Life Sciences Commons^™

Diversity In Darkness: Shedding Light On Cryptic Species In The Cave Beetle Genus Darlingtonea Valentine Using 3rad Sequencing And Consequences Of Climate Change Based On Thermal Tolerance And Dehydration Resistance, Athanasios S. Apostolopoulos

Masters Theses & Specialist Projects

The relationship between geographic distribution and phylogeny is pronounced in patchy, discontinuous habitats that limit gene flow, such as cave ecosystems. The isolating nature and selective pressures of cave environments can result in relatively high levels of endemism and cryptic speciation in cave faunas. Cryptic speciation exhibited in cave faunas is poorly studied, though it is an integral aspect to consider when discussing conservation efforts.

Our study employed the 3RAD genomic sequencing technique and robust population sampling across the distribution of a monotypic genus of cave beetle, Darlingtonea Valentine, that inhabits karst systems in a nearly 200 square mile range …

Delineation Of Undescribed, Morphologically Cryptic Cave Beetles Of The Pseudanophthalmus Pubescens Species-Group (Coleoptera: Carabidae: Trechinae), Jedidiah John Nixon

Masters Theses & Specialist Projects

The genus Pseudanophthalmus Jeannel is a widespread and extremely diverse taxon of troglobiont beetle endemic to the karst of eastern North America, with its distribution centered in Tennessee and Kentucky. Despite lying near the heart of this region, the pubescens species-group was thought to contain many undescribed species. In this study, the validities of several of the late Dr. Thomas Barr’s unpublished putative new species in the pubescens-group were tested both morphologically and molecularly. Body measurements (length and width for head, thorax, and abdomen), as well as male genital morphology were compared to see if they reflected theorized species limits.. …

Synthesis And Molecular Mechanics Study Of Platinum Triamine Complex With N-Acetyl-L-Methionine And Guanosine 5’Monophosphate, Nitin Tanajirao Katare

Masters Theses & Specialist Projects

Cisplatin and its analogues have been used and tested over decades for cancer treatment, but because of the serious side effects, it is important to find ways to overcome those side effect by synthesizing a novel platinum molecule. We have synthesized triamine complex and conducted its reaction with biomolecules such as 5’GMP and Methionine in the lab as well as conducted molecular mechanics (MM). MM methods are used for predicting and designing new structures and properties, and to model geometries of small molecules. MM and dynamics calculations have been used to study the 9 Ethylguanine and N-Ac-Lmethionine complexes with platinum …

An Intergrative Ecological And Taxonomic Assessment Of The Stoneflies (Insecta, Plecoptera) Of The Black Mountains, North Carolina, Usa, Madeline Lake Metzger

Masters Theses & Specialist Projects

The Black Mountains are a part of the Blue Ridge region of the Appalachian Mountains and include Mount Mitchell, the highest peak east of the Mississippi River at 2,037 m (= 6,684 ft). A comprehensive inventory of the stonefly fauna was conducted between 2014-2019 within Mt. Mitchell State Park and adjacent Pisgah National Forest. Collections occurred seasonally from 44 unique locations, covering a broad range of stream sizes and elevations in the region. Sequencing of the mitochondrial cytochrome c oxidase subunit I (mtCOI) gene was completed to assist with making positive determinations to species for females of taxonomically challenging genera …

Involvement Of Jak/Stat Signaling And A Basement Membrane-Associated Protein During Air Sac Primordium Development In Drosophila Melanogaster, Nathan Anthony Powers

Masters Theses & Specialist Projects

Tumor metastasis currently presents the greatest obstacle for effective cancer remediation. Metastatic growth necessitates both degradation of a specialized form of extracellular matrix (ECM) known as the basement membrane (BM) and the invasion of surrounding tissues thereafter. The thoracic air sacs of fruit flies (Drosophila melanogaster), which develop and operate in a fashion comparable to the human lung, provide a unique model for identifying and characterizing factors that contribute to its own development as well as tumoral invasion. We investigated the involvement of both Janus kinase (JAK)/Signal transducer and activator of transcription (STAT) signaling and a BMassociated protein during the …

Species Distribution And Conservation Genetics Of The Upland And Midland Chorus Frogs (Pseudacris) In Kentucky, Tucker Cambridge

Masters Theses & Specialist Projects

The upland (Pseudacris feriarum) and midland (P. triseriata) chorus frogs are closely related cryptic species that are best distinguished genetically. The distribution of these species within the Commonwealth of Kentucky has previously been defined by only a handful of genetic samples, making delineation of range limits for each species difficult. Accurate understanding of species distributions, and the genetic structure within them, are vitally important for conservation management of amphibian species. In this study, I have collected genetic samples from across the putative ranges of P. triseriata and P. feriarum in Kentucky and used next-generation sequencing technology to generate more fine-scale …

Transcriptomic Response To Immune Challenge In Zebra Finch (Taeniopygia Guttata) Using Rna-Seq, Cassandra Scalf

Masters Theses & Specialist Projects

Despite the convergence of rapid technological advances in genomics and the maturing field of ecoimmunology, our understanding of the genes that regulate immunity in wild populations is still nascent. Previous work to assess immune function has relied upon relatively crude measures of immunocompetence. However, with next-generation RNA-sequencing, it is now possible to create a profile of gene expression in response to an immune challenge. In this study, captive zebra finch (Taeniopygia guttata; adult males) were challenged with bacterial lipopolysaccharide (2 mg/Kg BW; dissolved in 0.9% saline) or vehicle (0.9% saline) to stimulate the immune system. Two hours after injection, birds …

Characterizing The Role Of Cp1 In Drosophila Melanogaster: Its Effects On Basement Membrane Degradation And Signaling, Dane Alan Flinchum

Masters Theses & Specialist Projects

CP1 is a well-conserved cathepsin L-like protease essential for proper growth and development in Drosophila melanogaster. Previous research has demonstrated that CP1 has the ability to break down the extracellular matrix. Using the UAS-GAL4 system, immunohistochemistry, and antibody-staining, this research attempts to characterize the role of CP1 and its effects on basement membrane degradation and signaling. These effects include actions at the cellular level and on a known signaling pathway. The genes involved in this pathway are known to be required for proper development of the wing disc into the adult wing. We have demonstrated the collagenase activity of CP1 …

Adaptive Variation In Tiger Salamander Populations, Meghan Parsley

Masters Theses & Specialist Projects

Amphibians face an unknown future in a time of rapid environmental change due to global climate perturbations. Since amphibians are perceived to be indicators of ecosystem health, understanding the causes of their declines can improve our perception of threats to other species. Molecular techniques have allowed us to explore how environmental change affects genetic variation and to predict evolutionary adaptive potential of amphibian populations. The identification of populations with the greatest potential to respond to changing environmental variables may be an important conservation strategy to aid in future management efforts. I utilized targeted exon capture sequencing to identify adaptive variation …

Landscape Genetics Of The California Tiger Salamander: Inferences From Multiple Methods, Samantha Gabrielle Thomas

Masters Theses & Specialist Projects

Landscape genetics is a rapidly growing field of study that compares patterns of gene flow among populations with habitat heterogeneity across a landscape to infer the interaction between dispersal of individuals and their physical environment. Empirical data generated from a landscape genetics study can inform conservation and management strategies, making the field increasing popular. However, concerns have arisen in the literature that the field is expanding faster than the analytic framework that supports it. Multiple methods for generating estimates of the association among habitat types and dispersal (i.e., least-cost paths and resistance surfaces) have been proposed, and there is a …

Sequence Extension Of The Tryptophan And Shikimate Operons In Clostridium Scatologenes Atcc 25775, Shawn Johnston Smiley

Masters Theses & Specialist Projects

3-Methylindole and 4-methylphenol are cytotoxic and malodorant compounds derived from tryptophan and tyrosine, respectively. Each is present in swine waste lagoons and contributes to malodorous emissions from agricultural facilities. Clostridium scatologenes ATCC 25775 produces both compounds and serves as a model organism to study their metabolism and function. Through the repeated assembly and annotation of the Clostridium scatologenes genome, we propose a novel pathway for tryptophan degradation and 3-methylindole production by this organism. The genome of Clostridium scatologenes was sequenced, and re-assembled into contigs. Key elements of the tryptophan and shikimate pathways were identified. Contigs containing these elements were extracted …

Consumer Knowledge, Perception And Attitudes Of Unlabeled Genetically Modified Foods Of An Educated Population In The State Of Kentucky, Aldious A. Waite

Masters Theses & Specialist Projects

Genetically modified (GM) foods technology is a novel idea for improving food and crop production, but the supposed health risk of GM foods, such as possible negative long-term health effects to humans, animals and the environment, have provoked the European Union to create assessment protocols to monitor and regulate the safety of GM foods and crops. This research investigates the perception and attitudes of unlabeled GM foods of the WKU faculty and staff. A survey was administered via WKU Qualtrics, and chi-square tests were performed to see how the benefits and disadvantages of GM foods may affect the purchasing decisions …

Functional And Expression Analysis Of A Novel Basement Membrane Degrader In Drosophila Melanogaster, Christopher J. Fields

Masters Theses & Specialist Projects

The Srivastava Lab is focused on the identification and characterization of genes that play a role in basement membrane remodeling. Previously, we identified putative basement membrane degraders through a genetic screen. One such gene has been suggested to play a role in the maintenance of the stem cell niche in Drosophila melanogaster, but no other information about the role this gene plays in development or disease has been published. Here, data are presented from experiments utilizing Drosophila genetics and immunohistochemistry that provide important insights on the biological role of this gene.

Collagenase activity was up-regulated upon overexpression of this gene, …

Single Nucleotide Polymorphisms Linked To Essential Hypertension In Kasigau, Kenya, Julia Carol Freeman

Masters Theses & Specialist Projects

Hypertension, or high blood pressure (BP), is an ever-growing epidemic in the developing world. Understanding the genetics behind essential hypertension (EH), or hypertension with no known cause, is especially important. In this study, three single nucleotide polymorphisms (SNPs) known to be linked to an increase in susceptibility to EH were quantified from a cohort of Kenyans living in the Kasigau region. The SNPs are located in three genes that are part of the renin angiotensin system, the primary regulatory pathway in humans controlling BP. They include: AGT (rs699), AGTR1 (rs5186), and HSD11β2 (rs5479). Overall, by using a fluorescent-based RT-PCR technique, …

Defining The Requirements For Early Gene Expression In Bacteriophage Hk639, Amanda L. Seaton

Masters Theses & Specialist Projects

Lambdoid phages suppress transcription termination to fully express their genes. Antitermination of early gene expression in most lambdoid phages is mediated by an interaction between the N protein and a number of host-encoded factors. Bacteriophage HK022 does not rely on a protein for antitermination. To promote full expression of early phage genes, the transcripts of the HK022 put sites interact directly with RNA polymerase to convert it to a termination resistant form. Bacteriophage HK639 also uses RNA-mediated antitermination. However, it only possesses a single put-like element in its left operon. Because most lambdoid phages, including HK022, have antiterminator elements in …

Development Of Tools To Assess The Effects Of Lunasin On Normal Development And Tumor Progression In Drosophila Melanogaster, Gillian E. Jones

Masters Theses & Specialist Projects

Soy contains many bioactive molecules known to elicit anti-cancer effects. One such peptide, Lunasin, has been shown to selectively act on newly transformed cells while having no cytotoxic effect on non-tumorigenic or established cancer cell lines. In this study we attempt to understand the developmental effects of Lunasin overexpression in vivo and create reagents that will help us understand Lunasin’s anti tumorigenic effects in an intact organism. cDNA encoding lunasin and EGFP-lunasin were cloned into pUAST and microinjected into Drosophila embryos. Tissue-specific overexpression of EGFP-Lun in the resulting transgenic lines was accomplished by crossing transgenics to various GAL4 driver lines. …

An Automatic Framework For Embryonic Localization Using Edges In A Scale Space, Zachary Bessinger

Masters Theses & Specialist Projects

Localization of Drosophila embryos in images is a fundamental step in an automatic computational system for the exploration of gene-gene interaction on Drosophila. Contour extraction of embryonic images is challenging due to many variations in embryonic images. In the thesis work, we develop a localization framework based on the analysis of connected components of edge pixels in a scale space. We propose criteria to select optimal scales for embryonic localization. Furthermore, we propose a scale mapping strategy to compress the range of a scale space in order to improve the efficiency of the localization framework. The effectiveness of the proposed …

The Torsion Angle Of Random Walks, Mu He

Masters Theses & Specialist Projects

In this thesis, we study the expected mean of the torsion angle of an n-step
equilateral random walk in 3D. We consider the random walk is generated within a confining sphere or without a confining sphere: given three consecutive vectors ^→e₁ , ^→e₂ , and ^→e₃ of the random walk then the vectors ^→e₁ and ^→e₂ define a plane and the vectors ^→e₂ and ^→e₃ define a second plane. The angle between the two planes is called the torsion angle of the three vectors. Algorithms are …

Contour Extraction Of Drosophila Embryos Using Active Contours In Scale Space, Soujanya Siddavaram Ananta

Masters Theses & Specialist Projects

Contour extraction of Drosophila embryos is an important step to build a computational system for pattern matching of embryonic images which aids in the discovery of genes. Automatic contour extraction of embryos is challenging due to several image variations such as size, shape, orientation and neigh- boring embryos such as touching and non-touching embryos. In this thesis, we introduce a framework for contour extraction based on the connected components in the gaussian scale space of an embryonic image. The active contour model is applied on the images to refine embryo contours. Data cleaning methods are applied to smooth the jaggy …

Assembly And Automated Annotation Of The Clostridium Scatologenes Genome, Jitesh Tiwari

Masters Theses & Specialist Projects

Clostridium scatologenes is an anaerobic bacterium that demonstrates some unusual metabolic traits such as the production of 3-methyl indole. The availability of genome level sequencing has lent itself to the exploration and elucidation of unique metabolic pathways in other organisms such as Clostridium botulinum. The Clostridium scatologenes genome, with an estimated length 4.2 million bp, was sequenced by the Applied Biosystems Solid method and the Roche 454 pyrosequencing method. The resulting DNA sequences were combined and assembled into 8267 contigs with an average length of 1250 bp with the Newbler Assembler program. Comparision of published subunits of csd gene …

The Genomic Sequence And Annotation Of Bacteriophage Hk239, Alice Ann Wright

Masters Theses & Specialist Projects

Bacteriophages are viruses that infect bacteria and they are the most numerous biological entities on Earth. Temperate phage can adopt two different lifestyles. In the lytic lifestyle, a phage injects its genome into the host and a controlled developmental program ensues. The phage DNA is replicated, phage genes are expressed and new viral particles are assembled. Ultimately, the host cell lyses and the phage particles are released into the environment. In the lysogenic lifestyle, a phage integrates its genome into the host chromosome, creating a prophage. The cell containing the prophage is known as a lysogen. Most prophage genes are …

Analysis Of The Clear Plaque Phenotype Of The Bacteriophage Hk75, Phani Chandrika Kunapuli

Masters Theses & Specialist Projects

The growth of bacteriophage HK75 is inhibited by specific mutations in the zinc binding domain of the host RNA polymerase beta prime subunit. It shares this rare property with bacteriophage HK022 and other phages that use RNA mediated antitermination to promote early gene expression. Recent genomic analysis of HK75 and HK022 has confirmed the relatedness of these two phages and place HK75 in the lambdoid family of bacteriophages. Lambdoid phages are temperate and can adopt a lytic or lysogenic lifestyle upon infection of a suitable host. However, HK75 only forms clear plaques and thus appears to be defective in its …

Appearance Based Stage Recognition Of Drosophila Embryos, Gopi Chand Nutakki

Masters Theses & Specialist Projects

Stages in Drosophila development denote the time after fertilization at which certain specific events occur in the developmental cycle. Stage information of a host embryo, as well as spatial information of a gene expression region is indispensable input for the discovery of the pattern of gene-gene interaction. Manual labeling of stages is becoming a bottleneck under the circumstance of high throughput embryo images. Automatic recognition based on the appearances of embryos is becoming a more desirable scheme. This problem, however, is very challenging due to severe variations of illumination and gene expressions. In this research thesis, we propose an appearance …

Development Of A Genetic Modification System In Clostridium Scatologenes Atcc 25775 For Generation Of Mutants, Prasanna Tamarapu Parthasarathy

Masters Theses & Specialist Projects

3-Methyl indole (3-MI) is a malodorant in food and animal waste and Clostridium scatologenes ATCC 25775 is the model organism for the study of 3-MI production. 3-MI is an anaerobic degradation product of L-tryptophan and can cause pulmonary disorders and death in cattle and goats. To elucidate the 3-MI biosynthesis pathway and the underlying genes, it is necessary to develop a system to allow genetic modification in Clostridium scatologenes ATCC 25775. Bacteriophages and transposons are useful tools to achieve this goal. Isolation of Clostridium scatologenes ATCC 25775 bacteriophage was attempted by prophage induction and enrichments using environmental sources. To induce …

The Effects Of Growth Hormone In The Inner Ear Of Zebrafish (Danio Rerio) During Hair Cell Regeneration, Chia-Hui Lin

Masters Theses & Specialist Projects

Although deafness is a universal problem, effective treatments have remained elusive. In order to develop potential treatments, an overall understanding of the cellular process of auditory hair cell regeneration, which occurs in fish but not mammals, must be established. A previous microarray analysis and qRT-PCR validation of noise-exposed zebrafish showed that growth hormone (GH) was significantly upregulated during the process of auditory hair cell regeneration. Thus, GH may play an important role during hair cell regeneration. However, cellular effects of exogenous GH in the zebrafish auditory hair cell regeneration have not been examined after noise exposure. To understand the effect …

Molecular Analysis Of Trypanosoma Cruzi Isolates Obtained From Raccoons (Procyon Lotor) In Warren And Barren Counties Of Kentucky, Lipeng Bi

Masters Theses & Specialist Projects

Trypanosoma cruzi, the etiologic agent of Chagas disease, infects a variety of wild mammals in the southern United States, but it has only recently been isolated from raccoons trapped in the state of Kentucky. The purpose of the present study was to use a molecular genotyping approach, followed by DNA sequencing to determine the genotypes (type I, or types IIa-IIe) of 15 of the Kentucky isolates. DNA samples were prepared from 15 T. cruzi- isolates using a Qiagen mini kit, and PCR amplification was performed using published primers for the 24S α rDNA sequence (D71 and D72), the …

Cloning Of "Animal Cryptochrome" Cdna From The Model Organism Chlamydomonas Reinhardtii For Functional Analysis Of Its Protein Product, Shobha Lavanya Silparasetty

Masters Theses & Specialist Projects

reinhardtii, a unicellular green alga, is a model organism to study the circadian clock. Cryptochromes are the blue light photoreceptors that entrain the clock in some organisms. The CPH1 protein of C. reinhardtii resembles the cryptochromes of the plant model Arabidopsis, but whether CPH1 entrains the circadian clock in C. reinhardtii is not yet known. Recent reports have suggested the existence of one more cryptochrome in C. reinhardtii, which resembles the cryptochromes of animals. However, the amino acid sequence of this protein shows even higher sequence similarity with the 6-4 DNA photolyase of Arabidopsis. DNA photolyases …

The Lyme Disease Spirochete, Borrelia Burgdorferi, In Tick Species Collected From Raccoons (Procyon Lotor) And Opossums (Didelphis Virginiana) Trapped In The Warren And Barren Counties Of South Central Kentucky, Kristina Tackett

Masters Theses & Specialist Projects

The incidence of tick-borne zoonoses such as Ehrlichiosis, Rocky Mountain Spotted Fever, and Lyme disease has steadily increased in the southeastern United States in recent years. According to the Centers for Disease Control and Prevention (CDC), the southeastern states accounted for 1,200 of the 27,000 total cases of Lyme disease reported in the U.S. in 2007. Although Ixodes scapularis is the most commonly recognized vector for the Lyme disease spirochete Borrelia burgdorferi, Dermacentor variabilis (a common vector for Rocky Mountain Spotted Fever) also has been shown to be a viable host for this pathogen. The purpose of the present …

Assessment Of The Endangered Species Podarcis Carbonelli On A Microgeographic Scale: A Molecular, Morphological And Physiological Approach, Maria Clara Figueirinhas Do Amaral

Masters Theses & Specialist Projects

The lizard Podarcis carbonelli is an endangered species endemic to the Iberian Peninsula. One location where this species occurs is at the Berlengas Natural Preserve, an Atlantic archipelago off the coast of Portugal. These island populations are geographically separated from nearby mainland populations. The fundamental question is, are these insular individuals distinct from the mainland populations? Four localities were chose for comparison: two island populations and two nearby coastal populations. We assessed this question using three distinct approaches: molecular, morphological and physiological approach. We sequenced the 12S RNA, the mtDNA Control Region and the 7th intron of the !-fibrinogen gene …

Adding Upstream Sequence And A Downstream Reporter To The Bile Acid Inducible Promoter Of Clostridium Scindens Vpi 12708, Bryan Patrick Mason

Masters Theses & Specialist Projects

Bile acids in the small intestines of animals serve to breakdown fats and fatsoluble vitamins. Most of the bile acids are reabsorbed into the enterohepatic circulation, but approximately five percent of these bile acids pass into the large intestine. These bile acids are swiftly deconjugated by the bacterial population, and then subjected to further intestinal bacterial chemical modifications. The most significant of these modifications are 7α-dehydroxylations which form secondary bile acids (deoxycholate and lithocholate). Much research has illuminated the 7α-dehydroxylation pathway: of particular interest is the bile acid inducible operon, for which Clostridium scindens VPI 12708 serves as the model …