Life Sciences Commons^™

The Purification Of Ribosomal Rna Gene Chromatin From Physarum Polycephalum, Sally A. Amero, Roy C. Ogle, John L. Keating, Vicky L. Montoya, Wendy L. Murdoch, Robert M. Grainger

School of Medical Diagnostics & Translational Sciences Publications

We have undertaken the purification of ribosomal RNA gene (rDNA) chromatin from the slime mold Physarum polycephalum, in order to study its chromatin structure. In this organism rDNA exists in nucleoli as highly repeated minichromosomes, and one can obtain crude chromatin fractions highly enriched in rDNA from isolated nucleoli. We first developed a nucleolar isolation method utilizing polyamines as stabilization agents that results in a chromatin fraction containing far more protein than is obtained by the more commonly used divalent cation isolation methods. The latter method appears to result in extensive histone loss during chromatin isolations. Two methods were then …

Host Selection In Subterranean Termites: Factors Affecting Choice (Isoptera: Rhinotermitidae), Deborah Ann Waller

Biological Sciences Faculty Publications

The effects of block size and species on preference were examined for the subterranean termite Reticulitermes sp. (Rhinotermitidae) in choice and no-choice tests. Termites significantly preferred Douglas fir over Ponderosa pine and red oak, irrespective of block size. The less-preferred woods were eaten in large amounts in no-choice tests, and all three woods appeared to be favorable for the termites and their gut protozoans. In single species tests, termites ate significantly more from larger blocks of Douglas fir, but ate similar amounts from large and small blocks of Ponderosa Pine and red oak. These results suggest that several factors can …

The Effects Of Siltation On Recruitment Of Spiny Lobsters, Panulirus Argus, William F. Herrnkind, Mark J. Butler, Richard A. Tankersley

Biological Sciences Faculty Publications

Fewer juvenile pinry lobsters were found in areas of the Florida Keys where their primary habitat was heavily silted as compared with similar, less silted habitat. Several hypotheses explaining this relationship were tested, and the time-to-metamorphosis was compared for settling pueruli within silted and nonsilted algae stands. Limited postlarval settlement and avoidance of silted algal habitats by juveniles, probably accounts for the paucity of young lobsters in heavily silted sites. In addition, although juvenile lobsters are nonselective predators, lower prey availability in silted algae probably promotes transciency which, in turn, causes increased mortality by predation while juveniles are exposed.

Evaluation Of Possible Reproductively Mediated Character Displacement In The Crayfishes, Orconectes Rusticus And O. Sanbornii, Mark J. Butler Iv

Biological Sciences Faculty Publications

Orconectes rusticus is replacing several species of crayfishes in north-central and northeastern North America, including O. sanbornii in Ohio. Recent evidence suggests that the species replacements may be driven by asymmetrical reproductive success favoring O. rusticus. Nonetheless, some sympatric associations appear locally persistent. Because crayfish demonstrate size assortative mating and there is a disparity in the sizes of the species, further divergence in the sizes of the species in sympatry could enhance reproductive isolation, ultimately providing a mechanism for character displacement. To test this hypothesis the size differentials between crayfish collected from allopatric and sympatric populations in east-central Ohio …

Consequences Of Thymidine Catabolism For Estimates Of Bacterial Production: An Example From A Coastal Marine Sediment, Kevin R. Carman, Fred C. Dobbs, James B. Guckert

OES Faculty Publications

Radioactively labeled thymidine (TdR) has been used extensively to measure bacterial production in aquatic environments, but critical assumptions of the TdR technique often have gone untested. In this study of a coastal marine sediment, the metabolic fate of methyl [³H]TdR and methyl [¹⁴C]TdR was at variance with the assumptions necessary for determining bacterial production. Only 2% of incorporated radioactivity was recovered in the DNA fraction of TCA-insoluble material following time-course incubations of l-300 min. At least the methyl group of TdR was extensively catabolized, as shown by copious production of ¹⁴C0₂. The temporal …

A Simple Solid-Phase Electrophoretic Procedure For The Separation Of Plasmid Dna, Linda Ann Simurra

Biological Sciences Theses & Dissertations

A method was developed for extraction of plasmid DNA from bacterial cells embedded in agarose blocks. Cell - containing blocks were treated with various lysing reagents and inserted into the wells of an agarose gel. Upon electrophoresis the plasmid DNA migrated out of the embedding block and into the gel leaving intact chromosomal DNA in the well. This method was tested with various organisms and found to be effective for plasmid isolation. In comparison to "traditional" procedures, this new method is less tedious since chemical separation of plasmids is not required prior to electrophoresis. Also, a higher yield of plasmid …

Preservation Of Atp In Hypersaline Environments, Bruce J. Tuovila, Fred C. Dobbs, Paul A. Larock, B. Z. Siegel

OES Faculty Publications

High concentrations of particulate ATP were found in the anoxic brines of the Orca Basin and East Flower Garden, Gulf of Mexico. Other measurements indicative of growth and respiration suggested that the microbial community in the brines was inactive, but somehow the ATP associated with the cells persisted. Conceivably, when cells growing just above the interface sank into the brine, the increased osmotic stress could elicit an osmoregulatory response resulting in increased ATP. It was also possible that hydrolytic enzymes were inactivated, resulting in the preservation of ATP. Experiments in which a culture of marine bacteria was suspended in menstrua …

Use Of Woody Vegetation By Beavers In Southeastern Virginia, John L. Echternach, Robert K. Rose

Biological Sciences Faculty Publications

Nonbreeding and sometimes transient beavers, Castor canadensis, cut a variety of woody plants throughout the year during a 15-month study at three sites in southeastern Virginia. Ten species of trees were cut in proportions greater than would be expected according to their availabilities. Beavers heavily used viburnum, alder, ironwood, bayberry, tulip poplar, and dogwood. Winter food caches were constructed but were only lightly used.

Phytoplankton Dynamics In The Very Low Salinity Region Of The James River Estuary, Virginia, U.S.A., Changho Moon

OES Theses and Dissertations

During summer and autumn discharge from the James River estuary, Virginia, was less than 120 _m³sec^-1. There was a peak phytoplankton biomass in the very low salinity region (defined as the location where surface salinity measured less than 0.5$ 0/00) and this peak represented five to ten times greater biomass than adjacent waters. The peak biomass occurred independent of the tidal state and the location of nutrient inputs. It disappeared during winter and spring, and nutrient limitation was not responsible for the low phytoplankton biomass, indicating there were physical, not chemical controlling factors.

The peak biomass …

Synthesis And Characterization Of N-Alkyl-Adenosine-5'-Diphosphates, Han-Ping Huang

Chemistry & Biochemistry Theses & Dissertations

n-Alkylphosphates and adenosine derivatives have been observed to be competitive inhibitors of coenzyme molecules in many NAD⁺ requiring enzymes. In earlier studies, adenosine derivatives such as AMP, ADP and ADP-ribose were found to be coenzyme competitive inhibitors of the Lactate Dehydrogenase catalyzed reaction. However, n-alkylphosphates, to the contrary, + were found to be noncompetitive inhibitors of NAD⁺.

Furthermore, it was observed previously in Dr. Yuan's laboratory that the effect of n-octylphosphate on the LDH catalyzed reaction was changed from noncompetitive to competitive in the presence of low concentration of AMP. Rossmann et al. reported that the binding …

Investigation Of Complex Formation By Oligomers Of Cytosine And Guanosine, Steven Roberts Davis

Chemistry & Biochemistry Theses & Dissertations

Duplex formation between oligo(C:G) _n where n=3 to 4 was shown not to occur under conditions favorable for duplex formation between poly G and poly C. Instead, a stable guano sine self-structure was found to form which a T_m of 50°C for (Gp)₃ and 80°C for (Gp)₄ at strand concentrations of 10^-5M in 1M NaCl. Neither a duplex nor a self-structure formed in the absence of salt.

Oligomers of guanosine and cytosine were obtained by basic hydrolysis and separated according to chain length using DEAE Sephadex column chromatography. Separation of cytosine oligomers with chain lengths …

Molting In The Mature Female Blue Crab, Callinectes Sapidus, Rathbun, Kirk J. Havens

OES Theses and Dissertations

The present study examines the hypothesis that mature female blue crabs, Callinectes sapidus, do not have a terminal anecdysis. Induced molting via eyestalk ablation, size frequency distributions and gonad and limb regeneration indices from field collected animals are used as indicators of potential post maturity molts.

Eyestalk ablation of mature females resulted in ecdysis approximately 51 days after ablation, indicating a physiological ability if mature females to complete ecdysis. Analysis of size frequency distributions indicates two distinct shifts from smaller to larger females, one in early spring and a second in early fall. While migration of larger females into …

A Comparison Of Potential Fishing Methods Associated With Fish Aggregating Devices Off The Northeast Coast Of Puerto Rico, Alan M. Friedlander

OES Theses and Dissertations

Six fish aggregating devices (FADs) were deployed in June and monitored through December 1986. Emphasis was placed on trolling, live-bait handlining and longlining.

There was a significant difference (x2 = 7.815, pCoryphaena hippurus) which made up 64.2% of the catch.

Handlining at night around inshore buoys for one boat yielded 82.7 kg per trip. Pelagic fish comprised 83.9% of the catch, with the majority (81.4%) being kingfish (Scomberomorous cavalla). Catch rates were 50% higher and the species diversity much greater on the FADs versus control during experimental nightfishing, but the difference was not significant.

Longlining resulted in 3.4 …

An Annual Study Of Phytoplankton Composition And Associated Environmental Conditions Of Lake Trashmore, Virginia Beach, Virginia, Seba B. Sheavly

Biological Sciences Theses & Dissertations

A 12 month study of Lake Trashmore was conducted on phytoplankton composition, inorganic nutrients and their seasonal associations. The dominant phytoplankton groups observed were diatoms, chlorophyceans and cyanobacteria. Phytoplankton assemblage patterns were coupled with pH, temperature, dissolved oxygen, nitrites and silicates. Diatoms dominated during periods of high nutrient concentrations, cooler temperatures and an unstable water column. Cyanobacteria forms dominated during periods of warmer temperatures, higher pH and decreased nitrogen and dissolved oxygen concentrations. A seasonal community shift occurred from cyanobacterial dominance to chlorophycean dominance when pH levels decreased. The lake is eutrophic based on indices of chlorophyll a concentrations, transparency …

Growth Potential Of Young Black Sea Bass, Centropristis Striata, In Artificial Environments, Joung Won Kim

OES Theses and Dissertations

The growth potential of the commercially important marine finfish, black sea bass (Centropristis striata), was examined for aquaculture under controlled conditions. The effect of food and body weight on growth was evaluated using juveniles and young black sea bass cultured with a natural diet and a commercial diet at various feeding levels.

Growth was affected by type of food, food consumption rate, and fish body weight. For fish having positive weight gain, absolute growth rate (gram/day) increased exponentially as food consumption rate (percent of body weight per day) and/or body weight increased. Instantaneous growth rate (percent of body weight per …

Synthesis Of Potential Long Acting Calcium Channel Antagonists: A Study Of 1,4-Dihydropyridine Analogs, Thomas E. Christos

Chemistry & Biochemistry Theses & Dissertations

An investigation was undertaken to synthesize and evaluate longer acting analogs to the antihypertensive agent nifedipine. Analogs that are capable of covalent bonding to the nifedipine receptor were considered. These compounds may show increased activity as they will remain at the receptor for a longer period of time.

During the course of this study, previously unreported and unexpected products were encountered and lead to a more intense examination of the Hantzsch synthesis. Of particular interest was the oxazolidine 25 which was a major component of the Hantzsch synthesis when ethanolamine was used instead of ammonium hydroxide. Also isolated during this …

Reactions Of Organic N-Chloramines In The Gastric Fluid Of The Rat, Kathryn E. Mazina

Chemistry & Biochemistry Theses & Dissertations

Using chlorine as a drinking water disinfectant may have potential health effects due to its reactivity with organic amino nitrogen compounds found in the stomach. Organic N-chloramines have been shown to form in the stomachs of laboratory rats. The possible reactions of N-chloramines in the stomach fluid were examined in this study using a model radiolabeled N-chloramine. ³⁶Cl-N-Chloropiperidine, was synthesized and purified to remove ³⁶Cl-chloride. Stomach fluid was obtained from Sprague-Dawley rats which had been first fasted for 24 or 48 hours and then administered 3 mL of deionized water. Different concentrations of radiolabeled chloramine were reacted with …

A Taxonomic Survey Of The Family Gyrinidae (Coleoptera) Of The State Of Virginia, Jerrold R. Harris

Biological Sciences Theses & Dissertations

The aquatic insect fanua of Virginia is not very well known. This is especially true of the order Coleoptera. There are few taxonomic studies on the families of Coleoptera found within the state, and one of the least known is the family Gyrinidae. It is the intent of this study to determine what species of Gyrinidae are found within the state and where they are found. Collections of the beetles were made throughout the state from March to December.

Twenty-one species of Gyrinidae were identified in Virginia, nine species of Dineutus and 12 species of Gyrinus. The family is distributed …

Evaluation Of Two Methods For Estimating Belowground Production In The Great Dismal Swamp, Millie Symbula

Biological Sciences Theses & Dissertations

Two methods were used to estimate belowground production in a maple-gum stand in the seasonally flooded Great Dismal Swamp of Virginia. The first was an auger method which involved sequential core sampling. The second method was an implanted soil mass technique. Nylon mesh bags were filled with peat and buried at the study site. New growth into these implants was taken to be production.

Mean annual lateral root biomass and necromass to 40 cm, determined from soil cores, were 583 g·m^-2 and 281 g·m^-2, respectively. Production estimates for roots ≤ 5 mm in diameter, using soil cores, …

Short Term Temporal And Spatial Variability Of Phytoplankton In The Elizabeth River, Robert L. O'Reilly

Biological Sciences Theses & Dissertations

This study was designed to evaluate short-term temporal and spatial components of phytoplankton variability within a tidal estuarine system, in reference to natural changes in the local environment.

Multiple water samples were collected weekly from October to December 1984 at 4 stations in different branches, or sections, of the Elizabeth River. Collection of chlorophyll~ data was structured to fit an analysis of variance, while the phytoplankton species and their environmental relationships were analyzed using multivariate techniques.

Lower chlorophyll a concentrations from 0.75 to 2.87 ug/1 were characteristic of the Main Stem and Southern Branch, while higher values of 1.27 …

Short-Term Feedback Regulation Of Camp By Accelerated Degradation In Rat Tissues, Tom W. Gettys, Peter F. Blackmore, J. Bruce Redmon, Stephen J. Beebe, Jackie D. Corbin

Bioelectrics Publications

A recent study showed that cAMP analogs lowered cAMP levels in rat hepatocytes. The present work demonstrates that cAMP analogs also lowered cAMP in a rapid, concentration-dependent manner in heart and fat cells. In order to determine if the cAMP-dependent protein kinase mediated this effect, techniques were developed to assay the protein kinase activity ratio in hepatocytes treated with cAMP analogs. The activation of protein kinase and phosphorylase in hepatocytes by 8-pClΦS-cAMP (where 8-pClΦS- indicates 8-parachlorothiophenyl-) was concentration-dependent and occurred in parallel to proportionate decreases in cAMP. More than 20% of the cAMP binding sites on the protein kinase were …

Variant Forms Of Ataxia Telangiectasia, A. M. R. Taylor, E. Flude, B. Laher, Michael W. Stacey, E. Mckay, J. Watt, S. H. Greens, A. E. Harding

Bioelectrics Publications

Two ataxia telangiectasia patients with unusual clinical and cellular features are described. Cultured fibroblasts and PHA stimulated lymphocytes from these two patients showed a smaller increase of radiosensitivity than cells from other A-T patients, as measured by colony forming ability or induced chromosome damage respectively, after exposure to ionising radiation. The response of DNA synthesis to irradiation of these cells was, however, the same as for other A-T patients. Cells from a third patient with some clinical features of A-T but with a very protracted course also showed low levels of radiation induced chromosome damage, but colony forming ability and …

Distribution And Current Status Of The Threatened Dismal Swamp Southeastern Shrew, Sorex Longirostris Fisheri, Robert K. Rose, Roger K. Everton, Thomas M. Padgett

Biological Sciences Faculty Publications

The Dismal Swamp southeastern shrew, Sorex longirostris fisheri, was given "Threatened" status by the U.S. Fish and Wildlife Service in 1986 because of its limited distribution and the potential threat of interbreeding with the nearby upland subspecies, Sorex l. longirostris. Known from about 20 specimens collected before 1980 and a few dozen taken since then, "fisheri" seems to have morphologically diverged from the smaller upland "longirostris" in association with the development of the Dismal Swamp. The detection of southeastern shrews that are intermediate in size between the two subspecies, coupled with the location of these collection sites on …

The Identification Of The Threatened Southeastern Shrew Using Multivariate Statistical Techniques, Thomas M. Padgett, Roger K. Everton, Robert K. Rose

Biological Sciences Faculty Publications

The threatened subspecies of the southeastern shrew, Sorex longirostris fisheri, is endemic to the Great Dismal Swamp of southeastern Virginia. Previous studies based on discriminant analysis of external measurements determined that intergrades with the upland form, Sorex l. longirostris, exist along the periphery of the Swamp. To better discriminate among these populations, a study of cranial morphology was initiated. Fifteen cranial measurements were taken, using 59 specimens of Sorex collected previously form southeastern Virginia, including the Dismal Swamp. Both Principal Component and Cluster Analyses revealed no significant cranial variation or morphometric patterns within the specimens examined, but a …

The Relationship Of Upwelling To Mussel Production In The Rias On The Western Coast Of Spain, Jack O. Blanton, Kenneth R. Tenore, F. Castillejo, Larry P. Atkinson, Franklin B. Schwing, Amy Lavin

CCPO Publications

We have calculated an upwelling index for each month over a 17-year period (1969-1985) for a point off the western coast of Spain. We interpret April through September values of the index to indicate the flux of nitrate-rich water in the Spanish Rias. The index representing the 6-month upwelling series has been correlated with an index representing the conditions of mussels grown during that season on rafts in Ria de Arosa. Two seasons represent extreme upwelling conditions over the 17-year sampling period: 1977 when the upwelling index was the highest, and 1983 when it was the lowest. A comparison of …

Validation Of The Otolith Increment Aging Technique For Striped Bass, Morone Saxatilis, Larvae Reared Under Suboptimal Feeding Conditions, Cynthia Jones, Edward B. Brothers

OES Faculty Publications

Striped bass, Morone saxatilis, larvae were reared in the laboratory for 97 days to validate the otolith increment aging technique for this species. Otolith-increment deposition rates were determined under optimal laboratory conditions for growth and under three conditions of restricted feeding and using both light and scanning electron microscopy (SEM). Under optimal laboratory conditions. increments were deposited daily from the fourth day after hatching through the first 2 months of life and were discernible with the light microscope. For larvae reared under restricted feeding regimes and readings done with the light microscope, counts did not reflect true age. Counts …

Is Growth Of Eelgrass Nitrogen Limited? A Numerical Simulation Of The Effects Of Light And Nitrogen On The Growth Dynamics Of Zostera Marina, Richard C. Zimmerman, Robert D. Smith, Randall S. Alberte

OES Faculty Publications

A numerical model of nitrogen uptake and growth was developed for the temperate seagrass Zostera marina L. Goals were to evaluate the relative effects of light and nitrogen availability on nitrogen uptake and partitioning between leaf and root tissue, and to estimate nitrogen concentrations in the sedment and water column required to saturate growth. Steady-state predictions are quite robust with respect to a range of parameter values justified by available data The calculations indicated that roots are probably more important in overall nitrogen acquisition in most light and nitrogen environments encountered in situ, but may contribute less than 50 …

Acceleration Of Nutrient Uptake By Phytoplankton In A Coastal Upwelling Ecosystem: A Modeling Analysis, Richard C. Zimmerman, James N. Kremer, Richard C. Dugdale

OES Faculty Publications

Studies of upwelling centers in the eastern Pacific suggest that maximum rates of nitrate uptake (light and nutrient saturated) increase, or shift-up, as newly upwelled water moves downstream. The rate of shift-up appears to be related to irradiance and the ambient concentration of limiting nutrient at the time of upwelling. A mathematical model was developed to evaluate effects of irradiance and initial nitrate concentration on temporal patterns of shift-up and subsequent time scales of nutrient utilization over a range of simulated upwelling conditions. When rates consistent with field studies were used, complete shift-up was possible only under certain conditions, and …

Biochemical And Histological Evaluation Of Normal Human Epidermis Vs Cultured Human Epidermis, Ngo Thi Bich Lien

Biological Sciences Theses & Dissertations

Cultured human keratinocytes were grown in vitro for transplantation purposes in case of severe burns. Single keratinocytes were harvested from human epidermis and grown in culture flasks such that multilayered sheets of keratinocyte cells were harvestable in approximately three weeks. In order to evaluate cultured epidermis for equivalency with normal human epidermis, biochemical and histological studies were performed. Thin sections of cultured epidermis were shown to be composed of cells that had undergone developmental differentiation, but not terminal differentiation. Biochemical analyses of keratin and prekeratin proteins isolated from cultured epidermis indicated that cultured keratinocytes have electrophoretic patterns similar to keratins/prekeratins …

Microsurgical Fertilization Of Mammalian Eggs: An Assessment Of Clinical Utilization, Susan E. Lanzendorf

Theses and Dissertations in Biomedical Sciences

Microsurgical fertilization is the technique in which a spermatozoon or sperm nucleus is injected into the cytoplasm of an egg. To establish a foundation for the use of microsurgical fertilization as a means of treating infertility, this study evaluated the procedure in hamster and human eggs. Hamster sperm nuclei were microinjected into hamster eggs to determine the rate of abnormal fertilization and to ultrastructurally assess cellular damage by transmission electron microscopy. Hamster eggs were also injected with human spermatozoa obtained from fertile and infertile men to evaluate the fertilizing potential of the sperm cells. In addition, human eggs donated by …