Alien Registration- Morin, Andrew (Brunswick, Cumberland County)

https://digitalmaine.com/alien_docs/31692/thumbnail.jp

Identification And Characterization Of DNA Repair Snf2/Swi2 Atpases In Tetrahymena Thermophila

The Snf2/Swi2 ATPases Rad5 and Rad16 have been shown to play vital roles in a number of DNA repair pathways. In both Saccharomyces cerevisiae and human cell lines, Rad5 homologs (SHPRH, HLTF) have been shown to function in DNA double strand break (DSB) repair along with pathways that repair damage after replication. The function of Rad16, unlike Rad5, has been found only in lower eukaryotes such as Saccharomyces, despite the fact that it plays an essential role in nucleotide excision repair (NER), and more specifically in the repair of silenced areas of the genome. In order to more fully understand the function of Rad16, this work focuses on using a model organism, Tetrahymena thermophila, to identify and characterize the functional aspects of both Rad5 and Rad16. To do this, qPCR analyses of the potential Rad5/16 homologs were conducted to determine their expression, while shRNA constructs were designed to inhibit their expression to assess the phenotypic consequences of DNA damage in deficient cells. Expression analyses showed that three of the potential homologs (Rad16, Rad5.2, and Rad5.1) have damage-depended expression, and that the levels of one can have substantial effects on levels of the others. Moreover, two of the homologs, Rad16NH and Rad5.1, show altered survival after genotoxic stress. The data showed that the functions of Rad16 and Rad5 homologs in Tetrahymena may diverge greatly from those in lower eukaryotes

Introductory Medical Microbiology

This Grants Collection for Introductory Medical Microbiology was created under a Round Nine ALG Textbook Transformation Grant. Affordable Learning Georgia Grants Collections are intended to provide faculty with the frameworks to quickly implement or revise the same materials as a Textbook Transformation Grants team, along with the aims and lessons learned from project teams during the implementation process. Documents are in .pdf format, with a separate .docx (Word) version available for download. Each collection contains the following materials: Linked Syllabus Initial Proposal Final Reporthttps://oer.galileo.usg.edu/biology-collections/1020/thumbnail.jp

Characterization of Cas9-Mediated microRNA Knockout

MicroRNAs are short regulatory RNAs that primarily operate at the posttranscriptional level, acting as part of the RISC complex to destabilize mRNA prior to, or during, translation. MicroRNAs are subject to several levels of processing by Drosha and Dicer prior to achievement of the active mature form, this processing being highly dependent on the secondary structure of the microRNA transcript. As such, microRNA biogenesis is highly sensitive to mutations. In this study, it was attempted to optimize Cas9-mediated genome editing techniques for microRNA knockout for two hematopoietic microRNAs, miR-142 and miR-223. While it has been previously demonstrated that Cas9 is capable of inhibiting microRNA biogenesis, the results have been characterized by variable and low-level efficiencies currently inherent to the system. This study attempted to rectify this shortcoming by bombarding the small pre-miR transcript with a combination of multiple independently targeted Cas9 complexes, using lentiviral delivery methods, measuring for evidence of impaired microRNA biogenesis by real-time quantitative PCR (RT-qPCR). The results suggest that there may be specific regions of the microRNA site in which mutations are more deleterious to function, but that further study is required. Additionally, it was determined that the use of multiple sgRNAs is not experimentally useful, proving unable to and in some cases, an impediment to successful microRNA knockout

Breaking the Cycle: Impact of Sterically-Tailored Tetra(pyrazolyl)lutidines on the Self-Assembly of Silver(I) Complexes

A improved preparation of the pentadentate ligand α,α,α′,α′-tetra(pyrazolyl)lutidine, pz4lut, and the syntheses of three new alkyl-substituted pyrazolyl derivatives pz4′4lut (pz4′ = 4-methylpyrazolyl), pz*4lut (pz* = 3,5-dimethylpyrazolyl), and pzDIP4lut (pzDIP = 3,5-diisopropylpyrazolyl) are described. The silver(I) complexes of these ligands were studied to ascertain the impact of pyrazolyl substitution, if any, on their binding modes and on solubility issues. In the solid state, [Ag(pz4lut)](BF4) (1), [Ag(pz4′4lut)](BF4) (2), and [Ag(pz*4lut)](BF4) (3) give cyclic dications as a result of two ligands sandwiching two silver centers where each ligand binds the metals through only pyrazolyl nitrogen donors. This cyclic motif is similar to those observed in the silver complexes of tetra(pyridyl)lutidine PY5-R derivatives (where the central pyridyl does not bind) and in related tetra(pyrazolyl)-m-xylene complexes. While suitable single crystals of [Ag(pzDIP4lut)](BF4) (4) could not be obtained, those of [Ag(pzDIP4lut)](OTf) (5) showed infinite polymeric chains secured via silver-bound μ-κ2Npz,κ2Npz- ligands. The different binding mode of the latter ligand versus the former three is likely due to unfavorable steric interactions between the bulky iso-propyl (pyrazolyl) substituents and the central pyridyl rings of hypothetical cyclic architectures. The combined electrospray ionization mass spectrometry (ESI(+)-MS), variable-temperature NMR (VT NMR), and diffusion pulsed field-gradient spin−echo (PFGSE) NMR data indicate that the solid state structures of each 1−5 are neither retained nor static in CD3CN, rather the cations are monomeric in solution

Variations in Modeled Dengue Transmission over Puerto Rico Using a Climate Driven Dynamic Model

Dengue fever is a mosquitoborne viral disease reemerging throughout much of the tropical Americas. Dengue virus transmission is explicitly influenced by climate and the environment through its primary vector, Aedes aegypti. Temperature regulates Ae. aegypti development, survival, and replication rates as well as the incubation period of the virus within the mosquito. Precipitation provides water for many of the preferred breeding habitats of the mosquito, including buckets, old tires, and other places water can collect. Because of variations in topography, ocean influences and atmospheric processes, temperature and rainfall patterns vary across Puerto Rico and so do dengue virus transmission rates. Using NASA's TRMM (Tropical Rainfall Measuring Mission) satellite for precipitation input, groundbased observations for temperature input, and laboratory confirmed dengue cases reported by the Centers for Disease Control and Prevention for parameter calibration, we modeled dengue transmission at the county level across Puerto Rico from 20102013 using a dynamic dengue transmission model that includes interacting vector ecology and epidemiological components. Employing a Monte Carlo approach, we performed ensembles of several thousands of model simulations for each county in order to resolve the model uncertainty arising from using different combinations of parameter values that are not well known. The top 1% of model simulations that best reproduced the reported dengue case data were then analyzed to determine the most important parameters for dengue virus transmission in each county, as well as the relative influence of climate variability on transmission. These results can be used by public health workers to implement dengue control methods that are targeted for specific locations and climate conditions

Assessing Disparities of Dengue Virus Transmission Risk across the US-Mexican Border Using a Climate Driven Vector-Epidemiological Model

Dengue fever is a mosquitoborne viral disease reemerging throughout much of the tropical Americas. Dengue virus transmission is explicitly influenced by climate and the environment through its primary vector, Aedes aegypti. Temperature regulates Ae. aegypti development, survival, and replication rates as well as the incubation period of the virus within the mosquito. Precipitation provides water for many of the preferred breeding habitats of the mosquito, including buckets, old tires, and other places water can collect. Although transmission regularly occurs along the border region in Mexico, dengue virus transmission in bordering Arizona has not occurred. Using NASA's TRMM (Tropical Rainfall Measuring Mission) satellite for precipitation input and Daymet for temperature and supplemental precipitation input, we modeled dengue transmission along a USMexico transect using a dynamic dengue transmission model that includes interacting vector ecology and epidemiological components. Model runs were performed for 5 cities in Sonora, Mexico and southern Arizona. Employing a Monte Carlo approach, we performed ensembles of several thousands of model simulations in order to resolve the model uncertainty arising from using different combinations of parameter values that are not well known. For cities with reported dengue case data, the top model simulations that best reproduced dengue case numbers were retained and their parameter values were extracted for comparison. These parameter values were used to run simulations in areas where dengue virus transmission does not occur or where dengue fever case data was unavailable. Additional model runs were performed to reveal how changes in climate or parameter values could alter transmission risk along the transect. The relative influence of climate variability and model parameters on dengue virus transmission is assessed to help public health workers prepare location specific infection prevention strategies