Medicine and Health Sciences Commons^™

Cellular Poly(C) Binding Proteins 1 And 2 Interact With Porcine Reproductive And Respiratory Syndrome Virus Nonstructural Protein 1Β And Support Viral Replication, Lalit Beura, Phat X. Dinh, Fernando A. Osorio, Asit K. Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

Porcine reproductive and respiratory syndrome virus (PRRSV) infection of swine results in substantial economic losses to the swine industry worldwide. Identification of cellular factors involved in PRRSV life cycle not only will enable a better understanding of virus biology but also has the potential for the development of antiviral therapeutics. The PRRSV nonstructural protein 1 (nsp1) has been shown to be involved in at least two important functions in the infected hosts: (i) mediation of viral subgenomic (sg) mRNA transcription and (ii) suppression of the host’s innate immune response mechanisms. To further our understanding of the role of the viral …

Empathy-Based Conservation: An Interdisciplinary Approach To Conservation Policy And Decision-Making, Kaitlyn Delashmutt

Department of Environmental Studies: Undergraduate Student Theses

In the late 20^th century, neuroscientists in Italy discovered a neuron in the brain capable of mentally mimicking the emotions derived from the actions of others (Rizzolatti and Craighero, 2004). It is the process that makes your elbow ache when someone else knocks their elbow on the counter or the uncontrollable smile that creeps up when someone smiles at you. No questions asked, people intuitively sense what others are feeling. The old school of thought was that humans deduced through logic and reason the actions of others and interpreted the emotions through a rational process (Carew et al, 2008). …

Species D Adenoviruses As Oncolytics Against B-Cell Cancers, Christopher Y. Chen, Julien S. Senac, Eric A. Weaver, Shannon M. May, Diane F. Jelinek, Philip Greipp, Thomas Witzig, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

Purpose: Oncolytic viruses are self-amplifying anticancer agents that make use of the natural ability of viruses to kill cells. Adenovirus serotype 5 (Ad5) has been extensively tested against solid cancers, but less so against B-cell cancers because these cells do not generally express the coxsackie and adenoviral receptor (CAR). To determine whether other adenoviruses might have better potency, we "mined" the adenovirus virome of 55 serotypes for viruses that could kill B-cell cancers.

Experimental Design: Fifteen adenoviruses selected to represent Ad species B, C, D, E, and F were tested in vitro against cell lines and primary patient B-cell cancers …

Surveillance Of Selected Diseases In Free-Ranging Elk (Cervus Elaphus Nelsoni) In Nebraska, 1995-2009, Michael A. Cover, Scott E. Hygnstrom, David W. Oates, Kit M. Hams, Kurt C. Vercauteren

Great Plains Research: A Journal of Natural and Social Sciences

Sera samples were collected from 21 free-ranging, captured female elk (Cervus elaphus nelsoni) in 1995- 96, and tissue and sera samples were collected from 415 hunter-harvested elk from 1995 to 2006 and tested for selected diseases. Titers for Anaplasma marginale were detected in 81 of 436 (19%) elk. Occurrence of antibodies to anaplasmosis increased from 4 to 40 elk from 2002 to 2006. Titers for bovine viral diarrhea virus (BVDV) were detected in 18 of 346 (5%) samples. Titers for Leptospira interrogans serovars were detected in 21 of 289 (7%) of samples from 1995 to 2004. Titers for …

Perceived Importance And Actual Intake Of Calcium And Vitamin D In Young Female Athletes, Miriam E. Zambrano

Department of Nutrition and Health Sciences: Dissertations, Theses, and Student Research

Calcium needs increase as a child grows. Actual intake often decreases as a child gets older, leaving his/her developing bones at risk for injury during childhood and increasing the risk of developing osteoporosis later in life. The purpose of this study was to investigate relationships between psychosocial factors influencing consumption of calcium and vitamin D-rich foods and actual intakes of these nutrients. We examined the dietary calcium and vitamin D intakes, determined serum vitamin D levels, and assessed nutrition knowledge and perception of a sample of 20 young girls participating in competitive cheerleading in Omaha, Nebraska. From this information we …

Antagonistic Effects Of Cellular Poly(C) Binding Proteins On Vesicular Stomatitis Virus Gene Expression, Phat X. Dinh, Lalit Beura, Debasis Panda, Anshuman Das, Asit K. Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

Immunoprecipitation and subsequent mass spectrometry analysis of the cellular proteins from cells expressing the vesicular stomatitis virus (VSV) P protein identified the poly(C) binding protein 2 (PCBP2) as one of the P protein-interacting proteins. To investigate the role of PCBP2 in the viral life cycle, we examined the effects of depletion or overexpression of this protein on VSV growth. Small interfering RNA-mediated silencing of PCBP2 promoted VSV replication. Conversely, overexpression of PCBP2 in transfected cells suppressed VSV growth. Further studies revealed that PCBP2 negatively regulates overall viral mRNA accumulation and subsequent genome replication. Coimmunoprecipitation and immunofluorescence microscopic studies showed that …

Acute Fatty Liver Of Pregnancy: An Update On Mechanisms, Sathish Kumar Natarajan, Kavitha R. Thangaraj, Ashish Goel, C. E. Eapen, K. A. Balasubramanian, Anup Ramachandran

School of Veterinary and Biomedical Sciences: Faculty Publications

Acute fatty liver of pregnancy (AFLP), characterized by hepatic microvesicular steatosis, is a sudden catastrophic illness occurring almost exclusively in the third trimester of pregnancy. Defective fatty acid oxidation in the fetus has been shown to be associated with this disease. Since the placenta has the same genetic makeup as the fetus and as AFLP patients generally recover following delivery, we hypothesized that the placenta might be involved in pathogenesis of this disease. In an animal model of hepatic microvesicular steatosis (using sodium valproate), we found that microvesicular steatosis results in mitochondrial structural alterations and oxidative stress in subcellular organelles …

Glutaredoxin 2 Prevents H2o2-Induced Cell Apoptosis By Protecting Complex I Activity In The Mitochondria, Hongli Wu, Kuiyi Xing, Marjorie F. Lou

School of Veterinary and Biomedical Sciences: Faculty Publications

Glutaredoxin 2 (Grx2) belongs to the oxidoreductase family and is an isozyme of glutaredoxin 1 (Grx1) present in the mitochondria, however its function is not well understood. The purpose of this study is to evaluate the potential anti-apoptotic function of Grx2 by examining its ability to protect complex I in the mitochondrial electron transport system using human lens epithelial cells as a model. We found that cells treated with 200 μM hydrogen peroxide (H2O2) for 24 h exhibited decreased viability and became apoptotic with corresponding Bax up-regulation, Bcl-2 down-regulation, caspase 3 activation and mitochondrial cytochrome c leakage. Grx2 over-expression (OE) …

Initial Mlu Predicts The Relative Efficacy Of Two Grammatical Treatments In Preschoolers With Specific Language Impairments, Paul J. Yoder, Dennis L. Molfese, Elizabeth Gardner

Center for Brain, Biology, and Behavior: Faculty and Staff Publications

Purpose—We sought to confirm predictions based on past findings that pre-treatment mean length of utterance (MLU) would predict which of two grammatical treatments would best facilitate generalized and maintained grammatical development in preschoolers with specific language impairment (SLI).

Method—The participants were 57 preschoolers with specific language impairment (SLI). A randomized group experiment was used. The two grammatical treatments were Broad Target Recasts (BTR) and Milieu Language Teaching (MLT). MLU was assessed at Time 1 in two conversational language samples. Growth rate of productive grammar was quantified using growth curve modeling on the Index of Productive Syntax (IPSyn) from …

Identification Of Host Proteins Required For Vesicular Stomatitis Virus Infection, Debasis Panda

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

Viruses usurp host cell pathways for different stages of their infection. Understanding virus-host interaction will be invaluable to elucidate molecular mechanisms of virus infection and to identify drug targets. In order to identify such critical cellular genes in vesicular stomatitis virus (VSV, a model non-segmented negative strand RNA virus) infection, we developed a stable cell line constitutively expressing replication proteins of VSV. Attempts to establish a cell line replicating a sub-genomic replicon was not successful because of induction of interferon response by replication of viral genomic analog. Subsequently, we used siRNA technology and conducted a genome-wide siRNA screen in HeLa …

Effect Of The Infection With Porcine Reproductive And Respiratory Syndrome Virus On The Regulation Of Cytokines - Tumor Necrosis Factor-Alpha And Interleukin-10, Sakthivel Subramaniam

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

Porcine reproductive and respiratory syndrome virus (PRRSV) causes late-term abortion in sows and pneumonia in growing piglets. PRRSV evades the host immune response by several mechanisms, including the modulation of cytokine secretions in infected pigs, which is the subject of this dissertation. Particularly, PRRSV reduces the secretion of the pro-inflammatory cytokine, tumor necrosis factor-α (TNF-α) but increases the secretion of the anti-inflammatory cytokine interleukin-10 (IL-10). The latter effect, however, is PRRSV strain-specific. In this dissertation, we have examined mechanisms by which PRRSV regulates TNF-α and IL-10 expressions. The pathogenic strain FL12, derived from a PRRSV infectious clone, consistently suppressed TNF-α …

Role Of Porcine Reproductive And Respiratory Syndrome Virus Non-Structural Protein 1 In Viral Replication And Pathogenesis, Lalit Beura

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

Porcine reproductive and respiratory syndrome virus (PRRSV) is an infectious agent of significant concern to the global swine industry. PRRSV infection of pigs is initiated by a long viremia after which the virus enters in an extended persistent phase of 3-4 months that ultimately is resolved by the immune system. The delayed and weak host adaptive response is responsible for the protracted convalescence period. An initial sub-optimal innate immune response is postulated to be the reason behind such meager adaptive immune response. A major focus of the studies undertaken in this dissertation was to identify the viral non-structural proteins (nsps) …

Effects Of Single Nucleotide Polymorphisms In The Human Holocarboxylase Synthetase Gene On Catalytic Activity, Shingo Esaki

Department of Nutrition and Health Sciences: Dissertations, Theses, and Student Research

Holocarboxylase synthetase (HCS) catalyzes the covalent binding of biotin to carboxylases and histones in eukaryotic cells. Biotinylated carboxylases play essential roles in the metabolism of fatty acids, amino acids, and glucose; biotinylated histones play essential roles in gene regulation and genome stability. HCS null individuals are not viable whereas HCS deficiency is linked to developmental delays and phenotypes such as short life span and low stress resistance. Greater than 2,500 single nucleotide polymorphisms (SNPs) have been reported for HCS, but the biological importance of these polymorphisms is unknown. We hypothesized that some of these SNPs impair catalytic activity and that …

Comparison Of Adenoviruses As Oncolytics And Cancer Vaccines In An Immunocompetent B Cell Lymphoma Model, Eric A. Weaver, Christopher Y. Chen, Shannon M. May, Mary E. Barry, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

We have screened human adenoviruses (Ads) for oncolytic activity against a variety of mouse and hamster cell lines and have found a number that are susceptible to a variety of Ad serotypes. A20 lymphoma is derived from BALB/c mice and is susceptible to infection and killing by a variety of human Ads. A20 is also a suitable cancer vaccine model, because these cells express a unique immunoglobulin variable region that can be targeted by vaccination. To compare Ads as cancer vaccines versus Ads as oncolytics, A20 tumors were initiated in immunocompetent BALB/c mice. Mice immunized with first-generation Ad5 expressing the …

Regulation Of Cytosolic Phospholipase A2 (Cpla2) And Its Association With Cell Proliferation In Human Lens Epithelial Cells, Yin Wang, Kui-Yi Xing, Marjorie F. Lou

School of Veterinary and Biomedical Sciences: Faculty Publications

PURPOSE. To investigate the molecular mechanism for cytosolic phospholipase A2 (cPLA2) regulation and its association to platelet-derived growth factor (PDGF)-induced cell proliferation.

METHODS. cPLA2 was examined using human lens epithelial (HLE) B3 cells. Reactive oxygen species (ROS) generation induced by PDGF was analyzed by luminescence assay. Cell proliferation was measured by cell counting and by BrdU assay. Human cPLA2 gene was cloned via RT-PCR followed by sitedirected mutagenesis to construct HLE B3 cells expressing either inactive cPLA2 enzyme with S228A mutation (S228A), or cPLA2 truncated at the calcium-binding C2 domain (C2D). Activity of cPLA2 was measured by arachidonic acid (AA) …

Generation Of A Kupffer Cell-Evading Adenovirus For Systemic And Liver-Directed Gene Transfer, Reeti Khare, Shannon M. May, Francesco Vetrini, Eric A. Weaver, Donna Palmer, Amanda Rosewell, Nathan Grove, Philip Ng, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

As much as 90% of an intravenously (i.v.) injected dose of adenovirus serotype 5 (Ad5) is absorbed and destroyed by liver Kupffer cells. Viruses that escape these cells can then transduce hepatocytes after binding factor X (FX). Given that interactions with FX and Kupffer cells are thought to occur on the Ad5 hexon protein, we replaced its exposed hypervariable regions (HVR) with those from Ad6. When tested in vivo in BALB/c mice and in hamsters, the Ad5/6 chimera mediated >10 times higher transduction in the liver. This effect was not due to changes in FX binding. Rather, Ad5/6 appeared to …

Intervention To Increase Knowledge And Consumption Of Folate-Rich Foods Based On The Health Belief Model, Lindsay M. Labrosse

Department of Nutrition and Health Sciences: Dissertations, Theses, and Student Research

Folate is important in the prevention of birth defects and in the maintenance of general health. Even after mandatory fortification, many people are still not consuming the RDA of 400 mcg/day. Consuming a diet rich in naturally high-folate foods, as opposed to supplementation, may offer additional health benefits and promote an overall healthy diet.

The purpose of this pilot study was to test the hypothesis that a learner-centered educational intervention based on the Health Belief Model (HBM) will successfully increase knowledge and consumption of folate-rich foods, while increasing positive beliefs about folate and health. A two group parallel control trial …

Interictal Magnetoencephalographic Findings Related With Surgical Outcomes In Lesional And Nonlesional Neocortical Epilepsy, Rui Zhang, Ting Wu, Yingying Wang, Hongyi Liu, Yuanjie Zou, Wen Liu, Jing Xiang, Chaoyong Xiao, Lu Yang, Zhen Fu

Center for Brain, Biology, and Behavior: Faculty and Staff Publications

Purpose: To investigate whether interictal magnetoencephalography (MEG) concordant with other techniques can predict surgical outcome in patients with lesional and nonlesional refractory neocortical epilepsy (NE).

Methods: 23 Patients with lesional NE and 20 patients with nonlesional NE were studied. MEG was recorded for all patients with a 275 channel whole-head system. Synthetic aperture magnetometry (SAM) with excess kurtosis (g2) and conventional Equivalent Current Dipole (ECD) were used for MEG data analysis. 27 Patients underwent long-term extraoperative intracranial video electroencephalography (iVEEG) monitoring. Surgical outcomes were assessed based on more than 1-year of post-surgical follow-up using Engel classification system.

Results: As we …

Understanding Of Functions Of Selenoproteins And Dietary Selenium By Using Animal Models, Marina V. Kasaikina

Department of Biochemistry: Dissertations, Theses, and Student Research

Selenium (Se) is a trace element that is incorporated into proteins in the form of the 21^st amino acid, selenocysteine (Sec). Se supplementation was reported to have beneficial roles in prevention of cardiovascular and muscle disorders, cancer prevention and enhancement of the immune function. However, recent studies also showed that excessive dietary Se increases the risk of development of type 2 diabetes mellitus. Thus, better understanding of Se and selenoprotein functions is required. We used three approaches to address this problem.

First, we used high-throughput sequencing to examine composition of the gut microflora in mice maintained on selenium-deficient, selenium-sufficient, …

Immune Evasion Of Porcine Reproductive And Respiratory Syndrome Virus Through Glycan Shielding Involves Both Glycoprotein 5 As Well As Glycoprotein 3, Hiep Vu, Byungjoon Kwon, Kyoung-Jin Yoon, William W. Laegreid, Asit K. Pattnaik, Fernando A. Osorio

School of Veterinary and Biomedical Sciences: Faculty Publications

Passive administration of porcine reproductive and respiratory syndrome virus (PRRSV) neutralizing antibodies (NAbs) can effectively protect pigs against PRRSV infection. However, after PRRSV infection, pigs typically develop a weak and deferred NAb response. One major reason for such a meager NAb response is the phenomenon of glycan shielding involving GP5, a major glycoprotein carrying one major neutralizing epitope. We describe here a type II PRRSV field isolate (PRRSV-01) that is highly susceptible to neutralization and induces an atypically rapid, robust NAb response in vivo. Sequence analysis shows that PRRSV-01 lacks two N-glycosylation sites, normally present in wild-type (wt) PRRSV strains, …

A Mark–Recapture Technique For Monitoring Feral Swine Populations, Matthew M. Reidy, Tyler A. Campbell, David G. Hewitt

USDA Wildlife Services: Staff Publications

Techniques to monitor populations of feral swine (Sus scrofa) relative to damage control activities are needed on rangelands. Our objectives were to describe and assess a mark–recapture technique using tetracycline hydrochloride (TH) for monitoring feral swine populations. We established bait stations at study sites in southern and central Texas. During 1 d, we replaced normal soured corn bait with bait containing TH and counted the number of feral swine that consumed bait with observers. We conducted feral swine removal using box-style traps and helicopters, at which time we collected teeth for TH analysis. In southern Texas, we estimated …

Regulation Of Immediate Early And Epithelial-Mesenchymal Transition Genes Expression By Adipocytokines In The Female Reproductive Tract, Zhufeng Yang

Department of Animal Science: Dissertations, Theses, and Student Research

Individuals who are overweight or obese are at increased risk for cancer. However, the mechanistic link between obesity and cancer is poorly defined. Adipose tissue produces hormones and pro-inflammatory cytokines with mitogenic properties. Many of these hormones and cytokines are altered in obese individuals and may lead to disruption of the normal balance between cell proliferation, differentiation, and apoptosis. Thus, the objective of this study was to determine how adipocyte-derived factors regulate the expression of genes that contribute to cell proliferation and migration. To compare immediate early gene (cell proliferation) and epithelial-mesenchymal transition (cell migration) gene expression between untreated and …

A New Single Workbout Test To Estimate Critical Power And Anaerobic Work Capacity, Haley Bergstrom

Department of Nutrition and Health Sciences: Dissertations, Theses, and Student Research

The purpose of the present study was to develop a new 3-min, single workbout test to estimate critical power (CP) and anaerobic work capacity (AWC) using the Monark cycle ergometer with the resistance based on the individual’s body weight. Twelve moderately-trained adults (6 men and 6 women; mean age ± SD = 23.2 ± 3.5 yr) performed an incremental cycle ergometer test to exhaustion. The CP and AWC were estimated from the original work limit (W_lim) versus time limit (T_lim) relationship (CP_PT) and a 3-min all-out test (CP_3min) against a fixed resistance …

Regulation Of Sterol Transport By Dietary Phytosterol Esters, Trevor J. Carden

Department of Nutrition and Health Sciences: Dissertations, Theses, and Student Research

LDL cholesterol is associated with the development of atherosclerosis and is therefore considered an important target for intervention to prevent cardiovascular diseases. The inhibition of cholesterol absorption in the small intestine is an attractive approach to lowering plasma cholesterol, one that is exploited by drug therapy as well as dietary supplementation with plant sterols. The mechanism of action of plant sterol esters (PSE) is still incompletely understood, therefore this study was conducted to test the hypothesis that hydrolysis of plant sterol esters is necessary for their cholesterol-lowering effects to be realized.

Male Syrian hamsters were fed diets containing no PSE, …

Human Papillomavirus Genotype Concordance Within Zambian Couples, Kgomotso Makhaola

School of Biological Sciences: Dissertations, Theses, and Student Research

Human Papillomavirus (HPV) is strongly associated with the development of cervical cancer, and has also been identified in other anogenital cancers such as penile, oral, and anal cancers. In regions like Sub Saharan Africa where the prevalence of HIV/AIDS is high, increased rates of HPV infections have also been observed, however the effect of HIV/AIDS on the transmission of HPV is not yet well understood. In this study specimens for HIV, HPV testing and pathology were collected from male participants and their female partners at a Urology department in Lusaka University Teaching Hospital, Zambia. Seventy four samples were collected but …

Molecular Structures Of Prrsv That Contribute To Prrsv Protective Immunity, Asit K. Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

One of the objectives of the proposal is to delineate the regions of the two glycoproteins (GP2 and GP4) of PRRSV that interact with CD163. Other objectives of the proposal are to generate antibodies to these small regions of the glycoproteins as well as to the full-length proteins for future studies to determine if any of these antibodies possess PRRSV neutralizing activity.

To carry out the studies in the proposed objectives, we generated a series of mutants of GP2 as well as GP4 proteins in which various regions were specifically removed by manipulating the plasmids encoding these proteins. We then …

Preschool Children’S Willingness To Try And Preference For Docosahexaenoic Acid Omega-3 Foods, Bethany Joann Bettenhausen

Department of Nutrition and Health Sciences: Dissertations, Theses, and Student Research

During the preschool years, obesigenic eating habits are formed which are strongly associated with risk of developing chronic diseases later in life particularly cardiovascular disease. Docosahexaenoic acid (DHA) omega-3 fatty acid is a polyunsaturated fat with known benefits for heart health, brain cell structure, and retinal development. Current intakes of DHA are below recommended levels. United States (US) Midwestern children may be particularly at risk for suboptimal intakes of DHA. However, foods fortified with life’sDHA^TM, a product of Martek Biosciences and omega-3 eggs may provide culturally acceptable sources of DHA to Midwestern children.

Very little is known about …

Neuromagnetic Measures Of Word Processing In Bilinguals And Monolinguals, Yingying Wang, Jing Xiang, Jennifer Vannest, Tom Holroyd, Daria Narmoneva, Paul Horn, Yinhong Liu, Douglas Rose, Ton Degrauw, Scott Holland

Center for Brain, Biology, and Behavior: Faculty and Staff Publications

Objective: This study aimed to use magnetoencephalography (MEG) to examine the question of whether Mandarin-English bilingual speakers recruit the same cortical areas or develop distinct language-specific networks without overlaps for word processing.

Methods: Eight healthy Mandarin-English bilingual adults and eight healthy English monolingual adults were scanned while single-word paradigms were audio-visually presented.

Results: Our results showed significantly stronger beta-band power suppression in the right inferior parietal lobe (IPL) covering the supramarginal gyrus (BA 40) and angular gyrus (BA 39) for bilinguals when processing Mandarin versus English. Moreover, there were no significant differences between bilinguals and monolinguals in the left inferior …

Advances And Future Challenges In Adenoviral Vector Pharmacology And Targeting, Reeti Khare, Christopher Y. Chen, Eric A. Weaver, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

Adenovirus is a robust vector for therapeutic applications, but its use is limited by our understanding of its complex in vivo pharmacology. In this review we describe the necessity of identifying its natural, widespread, and multifaceted interactions with the host since this information will be crucial for efficiently redirecting virus into target cells. In the rational design of vectors, the notion of overcoming a sequence of viral “sinks” must be combined with re-targeting to target populations with capsid as well as shielding the vectors from pre-existing or toxic immune responses. It must also be noted that most known adenoviral pharmacology …

Assessment Of Pathways For The Introduction And Spread Of Mycobacterium Bovis In The United States, Katie Portacci, Jason Lombard, Lauren Abrahamsen, Eric Bush, Charles Fossler, Robert Harris, Kamina Johnson, Ryan S. Miller, Dianna Mitchell, Randy Pritchard, Steven Sweeney, Todd Weaver

Other Publications in Zoonotics and Wildlife Disease

Bovine tuberculosis (TB) was responsible for more losses among U.S. farm animals in the early 20th century than all other infectious diseases combined. The Cooperative State-Federal Tuberculosis Eradication Program (established in 1917 and administered by APHIS, State animal health agencies, and U.S. livestock producers) has nearly eradicated bovine TB from the nation’s livestock population. However, despite the many accomplishments of the program, bovine TB remains a serious and costly disease of livestock in the United States. In 1992, VS conducted an assessment to identify pathways for the introduction and spread of bovine TB, in order to develop the most effective …