Genetics and Genomics Commons^™

Trail-Based High Throughput Screening Reveals A Link Between Trail-Mediated Apoptosis And Glutathione Reductase, A Key Component Of Oxidative Stress Response., Dmitri Rozanov, Anton Cheltsov, Eduard Sergienko, Stefan Vasile, Vladislav Golubkov, Alexander E Aleshin, Trevor Levin, Elie Traer, Byron Hann, Julia Freimuth, Nikita Alexeev, Max A Alekseyev, Sergey P Budko, Hans Peter Bächinger, Paul Spellman

Computational Biology Institute

A high throughput screen for compounds that induce TRAIL-mediated apoptosis identified ML100 as an active chemical probe, which potentiated TRAIL activity in prostate carcinoma PPC-1 and melanoma MDA-MB-435 cells. Follow-up in silico modeling and profiling in cell-based assays allowed us to identify NSC130362, pharmacophore analog of ML100 that induced 65-95% cytotoxicity in cancer cells and did not affect the viability of human primary hepatocytes. In agreement with the activation of the apoptotic pathway, both ML100 and NSC130362 synergistically with TRAIL induced caspase-3/7 activity in MDA-MB-435 cells. Subsequent affinity chromatography and inhibition studies convincingly demonstrated that glutathione reductase (GSR), a key …

2015 Oklahoma Research Day Full Program, Northeastern State University

Oklahoma Research Day Abstracts

This document contains all abstracts from the 2015 Oklahoma Research Day held at Northeastern State University.

Developmental Ethanol Exposure Leads To Dysregulation Of Lipid Metabolism And Oxidative Stress In Drosophila, Theresa Logan-Garbisch, Anthony Bortolazzo, Peter Luu, Audrey Ford, David Do, Payam Khodabakhshi, Rachael French

Faculty Publications, Biological Sciences

Ethanol exposure during development causes an array of developmental abnormalities, both physiological and behavioral. In mammals, these abnormalities are collectively known as Fetal Alcohol Effects (FAE) or Fetal Alcohol Spectrum Disorder (FASD). We have established a Drosophila melanogaster model of FASD, and have previously shown that developmental ethanol exposure in flies leads to reduced expression of insulin like peptides (dILPs) and their receptor. In this work, we link that observation to dysregulation of fatty acid metabolism and lipid accumulation. Further, we show that developmental ethanol exposure in Drosophila causes oxidative stress, that this stress is a primary cause of the …

Genome Engineering To Create Dominant Alleles In Caenorhabditis Elegans Using Crispr-Cas9 Technology, Abrar Sulaimani

Graduate School of Biomedical Sciences Theses and Dissertations

Many investigators have being using CRISPR-Cas9 as a method of genome engineering because it is easy, accurate and fast. This technique has been used to modify the genomes of a wide variety of organisms, including the nematode Caenorhabditis elegans (C. elegans). The short life cycle and ease of introducing exogenous plasmids make C. elegans an ideal system for advancing this technique. My thesis had two aims that focused on developing methods to create dominant alleles in C. elegans. Genetic modifications like precise deletion and insertions into a locus of chromosome are technically challenging. Additionally, although there are several ways of …

Subsistence Strategies In Traditional Societies Distinguish Gut Microbiomes, Alexandra J. Obregon-Tito, Raul T. Tito, Jessica Metcalf, Krithvivasa Sankaranarayanan, Jose C. Clemente, Luke K. Ursell, Zhenjiang Zech Xu, Will Van Treuren, Rob Knight, Morris Foster

Division of Research and Economic Development Faculty & Staff Publications

Recent studies suggest that gut microbiomes of urban-industrialized societies are different from those of traditional peoples. Here we examine the relationship between lifeways and gut microbiota through taxonomic and functional potential characterization of faecal samples from hunter-gatherer and traditional agriculturalist communities in Peru and an urban-industrialized community from the US. We find that in addition to taxonomic and metabolic differences between urban and traditional lifestyles, hunter-gatherers form a distinct sub-group among traditional peoples. As observed in previous studies, we find that Treponema are characteristic of traditional gut microbiomes. Moreover, through genome reconstruction (2.2-2.5 MB, coverage depth x26-513) and functional potential …

Early Initiation Of Antiretroviral Therapy Can Functionally Control Productive Hiv-1 Infection In Humanized-Blt Mice, Qingsheng Li, For Yue Tso, Guobin Kang, Wuxun Lu, Yue Li, Wenjin Fan, Zhe Yuan, Christopher J. Destache, Charles Wood

Nebraska Center for Virology: Faculty Publications

Background—Recent reports showed that functional control of HIV-1 infection for a prolonged time is possible by early anti-retroviral therapy (ART); however its underlying mechanism needs to be studied with a suitable animal model. Recently, humanized-BLT (bone marrow, liver and thymus) mouse (hu-BLT) was shown to be an excellent model for studying HIV-1 infection. We thus tested the feasibility of studying functional control of HIV-1 infection using hu-BLT mice.

Methods—Animals in three treatment groups (Rx-6h, Rx-24h, Rx-48h) and untreated group were infected with HIV-1, followed by ART initiation at 6, 24 or 48 hours post-infection and continued daily for two weeks. …

The Role Of Dendritic Cell Subsets And Innate Immunity In The Pathogenesis Of Type 1 Diabetes And Other Autoimmune Diseases, Jeffrey D. Price, Kristin V. Tarbell

Nebraska Center for Virology: Faculty Publications

Dendritic cells (DCs) are key antigen-presenting cells that have an important role in autoimmune pathogenesis. DCs control both steady-state T cell tolerance and activation of pathogenic responses. The balance between these two outcomes depends on several factors, including genetic susceptibility, environmental signals that stimulate varied innate responses, and which DC subset is presenting antigen. Although the specific DC phenotype can diverge depending on the tissue location and context, there are four main subsets identified in both mouse and human: conventional cDC1 and cDC2, plasmacytoid DCs, and monocyte-derived DCs. In this review, we will discuss the role of these subsets in …

Composition, Taxonomy And Functional Diversity Of The Oropharynx Microbiome In Individuals With Schizophrenia And Controls., Eduardo Castro-Nallar, Matthew L Bendall, Marcos Pérez-Losada, Sarven Sabuncyan, Emily G Severance, Faith B Dickerson, Jennifer R Schroeder, Robert H Yolken, Keith A Crandall

Computational Biology Institute

The role of the human microbiome in schizophrenia remains largely unexplored. The microbiome has been shown to alter brain development and modulate behavior and cognition in animals through gut-brain connections, and research in humans suggests that it may be a modulating factor in many disorders. This study reports findings from a shotgun metagenomic analysis of the oropharyngeal microbiome in 16 individuals with schizophrenia and 16 controls. High-level differences were evident at both the phylum and genus levels, with Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria dominating both schizophrenia patients and controls, and Ascomycota being more abundant in schizophrenia patients than controls. Controls …

Gene Function In Schistosomes: Recent Advances Toward A Cure, Arnon D. Jurburg, Paul J. Brindley

Microbiology, Immunology, and Tropical Medicine Faculty Publications

No abstract provided.

Further Description Of Hybridization In The Sea Stars Asterias Rubens And Asterias Forbesi Through The Application Of Species-Specific Genetic Markers, Callie Bateson

Honors Program Theses

Hybridization occurs when two separate species reproduce to form hybrid offspring. If the hybrid offspring backcross with individuals of their parental species, the subsequent offspring may have varying amounts of the parental genomes, if viable. Following the retreat of the Last Glacial Maxima 20,000 years ago, two sea star species, Asterias forbesi and A. rubens came into secondary contact in the Gulf of Maine where they began to hybridize. Previous research in our lab studied the frequency of hybridization at four locations along the east coast (Quebec, Nova Scotia, New Hampshire, and New York) using morphological identifications as well two …

A Protective Role Of Autophagy In A Drosophila Model Of Friedreich's Ataxia (Frda), Luan Wang

Wayne State University Dissertations

Friedreich’s ataxia (FRDA) is an inherited autosomal recessive neurodegenerative disease. It affects 1 in every 50,000 people in central Europe and North America. FRDA is caused by deficiency of Frataxin, an essential mitochondrial iron chaperone protein, and the associated oxidative stress damages. Autophagy, a housekeeping process responsible for the bulk degradation and turnover of long half-life proteins and organelles, is featured by the formation of double-membrane vacuoles and lysosomal degradation. Previous researches indicate that Danon’s disease, the inherited neural disorder disease that shares similar symptoms with FRDA, is due to the malfunction of autophagy. Based on this, we raise the …

Technical Note: Collection And Preparation Techniques Change Nutrient Composition Of Masticate Collected From Esophageally Fistulated Cattle, Jacqueline A. Musgrave, J. V. Judy, K. H. Jenkins, T. J. Klopfenstein, L. A. Stalker

Department of Animal Science: Faculty Publications

Two experiments determined effects of collection and preparation techniques on nutrient composition of masticate samples from esophageally fistulated cattle. In Exp. 1, 12 esophageally fistulated cattle were maintained on 2 precollection diets, high CP (24% CP; n = 6) or low CP (7.7% CP; n = 6), for 8 d. On d 9, the esophageal plug was removed, screen bottom bags were attached, and each cow was offered fresh grass. Immediately after fresh grass sample collection was complete, dry grass (hay) was offered and a sample was collected. Blood samples were collected and analyzed for serum urea nitrogen concentration. Masticate …

Comparative Proteomics Of Mouse Tears And Saliva: Evidence From Large Protein Families For Functional Adaptation, Robert C. Karn, Christina M. Laukaitis

Scholarship and Professional Work - LAS

We produced a tear proteome of the genome mouse, C57BL/6, that contained 139 different protein identifications: 110 from a two-dimensional (2D) gel with subsequent trypsin digestion, 19 from a one-dimensional (1D) gel with subsequent trypsin digestion and ten from a 1D gel with subsequent Asp-N digestion. We compared this tear proteome with a C57BL/6 mouse saliva proteome produced previously. Sixteen of the 139 tear proteins are shared between the two proteomes, including six proteins that combat microbial growth. Among the 123 other tear proteins, were members of four large protein families that have no counterparts in humans: Androgen-binding proteins (ABPs) …

Establishment Of Biotrophy By The Maize Anthracnose Pathogen Colletotrichum Graminicola: Use Of Bioinformatics And Transcriptomics To Address The Potential Roles Of Secretion, Stress Response, And Secreted Proteins, Ester Alvarenga Santos Buiate

Theses and Dissertations--Plant Pathology

Colletotrichum graminicola is a hemibiotrophic pathogen of maize that causes anthracnose leaf and stalk rot diseases. The pathogen penetrates the host and initially establishes an intracellular biotrophic infection, in which the hyphae are separated from the living host cell by a membrane that is elaborated by the host, apparently in response to pathogen signals. A nonpathogenic mutant (MT) of C. graminicola was generated that germinates and penetrates the host normally, but is incapable of establishing a normal biotrophic infection. The mutated gene is Cpr1, conserved in eukaryotes and predicted to encode a component of the signal peptidase complex. How …

Germline Mutations In Chek1 And Chek2 In Women With Ovarian, Peritoneal, Or Fallopian Tube Cancer, Maria Isabel Harrell

Walden Dissertations and Doctoral Studies

Ovarian cancer is the deadliest gynecological malignancy affecting women. Diagnosis often occurs late due to non-specific symptoms, but if detected early, there is excellent chance for survival. One of the most important risk factors is family history. Up to 24% of cases are due to inherited loss-of-function mutations in genes involved in the DNA damage response. The theory underlying hereditary cancers is Knudson's two-hit theory of cancer causation, where two hits are necessary for cancer to occur in an individual: one in the germline and one in the tissue. The genes, CHEK1 and CHEK2, are modulators of the DNA damage …

Investigating The Influence Of Chd1 On Gene Expression In Drosophila Melanogaster Using Position Effect Variegation, Phuongngan Thi Bui

Scripps Senior Theses

Position Effect Variegation (PEV) is the mosaic expression of a gene that has been moved out of its optimal environment and into a different area on the chromosome. Changing a gene’s environment may have profound effects on its eligibility for proper expression, which is a complicated process regulated by many factors. The PEV phenomenon is used as an assay to study gene expression as regulated by chromatin structure. In this study, the Drosophila melanogaster white gene was used as a reporter to study the various effects of CHD1, a chromatin regulating factor, on PEV gene expression. Inspired by preliminary data …

Proof-Of-Concept Of Environmental Dna Tools For Atlantic Sturgeon Management, Jameson Hinkle

Theses and Dissertations

Abstract

The Atlantic Sturgeon (Acipenser oxyrinchus oxyrinchus, Mitchell) is an anadromous species that spawns in tidal freshwater rivers from Canada to Florida. Overfishing, river sedimentation and alteration of the river bottom have decreased Atlantic Sturgeon populations, and NOAA lists the species as endangered. Ecologists sometimes find it difficult to locate individuals of a species that is rare, endangered or invasive. The need for methods less invasive that can create more resolution of cryptic species presence is necessary. Environmental DNA (eDNA) is a non-invasive means of detecting rare, endangered, or invasive species by isolating nuclear or mitochondrial DNA (mtDNA) from the …

High-Throughput Comparison, Functional Annotation, And Metabolic Modeling Of Plant Genomes Using The Plantseed Resource, Samuel M. D. Seaver, Svetlana Gerdes, Océane Frelin, Claudia Lerma-Ortiz, Louis M. T. Bradbury, Rémi Zallot, Ghulam Hasnain, Thomas D. Niehaus, Basma El Yacoubi, Shiran Pasternak, Robert Olson, Gordon Pusch, Ross Overbeek, Valérie De Crécy-Lagarde, Doreen Ware, Andrew D. Hanson, Christopher S. Henry

Publications and Research

There is a growing demand for genome-scale metabolic reconstructions for plants, fueled by the need to understand the metabolic basis of crop yield and by progress in genome and transcriptome sequencing. Methods are also required to enable the interpretation of plant transcriptome data to study how cellular metabolic activity varies under different growth conditions or even within different organs, tissues, and developmental stages. Such methods depend extensively on the accuracy with which genes have been mapped to the biochemical reactions in the plant metabolic pathways. Errors in these mappings lead to metabolic reconstructions with an inflated number of reactions and …

A Nuclear Fraction Of Turnip Crinkle Virus Capsid Protein Is Important For Elicitation Of The Host Resistance Response, Sung-Hwan Kang, Feng Qu, Thomas J. Morris

Nebraska Center for Virology: Faculty Publications

The N-terminal 25 amino acids (AAs) of turnip crinkle virus (TCV) capsid protein (CP) are recognized by the resistance protein HRT to trigger a hypersensitive response (HR) and systemic resistance to TCV infection. This same region of TCV CP also contains a motif that interacts with the transcription factor TIP, as well as a nuclear localization signal (NLS). However, it is not yet known whether nuclear localization of TCV CP is needed for the induction of HRT-mediated HR and resistance. Here we present new evidence suggesting a tight correlation between nuclear inclusions formed by CP and the manifestation of HR. …

A Novel Codon-Optimized Siv Gag-Pol Immunogen For Genebased Vaccination, Catherine M. Crosby, Eric A. Weaver, Reeti Khare, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

Simian immunodeficiency virus (SIV) is a robust pathogen used in non-human primates to model HIV vaccines. SIV encodes a number of potential vaccine targets. By far the largest and most conserved protein target in SIV is its gag-pol protein that bears many epitopes to drive multivalent immune T cell responses. While gag-pol is an attractive antigen, it is only translated after a frame shift between gag and pol with the effect that gag and pol are expressed at an approximate 10/1 ratio. The codon bias of native lentiviral genes are also mismatched with the abundance of tRNAs in mammalian cells …

Dehydrated Citrus Pulp Alters Feedlot Performance Of Crossbred Heifers During The Receiving Period And Modulates Serum Metabolite Concentrations Before And After An Endotoxin Challenge, J. T. Cribbs, B. C. Bernhard, T. R. Young, M. A. Jennings, N.C. Burdick Sanchez, J. A. Carroll, T. R. Callaway, T. B. Schmidt, B. J. Johnson, R. J. Rathmann

Department of Animal Science: Faculty Publications

English × Continental heifers (n = 180) were sourced in 2 loads (219.3 ± 16.0 and 221.4 ± 16.4 kg, respectively) from commercial auction barns to study the effects of feeding dehydrated citrus pulp (DCP) on feedlot performance of newly received heifers. A completely randomized block design was used with BW nested within arrival load and blocked by BW into 3 dietary treatments (36 pens, 5 heifers/ pen, 12 blocks, 3 pens/block, and 12 pens/treatment). Treatment diets contained 1) 0% DCP (control diet [CON]), 2) 10% DCP, or 3) 20% DCP on a DM basis. Diets containing DCP were exchanged …

Case Study: Producer Concerns And Perceptions Regarding The Effect Of Methane On Cattle Production And The Environment: A Survey Of Nebraska Producers, B. M. Boyd, A. Jones, Lisa Franzen-Castle, K. Jenkins, Richard J. Rasby, Matt K. Luebbe, Richard R. Stowell, Samodha C. Fernando, Galen E. Erickson

Department of Animal Science: Faculty Publications

Enteric methane production from cattle and its effect on climate change has been a topic of debate. Multiple studies have explored methods to reduce cattle enteric methane production while simultaneously improving performance. However, most strategies developed have not been widely implemented by cattle producers. Knowledge of producer concerns and perceptions on methane production from cattle and its effect on the environment may be limited. Therefore, the objectives of this survey were to determine what Nebraska producers know about methane production by cattle and how it affects performance and to determine whether different age groups, regions of Nebraska, and production size …

Development Of A Row-Crop Tractor Data Acquisition System, James Roeber, Santosh Pitla, Roger M. Hoy, Joe D. Luck

Department of Animal Science: Faculty Publications

Numerous advancements in machinery performance on row-crop agricultural tractors have been made in recent years. Currently, the Organisation for Economic Co-operation and Development (OECD) only tests predetermined points (E.g., max. power, max. torque) for drawbar and hydraulic power. This method of testing has been used for many years with the tractor at a steady state which is usually not typical of row-crop operations. A data acquisition system (DAQ) was designed that records implement energy consumption (E.g., drawbar loading, PTO power, and hydraulic power). The system used a LabVIEW program and a compact data acquisition system (cDAQ) to record data from …

An Application Of Mesh Enrichment Analysis In Livestock, Gota Morota, F. Moreno, James L. Petersen, Daniel C. Ciobanu, K. Tsuyuzaki, I. Nikaido

Department of Animal Science: Faculty Publications

An integral part of functional genomics studies is to assess the enrichment of specific biological terms in lists of genes found to be playing an important role in biological phenomena. Contrasting the observed frequency of annotated terms with those of the background is at the core of overrepresentation analysis (ORA). Gene Ontology (GO) is a means to consistently classify and annotate gene products and has become a mainstay in ORA. Alternatively, Medical Subject Headings (MeSH) offers a comprehensive life science vocabulary including additional categories that are not covered by GO. Although MeSH is applied predominantly in human and model organism …

Optimizing The Use Of Fibrous Residues In Beef And Dairy Diets, James C. Macdonald, G. E. Erickson, P. J. Kononoff, T. J. Klopfenstein

Department of Animal Science: Faculty Publications

Increased corn prices over the past decade have altered land use away from traditional forage in favor of corn. Accordingly, beef and dairy producers have had to adopt non-traditional forage resources into their production systems, many of which have become available as a result of increased corn production. Byproducts of the wet and dry milling industries have been used to replace both corn and forage in beef and dairy diets. Byproducts containing corn bran have large amounts of readily digestible hemicellulose. The use of byproducts may increase milk production, ADG, and G:F in dairy, beef growing, and beef finishing diets, …

Microbial Diversity And Activity In The Nematostella Vectensis Holobiont: Insights From 16s Rrna Gene Sequencing, Isolate Genomes, And A Pilot-Scale Survey Of Gene Expression, Jia Y. Har, Tim Helbig, Ju H. Lim, Samodha C. Fernando, Adam M. Reitzel, Kevin Penn, Janelle R. Thompson

Department of Animal Science: Faculty Publications

We have characterized the molecular and genomic diversity of the microbiota of the starlet sea anemone Nematostella vectensis, a cnidarian model for comparative developmental and functional biology and a year-round inhabitant of temperate salt marshes. Molecular phylogenetic analysis of 16S rRNA gene clone libraries revealed four ribotypes associated with N. vectensis at multiple locations and times. These associates include two novel ribotypes within the ε-Proteobacterial order Campylobacterales and the Spirochetes, respectively, each sharing 99% 16S rRNA identity with Endozoicomonas elysicola and Pseudomonas oleovorans, respectively. Species-specific PCR revealed that these populations persisted in N. vectensis asecually propagated under laboratory …

Sheep Updates 2015 - Moora, Bruce Mullen, Kate Pritchett, Kimbal Curtis, Daniel Real, Jaq Pearson, Lynne Bradshaw, Geoff Lindon, Katherine Davies, Kevin Hepworth, Stephen Lee, Dawson Bradford, Lucy Anderton, Jackie Jarvis, Ben Patrick

Sheep Updates

This session covers thirteen papers from different authors:

1. The Sheep Industry Business Innovation project, Bruce Mullan, Sheep Industry Development Director, Department of Agriculture and Food, Western Australia

2. Western Australian sheep stocktake, Kate Pritchett and Kimbal Curtis, Research Officers, Department of Agriculture and Food, Western Australia

3. Tedera - a perenial forage legume to reduce your supplementary feeding in summer and autumn, Dr. Daniel Real, Senior Plant Breeder, Department of Agriculture and Food, Western Australia

4. National Livestock Identification System (NLIS) for sheep and goats - what is the NLIS database? Jac Pearson, Biosecurity Officer, Department of Agriculture and …

Sheep Updates 2015 - Katanning, Bruce Mullan, Kate Pritchett, Kimbal Curtis, Chris Wilcox, Mike Hyder, Leigh Sonnerman, Lynne Bradshaw, Geoff Lindon, Katherine Davies, Joe Young, Stephen Lee, Ian Robertson, Lucy Anderton, Hayley Norman, Ed Barrett-Lennard, Jackie Jarvis, Ben Patrick

Sheep Updates

This session covers fourteen papers from different authors:

1. The Sheep Industry Business Innovation project, Bruce Mullan, Sheep Industry Development Director, Department of Agriculture and Food, Western Australia

2. Western Australian sheep stocktake, Kate Pritchett and Kimbal Curtis, Research Officers, Department of Agriculture and Food, Western Australia

3. Wool demand and supply - short term volatility, long term opportunities, Chris Wilcox, Principal of Poimena Analysis

4. Lifetime management for maternal ewes, Mike Hyder, Research Officer, Department of Agriculture and Food, Western Australia

5. National Livestock Identification System (NLIS) for sheep and goats - what is the NLIS database? Leigh Sonnermann, …

Cellular Defects Caused By Hypomorphic Variants Of The Bloom Syndrome Helicase Gene Blm, Vivek M. Shastri, Kristina H. Schmidt

Molecular Biosciences Faculty Publications

BACKGROUND: Bloom syndrome is an autosomal recessive disorder characterized by extraordinary cancer incidence early in life and an average life expectancy of ~27 years. Premature stop codons in BLM, which encodes a DNA helicase that functions in DNA double-strand-break repair, make up the vast majority of Bloom syndrome mutations, with only 13 single amino acid changes identified in the syndrome. Sequencing projects have identified nearly one hundred single nucleotide variants in BLM that cause amino acid changes of uncertain significance.

METHODS AND RESULTS: Here, in addition to identifying five BLM variants incapable of complementing certain defects of Bloom syndrome cells, …

Exogenous Administration Of Lipids To Steers Alters Aspects Of The Innate Immune Response To Endotoxin Challenge, Nicole C. Burdick Sanchez, Jeffery A. Carroll, Janet R. Donaldson, Joe O. Buntyn, Ty B. Schmidt

Department of Animal Science: Faculty Publications

This study examined the effects of increasing energy availability from both dextrose and lipid treatments on the proinflammatory response to LPS in Holstein steers. Steers were randomly assigned to one of three groups: saline at 0.5 ml/ kg body weight (Control) or 50% dextrose [0.5 ml/kg body weight (Dextrose) to mimic calm cattle’s response to LPS] administered immediately prior to LPS (0.5 mg/kg body weight at 0 h) or continuous lipid emulsion infusion from –1 to 6 h [Intralipid 20% (Baxter, Deerfield, IL USA); 0.5 ml/kg/hr (Lipid) to mimic temperamental cattle]. Concentrations of non-esterified fatty acids (NEFA) were greater in …