Medicine and Health Sciences Commons^™

Effects Of Shielding Adenoviral Vectors With Polyethylene Glycol On Vector-Specific And Vaccine-Mediated Immune Responses, Eric A. Weaver, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

Many individuals have been previously exposed to human adenovirus serotype 5 (Ad5). This prior immunity has long been known to hinder its use for gene therapy and as a gene-based vaccine. Given these immunogenicity problems, we have tested whether polyethylene glycol (PEG) can blunt immune effects against Ad5 during systemic and mucosal vaccination. Ad5 vectors were covalently modified with 5-, 20-, and 35-kDa linear PEG polymers and evaluated for their ability to produce immune responses against transgene antigen products and the vector itself. We show that shielding Ad5 with different-sized PEGs generally reduces transduction and primary antibody responses by the …

Ecological Niche Of The 2003 West Nile Virus Epidemic In The Northern Great Plains Of The United States, Michael Wimberly, Michael B. Hildreth, Stephen P. Boyte, Erik Lindquist, Lon Kightlinger

Public Health Resources

Background: The incidence of West Nile virus (WNv) has remained high in the northern Great Plains compared to the rest of the United States. However, the reasons for the sustained high risk of WNv transmission in this region have not been determined. To assess the environmental drivers of WNv in the northern Great Plains, we analyzed the county-level spatial pattern of human cases during the 2003 epidemic across a seven-state region.
Methodology/Principal Findings: County-level data on WNv cases were examined using spatial cluster analysis, and were used to fit statistical models with weather, climate, and land use variables as predictors. …

How Does A Riverine Setting Affect The Lifestyle Of Shellmound Builders In Brazil?, Sabine Eggers, C. C. Petronilho, K. Brandt, C. Jericó-Daminello, J. Filippini, Karl Reinhard

Karl Reinhard Publications

The contact of inland and coastal prehistoric groups in Brazil is believed to have been restricted to regions with no geographical barrier, as is the case in the Ribeira de Iguape valley. The inland osteological collection from the riverine shellmound Moraes (5800–4500 BP) represents a unique opportunity to test this assumption for this region. Despite cultural similarities between riverine and coastal shellmounds, important ecological and site distribution differences are expected to impact on lifestyle. The purpose of this study is thus to document and interpret health and lifestyle indicators in Moraes in comparison to coastal shellmound groups. Specifically we test …

Intraretinal Signaling By Ganglion Cell Photoreceptors To Dopaminergic Amacrine Neurons, Dao-Qi Zhang, Kwoon Y. Wong, Patricia J. Sollars, David M. Berson, Gary E. Pickard, Douglas G. Mcmahon

School of Veterinary and Biomedical Sciences: Faculty Publications

Retinal dopaminergic amacrine neurons (DA neurons) play a central role in reconfiguring retinal function according to prevailing illumination conditions, yet the mechanisms by which light regulates their activity are poorly understood. We investigated the means by which sustained light responses are evoked in DA neurons. Sustained light responses were driven by cationic currents and persisted in vitro and in vivo in the presence of L-AP4, a blocker of retinal ON-bipolar cells. Several characteristics of these L-AP4-resistant light responses suggested that they were driven by melanopsin-expressing intrinsically photosensitive retinal ganglion cells (ipRGCs), including long latencies, marked poststimulus persistence, and a peak …

Regulation Of The Bioavailability Of Thioredoxin In The Lens By A Specific Thioredoxin-Binding Protein (Tbp-2), Namal P.M. Liyanage, M. Rohan Fernando, Marjorie F. Lou

School of Veterinary and Biomedical Sciences: Faculty Publications

Thioredoxin (TRx) is known to control redox homeostasis in cells. In recent years, a specific TRx binding protein called thioredoxin binding protein-2 (TBP-2) was found in other cell types and it appeared to negatively regulate TRx bioavailability and thereby control TRx biological function. In view of the sensitivity of lens transparency to redox status, proper regulation of TRx bioavailability is of the utmost importance. This study was conducted to examine the presence and function of TBP-2 in human lens epithelial cells (HLE B3). We cloned human lens TBP-2 from a human cDNA library (GenBank accession number AY 594328) and showed …

Transmembrane Domain Length Of Viral K+ Channels Is A Signal For Mitochondria Targeting, Jorg Balss, Panagiotis Papatheodorou, Mario Mehmel, Dirk Baumeister, Brigitte Hertel, Nicolas Delaroque, Frank C. Chatelain, Daniel L. Minor Jr., James L. Van Etten, Joachim Rassow, Anna Moroni, Gerhard Thiel

James Van Etten Publications

K⁺ channels operate in the plasma membrane and in membranes of organelles including mitochondria. The mechanisms and topogenic information for their differential synthesis and targeting is unknown. This article describes 2 similar viral K⁺ channels that are differentially sorted; one protein (Kesv) is imported by the Tom complex into the mitochondria, the other (Kcv) to the plasma membrane. By creating chimeras we discovered that mitochondrial sorting of Kesv depends on a hierarchical combination of N- and C-terminal signals. Crucial is the length of the second transmembrane domain; extending its C terminus by >2 hydrophobic amino acids redirects Kesv …

Tendinopathy Discrimination By Use Of Spatial Frequency Parameters In Ultrasound B-Mode Images, Gregory R. Bashford, Nicholas Tomsen, Shruti Arya, Judith M. Burnfield, Kornelia Kulig

Biomedical Imaging and Biosignal Analysis Laboratory

The structural characteristics of a healthy tendon are related to the anisotropic speckle patterns observed in ultrasonic images. This speckle orientation is disrupted upon damage to the tendon structure as observed in patients with tendinopathy. Quantification of the structural appearance of tendon shows promise in creating a tool for diagnosing, prognosing, or measuring changes in tendon organization over time. The current work describes a first step taken towards this goal—classification of Achilles tendon images into tendinopathy and control categories. Eight spatial frequency parameters were extracted from regions of interest on tendon images, filtered and classified using linear discriminant analysis. Resulting …

In Situ Measurement Of Three-Dimensional Ion Densities In Focused Femtosecond Pulses, James Strohaber, Cornelis J. Uiterwaal

Department of Physics and Astronomy: Dissertations, Theses, and Student Research

We image spatial distribution of xenon ions in the focus of a laser beam of ultrashort, intense pulses in all three dimensions, with a resolution of three by twelve microns in the two transverse directions. This allows for studying ionization processes without spatially averaging ion yields. Our in situ ion imaging is also useful to analyze focal intensity profiles and to investigate the transverse modal purity of tightly focused beams of complex light. As an example, the intensity profile of a Hermite-Gaussian beam mode HG recorded with ions is found to be in good agreement with optical images.

Differential Role Of Nadp+ And Nadph In The Activity And Structure Of Gdp-D-Mannose 4,6-Dehydratase From Two Chlorella Viruses, Floriana Fruscione, Laura Sturla, Garry Duncan, James L. Van Etten, Paola Valbuzzi, Antonio De Flora, Eleonopra Di Zanni, Michela Tonetti

James Van Etten Publications

GDP-D-mannose 4,6-dehydratase (GMD) is a key enzyme involved in the synthesis of 6-deoxyhexoses in prokaryotes and eukaryotes. Paramecium bursaria chlorella virus-1 (PBCV-1) encodes a functional GMD, which is unique among characterized GMDs because it also has a strong stereospecific NADPH-dependent reductase activity leading to GDP-D-rhamnose formation (Tonetti, M., Zanardi, D., Gurnon, J., Fruscione, F., Armirotti, A., Damonte, G., Sturla, L., De Flora, A., and Van Etten, J.L. (2003) J. Biol. Chem. 278, 21559–21565). In the present study we characterized a recombinant GMD encoded by another chlorella virus, Acanthocystis turfacea chlorella virus 1 (ATCV-1), demonstrating that it has the expected dehydratase …

Development Of An Immunofluorescence Assay Using Recombinant Proteins Expressed In Insect Cells To Screen And Confirm Presence Of Human Herpesvirus 8-Specific Antibodies, Veenu Minhas, Lynsey N. Crosby, Kay L. Crabtree, Saul Phiri, Tendai J. M'Soka, Chipepo Kankasa, William J. Harrington, Charles D. Mitchell, Charles Wood

Nebraska Center for Virology: Faculty Publications

Human herpesvirus 8 (HHV-8), or Kaposi’s sarcoma (KS)-associated herpesvirus, has been linked to all forms of KS. The results of most current serological assays for the detection of HHV-8-specific antibodies have low levels of concordance among themselves. To establish a sensitive and specific testing strategy that can be used to screen for HHV-8-specific antibodies, three HHV-8 proteins, ORF65, ORF73, and K8.1A, were expressed by using baculoviral vectors in insect cells and incorporated into a monoclonal antibodyenhanced immunofluorescence assay (mIFA) termed the Sf9 three-antigen mIFA. The results obtained by this mIFA were compared to those obtained by a standard mIFA with …

Effects, Uptake, And Fate Of 2,4,6-Trinitrotoluene Aged In Soil In Plants And Worms, Elly Ph Best, Henry E. Tatem, Kaaren N. Geter, Melissa L. Wells, Brian K. Lane

US Army Corps of Engineers

The present study was aimed at providing data to be used at predicting exposure-based effects of 2,4,6-trinitrotoluene (TNT) aged in soil on endpoint organisms representing two trophic levels. These data can be used to define criteria or reference values for environmental management and conducting specific risk assessment. Long-term exposure tests were conducted to evaluate sublethal toxicity and uptake of aged soil-based explosives, with TNT as the main contaminant. In these tests, plants were exposed for 55 d, and biomass and explosives residues were determined. Worms were exposed for 28 and 42 d, and biomass, number, and tissue residues were determined. …

Small-Molecule Cd4 Mimics Interact With A Highly Conserved Pocket On Hiv-1 Gp120, Navid Madani, Arne Schön, Amy M. Princiotto, Judith M. Lalonde, Joel R. Cpurter, Takahiro Soeta, Danny Ng, Liping Wang, Evan T. Brower, Shi-Hua Xiang, Young Do Kwon, Chih-Chin Huang, Richard Wyatt, Peter D. Kwong, Ernesto Freire, Amos B. Smith Iii, Joseph Sodroski

Nebraska Center for Virology: Faculty Publications

Human immunodeficiency virus (HIV-1) interaction with the primary receptor, CD4, induces conformational changes in the viral envelope glycoproteins that allow binding to the CCR5 second receptor and virus entry into the host cell. The small molecule NBD-556 mimics CD4 by binding the gp120 exterior envelope glycoprotein, moderately inhibiting virus entry into CD4-expressing target cells, and enhancing CCR5 binding and virus entry into CCR5-expressing cells lacking CD4. Studies of NBD-556 analogues and gp120 mutants suggest that: 1) NBD-556 binds within the Phe 43 cavity, a highly conserved, functionally important pocket formed as gp120 assumes the CD4- bound conformation; 2) the NBD-556 …

The Human Immunodeficiency Virus Type 1 Envelope Confers Higher Rates Of Replicative Fitness To Perinatally Transmitted Viruses Than To Nontransmitted Viruses, Xiaohong Kong, John T. West, Hong Zhang, Danielle M. Shea, Tendai J. M’Soka, Charles Wood

Nebraska Center for Virology: Faculty Publications

Selection of a minor viral genotype during perinatal transmission of human Immunodeficiency virus type 1

(HIV-1) has been observed, but there is a lack of information on the correlation of the restrictive transmission

with biological properties of the virus, such as replicative fitness. Recombinant viruses expressing the enhanced

green fluorescent protein or the Discosoma sp. red fluorescent (DsRed2) protein carrying the V1 to V5

regions of env from seven mother-infant pairs (MIPs) infected by subtype C HIV-1 were constructed, and

competition assays were carried out to compare the fitness between the transmitted and nontransmitted

viruses. Flow cytometry was used to …

Varying Efficiency Of Long-Term Replication Of Papillomaviruses In Saccharomyces Cerevisiae, Adam J. Rogers, Malte Loggen, Karen Lee, Peter C. Angeletti

Nebraska Center for Virology: Faculty Publications

Human papillomaviruses (HPVs) replicate in mitotically active basal keratinocytes. Two virally encoded proteins, E1, a helicase, and E2, a transcription factor, are important players in replication and maintenance of HPV episomes. We previously showed that HPV16 could replicate stably in Saccharomyces cerevisiae [Angeletti, P.C., Kim, K., Fernandes, F.J., and Lambert, P.F. (2002)] and we identified cis-elements that mediate replication and maintenance [J. Virol. 76(7), 3350-3358.; Kim, K., Angeletti, P.C., Hassebroek, E.C., and Lambert, P.F. (2005)]. Here, we demonstrate that although multiple HPV genomes replicate stably in yeast, they do so with differing long-term efficiency; HPV6-Ura3 is replicated at the …

A Versatile Assay For The Identification Of Rna Silencing Suppressors Based On Complementation Of Viral Movement, Jason G. Powers, Tim L. Sit, Feng Qu, T. Jack Morris, Kook-Hyung Kim, Steven A. Lommel

Nebraska Center for Virology: Faculty Publications

The cell-to-cell movement of Turnip crinkle virus (TCV) in Nicotiana benthamiana requires the presence of its coat protein (CP), a known suppressor of RNA silencing. RNA transcripts of a TCV construct containing a reporter gene (green fluorescent protein) (TCV-sGFP) in place of the CP open reading frame generated foci of three to five cells. TCV CP delivered in trans by Agrobacterium tumefaciens infiltration potentiated movement of TCV-sGFP and increased foci diameter, on average, by a factor of four. Deletion of the TCV movement proteins in TCV-sGFP (construct TCVΔ92-sGFP) abolished the movement complementation ability of TCV CP. Other known suppressors of …

Effects Of Shielding Adenoviral Vectors With Polyethylene Glycol On Vector-Specific And Vaccine-Mediated Immune Responses, Eric A. Weaver, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

Many individuals have been previously exposed to human adenovirus serotype 5 (Ad5). This prior immunity has long been known to hinder its use for gene therapy and as a gene-based vaccine. Given these immunogenicity problems, we have tested whether polyethylene glycol (PEG) can blunt immune effects against Ad5 during systemic and mucosal vaccination. Ad5 vectors were covalently modified with 5-, 20-, and 35-kDa linear PEG polymers and evaluated for their ability to produce immune responses against transgene antigen prod- ucts and the vector itself. We show that shielding Ad5 with different-sized PEGs generally reduces transduction and primary antibody responses by …

Pathoecology And The Future Of Coprolite Studies In Bioarchaeology, Karl J. Reinhard, Vaughn M. Bryant Jr.

Karl Reinhard Publications

Human coprolites currently provide an expanding array of information about the diet, health, and ecology of prehistoric people in the Southwest, but for many years coprolites were not recognized or preserved, or they were not considered important and thus were not saved (Bryant and Dean 2006). With the expansion of archaeological field work during the last half of the twentieth century archaeologists have increasingly explored the "complete" potentials of sites, including the collection and analysis of geomorphologic, botanical, and faunal data. In some ideal habitats (e.g., very dry or frozen) this includes exploring the scientific potential of human coprolite studies. …

Pathoecology Of Two Ancestral Pueblo Villages, Karl Reinhard

Karl Reinhard Publications

Pathoecology is the study of the biotic, abiotic, and cultural environments of disease (Martinson et al. 2003). A parasitic infection is the result of the pathoecological interaction of host behavior, parasite life cycle, the environment in which both life forms live, the nutritional status of the host, and host physiological responses to all of these factors. Parasites contribute to anemia in many ways. Some, such as hookworm, actually consume blood and cause iron loss through their activities. For other parasites, symptoms such as profuse diarrhea reduce intestinal absorption of nutrients. Others, such as certain fish tapeworms, actually compete for absorption …

Parasite Pathoecology Of Salmon Pueblo And Other Chacoan Great Houses: The Healthiest And Wormiest Ancestral Puebloans, Karl Reinhard

Karl Reinhard Publications

Two fields of paleopathological investigation originated in the Southwest. Archaeoparasitology is the study of ancient parasite infection (Reinhard 1990, 1992b). It includes comparisons between time periods of single societies as well as comparisons of parasitism between different, contemporaneous cultures. For example, Fry (1980) compared Fremont and Anasazi parasitism, and also Archaic hunter-gatherer and ancestral Pueblo parasitism. All of these studies fall into the definition of archaeoparasitology.

By contrast, pathoecology is the reconstruction of relationships among behavior, environment, and disease organisms in the development of illness (Martinson et al. 2003; ReinhardandBuikstra2003; Reinhardet al. 2003; Santoro et al. 2003).1his field developed from …

Cranial Deformation As The Cause Of Death For A Child From The Chillon River Valley, Peru, Shelia M.F. Medoncade Souza, Karl J. Reinhard, Andrea Lessa

Karl Reinhard Publications

Two small mummy bundles, found in a tomb at the Chillon River Valley, Lima Plains, Peru exist in the collections of the Museu Nacional, Rio de Janeiro, Brazil. They were donated to the collections at the beginning of the 20th century. A multidisciplinary team is now curating and studying them. CT scans confirmed both skeletons were of individuals less than one year old at death. One of the bundles is intact and will be preserved and displayed. The other one was partly decomposed and the authors received permission to unwrap it to analyze the bones in details. Mites and pollen …

A Child And His Fate: Tuberculosis, Perimortem Trauma And Mummification, Sheila M. M.De Souza, Karl J. Reinhard, Bernardo Tessarollo, Jose Fernando Cardona Zannier, Adauto Araujo

Karl Reinhard Publications

A male child, sitting, 7 years, circumferential deformation, skin marked by coiled fibers, nostril plugs, consistent with Aymara practices was CT scanned. Puppae in the skin suggest exposition of the body previous to the funeral. Dark brown color at the skin suggest blood perfusion. A broken area at the right frontal with the skin partially inside is suggestive of perimorten trauma. There were no bone debris inside the skull and the CT images show two confluent fractures defined by radiating lines extending beyond the missing bone surface. The right eyeball is projected out, the tongue is outside the mouth, the …

Enterobius Vermicularis Ancient Dna In Pre-Columbian Human Populations, Alena Mayo Iniguez, Karl J. Reinhard, Luiz Fernando Ferreira, Adauto Araujo, Ana Carolina, Paulo Vincente

Karl Reinhard Publications

In prehistoric populations the paleoparasitological findings show an Enterobius vennicularis homogeneous distribution among North American hunter-gatherers intensified with the advent of agriculture. The same occurred in the transition from nomad hunter-gatherers to sedentary farmers in South America, although E. vermicularis infection encompasses only the ancient Andean peoples. Since molecular techniques are sensitive in detecting ancient DNA (aDNA), in this work we have performed a molecular paleoparasitological study of E. vermicularis. aDNA was recovered from North and South American coprolites (4110 BC-AD 900). Human (cox 2 and HVR) and pinworm (5S rRNA spacer) sequences were determined. The sequence analysis confirmed E. …

Paleoparasitology: Perspectives With New Techniques, Adauto Araújo, Karl Reinhard, Otilio M. Bastos, Ligia C. Costa, Claude Pirmez, Alena Iñighez, Ana Carolina Vicente, Carlos M. Morel, Luiz Fernando Ferreira

Karl Reinhard Publications

Paleoparasitology is the study of parasites found in archaeological material. The development of this field of research began with histological identification of helminth eggs in mummy tissues, analysis of coprolites, and recently through molecular biology. An approach to the history of paleoparasitology is reviewed in this paper, with special reference to the studies of ancient DNA identified in archaeological material.

Paleoparasitologia: perspectivas com novas técnicas
Paleoparasitologia é o estudo de parasitos encontrados em material arqueológico. O desenvolvimento deste campo da pesquisa teve início com a identificação de ovos de helmintos em tecidos mumificados, análise de coprólitos e, recentemente, através da …