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- Chemotactic Response of Lumbricus terrestris Coelomocytes to Larval and Adult Stages of Rhabditis pellio
- Experiments were performed to assess the suitability of Rhabditis pellio, a nematode found in earthworms, as a challenge organism for use in development of a biomarker assay to determine the potential of chemicals to suppress the immunocompetence of the non-specific immune system. To accomplish this goal, information on the life cycle of R. pellio was determined; including effects of incubation time and temperature on growth rates; along with information on the immune response elicited in the earthworm, Lumbricus terrestris. Immune parameters measured were coelomocyte migration toward and attachment to R. pellio larvae and adults. Preliminary background information showed that R. pellio has potential as a challenge organism for development of a biomarker assay.
- Determination of Dissociation Constants for GABAA Receptor Antagonists using Spontaneously Active Neuronal Networks in vitro
- Changes in spontaneous spike activities recorded from murine frontal cortex networks grown on substrate-integrated microelectrodes were used to determine the dissociation constant (KB) of three GABAA antagonists. Neuronal networks were treated with fixed concentrations of GABAA antagonists and titrated with muscimol, a GABAA receptor agonist. Muscimol decreased spike activity in a concentration dependent manner with full efficacy (100% spike inhibition) and a 50% inhibitory concentration (IC50) of 0.14 ± 0.05 µM (mean ± SD, n=6). At 10, 20, 40 and 80 µM bicuculline, the muscimol IC50 values were shifted to 4.3 ± 1.8 µM (n=6), 6.8 ± 1.7 µM (n=6), 19.3 ± 3.54 µM (n=10) and 43.5 µM (n=2), respectively (mean ± SD). Muscimol titration in the presence of 10, 20, 40 µM of gabazine resulted in IC50s values of 20.1 (n=2), 37.17 (n=4), and 120.45 (n=2), respectively. In the presence of 20, 80, and 160 µM of TMPP (trimethylolpropane phosphate) the IC50s were 0.86 (n=2), 3.07 (n=3), 6.67 (n=2) µM, respectively. Increasing concentrations of GABAA antagonists shifted agonist log concentration-response curves to the right with identical efficacies, indicating direct competition for the GABAA receptor. A Schild plot analysis with linear regression resulted in slopes of 1.18 ± 0.18, 1.29 ± 0.23 and 1.05 ± 0.03 for bicuculline, gabazine and TMPP, respectively. The potency of antagonists was determined in terms of pA2 values. The pA2 values were 6.63 (gabazine), 6.21 (bicuculline), and 5.4 (TMPP). This suggests that gabazine has a higher binding affinity to the GABAA receptor than bicuculline and TMPP. Hence, using spike rate data obtained from population responses of spontaneously active neuronal networks, it is possible to determine key pharmacological properties of drug-receptor interactions.
- Evaluation of virulence in wild type and pyrimidine auxotrophs of Pseudomonas aeruginosa using the eukaryotic model system Caenorhabditis elegans.
- The human opportunistic pathogen, Pseudomonas aeruginosa PAO1, has been shown to kill the nematode Caenorhabditis elegans. C. elegans has been a valuable model for the study of bacterial pathogenesis, and has reinforced the notion that common virulence and host defense mechanisms exist. Recently, the pyrimidine pathway was shown to regulate virulence levels. Therefore, mutations in the pyrimidine pathway of PAO1 showed decrease virulence in the nematode. When starving the nematode, bacterial resistance was also shown to increase. It was hypothesized that starvation induced the DAF pathway, which regulates the transcription of genes involved with the antibacterial defense mechanism. Further research will be conducted to test this theory by performing RNAi experiments for the genes functioning in the antibacterial defense mechanism.
- Evidentiary Value of Condoms: Comparison of Durable Physical and Chemical Characteristics of Condoms
- Condom trace evidence must not be overlooked in sexual assault cases; understanding the chemical and physical characteristics of condoms is imperative if condoms are to be useful evidence. Previous research shows that condom identification is possible, but it is equally important to evaluate durability of condom residues versus time. Using FT-IR, this study examined vaginal swabs from subjects who self-sampled at intervals for up to 72 hours after having intercourse with a condom. This study investigated whether age and the stage of the menstrual cycle affected the durability of residues in the vagina over time. This study revealed that condoms containing nonoxynol-9, silicone-based lubricants, and particulates provide valuable information for identification, and that nonoxynol-9 specifically withstands the vaginal environment for up to 72 hours. Additionally, age and menstrual cycle both appeared to have an effect on the durability of residues although larger sample size is desirable.
- General Biology Lecture and Laboratory Curriculum Outline in a Two or Four-Year College
- In July of 1999, I wrote to 24 Texas junior and community colleges (and one four-year institution) describing my thesis agenda of a general biology lecture and laboratory syllabus for introductory biology students. I requested the titles and authors of the general biology textbooks and laboratory manuals they were currently using, the publishers of these texts, and the edition of said texts. I then contacted publishers of the various textbooks who, in turn, directed me to the Dallas-area representatives for further inquiries. I assimilated the various authors' general biology topics into a two-semester syllabus of lecture and one semester of laboratory. The document is not a text manuscript, but an all-inclusive listing of a general biology syllabus broken down by subject.
- Inquiry-based science for high school students: a forensic unit
- This project constitutes an instructional unit for honors biology that involves the use of science in the field of criminal investigation and forensics. Before beginning the unit, the learners should have mastered basic laboratory skills, including use of the microscope. They should also have an understanding of the basic structure and function of DNA and its role in heredity and protein synthesis. The standard time frame is 24 days with 70-minute periods, but can be easily adjusted to meet classroom needs. Several instructional strategies enhance student learning and make science fun. The unit is inquiry-driven and activity-based. Students are surprised by the crime, gather and analyze evidence, and work towards proposing an explanation. This real world problem involves the use of cooperative learning and a variety of assessment techniques.
- Integrating Concepts in Modern Molecular Biology into a High School Biology Curriculum
- More so than any other science in the past several decades, Biology has seen an explosion of new information and monumental discoveries that have had a profound impact on much more than the science itself. Much of this has occurred at the molecular level. Many of these modern concepts, ideas, and technologies, as well as their historical context, can be easily understood and appreciated at the high school level. Moreover, it is argued here that the integration of this is critical for making biology relevant as a modern science. A contemporary high school biology curriculum should adequately reflect this newly acquired knowledge and how it has already has already begun to revolutionize medicine, agriculture, and the study of biology itself. This curriculum provides teachers with a detailed framework for integrating molecular biology into a high school biology curriculum. It is not intended to represent the curriculum for an entire academic year, but should be considered a significant component. In addition to examining key concepts and discoveries, it examines modern molecular techniques, their applications, and their relevance to science and beyond. It also provides several recommended labs and helpful protocols.
- 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 Case Building Behaviors of Phylloicus ornatus (Banks) (Trichoptera: Calamoceratidae) In Two Spring Fed Tributaries in the Central Edwards Plateau Bioregion of Texas
- The life history and case-making behaviors of Phylloicus ornatus from two springfed first order streams in the Edwards Plateau Bioregion of Texas were studied from January 1998 to November 1999. Field larval, pupal and adult samples and laboratory rearings indicated a multivoltine cycle. First instars differ from late instars in number of labral setae and in having a unique spur-like claw on each lateral hump. Larval development was asynchronous with second through fifth instars and pupae present most months. First instars were present April through July, October and November. Case making of first instar and case reconstruction of later instars extracted from their cases was documented by videophotography.
- Macroinvertebrate Community Structure as an Indicator of Watershed Health in the Upper Trinity River Basin, North Central Texas
- This study describes macroinvertebrate community structure and assesses its potential in detecting point and non-point sources of disturbance associated with rural and urban areas in the Upper Trinity River Basin. Geospatial techniques were used to quantify landuse within the watershed in a GIS. At rural sites near the headwaters of the Trinity River, collector-gathering burrowers that are adapted to minimal flow comprised the majority of taxa. Destinies of taxa compositions at downstream sites increased and shifted toward psammophilic and rheophilic invertebrates, including primarily collector-filtering clingers, that are characteristic of shifting sand habitats in large prairie rivers. Benthic community structure generally benefited from point source impacts including wastewater treatment plant effluents that maintained higher flow. Community indices were negatively associated with forest landuse and positively associated with urban landuse. Partial CCA determined that flow and landuse contributed equally to species dispersions. Comparisons with historical biomonitoring studies in upper Trinity River Basin indicate improved watershed health.
- A morphological study of the avian (Gallus domesticus) ductus arteriosi during hatching.
- The ductus arteriosi (DA) are two blood vessels connecting the pulmonary arteries to the descending aorta in the avian embryo. Following hatching, the DA closes, separation of the systemic and pulmonary circulation. I present the morphological changes that occur in the chicken DA during prepipping, internal pipping, external pipping, and hatching. The avian DA consists of two distinct tissue types, a proximal and a distal portion. Histological examination shows developmental differences between the proximal and distal portions of the DA with regard to lumen occlusion, endothelial cells, smooth muscle and elastin. Endothelial cell proliferation begins to occur as early as external pipping, with the lumen almost completely occluded by the 3rd day of post-hatching life. Expression of vascular endothelial growth factor (VEGF) increases in avian endothelial cells during hatching. I provide a morphological timeline of changes in the DA as the chicken develops from embryo to hatchling.
- A Multimedia Atlas of Dissection for Comparative Anatomy of the Vertebrates
- Traditional methods of teaching the laboratory course for Comparative Anatomy of the Vertebrates could be improved by applying current computer technology to construct an interactive, multimedial atlas of dissection. Five specimens used in comparative anatomy courses at most institutions were chosen as representative members of the Phylum Chordata: amphioxus, lamprey, dogfish shark, mud puppy, and cat. Specimens were dissected according to the modified method of Wischnitzer, 1993, and each stage was photographed with a Kodak DC120 digital zoom camera. These images were processed on a Power Macintosh 7600 computer with Adobe Photoshop v. 5.0. The atlas was constructed from these images using Macromedia Authorware v. 4.0.3. Each image contains a series of interactive objects that display a highlight and descriptive text as the cursor passes over each object.
- Phenotype Analysis of the CISD Gene Family Relative to Mitochondrial Function in Caenorhabditis elegans
- NEET proteins belong to a unique class of [2Fe-2S] cluster proteins that have been shown to participate in various biological processes such as regulating iron, reactive oxygen species and apoptosis within the cell and are localized to the mitochondria. Disruption of the mitochondrial NEET proteins are associated with different human diseases such as obesity, neurodegeneration, cancer and diabetes. In humans, a missense mutation in the CISD2 gene results in a heritable multisystem disorder termed Wolfram syndrome 2 (WFS2), a disease which displays an early onset of juvenile diabetes and various neuropsychiatric disorders. The C. elegans genome contains three previously uncharacterized cisd genes: cisd-1, which has homology to the human CISD1 and CISD2, and cisd-3.1 and cisd-3.2, both of which have homology to the human CISD3. Disruption of the cisd-3 gene(s) function results in mis-regulation of proteostasis in the mitochondria, whereas cisd-1 and cisd-3.1 disruption impacts proteostasis in the endoplasmic reticulum. Reduction of cisd-3.2 gene function also leads to a developmental delay in C. elegans. A knockout mutation of the cisd-3.2(pn68) gene function results in various germline defects including delayed development progression and morphological defects. Furthermore, I show the cisd gene(s) and protein expression profiles is present relative to sex, tissue type and developmental stages. This work is significant because it provides further insight of the essential role of CISD-3 relative to C. elegans. Furthermore, my studies can contribute to new genetic discoveries that will widen the scientific research relative to NEET protein family studies.
- A Physiological Age-Grading System for Female Hydrellia pakistanae Deonier (Diptera: Ephydridae)
- Conflicting opinions about the effectiveness of H. pakistanae as a biological control agent for hydrilla prompt researchers to find a method for assessing the fly's success. Developing a physiological age-grading system for the fly using ovarian morphology to detect changes in reproductive activity is useful for evaluating reproductive status of the fly in field populations. Changes in the appearance of follicular relics in ovaries with oviposition provide a reliable method to estimate fecundity. Characteristics of follicular relics were used to develop a system with eight physiological age classes, three nulliparous and five parous. Changes that occur in the fat body were used to assist in classification of nulliparous females or those with low egg counts.
- Use of Geographic Information System and Remote Sensing Technologies to Describe Mosquito Population Dynamics in the Ray Roberts Greenbelt, Denton County, Texas
- A population survey was conducted from April through September 2002 on mosquito species occurring on the Ray Roberts Greenbelt, a riparian corridor used for public recreation on the Elm Fork of the Trinity River, in Denton County, Texas. ArcGIS software was used to set up a stratified random sampling design based on habitat parameters. Multivariate analyses of sampling data and climatic variables were used to describe spatial and temporal patterns of mosquito species. A total of 33 species were collected during this study belonging to the following genera: Aedes, Anopheles, Coquillettidia, Culex, Mansonia, Ochlerotatus, Orthopodomyia, Psorophora, Toxorhynchites, and Uranotaenia. Seasonal distributions of the dominant species revealed population fluctuations. Aedes vexans was the primary species collected in April and May, occurring in low numbers throughout the rest of the sampling period. Psorophora columbiae reached its highest population density in June, with a smaller peak occurring in late July. Present from May through the end of September, Culex erraticus was the most abundant species collected with major peaks in mid-June and the end of July. Abundance of Culex salinarius followed the same general trend as that for Cx. erraticus, but with smaller numbers. The specimens were tested for a variety of arboviruses by the Texas Department of Health. One pool of Cx. erraticus and Cx. salinarius, collected in August 2002, tested positive for West Nile virus. Variables that were important factors for determining dominant species abundance were temperature, wind speed, rain accumulation occurring one-week and two-weeks prior to sampling, number of day since last rain event, dew point, and average canopy coverage.
- 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.
- Utilization of Corridor Habitat by White-tailed Deer (Odocoileus virginianus) in Denton County, Texas
- White-tailed deer (Odocoileus virginianus) (N=15) movements were determined with use of radio telemetry techniques to determine the utilization of corridor habitat on the Lake Ray Roberts Greenbelt Corridor (RRGC) in north central Texas. Home ranges were calculated using three estimation types. Male white-tailed deer tend to have home ranges twice that of female home ranges. Seasonal home ranges were largest during spring (Feb. - April) and fall (Aug. - Oct.) seasons. Males had greater seasonal variation in utilization than females. No statistically significant difference (p=0.24) between white-tailed deer locations when the RRGC experiences heavy human traffic compared to days when there is light human traffic. Linearity indices indicated home ranges less linear than expected (LI = 3.02). The RRGC should be maintained at its current status to provide a variety of vegetational types and protective cover for white-tailed deer and other wildlife of Denton County.