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Agricultural Research Division 103rd Annual Report, July 1, 1988, To June 30, 1989

Agricultural Research Division: News and Annual Reports

It is a pleasure to provide the 103rd Annual Report of the UNL Agricultural Research Division. This report contains lists of current faculty, active projects, refereed journal articles, books and book chapters, germ plasm releases, brief descriptions of research in selected areas, and the financial report for the period of July 1, 1988, through June 30, 1989. This report was compiled in compliance with the intent of the law of the State of Nebraska that established the Nebraska Agricultural Experiment Station on March 31, 1887.

Foreword ... v

Administrative Personnel ... vii

Administrative Units ... viii

Organizational Chart ... viii …

The Effects Of Retinoic Acid And Butyric Acid On In Vitro Migration By Murine B16a Cells: A Quantitative Scanning Electron Microscopic Study, T. W. Mcgarvey, B. Persky

Scanning Microscopy

Retinoic acid (RA) and butyric acid (BA) were investigated for their effect on in vitro migration of highly metastatic murine B16a melanoma cells. These potential antitumor agents are known to alter the cytoskeleton. Our initial studies determined the 72 h cytostatic /cytotoxic concentration of RA (1 X 10^-6 M / > 1 X 10^-5M) and BA (1.5 mM) / > 2.0 mM). Cytostasis by RA and BA was confirmed by autoradiography and radioisotope incorporation. For migration assays, cells were plated on 3 and 5 μm diameter pore polycarbonate membranes. Complete media was added containing RA or BA at time …

Fault Tolerance In A Very Large Database System: A Strawman Analysis, Amit P. Sheth

Kno.e.sis Publications

A simple model is used to study the effect of fault-tolerance techniques and system design on system availability. A generic multiprocessor architecture is used that can be configured in different ways to study the effect of system architectures. Important parameters studied are different system architectures and hardware fault-tolerance techniques, mean time to failure of basic components, database size and distribution, interconnect capacity, etc. Quantitative analysis compares the relative effect of different parameter values. Results show that the effect of different parameter values on system availability can be very significant. System architecture, use of hardware fault tolerance (particularly mirroring), and data …

Identification Of Cyclic Intermediates In Azorhizobium Caulinodans Nicotinate Catabolism, Christopher Kitts, Lisa E. Schaechter, Ross S. Rabin, Robert A. Ludwig

Biological Sciences

In wild-type Azorhizobium caulinodans ORS571, nicotinate served both as anabolic substrate for NAD⁺ production and as catabolic substrate for use as the N source. Catabolic enzyme activities were greatest from cultures grown with nicotinate as the N source and least when cultures were grown with ammonium as the N source. Vector insertion mutants unable to catabolize nicotinate (nic::Vi mutants) still required micromolar quantities of this compound for growth. Therefore, A. caulinodans wild type is NAD⁺ auxotrophic. As the first two intermediates in A. caulinodans nicotinate catabolism, two cyclic compounds, 6-hydroxynicotinate and 1,4,5,6-tetrahydro-6-oxonicotinate, were identified. These compounds …

Flora Of Richmond National Battlefield Park, Virginia, W. John Hayden, Melanie Lynn Haskins, Miles F. Johnson, James M. Gardner

Biology Faculty Publications

An inventory of the vascular flora of nine of eleven units of Richmond National Battlefield Park was compiled from 1985 to 1987. Each site was visited during the growing season in two to four week intervals; plant species were identified and recorded in the field and/or collected for later study. A total of 761 different species were identified in the surveyed units, and 2487 individual records of species per particular park unit were noted. Twenty-three percent of the flora consists of exotic species, largely from Eurasia. Voucher specimens are housed in the herbaria of the University of Richmond and Virginia …

Molecular Analysis Of N6-Methyladenine Patterns In Tetrahymena Thermophila Nuclear Dna, Elizabeth E. Capowski, John M. Wells, Gail Singer Harrison, Kathleen M. Karrer

Biological Sciences Faculty Research and Publications

We have cloned two DNA fragments containing 5'-GATC-3' sites at which the adenine is methylated in the macronucleus of the ciliate Tetrahymena thermophila. Using these cloned fragments as molecular probes, we analyzed the maintenance of methylation patterns at two partially and two uniformly methylated sites. Our results suggest that a semiconservative copying model for maintenance of methylation is not sufficient to account for the methylation patterns we found during somatic growth of Tetrahymena. Although we detected hemimethylated molecules in macronuclear DNA, they were present in both replicating and nonreplicating DNA. In addition, we observed that a complex methylation …

Regulation Of Epidermal Growth Factor Receptor Synthesis By Retinoic Acid And Dexamethasone, Kerby C. Oberg

Loma Linda University Electronic Theses, Dissertations & Projects

Fetal rat lung (FRL) cells incubated in the presence of retinoic acid or dexamethasone exhibit altered ¹²⁵I-epidermal growth factor (EOF) binding capacity. Retinoic acid enhances ¹²⁵I-EGF binding 3-fold following 24 hours of 37° incubation. In contrast, FRL cells incubated in the presence of dexamethasone display a 2-fold reduction in ¹²⁵I-EGF binding capacity. The change in binding capacity, induced by either hormone, was due to alterations in the average number of receptors per cell as determined by Scatchard analysis of ¹²⁵I-EGF binding. In the absence of retinoic acid or dexamethasone, FRL cells were calculated to have approximately …

Isolation, Growth Characteristics, And Long-Term Storage Of Fungi Cultivated By Attine Ants, Jerome J. Howard

Biological Sciences Faculty Publications

Seven pure-culture strains of fungi cultivated by attine ants (ant-garden fungi) were isolated from locally maintained leaf-cutting ant colonies. An ant-garden fungus strain obtained from an Atta cephalotes colony, when offered to ants of the colony from which the fungus was isolated, was accepted as their own. Young fungus cultures were harvested and incorporated into the fungus garden, and cultures of intermediate age were used to begin a new fungus garden; old cultures were simply harvested. To facilitate further research on this fungus, growth characteristics of the different isolates were studied under a variety of conditions. They grew better at …

Modulation Of Macrophage Chemiluminescence By Biological Response Modifiers And Neuroendocrine Hormones, Jeffrey M. Tosk

Loma Linda University Electronic Theses, Dissertations & Projects

Neuroendocrine mediators have been shown to influence a broad spectrum of immune functions suggesting the existence of a neuroimmune regulatory axis effecting feedback control of the immune response. Most studies in support of this paradigm have employed either intact animals or immune cells derived from blood and other tissues. Under these conditions it becomes difficult to cope with numerous variables associated with the complex milieu of the immune system and the interpretation of data. An in vitro model using a murine macrophage cell line, J774, was developed to circumvent these problems. Using this model, macrophages activated by exposure to the …

Microbial Competition In The Biodegradation Of Phenol, Mary Ellen Frank

Theses

Three phenol degraders were isolated from the mixed liquor of the Livingston, NJ municipal treatment plant. Shaker flask experiments using one of the degraders (a Pseudomonas sp.) resulted in the determination of its kinetic rate constants. The second phenol degrader (a Xanthamonas) lost its phenol degrading ability while being stored, and its kinetic rate constants could not be determined. The third phenol degrader (a Pseudomonas cepacia) formed flocs which interfered with the spectrophotometric determination of biomass and therefore its kinetic rate constants also could not be determined.

Mixed culture experiments were carried out using a the Pseudomonas sp. and a …

Thermal Desorption Of Hazardous And Toxic Organic Compounds, Manuel S. Nolau

Theses

The purpose of this thesis is to study the thermal desorption behavior of toxic organic compounds, specifically o-xylene, p-xylene, ethyl benzene, p-chlorotoluene, and anthracene, from soil with respect to temperature and time, and to develop a mathematical model that will describe this desorption behavior. The model will allow prediction of the concentration of an organic contaminant in commercial soil desorption system over time given a known set of parameters of the specific chemical as well as flow rate and temperature. For this purpose, two sets of experiments were done: These are identified as 1. Thermal desorption and 2. plug flow …

Study Of An Analytical Method For Benzo(A)Pyrene Metabolites In Human Urine, Zheng Ouyang

Theses

A method developed by Becher and Bjorseth for analysis of PAH metabolites in urine was employed to investigate human exposure to benzo(a)pyrene (BaP) - an ubiquitous environmental carcinogen. Preliminary results are presented showing the relationship between exposure to BaP and urinary elimination. Although the correlation between the two variables is not statistically significant, there appears to be a positive association with selected exposure varibles such as smoking. The identification of an association may establish urinary BaP as a marker of exposure.

However, detailed study of the analytical procedure indicated that recoveries with this method were as low as 3-8% for …

Annual Variation Of Estuarine And Oceanic Oyster Crassostrea Virginica Gmelin Hemocyte Capacity, William S. Fisher, Marnita M. Chintala, Mark A. Moline

Biological Sciences

Defense-related hemocyte activities of American oysters Crassostrea virginica Gmelin from an oceanic and an estuarine habitat were monitored from spring to winter of 1987 at a constant temperature (15 °C). The ability of cells to spread to an ameboid shape in vitro at ambient salinity and after an acute change in salinity was much reduced from late spring to early fall. This summer-stress period was also indicated by laboratory experiments. Stress may have been due to high water temperatures but spawning efforts may have also contributed. Reduced hemocyte activity occurred at a time when infections by two major oyster parasites …

Isolation & Characterization Of A Type I Topoisomerase From The Hypocotyls Of Etiolated Soybeans, Rick Dye

Masters Theses & Specialist Projects

Topoisomerases are enzymes of critical biological significance. Despite this fact, little data specific to the topoisomerases of higher plants has been published. This research was undertaken to isolate and characterize a higher plant topoisomerase from soybean hypocotyls to further the understanding of the structure and function of these enzymes.

Nuclei were isolated from frozen hypocotyls of 4 day old etiolated soybeans by homogenization and centrifugation, then lysed by gentle stirring in the presence of 1.5 mM ammonium sulfate. The resultant extract was desalted and purified by column chromatography on DEAF sepharose, 5-200 sephacryl and CM cellulose. The components of the …

Animal Companions--The Promise Kept; The Problems Broken

Close Up Reports

The powerful bond between people and dogs and cats is accompanied by serious ethical obligations on our part.

Export And Intercellular Transfer Of Dna Via Membrane Blebs Of Neisseria-Gonorrhoeae, David W. Dorward, Claude F. Garon, Ralph C. Judd

Biological Sciences Faculty Publications

Naturally elaborated membrane bleb material is frequently observed in cultures of Neisseria gonorrhoeae. This material was purified and analyzed for protein, lipopolysaccharide, and nucleic acid content. The electrophoretic protein profiles of two bleb-rich fractions, called BI and BII, were distinct, with only BII containing lipopolysaccharide and outer membrane proteins I and III. Both fractions contained RNA, circular DNA, and linear DNA. Exogenous pancreatic DNase I appeared to hydrolyze all bleb-associated DNA in fraction BI and the linear DNA in fraction BII. The circular DNA molecules associated with fraction BII resisted digestion. Electron microscopy of the bleb fractions verified their DNA …

Immunological Characterization Of An Sds/Kcl Isolated Total Protein Antigen Of Bluetongue Virus Serotype 13, Vicki L. Shore

Undergraduate Honors Capstone Projects

Because of the damaging effects of bluetongue disease on the livestock industry, interest exists in developing a means of inducing passive immunity in infected animals. Antibodies against VP2, the polypeptide responsible for immunoantigenicity in bluetongue virus (BTV), have been found to elicit neutralizing antibodies; however, production of this purified antigen is very costly and thus impractical for commercial purposes. Total viral protein, prepared by a simple, rapid, highly reproducible SDS/KCl method, was injected into rabbits and found to produce antibodies which are useful in immunoblots. However, plaque neutralization assays failed to detect any neutralization activity by these antibodies. Denaturation of …

Effect Of Proteolytic Activity Of The Lactic Cultures On Mozzarella Cheese Quality, Wen-Hsu Amos Wang

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The Mozzarella cheese market is growing rapidly. Major concerns with cheese meltability and color have arisen in the fast food industry. Pre starter culture was used in this study to improve the physical properties of Mozzarella cheese. Three tests (stretch test, melt test, and browning test) were modified to evaluate the quality of cheese.

A stretch test using the Brookfield helipath viscometer to stretch the cheese sample at 60°C was successful in distinguishing cheeses from different make procedures and from different proteolytic strains. A melt test using a glass tube to hold the cheese flow at 110°C for 60 min …

Energy-Dependent, High-Affinity Transport Of Nickel By Thermophilic Acetogenic Bacteria, Hsiuchin Yang

Electronic Theses and Dissertations

Nickel plays an essential role in acetogenesis at the level of acetyl-CoA synthetase, and the mechanism of nickel transport is therefore central to the metabolic capacities of acetogens. The mechanism of nickel transport by the thermophilic acetogenic bacteria Clostridium thermoaceticum ATCC 39073 and Acetogenium kivui ATCC 33488 was investigated using ⁶³NiCl₂ and an anaerobic microfiltration transport assay. Nickel transport by C. thermoaceticum was optimal at pH 7-7.5 and 65°C; nickel transport by A. kivui was optimal at pH 6.4 and 65°C. Cells during the early stages of growth displayed the highest propensity for nickel transport. Nickel transport was …

Variants And Polymorphisms Of Three Repetitive Dna Families In The Human Genome, Robert M. Roudabush

Electronic Theses and Dissertations

A novel 0.6 kb LINE family in human DNA, designated L2Hs, has been described (Musich and Dykes 1986). Studies employing clone N6.4, containing three 0.6 kb segments of this family, indicate that these sequences are interspersed and moderately repetitive. Two additional variant sequences of the L2Hs family, N6.1 and N6.3, have been identified. Restriction mapping of each cloned segment indicates similarities among N6.4, N6.3 and N6.1. When the cloned DNAs were cleaved with restriction enzymes and subjected to cross-hybridization, each cloned insert produced a pattern indicating that the sequences contained in N6.1 and N6.3 are represented in at least one …

The T(11;14)(P15;Q11) In A T-Cell Acute Lymphoblastic Leukemia Cell Line Activates Multiple Transcripts, Including Ttg-1, A Gene Encoding A Potential Zinc Finger Protein, Elizabeth A. Mcguire, Richard D. Hockett, Karen M. Pollock, Marty F. Bartholdi, Stephen J. O'Brien, Stanley J. Korsmeyer

Biology Faculty Articles

Interchromosomal translocations within lymphoid neoplasms frequently involve the antigen receptor genes. We cloned the breakpoints of the t(11;14)(p15;q11) in a CD3-negative T-cell acute lymphoblastic leukemia cell line (RPMI 8402) in order to identify new genes potentially involved in T-cell neoplasia. An extensive comparison of both breakpoints and their germ line counterparts indicated that an inadvertant recombinase-mediated break at chromosome segment 11p15 recombined with the delta T-cell receptor at 14q11. The derivative 11 breakpoint resembles a coding joint in which 11p15 rather than a variable region was introduced 5' to a D_δ1D_δ2J_δ1 intermediate rearrangement. Conversely, the …

Immunologic, Hematologic, And Endocrine Responses To Subacute And Subchronic Exposures To Graded, Subanesthetic Levels Of Nitrous Oxide In Cd-1 Mice, Charles E. Healy

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Nitrous oxide (N₂O) oxidizes vitamin B12. disrupting deoxyribonucleic acid (DNA) synthesis. Occupational exposures to subanesthetic levels of the gas have been documented that may result in suppressed proliferative cell activities. Male CD-I mice were exposed to 0, 50, 500, and 5000 parts of N₂O per million parts of air (ppm) for 6 hr/day, 5 days/week for 2 and 13 weeks. Splenic lymphocytes were assayed for responsiveness to mitogens and for the ability to produce interleukin-2 (lL-2) . Tritiated-thymidine ([³H]-TdR) uptake was measured in CD-I splenic lymphocytes cultured in a mixed-lymphocyte culture (MLC). Cytolytic cell …

Peptide Mapping Analysis Of Vp5 And Vp7 From Five Bluetongue Viruses Isolated In The United States, Yi-Yuan Yang

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

BTV-2, 10, 11, 13 and 17 were purified by three discontinuous sucrose gradients. Viral proteins were separated from the genomic double-stranded RNA using a newly developed KCI-SDS (sodium dodecyl sulfate) precipitation method. Cyanogen bromide (CNBr) peptide mapping of VP5 and VP7 from bluetongue virus (BTV) serotypes 2, 10, 11, 13 and 17 was then done using silver-stained SDS-polyacrylamide gels. The antigenic epitopes of VP5 and VP7 digested with CNBr were also investigated with polyclonal antibodies produced against KCl-SCS precipitated BTV-13 viral proteins. The peptide maps and the antigenic epitopes of VP7 were similar among the five American serotypes with very …

Effects Of Methylene Chloride On Immune Function In Mice And The In Vitro Effect Of Methylene Chloride In Immunologic Assays, Man-Ping Wang

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

A number of toxicities associated with methylene chloride have been found in both human subjects and mice. However, relatively few studies have probed immunotoxicities of methylene chloride. In order to examine possible immunotoxicities or immunomodulating effects of methylene chloride, several tests of cellular immune function were performed using both human in in vitro studies and a mouse model.

Body weights and specific organ weights of thymus, spleen, liver, and kidney were normal in CD-1 mice given various concentrations of methylenechloride. However, a significantly reduced mitogenic response to phytohemagglutinin (PHA} and reduced interleukin-2 (IL-2} production was found in these methylene-chloride-treated mice. …

The Influence Of Embryo Cell Culture Systems On Pretransfer Development Of Early Ovine Embryos, Pablo Gabriel Daniel

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The complete requirements for early embryo development in vitro of the ovine and other domestic species remain unknown. Many studies have concentrated on new media, supplementation, and gas atmosphere formulations.

A newer approach is to coculture early embryos with different cell types to provide the physico-chemical requirements for their development.

In this study, oviduct epithelial (OEC) and dissociated embryo cell (DEC) growth were tested in minimum essential media (MEM) and RPMI. Media were supplemented with fetal calf serum (FCS) and equine derived serum (EDS). Fetal calf serum supported maximum cell confluence in OEC collected on day 3 and day 13 …

Phenotypic Characterization Of Actinobacillus And Actinobacillus-Like Species Isolated From Ram Lambs With Epididymitis, Yen-Bou Liu

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The purpose of this study was to characterize phenotypically Actinobacillus and Actinobacillus-like species cultured from ram lambs with epididymitis (lamb epididymitis), so as to better understand the species and strains of Actinobacillus responsible for causing the disease. Two American Type Culture Collection (ATCC) type species of Actinobacillus (A actinomycetemcomitans and A seminis) and 24 field isolates were tested in this study. The field isolates were cultured from either lesions or semen of ram lambs with clinical epididymitis and had been identified as members of the genus Actinobacillus, based on cultural and morphological criteria. When analyzed by …

Characterization Of Anti-Pichinde Virus Monoclonal Antibodies For The Directed Delivery Of Antiviral Drugs And Toxins, Noah Jefferson Burns Iii

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Mouse monoclonal antibodies directed against Pichinde virus (PCV) were produced to evaluate their application as vehicles for the delivery of antiviral drugs or toxins to virus-infected cells. Four monoclonal antibodies, PC4.9A6, PC4.9D3, PC4.7C2, and PC4.8D3, were of the IgG2a subisotype and reacted with acetone-fixed and live PCV-infected Vero-76 cells. In vivo stained splenic macrophages derived from PCV-infected hamsters that had been injected with fluorescein-labeled PC4.9A6 (FITC9A6) demonstrated a 400% increase in total fluorescence over similarly treated, non-infected cells when analyzed by flow cytometry. This is an indication that FITC-9A6 does have some ability to specifically target PCV infected cells in …

Morphological Effects Of Lonidamine On Two Human-Tumor Cell Culture Lines, Joseph G. Szekely, Anya U. Lobreau, Sabine Delaney, G. Peter Raaphorst, Mark Feeley

Scanning Microscopy

Lonidamine, 1-(2-4-dichlorobenzyl)-1-H-indazol-3-carboxylic acid. is an anticancer drug that has its primary action on cellular metabolism rather than cell division. Since lonidamine is not effective in all tumor cells, we have tested it in two human-tumor cell culture lines: MOLT-4, a T-leukemia and U-87 MG. a glioma. Lonidamine exposure of MOLT-4 cells at 50 μg/mL and pH 6.7 disrupted the mitochondria within 1 h of treatment. The mitochondria were swollen and the cristae were disrupted. When the treated cells were re-incubated in fresh medium at pH 7.4 the mitochondria rapidly returned to their normal morphology. The U-87 MG glioma cells did …

Salinity Tolerance In Plantago Species: An Ultrastructural And X-Ray Microanalytical Investigation Of Differences In Ion Transport In The Roots, Diana M. R. Harvey

Scanning Microscopy

Root ultrastructure and elemental distributions in regions 3-4 mm behind the apex were examined by transmission electron microscopy and x-ray microanalysis in Plantago maritima ( grown ± 200 mM NaCl) and Plantago media (grown± 20 mM NaCl). In Plantago media vacuolation was incomplete in the cortical and xylem parenchyma cells. Epidermal transfer cells were observed in plants grown in the presence of NaCl. In Plantago maritima grown in the presence of NaCl, vacuolation was complete. In the xylem parenchyma cells of Plantago maritima controls the mitochondria were clustered adjacent to the vessels, rather than evenly distributed throughout the cytoplasm. X-ray …

Habitat Use And Diet Analysis Of Breeding Common Barn Owls In Western Nebraska, Joseph A. Gubanyi M.S.

School of Natural Resources: Dissertations, Theses, and Student Research

I studied barn owl (Tyto alba) breeding biology in western Nebraska 1984-1986. I had greatest success capturing males (56% success) at night using trap doors at nest sites and females (91 % success) using hoop nets at nest sites during the day. Barn owls removed 16 of 23 tail-mounted radios. Eight birds were radio-tracked for 7-14.5 hours. The mean foraging range was 198 ha (32- 299 ha, n = 8) with < 1 % overlap among birds from adjacent nest sites. Field-tested telemetry error was high (mean displacements of radio-locations for 2 birds were 208 and 241 m). I found no relationship between percent cover in foraging habitat and reproductive success. I identified 10,140 prey items from 15 nest sites and found both annual and seasonal variation in barn owl diets. Microtus ochrogaster occurred most frequently (32.7%) and increased in the diet from 17.6 to 27.2 to 43.5% 1984-1986. M. ochrogaster and Perognathus hispidus annual frequencies were …