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Autonomic Reflexes of the Heart During Acute Myocardial Ischemia
This study investigated whether acute myocardial ischemia of the anterior left ventricular wall induced an increase in cardiac sympathetic efferent nerve activity and thereby affected regional myocardial blood flow and contractile function.
The Role of Gabergic Inhibition in Modulating Receptive Field Size of Cuneate Neurons
A blockade of GABAergic inhibition increases the receptive field(RF) size of most somatosensory cortex (SI) and some ventrobasal thalamus (VB) neurons. The results suggest RF size of cuneate neurons may be modulated through GABAa and GABAb receptors, independent of firing frequency.
Head Trauma Release of Histamine from Dural Mast Cells Alters Blood-Brain Barrier: Attenuation with Zolantidine
This study employed a new model of mild-to-moderate head trauma to specifically identify the role of dural mast cell (MC) histamine in trauma-induced increased permeability in the blood-brain barrier (BBB). A single line was scored partially through the left dorsal parietal skull. Immediately following the trauma, degranulation was seen in 39% of the MCs on the left and in 2% on the right. After a 20 min survival period, left duras showed 55% with MC degranulation (fewer with complete degranulation) compared to 34% on the right. In the other experiments two parallel lines were scored following the injection of Evan's blue. Histamine assay showed histamine increased in the left cortex to 154% at 5 min, 174% at 10 min, and 151% at 20 min. Fluorescent quantitation of extravasated Evan's blue at 20 min following the trauma gave an increase of 1385% over the value measured for the right cortex. Zolantidine, a selective histamine H2 receptor antagonist, administered at 10- and 20- mg/kg 30 min before the trauma blocked 65% of the Evan's blue extravasation compared with the control and 2.5 mg group.
Characterization of Arthrobacter Globiformis Aspartate Transcarbamylase Concentrations of Substrates
This thesis consists of one major section with two subsections. The first subsection investigates the activity of Arthrobacter globiformis aspartate transcarbamylase's specific activity with increasing concentrations of the enzyme's substrate. Dihydroorotase (DHOase) activity was also measured with increasing concentrations of the substrate dihydroorotate. The second subsection collected data in order to classify the enzyme, resulting in a classification into the category of class A ATCases with bifunctional ATCase-DHOase complexes. The thesis provides evidence to broaden understanding of the ATCase and DHOase enzymes for members of the family that Arthrobacter belongs to.
Bioconcentration and Morphological Effects of Triclosan on Three Species of Wetland Plants
Triclosan (TCS) is an antimicrobial compound found in several types of common household products. After being washed down the drain, TCS will then end up in the local watershed. Although numerous studies have been conducted to evaluate the fate and effects of TCS in aquatic environments, there have been no studies evaluating the role arbuscular mycorrhizal fungi (AM) play in a plants response to TCS exposure. Three species of wetland plants native North Texas were inoculated with AM spores and exposed to 0, 0.4 g/L and 4.0 g/L TCS concentrations. Root morphology of E. prostrata and S. herbacea showed AM and exposure responses. S. herbacea produced the greatest amounts biomass and TCS bioaccumulation, in all but one treatment. It also displayed opposing results to E. prostrata in measures of root length, root surface area, relative root mass, relative shoot mass and shoot:root ratio. TCS root tissue concentrations increased with increased exposures for both E. prostrata and S. herbacea. Even though E. prostrata had the lowest levels in each measure of biomass production, it had the highest amount of root TCS bioaccumulation in the AM inoculated 4.0 g/L treatment. H. laevis was between the other two species in terms of biomass production, and did not demonstrate any exposure or inoculation effects in morphology or TCS accumulation. Overall, no clear patterns were detected, which highlights the fact that further study is required to completely understand the effects compounds like TCS can have on plant community structure, and ultimately ecosystem function.
Exploring the Evolutionary History of North American Prairie Grouse (Genus: Tympanuchus) Using Multi-locus Coalescent Analyses
Conservation biologists are increasingly using phylogenetics as a tool to understand evolutionary relationships and taxonomic classification. The taxonomy of North American prairie grouse (sharp-tailed grouse, T. phasianellus; lesser prairie-chicken, T. pallidicinctus; greater prairie-chicken, T. cupido; including multiple subspecies) has been designated based on physical characteristics, geography, and behavior. However, previous studies have been inconclusive in determining the evolutionary history of prairie grouse based on genetic data. Therefore, additional research investigating the evolutionary history of prairie grouse is warranted. In this study, ten loci (including mitochondrial, autosomal, and Z-linked markers) were sequenced across multiple populations of prairie grouse, and both traditional and coalescent-based phylogenetic analyses were used to address the evolutionary history of this genus. Results from this study indicate that North American prairie grouse diverged in the last 200,000 years, with species-level taxa forming well-supported monophyletic clades in species tree analyses. With these results, managers of the critically endangered Attwater's prairie-chicken (T. c. attwateri) can better evaluate whether outcrossing Attwater's with greater prairie-chickens would be a viable management tool for Attwater's conservation.
Aging Is a Determinant in Anoxia Stress Tolerance in Caenorhabditis Elegans
Oxygen availability is critical for survival for most organisms. The nematode, C. elegans, has been useful for studying genetic regulation of anoxia tolerance due to the oxygen deprivation response mechanisms shared with other metazoans. Studies examining long-term anoxia (72h, LTA) tolerance have only been conducted at adult day 1. To investigate the effect of aging on anoxia tolerance wild-type and mutant strains were exposed to LTA between adult day 1 and day 9. Wild-type isolates and daf-16(mu86) (FOXO transcription factor regulated by insulin-signaling) and aak-2(gt33) (catalytic subunit of AMP-activated protein kinase) strains were anoxia sensitive at day 1 and displayed increased LTA tolerance with aging correlated with reproductive senescence followed by a decline in survivorhsip through day 9. The daf-2(e1370) (insulin receptor homologue of C. elegans), glp-1(e2141) (a lin-12/Notch receptor) and fog-2(q71) (required for spermatogenesis) strains were LTA-tolerant through day 5. I conclude that aging influences LTA-tolerance in a strain- and age-dependent manner. In addition to being LTA-tolerant the daf-2(e1370) and glp-1(e2141) strains have a longevity phenotype that is suppressed by loss of kri-1 or daf-12. While loss of kri-1 did not suppress the LTA-tolerant phenotype of glp-1(e2141) at day 1 the portion of impaired survivors increased at day 3 and by day 5 tolerance was suppressed. Similarly, when exposed to 4 days of anoxia the glp-1(e2141);daf-12(rh41rh611) double mutant had a reduced survivor rate at all ages analyzed compared to glp-1(e2141) controls. To better understand formation of an anoxia-tolerant physiology I exposed adults to one or more 24h bouts. Recurrent bouts increased LTA tolerance in wild-type hermaphrodites in a dose-dependent manner. Bout-treated daf-16(mu86) animals had increased survival rate compared to controls yet maximum survival remained below age-matched wild-type. Anoxia bouts decreased LTA-tolerance in aak-2(gt33) mutants, indicating the requirement for ATP regulation in establishing an LTA-tolerant phenotype. These data support the …
Evaluating the Habitat Requirements of the Golden Orb Mussel (Quadrula Aurea) for Conservation Purposes
Many freshwater mussels are imperiled, due to a number of interrelated factors such as habitat alteration, degradation of water quality, and impoundments. The Golden Orb mussel (Quadrula aurea, I. Lea, 1859) is endemic to the state of Texas and is currently a candidate for the endangered species list, as the number of known populations has been declining in recent years. Little is currently known about Q. aurea aside from basic distribution data. This study is focused on evaluating a combination of macro-habitat and micro-habitat variables to determine their influence on the distribution and density of this species. Macro-habitat variables, including dominant land cover, surface geology, and soil erodibility factor, did not have a significant relationship with mussel distributions. The best model of micro-habitat variables that impacts the Q. aurea distributions is comprised of relative substrate stability (RSS) at moderate flows and current velocity at low flows. For all mussel species in this study, current velocity at low flows is the primary variable that influences distribution. Q. aurea are associated with habitats where larger sediment particles (large gravel and cobble) help to stabilize the substrate in areas with higher current velocities. An understanding of the preferred habitats for Q. aurea can be used to help focus conservation efforts and practices.
Primary Cilia in the Oligodendrocyte Lineage
oligodendrocytes migrate from the corpus callosum into the overlying cortex. The incidence of cilia did not change markedly across age groups, and did not vary consistently with the number of processes per cell, which was used as an indication of the maturation stage of OPCs and young OLs. The mean percent of Olig1 immunopositive (Olig1+) cells having cilia across ages was 33.1% + 16.5%, with all ages combined. In O4+ cells of these mice, 56.7 + 3.6% had primary cilia. If it is the case that adult OLs do not have cilia, the point in the lineage when primary cilia are lost is still unknown. Adult mice that had been injected with cyclopamine to block cilia-dependent Shh signaling were examined to determine whether the rate of generating new OPCs was influenced. In the CC of control mice, the numerical density of Olig1+/BrdU+ cells was 1.29 + 0.07/mm2 was reduced to 0.68 + 0.38/mm2 in the cyclopamine-injected group, and the numerical density of all BrdU+ cells (including both Olig1+ and Olig1- cells) of 4.55 + 1.50/mm2 in the control group was reduced to 3.14 + 1.27/mm2 in the cyclopamine-injected group. However, there were only 2 mice in each group and the differences were not statistically significant.
Examination of the Relationship Between Glucuronic Acid and Vascular Damage in Rats
The goal of this experiment was to examine the role of glucuronic acid in the development of vascular damage in the kidneys and retinas of diabetic individuals. Glucuronic acid was provided to rats in their water at various concentrations in order to increase plasma levels of the compound. Kidneys and retinas were excised and compared to control specimens using microscopy to determine the effect of elevated blood glucuronic acid levels on the occurrence of microaneurysms in renal capillary networks. No differences were seen between the treatment and control groups. Further study needs to be conducted to determine a more suitable time frame for this experiment.
Genetic Interest Assessment
Genetics is becoming increasingly integrated into peoples' lives. Different measures have been taken to try and better genetics education. This thesis examined undergraduate students at the University of North Texas not majoring in the life sciences interest in genetic concepts through the means of a Likert style survey. ANOVA analysis showed there was variation amongst the interest level in different genetic concepts. In addition age and lecture were also analyzed as contributing factors to students' interest. Both age and lecture were evaluated to see if they contributed to the interest of students in genetic concepts and neither showed statistical significance. The Genetic Interest Assessment (GIA) serves to help mediate the gap between genetic curriculum and students' interest.
The Developmental Physiology of the Zebrafish: Influence of Environment and Cardiovascular Attributes
Temperature effects on the development of the zebrafish embryos and larvae and adults were examined. It was found that the earlier in development a temperature change was performed on an embryo, the more significant the change in survival and/or subsequent development. Thus, viable temperature ranges for zebrafish widened significantly as development proceeded. Adults reared and bred at 25oC produced embryos that were significantly more successful at the lower range of rearing temperatures compared to embryos produced from adults reared at 28oC. The majority of this study focused on the physiological effects of swim training during development in the zebrafish. The earlier in development the zebrafish larvae were trained, the greater the mortality. Trained free swimming larvae had a significantly higher routine oxygen consumption after 11 days of training, and a higher mass specific routine metabolic rate after 8 and 11 days of training. Trained free swimming larvae consumed significantly less oxygen during swimming and were more efficient at locomotion, compared to control larvae. Training enhanced survival during exposure to extreme hypoxia in all age groups. Performance aspects of training were investigated in attempt to quantify training effects and in most cases, trained fish performed significantly better than controls. As blood vessels formed during development, they decreased in cross sectional area from days two to six. It was also shown that the variability in visual stroke volume measurements could be reduced significantly by using a third dimension in the analysis with a more accurate volume equation. Finally, the ontogeny of cardiac control was evaluated. The adrenergic receptors were the first to respond to pharmacological stimulation but were closely followed by cholinergic pharmacological stimulation a few days later. There was a significant cholinergic tone present in day 15 zebrafish larvae which persisted. Although an adrenergic tone was not documented in this study, …
Comparative Morphology of Sensilla Styloconica on the Proboscis of North American Nymphalidae and Other Selected Taxa: Systematic and Ecological Considerations.
Sensilla styloconica on the proboscis of 107 species of North American and tropical butterflies were comparatively studied using the scanning electron microscope. Focus was on 76 species of North American Nymphalidae representing 45 genera and 11 subfamilies. Nomenclature for generalized and specific types of nymphalid sensilla is proposed. Written descriptions and micrographs are presented for each species studied. Morphological features were generally consistent for all or most species within genera and sometimes within subfamilies, with specified exceptions. Statistical analysis revealed significant differences for six of eight variables tested between two distinct feeding guilds of North American Nymphalidae. Average number, density, extent of proboscis coverage with sensilla, their total length, and shoulder spine length were all significantly greater in the non-nectar feeding guild than in nectar feeders, and may indicate adaptation for greater efficiency in feeding on flat surfaces. The greater frequency of apical shoulder spines in non-nectar feeders may represent adaptation for protection of sensory pegs from mechanical abrasion during feeding, or for anchoring the flexible proboscis tip to the surface. Correlation analysis revealed 9 out of 28 positive correlations in nectar feeders and 5 out of 28 in non-nectar feeders. Results of preliminary cladistic analysis were not considered to be meaningfully robust due to few available characters. The stylar characters identified in this study should be more useful in future analyses when included with characters from other lines of evidence. The presence of sensilla styloconica in all subfamilies of Nymphalidae, except Danainae, largely supports Ehrlich's (1958) higher classification concept for the family. The presence of less conspicuous sensilla in the Danainae, and other characteristics are presented as further evidence that they should be reconsidered for full family status. Sensilla styloconica in nymphalid butterflies appear to function as extensions that provide greater sensory reach during feeding. The role of these …
A Novel Phytoestrogen that Acts as an Agonist for Human Estrogen Receptors.
Estrogen is the natural agonist of the estrogen receptor (ER). However, certain plant-derived compounds or phytoestrogens have been identified that mimic estrogens and act as agonists and/or antagonists of ERs, depending on subtype and target tissue. Understanding how phytoestrogens interact with ERs, and therefore effect the estrogenic response, may prove beneficial in hormone replacement therapy and in the prevention and treatment of hormone-related diseases. Using Thin Layer Chromatography, gas chromatography/mass spectrometry (GC/MS), and proton nuclear nagnetic resonance (HNMR), I identified 4-ethoxymethylphenol (4EM) found in Maclura pomifera. While most phytoestrogens are heterocyclic compounds, 4EM is a simple phenol that acts as an agonist of ER-alpha and -beta in HeLa and MCF-7 cells. To study the effect of 4EM on ER-alpha and -beta activity, I performed transient transfection assays and showed that 4EM activates ER dependent gene transcription in a dose dependent manner in both ER subtypes. Further, 4EM- mediated transcription in ER-alpha, like estrogen, was enhance in the presense of co-activators, SRC-1 (steroid receptor coactivator-1), CBP (CREB binding proteins), and E6-AP (E6-associated protein) and inhibited by trans-4- hydroxytamoxifen (4HT). I found that 4EM was specific for ER and did not activate transcription of the progesterone receptor in HeLa cells.
Investigations of neuronal network responses to electrical stimulation in murine spinal cultures.
Spontaneous activity in neuronal networks in vitro is common and has been well documented. However, alteration of spontaneous activity in such networks via conditioning electrical stimulation has received much less experimental attention. Two different patterns of electrical stimulation were used to enhance or depress the level of spontaneous activity in spinal cord cultures. High-frequency stimulation (HFS), a method routinely shown to increase the efficacy of synaptic transmission, was employed to augment spontaneous activity. Low-frequency stimulation (LFS), the technique often applied to depress synaptic efficacy, was employed to decrease spontaneous activity. In addition, LFS was used to reverse the effect of HFS on spontaneous activity. Likewise, HFS was applied to counter the effect of LFS. Because these networks were grown on multi-microelectrode plates (MMEPs), this allowed the simultaneous stimulation of any combination of the 64 electrodes in the array. Thus, the possible differences in response to single versus multi-electrode stimulation were also addressed. Finally, test-pulses were delivered before and after the conditioning stimulation on the same stimulation electrode(s) in order to assess the change in mean evoked action potentials (MEAPs). Dissociated spinal tissue from embryonic mice was allowed to mature into self-organized networks that exhibited spontaneous bursting activity after two weeks of incubation. Spontaneous activity was monitored from up to 14 recording channels simultaneously. Although uniform responses to stimulation across all recording electrodes were rarely observed, a large majority of the recording channels had similar responses. Spontaneous activity was increased in 52% of 89 HFS trials, whereas activity was decreased in 35% of 75 LFS trials. The duration of most of these increases was less than 5 minutes. When there were substantial and long-term (> 15 min) changes in spontaneous activity, the opposing stimulation pattern successfully reversed the effect of the previous stimulation. The percent change in MEAPs following conditioning stimulation …
A laboratory course in experimental genetics for the biology major.
This manual has been designed for a class of twenty- four students concurrently enrolled in the lecture course. The laboratory aids in the learning process and fosters an interest in the science of genetics. This manual and the experiments contained within are both informative and fun. The manual correlates with and expands upon the genetics course. Each investigation, with the exception of the Drosophila melanogaster project, can be completed in a 3-4 hour timeframe. This manual provides a “hands on” experience of theories simply discussed in the lecture course. This manual is intended to be a one-source manual where each investigation is designed to include an adequate introduction. Special attention has been given for each investigation with both the student and instructor in mind.
Life History and Secondary Production of Caenis latipennis Banks (Ephemeroptera: Caenidae) in Honey Creek, Oklahoma
A study of the life history and secondary production of Caenis latipennis, a caenid mayfly, was conducted on Honey Creek, OK. from August 1999 through September 2000. The first instar nymph was described. Nymphs were separated into five development classes. Laboratory egg and nymph development rates, emergence, fecundity, voltinism, and secondary production were analyzed. C. latipennis eggs and nymphs take 132 and 1709 degree days to develop. C. latipennis had an extended emergence with five peaks. Females emerged, molted, mated, and oviposited in an estimated 37 minutes. Mean fecundity was 888.4 ± 291.9 eggs per individual (range 239 -1576). C. latipennis exhibited a multivoltine life cycle with four overlapping generations. Secondary production was 6,052.57 mg/m2/yr.
Unique applications of cultured neuronal networks in pharmacology, toxicology, and basic neuroscience
This dissertation research explored the capabilities of neuronal networks grown on substrate integrated microelectrode arrays in vitro with emphasis on utilizing such preparations in three specific application domains: pharmacology and drug development, biosensors and neurotoxicology, and the study of burst and synaptic mechanisms. Chapter 1 details the testing of seven novel AChE inhibitors, demonstrating that neuronal networks rapidly detect small molecular differences in closely related compounds, and reveal information about their probable physiological effects that are not attainable through biochemical characterization alone. Chapter 2 shows how neuronal networks may be used to classify and characterize an unknown compound. The compound, trimethylol propane phosphate (TMPP) elicited changes in network activity that resembled those induced by bicuculline, a known epileptogenic. Further work determined that TMPP produces its effects on network activity through a competitive inhibition of the GABAA receptor. This demonstrates that neuronal networks can provide rapid, reliable warning of the presence of toxic substances, and from the manner in which the spontaneous activity changes provide information on the class of compound present and its potential physiological effects. Additional simple pharmacological tests can provide valuable information on primary mechanisms involved in the altered neuronal network responses. Chapter 3 explores the effects produced by a radical simplification of synaptic driving forces. With all synaptic interactions pharmacologically limited to those mediated through the NMDA synapse, spinal cord networks exhibited an extremely regular burst oscillation characterized by a period of 2.9 ± 0.3 s, with mean coefficients of variation of 3.7, 4.7, and 4.9 % for burst rate, burst duration, and inter-burst interval, respectively (16 separate cultures). The reliability of expression of this oscillation suggests that it may represent a fundamental mechanism of importance during periods of NMDA receptor dominated activity, such as embryonic and early postnatal development. NMDA synapse mediated activity produces a precise …
Autoradiographic Localization of Carbachol-Induced Second Messenger Response in the Rat Spinal Cord Following Inflammation.
This study examined central mechanisms of persistent pain using an autoradiographic technique to localize phosphoinositide hydrolysis (PI) in the rat spinal cord dorsal horn. The lateral half of laminae I-II showed the highest levels of baseline PI turnover and carbachol-stimulated PI turnover in normal animals as well as after inflammation. Inflammation resulted in increased baseline PI turnover in this region of the ipsilateral (76%) and contralateral (65%) dorsal horns. Carbachol increased PI turnover in this region in normal rats (55%) and following inflammation (ipsilateral: 46%, contralateral: 45%). The absolute magnitudes of these increases were 1.85, 2.71, and 2.51 nCi/mg, respectively. The results of this study demonstrate the involvement of PI turnover in neural mechanisms of persistent pain, and provide evidence for the involvement of cholinergic systems in this process. Because spinal cholinergic systems have been reported to be anti-nociceptive, the present results appear to reflect an upregulation of anti-nociceptive activity in response to inflammation. Thus, the spinal cholinergic system may be a regulatory site within the anti-nociceptive pathway, and may provide an attractive target for the development of new therapeutic agents.
A Correlation of Humus with Water Content and Crop Rotation in San Saba and Denton Clays, Denton County, Texas
A study of the humus content and the water content of San Saba clay and Denton clay. Studies show that the physical properties of soil are conducive to moisture retention and the availability of moisture to plants.
Neuronal Network Analyses in vitro of Acute Individual and Combined Responses to Fluoxetine and Ethanol
Embryonic murine neuronal networks cultured on microelectrode arrays were used to quantify acute electrophysiological effects of fluoxetine and ethanol. Spontaneously active frontal cortex cultures showed highly repeatable, dose-dependent sensitivities to both compounds. Cultures began to respond to fluoxetine at 3 µM and were shut off at 10-16 µM. EC50s mean ± S.D. for spike and burst rates were 4.1 ± 1.5 µM and 4.5 ± 1.1 µM (n=14). The fluoxetine inhibition was reversible and without effect on action potential wave shapes. Ethanol showed initial inhibition at 20 mM, with spike and burst rate EC50s at 52.0 ± 17.4 mM and 56.0 ± 17.0 mM (n=15). Ethanol concentrations above 100 -140 mM led to cessation of activity. Although ethanol did not change the shape and amplitude of action potentials, unit specific effects were found. The combined application of ethanol and fluoxetine was additive. Ethanol did not potentiate the effect of fluoxetine.
Use of Satellite Imagery and GIS to Model Brood-Rearing Habitat for Rio Grande Wild Turkey Populations Occurring in the Western Cross Timbers Region of Texas
Remote sensing and GIS have become standard tools for evaluating spatial components of wildlife habitats. These techniques were implemented to evaluate Rio Grande wild turkey (Meleagris gallopavo) poult-rearing habitat in the Western Cross Timbers region of Texas. Texas Parks and Wildlife (TPWD) random roving turkey counts for 1987-1989 and 1998-2000 were selected, indicating locations where hens with poults were observed. Satellite imagery from 1988 and 1999 was classified and then processed with Patch Analyst. To add robustness, stream, road and census population densities were also evaluated for each turkey location. Analysis of the 1988 canopy cover image, comparing observed locations with randomly-selected habitat cells (N = 20) indicated significant differences (p <.05) for patch edge variables. Mean patch edge was significantly greater for habitat locations where hens with poults were observed than for those selected at random. Spatial data for 1999 did not indicate a significant difference (p < .05) between sampling groups (observed vs. random, N = 30). Significant differences (p <.05) did occur for turkey locations observed in both 1988 and 1999 (N = 7). This demonstrates the adaptability of wild turkey hens, as habitats change over time, hens continued to visit the same locations even though the habitat had significantly changed for select spatial variables.
Functional Neural Toxicity and Endocrine Responses in Mice Following Naphthalene Exposure
Polycyclic aromatic hydrocarbons (PAHs) are a well studied and diverse class of environmental toxicants. PAHs act via the aryl hydrocarbon receptor (AhR), and studies have suggested that PAHs may elicit neurological and estrogenic effects. Doses of PAHs between 50 to 150 ppm may elicit neurotoxicity in rodent models. The present study investigated the effects of naphthalene on in vivo steroidogenesis in Swiss Webster male mice, and in vitro neural function of Balb-C/ICR mice frontal cortex neurons. These data suggest that naphthalene may not elicit steroidogenic effects at concentrations ranging from 0.2 to 25 mg/kg/day, following a 7 day subcutaneous dosing regime. In addition, naphthalene may cause functional toxicity of frontal cortex neurons at concentrations of 32 to 160 ppm naphthalene.
College Freshman Biology Two Semester Course: Integrating Deep Processing Teaching Techniques
Development of a college level freshman biology course was undertaken in response to government reports that American students have fallen behind students of other countries in the area of the sciences. Teaching strategies were investigated to accomplish two objectives, to define essential academic material to include in the course and to investigate teaching techniques that would increase deep processing of the information. An active process that consisted of applying the cognitive information to solving problems or developing answers to questions was defined as critical thinking. Critical thinking was incorporated into the course by the use of case studies.
The Use of Genetic Polymorphisms and Discriminant Analysis in Evaluating Genetic Polymorphisms as a Predictor of Population
Discriminant analysis is a procedure for identifying the relationships between qualitative criterion variables and quantitative predictor variables. Data bases of genetic polymorphisms are currently available that group such polymorphisms by ethnic origin or nationality. Such information could be useful to entities that base financial determinations upon predictions of disease or to medical researchers who wish to target prevention and treatment to population groups. While the use of genetic information to make such determinations is unlawful in states and confidentiality and privacy concerns abound, methods for human “redlining” may occur. Thus, it is necessary to investigate the efficacy of the relationship of certain genetic information to ethnicity to determine if a statistical analysis can provide information concerning such relationship. The use of the statistical technique of discriminant analysis provides a tool for examining such relationship.
Molecular systematics of Baird's pocket gopher (Geomys breviceps)
Baird's pocket gopher (Geomys breviceps) is found in eastern Texas, eastern Oklahoma, central and western Arkansas, and western Louisiana. The cytochrome-b gene was sequenced and analyzed for 16 pocket gophers from throughout the range of the species. Similar phylogenetic trees were obtained using maximum-parsimony, maximum-likelihood, neighbor-joining, and Bayesian analyses. Two major clades were formed with northern individuals belonging to clade I and southern individuals belonging to clade II. G. b. sagittalis was paraphyletic in relation to G. b. breviceps in all analyses. Based on inconsistencies between the taxonomic classification and systematic relationships within Baird's pocket gopher, a taxonomic restructuring appears warranted.
The Influence of in Vitro Gill and Liver Metabolism of Xenobiotics on Fish Bioconcentration
This dissertation examines the ability of in vitro biotransformation assays to provide an indication of metabolic potential. The potential for xenobiotic compounds to bioconcentrate in aquatic organisms is expressed through the bioconcentration factor (BCF). The metabolic loss of ibuprofen, norethindrone and propranolol was measured using rainbow trout (Oncorhynchus mykiss) and channel catfish (Ictalurus punctatus) gill and liver S9 fractions, microsomes and cell suspensions. Metabolic transformation rates (kM) were extrapolated from in vitro intrinsic clearance of parent compound (CLm) and integrated into a refined BCF model. In general, CLm of test compounds was greater in liver S9 fractions and hepatocytes. However, the influence of hepatic metabolism on kM and BCF was limited by hepatic blood flow (20-25%) compared to gill blood flow (~100%). A significant difference was noted between BCF solely based on KOW and BCF including kM. These studies indicate that the inclusion of kM in BCF models can bring predicted bioconcentration estimates closer to in vivo values. Primary cell suspensions are preferred over subcellular fractions as cell suspensions possess both phase I and phase II enzyme activity. Further study was conducted on ibuprofen biotransformation pathways. As fish do not contain the same cytochrome P450 (CYP) 2C homologs known to metabolize ibuprofen in mammals, it cannot be assumed that piscine biotransformation is similar. Metabolite analysis found 2-hydroxy-ibuprofen as the major metabolite in S9 and microsomal fractions. Additional assays involving the induction and inhibition of specific CYP isozymes support CYP1A2 as an alternative metabolic pathway.
A Study of the Pyrimidine Biosynthesis Pathway and its Regulation in Two Distinct Organisms: Methanococcus jannaschii and Pseudomonas aeruginosa
Methanococcus jannaschii is a thermophilic methane producing archaebacterium. In this organism genes encoding the aspartate transcarbamoylase (ATCase) catalytic (PyrB) and regulatory (PyrI) polypeptides were found. Unlike Escherichia coli where the above genes are expressed from a biscistronic operon the two genes in M. jannaschii are separated by 200-kb stretch of genome. Previous researchers have not been able to show regulation of the M. jannaschii enzyme by the nucleotide effectors ATP, CTP and UTP. In this research project we have genetically manipulated the M. jannaschii pyrI gene and have been able to assemble a 310 kDa E. coli like enzyme. By using the second methionine in the sequence we have shown that the enzyme from this organism can assemble into a 310 kDa enzyme and that this enzyme is activated by ATP, CTP and inhibited by UTP. Thus strongly suggesting that the second methionine is the real start of the gene. The regulation of the biosynthetic pathway in Pseudomoans aeruginosa has previously been impossible to study due to the lack of CTP synthase (pyrG) mutants. By incorporating a functional uridine (cytidine) kinase gene from E. coli it has been possible to isolate a pyrG mutant. In this novel mutant we have been able to independently manipulate the nucleotide pools and study its effects on the enzymes in the biosynthetic pathway. The enzyme asapartate transcarbamoylase was repressed 5-fold when exogenous uridine was high and cytidine was low. The enzyme dihydroorotate was repressed 9-fold when uridine was high. These results suggest that a uridine compound may be the primary repressing metabolite for the enzymes encoded by pyrB and pyrC. This is the first study to be done with the proper necessary mutants in the biosynthetic pathway of P aeruginosa. In the past it has been impossible to vary the internal UTP and CTP …
Bacterial Challenge in Lumbricus Terrestris: A Terrestrial Invertebrate Immunotoxicity Model.
A bacterial challenge assay was developed utilizing the earthworm, Lumbricus terrestris, in order to assess potential immunotoxic effects from exposure to specific polychlorinated biphenyl congeners. Earthworms were inoculated with Aeromonous hydrophila, establishing a 10-day LD50. In vitro assays for effects of PCBs on phagocytosis agreed with mammalian studies, demonstrating potent suppression of phagocytosis by the non-coplanar PCB congener 138 and no suppression by the coplanar congener 126. However, when the effects of the two PCB congeners were evaluated for suppression of resistance to a whole animal infection challenge assay, coplanar PCB 126 decreased the ability of L. terrestris to withstand infection while non-coplanar PCB 138 did not.
Influence of parental swimming stamina on the cardiac and metabolic performance of larval zebrafish (Danio rerio).
Superior swimming stamina in adult fish is presumably passed on to their offspring, but the ontogeny of the appearance of superior stamina and the requisite enhanced cardio-respiratory support for locomotion in larval fishes has not been determined. Is the expression of the suite of parental traits enabling superior swimming stamina in their offspring dependent upon their achieving juvenile/adult morphology, or does it appear earlier in their larvae? To answer this, adults were classified into three groups based on swimming stamina, followed by measurement of length, mass, and width. Larval offspring from the two parental groups -high stamina larvae (HSL) and low stamina larvae (LSL)- were reared at 27°C in aerated water (21% O2). Routine and active heart rate, routine and active mass specific oxygen consumption were recorded through 21dpf, and cost of transport (COT) and factorial aerobic scope were derived from oxygen consumption measurements. Routine heart rate at 2dpf of LSL was 164 ± 1 b·min-1, compared to only 125 ± 2 b·min-1 for HSL. Routine heart rate subsequently peaked at 203 ± 1 b·min-1 at 5dpf in the HSL group, compared to 207 ± 1 b·min-1, at 4dpf in the LSP larvae. Active heart rate at 5 dpf of LSL was 218 ± 2 b·min-1 compared to 216 ± 2 b·min-1 for HSL. Active heart rate increased slightly to 227 ± 2 b·min-1 for LSL before decreasing again, while active heart rate remained relatively constant for HSL. Routine O2 consumption at 2dpf of HSL was 0.09 μmol·mg-1·hr-1, compared to 0.03 μmol·mg-1·hr-1 in LSL. Routine O2 consumption subsequently peaked at 0.70 μmol·mg-1·hr-1 at 9dpf in the HSL, compared to 0.71 μmol·mg-1·hr-1, at 9dpf in the LSL. These values dramatically decreased before leveling off at around 0.20 μmol·mg-1·hr-1 and 0.15 μmol·mg-1·h-1, respectively. Active O2 consumption at 5dpf for HSL was 0.38 …
A Contravention of Established Principles of Interspecific Allometric Metabolic Scaling in Developing Silkworms, Bombyx Mori.
Established interspecific metabolic allometric relationships do not adequately describe the complexity and variable physiological states of developing animals. Consequently, intraspecific allometric relationships of oxygen consumption and carbon dioxide production as a function of body mass; the respiratory quotient; the function of the silk cocoon; and body composition were investigated for each distinct developmental stage of the silkworm, Bombyx mori. Whole animal O2 consumption in Bombyx ranged from 0.00064 + 0.000047 ml O2 .hr-1 at larval instar I to 0.77 + 0.06 ml O2 .hr-1 in pre-pupal, falling to 0.21+ 0.01 ml O2 .hr-1 in the pupae. Those instars having a significant relationship between O2 consumption as a function of body mass, the slope of the line relating O2 consumption to body mass varied between 0.99 and 1.02, while across all instars the slope was 0.82. Developmental allometry should be presented for individual developmental stages because the individual allometric exponents of the stages can be significantly different from the overall allometric exponent throughout development and in some cases, the overall allometric exponent can be a statistical artifact. The first larval instar of Bombyx mori has the lowest cross sectional area of high metabolic tissue of the midgut (27%) and had one of the highest percentages of some metabolically inert tissues (i.e. lipid, 7.5%). Body composition of the first instar does not support the idea that smaller mass animals having the highest O2 consumption are composed of a greater percentage of metabolically active organs when compared to larger animals. However, this developmental stage has the highest percentage of the mitochondrial marker cytochrome oxidase, which correlates well with the high O2 consumption rate of the smaller mass. Therefore, established interspecific principles should not be assumed to function as valid models for intraspecific developmental relationships of metabolism as a function of body mass. Developmental …
Establishing genetic and physiological baselines for the black-tailed prairie dog (Cynomys ludovicianus).
The black-tailed prairie dog (Cynomys ludovicianus) has experienced dramatic declines over much of its historical range due to habitat loss, plague, poisonings, and shootings. Many populations now occur as isolated genetic relicts. A multiple locus genetic profile was obtained using microsatellite analyses of six polymorphic nucleotide repeats from 319 black-tailed prairie dogs collected from 16 colonies throughout the state of Texas. This assessment revealed that existing populations have sufficient variation at all six loci to verify the usefulness of this approach as a primary genetic tool in conservation and preservation. The data reveals regional-dependent frequency patterns as well as support for founder/bottleneck effects for several of the 16 sites. Although the prairie dog population in Texas as a whole may appear genetically diverse, considerable genetic divergence has already occurred among the subpopulations (FST = 0.164). Isolation by distance is supported by genic differentiation analysis (P < 0.001) and pairwise correlation analysis between genetic distance and geographic distance (P < 0.001). Prairie dogs from six (COC, LUBA, LUBC, LUBD, LUBE, and TAR) of the original 16 sites have been relocated or exterminated, or were in the process of being relocated. Results indicated the following colonies (COT, DAL, HOW, and HUD) are of sufficient size and possess ample genetic diversity to be characterized as candidate foundation populations for future preservation efforts. The proximity of small colonies (< 20 hectares) such as HEMB, LUBB, and PEC, to other colonies should be examined to determine if they are isolated or part of a metapopulation. Colonies (HAR, HEMA, and SCH) with low genetic diversity would be ideal candidates for supplementation. Alternatively, these colonies could be relocated or blended with other similar but genetically distinct colonies. Baselines for healthy, pet prairie dog hematology and blood chemistries were also established. Results signify that data gathered from pet …
New Descriptions, Intraspecific Variation and Systematic Importance of Drumming Behavior in Selected North American Plecoptera
Drumming behavior is described for the first time in 33 North American Plecoptera species, and signals of an additional five species, Pteronarcys pictetii, Acroneuria abnormis, Paragnetina media, Clioperla clio and Isogenoides zionensis, are further detailed. An out-group comparison of behavioral characters in all 104 world species whose drumming is known showed that the behavior is more advanced in the Arctoperlaria Group Systellognatha than in the Group Euholognatha. In general, tapping, monophasy, touching, sequenced exchange and less than 50 taps/answer are ancestral states, and rubbing, grouping, phasing, tremulation, interspersed exchange and equal or more than 50 taps/answer are derived states. There has been some co-evolution between abdominal structure and drumming behavior. Scanning Electron Micrographs of 30 species showed that the primitive state of tapping is ascociated with three classes of abdominal structure: (1) absence of derived structures, (2) lobes or vesicles, and (3) hammers. The derived behavior of rubbing, however, occurs only in species with derived structures, and is predominant in species having vesicles and hammers. Drumming can be used as a line of evidence to aid in defining genera and species, since the behavior has a variable degree of specificity or exclusiveness in all species, particularly in groups of species I have studied in the genera Isoperla, Pteronarcys and Taeniopteryx. Typical and variant computer-synthesized male calls of three stonefly species were tested with live females. They responded at high levels in such a way that the important informational content conveyed was identified as: (1) a minimal threshold of beat numbers, and (2) a discriminant window of beat intervals. Rub frequency and bibeat calling were critical informational parameters in two species.
Methylglyoxal Effects on Cell Division of Scenedesmus quadricauda (Scenedesmaceae)
Cell division of ggeneflesmus quadricauda (Turp.) Breb. (Scenedesmaceae) is enhanced by methylglyoxal, a general inhibitor of cell division, at threshold concentration in conjunction with treatment timing related to growth stage of batch cultures. At 0.5 mM methylglyoxal concentration, cell division was significantly enhanced in algae treated in the logarithmic phase. Specific growth rates of methylglyoxal-treated cultures were rapidly increased at the beginning of logarithmic phase. Cultures inoculated with high cell numbers were less sensitive, but still showed high specific growth rates in logarithmic phase. Cell division in cultures which had low cell numbers was inhibited by 0.5 mM methylglyoxal treatment. Both specific activity of Glyoxalase I and the ratio of Glyoxalase I to Glyoxalase II of methylgloxal-treated cultures were higher than those of controls (1.3 and 2.1- fold, respectively). Pyruvate concentration in treated cultures was increased after methylglyoxal treatment.
Fitness-Related Alterations in Blood Pressure Control: The Role of the Autonomic Nervous System
Baroreflex function and cardiovascular responses to lower body negative pressure during selective autonomic blockade were evaluated in endurance exercise trained (ET) and untrained (UT) men. Baroreflex function was evaluated using a progressive intravenous infusion of phenylephrine HCL (PE) to a maximum of 0.12 mg/min. Heart rate, arterial blood pressure, cardiac output and forearm blood flow were measured at each infusion rate of PE. The reduction in forearm blood flow and concomitant rise in forearm vascular resistance was the same for each subject group. However, the heart rate decreases per unit increase of systolic or mean blood pressure were significantly (P<.05) less in the ET subjects (0.91 ± 0.30 versus 1.62 ± 0.28 for UT). During progressive lower body negative pressure with no drug intervention, the ET subjects had a significantly (P<.05) greater fall in systolic blood pressure (33.8 ± 4.8 torr versus 16.7 ± 3.9 torr). However, the change in forearm blood flow or resistance was not significantly different between groups. Blockade of parasympathetic receptors with atropine (0.04 mg/kg) eliminated the differences in response to lower body negative pressure. Blockade of cardiac sympathetic receptors with metoprolol (0.02 mg/kg) did not affect the differences observed during the control test. It was concluded that the ET subjects were less effective in regulating blood pressure than the UT subjects, because of 1) an attenuated baroreflex sensitivity, and 2) parasympathetic-mediated depression of cardiac and vasoconstrictive responses to the hypotensive stress.
Endocannabinoid System in a Planarian Model
In this study, the presence and possible function of endocannabinoid ligands in the planarian is investigated. The endocannabinoids ananadamide (AEA) and 2-arachidonoylglycerol (2-AG) and entourage NAE compounds palmitoylethanolamide (PEA), stearoylethanolamide (SEA) and oleoylethanolamide (OEA) were found in Dugesia dorotocephala. Changes in SEA, PEA, and AEA levels were observed over the initial twelve hours of active regeneration. Exogenously applied AEA, 2-AG and their catabolic inhibition effected biphasic changes in locomotor velocity, analogous to those observed in murines. The genome of a close relative, Schmidtea mediterranea, courtesy of the University of Utah S. med genome database, was explored for cannabinoid receptors, none were found. A putative fatty acid amide hydrolase (FAAH) homolog was found in Schmidtea mediterranea.
Life Histories Behavior and Space Partitioning in Selected Species of Western North American Plecoptera
Five species of stoneflies (Zapada haysi, Plumiperla diversa, Taenionema pacificum, Isoperla petersoni, Arcynopteryx compacta) from the North Slope and Interior of Alaska were examined for seasonal patterns of emergence of adults and growth of nymphs. Generally growth was retarded during the winter in this region, and all species except I. petersoni completed growth prior to January. The life cycles of six stonefly species (Prostoia besametsa, Triznaka signata, Sweltsa coloradensis. Isoperla fulva, Skwala parallela, Claassenia sabulosa) are described from northern New Mexico. In this region growth was generally less retarded during the winter than in Alaska; P. besametsa completed all nymphal growth during late fall and winter. Drumming behavior of a Colorado population of Pteronarcella badia was described using an evolutionary framework to explain the maintenance of signal variation in this species. Laboratory experiments were used to explore the effect of intraspecific and interspecific interactions on spatial partitioning in P. badia and Claassenia sabulosa. P. badia exhibited clumping and distributed itself as the surface area of substrate in low densities; however, in the presence of C. sabulosa its distribution was random and different from available surface area. A field study was used to examine spatial partitioning by three New Mexico stonefly species (I_. fulva, P. besametsa, T. signata) and to ascertain patterns of microdistribution relating to several abiotic and biotic factors. Generally, there was an interaction of the measured abiotic parameters (current, water temperature, time) with nymphal size. Additionally, void space and sample volume were successfully used to compare biotic densities among leaf and mineral substrates, which were higher in leaf packs than in mineral substrates.
Determination of Bioconcentration Potential of Selected Pharmaceuticals in Fathead Minnow, Pimephales promelas, and Channel Catfish, Ictalurus punctatus
The primary objective of this work was to determine the tissue-specific bioconcentration factors (BCFs) of the selected pharmaceuticals: norethindrone (NET), ibuprofen (IBU), verapamil (VER), clozapine (CLZ) and fenofibrate (FFB) in two freshwater fishes: fathead minnow and channel catfish. BCF tests on fathead followed the standard OECD 42-day test while a 14-day abridged test design was used in catfish exposures. Additional objectives included a) comparing the measured BCFs to the US EPA's BCFWIN model predicted values, b) comparing the BCF results from the standard and reduced tests, and c) prediction of chronic risk of the pharmaceuticals in fish using the human therapeutic plasma concentrations. Each test included uptake and depuration phases to measure tissue-specific kinetic BCFs. The results indicated that all the pharmaceuticals, except IBU, have the potential for accumulation in fish. Estimated BCFs for NET, VER and FFB may not be significant in view of the current regulatory trigger level (BCF &#8805; 2000); however, CLZ's BCF in the liver had approached the criterion level. Significant differences were noticed in the tissue-specific uptake levels of the pharmaceuticals with the following general trend: (liver/kidney) > (gill/brain) > (heart/muscle) > plasma. IBU uptake was highest in the plasma. When compared to the measured BCFs, predicted values for NET, IBU, VER and FFB were slightly overestimated but did not differ largely. However, the measured BCF of CLZ in the liver was approximately two-orders of magnitude higher than the predicted level. The tissue-BCFs for the two species were not widely different indicating the potential usefulness of the reduced BCF test. Comparison of fish and human plasma levels indicated that NET, CLZ and VER have the potential to cause chronic effects in fish.
Interactions between Carotid and Cardiopulmonary Baroreceptor Populations in Men with Varied Levels of Maximal Aerobic Power
Reductions in baroreflex responsiveness have been thought to increase the prevalence of orthostatic hypotension in endurance trained athletes. To test this hypothesis, cardiovascular responses to orthostatic stress, cardiopulmonary and carotid baroreflex responsiveness, and the effect of cardiopulmonary receptor deactivation on carotid baroreflex responses were examined in 24 men categorized by maximal aerobic power (V02max) into one of three groups: high fit (HF, V0-2max=67.0±1.9 ml•kg^-1•min^-1), moderately fit (MF, V0-2max=50.9±1.4 ml•kg^-1•min^-1), and low fit (LF, V0-2max=38.9±1.5 ml•kg^-1•min^-1). Orthostatic stress was induced using lower body negative pressure (LBNP) at -5, -10, -15, -20, -35, and -50 torr. Cardiopulmonary baroreflex responsiveness was assessed as the slope of the relationship between forearm vascular resistance (FVR, strain gauge plethysmography) and central venous pressure (CVP, dependent arm technigue) during LBNP<-35 torr. Carotid baroreflex responsiveness was assessed as the change in heart rate (HR, electrocardiography) or mean arterial pressure (MAP, radial artery catheter) elicited by 600 msec pulses of neck pressure and neck suction (NP/NS) from +40 to -70 torr. Pressures were applied using a lead collar wrapped about the subjects' necks during held expiration. Stimulus response data were fit to a logistic model and the parameters describing the curve were compared using two-factor ANOVA. The reductions CVP, mean (MAP), systolic, and pulse pressures during LBNP were similar between groups (P<0.05). However, diastolic blood pressure increased during LBNP m all but the HF group. (P<0.05). The slope of the FVR/CVP relationship did not differ between groups, nor did the form of the carotid-cardiac baroreflex stimulus response curve change during LBNP. changes in HR elicited with NP/NS were not different between groups (£>0.05). The range of the MAP stimulus response curve, however, was significantly less in the HP group compared to either the MP or LF group (£<0.05). These data imply that carotid baroreflex control of HR is unaltered …
Neuropharmacological Characteristics of Tolerance for Cocaine Used as a Discriminative Stimulus
The main purpose of this research was to investigate the phenomenon of tolerance to cocaine. Tolerance is operationally defined as a decreased drug effect due to prior history of drug administration. The animal model that was chosen to investigate tolerance to cocaine was the drug discrimination model, which is an animal analogue of human subjective drug effects. In the drug discrimination procedure, animals are trained to emit one behavior when injected with saline. In the present experiments, rats were trained to press one lever when injected with cocaine, 10 mg/kg, and a different lever when injected with saline for food reinforcement. Once rats are trained, they can accurately detect the cocaine stimulus greater than 95% of the time.
Testing and Evaluation of Environmental Fate Models Using Aquatic Microcosms and Three Organic Chemicals
Two compartment (sediment and water) flow-through model ecosystems were constructed to investigate the compartmentalization of different organic chemicals. Lindane, naphthalene, and mirex were pumped into the systems and resultant compartmental chemical concentrations determined. Steady state concentrations were compared to those predicted by two environmental fate models - EXAMS (Exposure Analysis Modeling System) and SLSA (Simplified Lake and Stream Analysis) which were developed by EPA-Athens, Georgia and HydroQual, Inc., respectively.
Induced Bradycardia Effects on Angiogenesis, Growth and Development in Early Development in Chicken Embryos, Gallus Domesticus
Cardiac performance, angiogenesis and growth was investigated during early chicken development. Heart rate, and thus arterial pulse pressure and cardiac output, were altered with the bradycardic drug ZD7288. Heart rates at 72 h of development of control embryos and those dosed with chicken Ringer were not different at 171 bpm. Acute and chronic application of ZD7288 caused significant bradycardia. Chronic dosing of Ringer and ZD7288 changed neither eye diameter nor development rate. Chronic dosing of ZD7288 did not significantly alter CAM vessel density close to the embryo (2, 3 and 4 mm) but at farther distances (5 and 6 mm) chronic dosing with both Ringer and ZD7288 decreased vessel density by 13 - 16%. Chronic dosing with ZD7288 also reduced body mass by 20%. Thus, lowered heart rate and cardiac output had little effect on vessel density or developmental stage, but did reduce embryo growth.
Thermal Identification of Clandestine Burials: A Signature Analysis and Image Classification Approach
Clandestine burials, the interred human remains of forensic interest, are generally small features located in isolated environments. Typical ground searches can be both time-consuming and dangerous. Thermal remote sensing has been recognized for some time as a possible search strategy for such burials that are in relatively open areas; however, there is a paucity of published research with respect to this application. This project involved image manipulation, the analyses of signatures for "graves" of various depths when compared to an undisturbed background, and the use of image classification techniques to tease out these features. This research demonstrates a relationship between the depth of burial disturbance and the resultant signature. Further, image classification techniques, especially object-oriented algorithms, can be successfully applied to single band thermal imagery. These findings may ultimately decrease burial search times for law enforcement and increase the likelihood of locating clandestine graves.
Life History Energetics of The Red-Eared Turtle, Pseudemys scripta in North Central Texas
A population of the red-eared slider, Pseudemys scripta, in north central Texas was studied from 1975 to 1980. A life history energy budget was developed for a typical individual in the population and the population dynamics were estimated. A growth model relating growth rate to mean plastron length (PL) was developed from recapture data and used to 1) establish age classes and 2) age individuals. Growth rate was highly variable in both sexes. Females grew more rapidly than males and attained a larger maximum size (230 mm and 195 mm PL in females and males respectively). Females reached sexual maturity in their ninth year at a PL of 185-190 mm. Males matured in their sixth year at a PL of 90-100 mm. Females produced three clutches annually; clutch size ranged from 7 to 14 (X=10.3; N=20). Ova were enlarged in the early spring and ovulation began in late April and early May. Egg laying occurred from mid- May through June. Both egg size and clutch size increased with female body size. Lipid levels were variable within and among seasons. No annual lipid cycling pattern was evident in females. The proportion of assimilated energy devoted to reproduction, a measure of reproductive effort (RE), by females, was 13 per cent the first year of maturity (9 y) and increased to 20 per cent by their twentieth year. Lifetime RE was 16 per cent. Population density was estimated as 51 males and 50 females per ha in a 8.2-ha area of the lake. Young juveniles (less than 2 y) were not present although other subadult age classes were about equally represented. Fewer adults were encountered. An annual total production of 4000 eggs was estimated for the population present in 1977. A survival rate of from 2 to 18 per cent was estimated for …
Factors Inhibiting Dissociation Of Neisseria gonorrhoeae Cells
The initial studies reported in this dissertation were attempts to induce mutations in those genes which control dissociation in cells of Nei sseria gonorrhoeae. These studies led to an investigation of survival curves of cells grown in liquid media. Instead of survival curves reflecting the diploid nature of gonococci, multiple cell kinetics were observed. It was found that large clumps contained a predominance of cells of the T2 type and that when these clumps were dispersed by DNAase, it appeared that dissociation of T2 was inhibited. The notion of a mechanism of T2 to T4 dissociation being due to genetic transformation was disspelled by these data.
Some Responses of Bacillus subtilis Spores to Glutaraldehyde
Bacillus subtilis (ATCC 19659) were damaged by exposure to various concentrations of glutaraldehyde, as shown by decreased germination rates. The damage caused was repaired or otherwise obviated by the presence of sodium lactate in the holding medium. When two different salts of lactic acid were compared for ability to overcome the effect of glutaraldehyde, it was found that calcium salt of lactate was more effective than the sodium salt. The damage repair system involved l-alanine, lactate and either the sodium or calcium ions. The study involved in determining the difference in efficiency of spore repair was due to an organic or an amino acii snowed that the presence of two carboxylic functional groups did not effectively alter the reactivity.
The Stoneflies (Plecoptera) of the Ozark and Ouachita Mountains
Collections of stoneflies (Plecoptera) were made at 603 stream sites from Nov. 1983 - May 1988 in the Ozark-Ouachita Mountain region, in relation to physiographic and vegetational characteristics. Examination of approximately 9000 vials from these collections, supplemented with material from major museums and other collectors, revealed 88 stonefly species in 8 families and 24 genera. Pearson's measure of association (R) showed there was a significant association between species present and each of the tested variables.
Studies on the Drumming Behavior of North American Stoneflies (Plecoptera)
Drumming behavior is described for the first time in 16 North American species of Plecoptera, and signals of a 17th species, Isogenoides zionensis, are further detailed. The effective distance over which drumming signals may be transmitted was tested for four communication modes. Results indicate that substratum vibrations are far superior to sound in the transmission of drumming signals, and that dense substrates such as rocks are poor channels for signal transfer. Long communication periods between stonefly pairs of Taeniopteryx burksi resulted in some alterations from initial signaling characteristics.
Fungal Antigens and Fungal Disease: An Alkali-Soluble, Water Soluble Antigen from Coccidioides immitis and Coccidioidomycosis
Diagnostic medical mycology has been slow to advance due to a lack of species specific antigens in organisms which cause serious diseases in man. Toward this end, an HPLC analysis was done of the following fungal antigens: histoplasmins HKC-43 and H-42, blastomycin KCB-26, an alkali-soluble, water soluble antigen from Blastomyces dermatitidis (b-ASWS), a coccidioidin prepared from a toluene lysate of the mycelial-arthroconidia phase of Coccidioides immitis, and an alkali-soluble, water-soluble antigen from Coccidioides immitis (c-ASWS). The HPLC survey included size-exclusion chromatography (SEC), ion exchange chromatography (HPIEC), and reversephase chromatography (RP). Resolution was poor with both SEC and HPIEC but was excellent with RP chromatography. The use of RP will allow sufficient separation for further antigenic and structural analysis.
Biology and Production of Net-Spinning Caddisflies (Hydropsychidae And Philopotamidae) in a Regulated Portion Of The Brazos River, Texas
Four species of net-spinning caddisflies, Hydropsyche simulans Ross, Cheumatopsyche lasia Ross, Cheumatopsyche campyla Ross and Chimarra obscura (Walker) are common in the regulated portions of the Brazos River. Hydropsyche simulans spun capture nets with the largest meshdimensions; the two Cheumatopsyche species' nets had the next largest meshes, and Chimarra obscura spun nets with the smallest dimensions. Cheumatopsyche lasia and C. campyla constructed nets with similar sized meshes. The number of individuals m~2 and standing crop biomass were not significantly different among low, medium, and high velocities. Early hydropsychid instars fed on detritus while later instar H. simulans and C. campyla had larger proportions of animal material. Guts of later instar C. lasia individuals had a greater percentage of algae and diatoms.
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