to develop an immersive on-boarding process which balances the desired efficiency with the large volume of technical knowledge learned. Description: Due to a recent shift in the complexity of client needs, the technical knowledge required by contractor employees has greatly increased. Because many new employees do not have a background in toxicology, an efficient yet thorough education in human toxicology is essential during the on-boarding process. A small team has been charged with re-structuring the on-boarding process from a series of disjointed, overwhelming presentations into an interactive learning experience which creates confidence. I have created or re-designed some presentations, including topics such as risk assessment and an introduction to the position. Within the context of the group, I have given input and made changes to multiple other trainings, discussing core concepts as well as internal client databases and systems. Additionally, the group has developed a series of exercises which build on one another to produce hazard assessments, risk assessments, and mock client requests. Dec. 2010 — May 2012 Database Design Project, Ball State University, Muncie, IN
  3. 3. Adviser: Dr. C. Ann Blakey. Purpose: The end product of this project is the creation of the Breeds of Rare & Endangered Database System (BREEDS), which will allow breeders to make informed decisions for the breeding of endangered horses and ponies. Description: Rare and endangered breeds often suffer from substantial levels of inbreeding. Due to their dwindling numbers and human mis-communication, horses, ponies, and jackstock can be accidentally bred with relatives or with those in their breed which share specific detrimental genes. In each case, anomalies such as developmental disorders, sub-standard offspring, and death and can occur. In order to combat the worst inbreeding decisions, we seek to create the BREEDatabase System. This system will incorporate information on specific horse and pony lineages to help reduce familial matings, as well as provide specific genetic information to breeders so that breed characteristic phenotypes may be maintained. For my contribution, I designed the entire initial draft of the system conceptual schema and database schema for a model critically endangered horse breed. Due to the differing characteristics, population sizes, and organizations associated with each breed, the database schema will then be optimized for each breed involved. These schemas are currently assisting the database creators. I also developed an algorithm for determining and predicting horse coat colors and the associated genotypes based on physical characteristics and lineage. This algorithm is rooted in current research on classic and molecular genetics. In addition to these projects, I directly worked with other team members to strategize and complete component arrays of data entry including data input approach, categorization, and format changes. All aspects of this project adhered to the privacy agreements made with those who own or are responsible for the data used. Sept. — Dec. 2011 Riboswitch Silencing Project, Ball State University, Muncie, IN Class: Biochemistry Laboratory Techniques, CHEM 465, Dr. Scott Pattison. Thesis: The binding affinity of Mutant ykkCD toxin sensor riboswitch will have a low affinity for tetracycline antibiotics. Description: Riboswitches are segments of RNA which interact with molecules in the cell for regulation purposes, such as gene regulation. The bacterial riboswitch ykkCD is sensitive to the antibiotic tetracycline, and marks it for transport out of the cell. In order to study the sensitivity of a mutant conformation, site-directed mutagenesis was performed to delete a cytosine from a riboswitch at a position important to correct folding. Our primers were designed accordingly. Quikchange PCR was used to generate the mutant ykkCD. The sample was then propagated using E. coli transfection and isolation of DNA plasmid, to produce a mutant ykkCD riboswitch. The plasmid was linearized and isolated via chloroform extraction. Finally, RNA was synthesized from the linear RNA sequence and segregated from the enzymes using denaturing urea PAGE. The binding affinity of the resultant mutant riboswitch to tetracycline was assessed to determine the mutation’s effect. Oct. — Dec. 2011 Protein Expression Project, Ball State University, Muncie, IN Class: Protein Isolation and Analysis, BIOT 492, Dr. Susan McDowell. Thesis: Human gene ACTG1 can be transfected into HEK 293 cells for protein isolation.
  4. 4. Description: ACTG1 is one of six genes which code for actin gamma 1, which is essential in cellular development and survivability in many species. The gene, ACTG1, was transferred to the destination vector, pDEST26, using LR recombination. The plasmid was then used to transform E. coli. Replicated plasmid was isolated from colonies and assessed for orientation. Plasmids of correct orientation were then used in HEK 293 cell transfection. Total protein from HEK 293 cells were isolated and prepared for western blot analysis. Jan. — May 2011 Bioinformatics Project, Ball State University, Muncie, IN Class: DNA Sequencing and Bioinformatics, BIOT 495, Dr. Susan McDowell. Thesis: Downregulation in Homogentisate 1,2 Dioxygenase in fibroblast cells is directly related to breast cancer. Description: Homogentisate 1,2 dioxygenase (HGD) is the third in a pathway of five enzymes which metabolize tyrosine into acetoacetate and fumarate. The HGD gene is highly conserved throughout most living organisms. In a 2010 study, gene expression in MCF7 beast cancer tumor and CCL171 lung stromal cells with and without insulin growth factor I (IGF-I) was assessed in duplicate using microarrays. All CCL171 cultures showed increased expression of HGD, while one MCF7 IGF+ culture showed increased expression. CCL171 IGF+ cultures had a higher expression than IGF- cultures. Utilizing NCBI services such as BLAST, the links between HGD and cancer were researched. Based on the analyses, the expression of HGD is not directly related to cancer in fibroblast or tumor cells. However, by down-regulating all pathways associated with tyrosine degradation, tyrosine in its former state may be conserved. An example of such a state is a receptor tyrosine kinase. For cancerous cells, turning off a pathway which degrades receptor tyrosine kinases may be beneficial. To verify such a hypothesis, experiments studying the relationship between tyrosine and cancer as well as experiments determining the link between receptor tyrosine kinase degradation and tyrosine in the HGD pathway should be conducted. Oct. — Dec. 2010 T4 DNA Ligase Cloning Project, Ball State University, Muncie, IN Class: Introduction to Recombinant DNA and RNA, BIOT 490, Dr. Susan McDowell. Thesis: Mouse gene Tardbp can be cloned into vector CMV6-A-Puro using a T4 DNA ligase cloning procedure, then transformed into E. coli. Description: Tardbp is a gene that can become mutated to cause amyotrophic lateral sclerosis and frontotemporal lobar degeneration. Tardbp codes for a TAR DNA-binding protein that is responsible for the correct splicing of the cystic fibrosis transmembrane conductance regulator. Very interested in neural diseases, we chose to clone this gene with the goal of making it available to our school. Using T4 DNA ligase, we cloned Tardbp from pCMV6-Kan/Neo into CMV6-A-Puro. The resulting plasmid was transformed into competent E. coli. Plasmid samples were extracted from colonies. Agarose gel electrophoresis revealed Tardbp presence in two of four samples. Colonies containing the gene were frozen for future use. Aug. — Dec. 2009 Ecology Research Project, Ball State University, Muncie, IN Class: Methods in Ecology, BIO 217, Dr. David LeBlanc.
  5. 5. Thesis: The density of earthworm types at Cooper Farm, Muncie, Indiana, is based on differing soil compositions. Description: Earthworms are important to the structure and composition of soil and can help determine plant communities and nutritional dynamics. Earthworm samples were obtained from three areas in Cooper Farm: a wet prairie, a comparatively arid prairie, and a soil type rich in organic matter. For three nights, ten soil blocks (0.17m x 0.17m x 0.17m) were dug in each area. Worms were hand sorted, counted from these plots, and statistical analysis performed. Mean density of earthworms in the organic site was 15.3 ± 8.51 worms/m2 greater than the mean density of earthworms in the wet prairie and 10.6 ± 8.51 worms/m2 greater than the mean density of earthworms in the arid prairie. Mean density did not differ between the wet prairie and arid prairie sites. The proportion of species types and age was not significantly different among the sites tested. These results suggest that additional organic matter is significantly better for earthworm sustainability. Jan. — May 2009 Center for Authentic Science Practice in Education (CASPiE) Program, Ball State University, Muncie, IN Class: CASPiE Program in General Chemistry II, CHEM 112, Dr. Jason Ribblett. Thesis: Different beverages contain varying levels of antioxidants based on inhibition levels of ABTS-+ . Description: This was an NSF funded grant project to expose exceptional students to research design and execution in chemistry using antioxidant experiments. Antioxidants neutralize molecules called oxidants, which damage cells and tissues, ABTS-+ (an oxidant solution) is commonly used to test the amount of antioxidants in beverages and foods in the United States. Based on the CASPiE protocol, ABTS-+ was added to the beverages of our choice. The antioxidant level was determined via spectrophotometry based on how much ABTS-+ remained in the sample. Amongst V-8 (regular), Welch’s Grape Juice, Welch’s Apple Juice, Snapple Green Tea, Snapple Lemonade, Capri Sun Grape, and Capri Sun Orange, the V-8 showed the highest level of antioxidants with the rest following respectively in the order above. These results suggest that higher levels of vegetable and fruit content can increase the active antioxidant level in a beverage.
  6. 6. Thesis: The density of earthworm types at Cooper Farm, Muncie, Indiana, is based on differing soil compositions. Description: Earthworms are important to the structure and composition of soil and can help determine plant communities and nutritional dynamics. Earthworm samples were obtained from three areas in Cooper Farm: a wet prairie, a comparatively arid prairie, and a soil type rich in organic matter. For three nights, ten soil blocks (0.17m x 0.17m x 0.17m) were dug in each area. Worms were hand sorted, counted from these plots, and statistical analysis performed. Mean density of earthworms in the organic site was 15.3 ± 8.51 worms/m2 greater than the mean density of earthworms in the wet prairie and 10.6 ± 8.51 worms/m2 greater than the mean density of earthworms in the arid prairie. Mean density did not differ between the wet prairie and arid prairie sites. The proportion of species types and age was not significantly different among the sites tested. These results suggest that additional organic matter is significantly better for earthworm sustainability. Jan. — May 2009 Center for Authentic Science Practice in Education (CASPiE) Program, Ball State University, Muncie, IN Class: CASPiE Program in General Chemistry II, CHEM 112, Dr. Jason Ribblett. Thesis: Different beverages contain varying levels of antioxidants based on inhibition levels of ABTS-+ . Description: This was an NSF funded grant project to expose exceptional students to research design and execution in chemistry using antioxidant experiments. Antioxidants neutralize molecules called oxidants, which damage cells and tissues, ABTS-+ (an oxidant solution) is commonly used to test the amount of antioxidants in beverages and foods in the United States. Based on the CASPiE protocol, ABTS-+ was added to the beverages of our choice. The antioxidant level was determined via spectrophotometry based on how much ABTS-+ remained in the sample. Amongst V-8 (regular), Welch’s Grape Juice, Welch’s Apple Juice, Snapple Green Tea, Snapple Lemonade, Capri Sun Grape, and Capri Sun Orange, the V-8 showed the highest level of antioxidants with the rest following respectively in the order above. These results suggest that higher levels of vegetable and fruit content can increase the active antioxidant level in a beverage.

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