2004 Projects
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Item Open Access Assessing genetic diversity and phenotypic plasticity in Cirsium arvense: evaluation with greenhouse trials and ISSR(Colorado State University. Libraries, 2004) Gaines, Todd A., author; Brown, Cynthia S., author; Hufbauer, Ruth A., authorInvasive plant species cause damage to ecosystems and economic loss to land managers. One particularly invasive plant species is Canada thistle (Cirsium arvense). This plant is very difficult to control and is a successful invader in many diverse ecosystems, including cropping systems and non-cultivated lands. If C. arvense is successful because it has adapted genetically to new environments, then these differences may be quantified using experimental observations. The objective of this project is to quantify genetic diversity and site-specific genetic differentiation based on phenotypic responses in common garden trials. Canada thistle specimens from crop and non-crop habitats in two different biogeographic regions of Colorado were used. A second objective is to correlate data from genetic markers with phenotypic data to further quantify genetic diversity in specimens from crop and non-crop habitats in three different regions of Colorado. The greenhouse experiments compared the phenotypic responses of the collected plants to variations in soil fertility and water stress. Leaf tissue DNA collected from the sites was amplified with the inter-simple sequence repeat (ISSR) method and the resulting banding patterns were analyzed for genetic variation. Data from the greenhouse trials indicate a greater response to treatments only in Larimer County non-crop populations. Data from genetic analyses indicate a high level of diversity in the sampled genomes. These data are consistent with the invasive characteristics of C. arvense but indicate that Colorado populations have not genetically differentiated within the sampled ecosystems. More study is warranted to further investigate this question.Item Open Access Conocimiento e impedimentos de metodos anticonceptivos: las mujeres del Centro de Salud en Cerro Verde, Cochambamba, Bolivia(Colorado State University. Libraries, 2004) King, Erin M., author; Yarrington, Douglas, authorThe purpose of this project was to measure the level of knowledge regarding contraception and family planning and the actual and potential impediments to women's ability to plan the number of children in their families. Methodology: The information was obtained through semi-structured interviews with NGO program directors, health center doctors, and the women who utilized the services of the Cerro Verde health center. Research conducted previous to the interviews provided a hypothesis, guided the structure of the interviews, and later supplemented the findings. Findings: There are several factors that influence women's access to and decision to use contraceptives: availability of information, presentation of the information, accessibility of contraceptives, economic situation of the family, and the opinion of the husband. Implications: Increasing the number of community programs, sponsored by the health centers and NGOs, aimed at both women and men, and teenaged girls and boys could potentially increase familiarity with methods of family planning, including contraception thereby expelling any misunderstandings or rumors about the effects of such methods.Item Open Access Copper chaperones in Arabidopsis thaliana. Intra cellular copper trafficking: uptake, delivery and regulation(Colorado State University. Libraries., 2004) Bodecker, Jared, author; Abdel-Ghany, Salah E., author; Burkhead, Jason, author; Pilon, Marinus, authorThe chloroplast is the site of photosynthesis. Copper is an essential element to chloroplast function as a co-factor of superoxide dismutase (SOD) and plastocyanin, which function in photosynthesis. Chloroplasts have a complex structure due to the presence of three membranes, so how then is Copper delivered throughout the complex internal structure of plant chloroplasts? In plant chloroplasts, membrane transporters have been identified that transport Copper across these membranes. In microbes, a family of small cytoplasmic proteins called 1cmetallochaperones 1d or Copper Chaperones carries out the delivery of copper from transporters to targets. After these chaperones bind to copper, they deliver and insert the copper ions into an active site of a specific partner, a copper dependent enzyme or another transporter. In the genome of A. thaliana possible genes encoding for copper chaperones have been identified based on the similarity of sequences with microbial chaperones. We named these, CpCCS (Chloroplastic Copper Chaperone for SOD1), ATX2 (similar to yeast Antioxidant protein) and CpCopZ. These three A. thaliana Cu Chaperone proteins may be required for Cu transport to the internal compartments of the chloroplast. To test the function of the putative copper chaperones in copper delivery two approaches are taken. First we test complementation of yeast mutants, deficient in copper chaperones. Second, we analyze the function and need for these proteins in plants with the insertions (KO-mutants) in each copper chaperone gene.Item Open Access Creation of an endA mutant strain in Pseudomonas aeruginosa PAO1 using gene replacement(Colorado State University. Libraries, 2004) Olsen, Cassie J., author; Karkhoff-Schweizer, Roxann R., authorEndonuclease I is an enzyme encoded by the endA gene. This nuclease degrades double stranded DNA. Many Escherichia coli common laboratory strains contain a mutation in the endA gene that inactivates the DNA-specific endonuclease I. A mutation in this gene greatly increases plasmid DNA yields in such E. coli strains as well as improves the quality of DNA that is isolated. The purpose of this research is to create an endA mutant strain in Pseudomonas aeruginosa PAO1 using gene replacement, thereby leading to the development of a useful laboratory Pseudomonas strain for use as a cloning strain. To accomplish this, chromosomal DNA from P. aeruginosa PAO1 was isolated, and the endA gene was then amplified by PCR using specific primers designed to the flanking upstream and downstream sequence of the endA coding region. The resulting amplified 1100 bp DNA fragment containing the endA gene was cloned into pCR2.1. This newly created plasmid was named pCR2.1-endA. In order to create an insertionally inactivated endA gene, a GmR encoding cassette from pPS856 needed to be inserted into the SalI sites of the cloned endA gene. The pCR2.1-endA plasmid was digested using SalI restriction enzyme. A 4500 bp SalI fragment of pCR2.1-endA was isolated and then religated by T4 DNA ligase. The new plasmid created was called pCR2.1-endASalID. This plasmid was digested with SalI, and blunt ends were created with T4 DNA polymerase. Inactivation of the endA gene was accomplished by insertion of a blunt-ended, GmR encoding gene into the blunt-ended SalI site of the endA coding sequence. The resulting recombinant plasmid was called pCR2.1-endASalID(Gm1). A 1700 bp HindIII x PstI DNA fragment from pCR2.1-endASalID(Gm1), containing the insertionally inactivated endA gene, was isolated and cloned into the similarly digested pEX18Ap plasmid.Item Open Access Development and characterization of an anti-bat antisera and its implementation in screening for rabies antibody in bats via ELISA and IFA(Colorado State University. Libraries, 2004) Schmidt, Kelsey M., author; Gordy, Paul W., author; Bowen, Richard A., authorBats have long been associated with the transmission of a number of zoonotic agents, including human rabies. Comprehensive surveys of wild bat populations to characterize the seroprevalance rate of the population could prove helpful in implementing control programs designed to reduce the number of human and veterinary cases resulting from bat-associated rabies. The antiserum was developed by immunizing rabbits with purified Eptesicus fuscus (big brown bat) IgG. Following immunization, the rabbits were bled and the anti-bat antiserum was purified and characterized. The antisera is currently being used to develop an indirect enzyme-linked immunosorbent assay (ELISA) and an indirect immunofluorescent antibody test (IFA), which will be used to screen for rabies-specific IgG antibody in bats. While these tests do not detect active viral infection or neutralizing antibody, they have clear advantages over the rapid fluorescent focus inhibition test (RFFIT), fluorescent antibody virus neutralization test (FAVN) and mouse inoculation test. The proposed methods do not require the use of live virus for the assay and are inexpensive. They also confer a clear advantage in that the amount of serum required for the assay is considerably less than that which is required for RFFIT or FAVN. We describe the development of the rabbit anti-bat IgG antibody and report preliminary results on our ELISA and IFA assays.Item Open Access Employing popular children's literature to teach elementary school chemistry: an engaging outreach program(Colorado State University. Libraries, 2004) Wally, Laura M., author; Levinger, Nancy E., author; Grainger, David W., authorPopular cultural themes or fads can provide an effective vehicle to enthuse students about science. The program described here uses current children's literature in a versatile chemical education activity. This activity generates excitement in elementary school students because of the integration of the popular Harry Potter literature series with hands-on experiments. Elementary school student participants are prompted to explore three main scientific topics during this activity: properties of solids, liquids, and gases ; ethics in science ; and the scientific method. Undergraduate and high school student volunteer mentors are readily trained to conduct the activity within elementary schools via an introduction to the philosophy of the exercise in conjunction with working through the activity themselves. As an outreach activity it serves to connect undergraduate and high school students both with faculty and their community.Item Open Access Regulation of Sarco/Endoplasmic Reticulum Ca2+ ATPase mRNA as a mechanism to prevent Ca2+ cytotoxicity(Colorado State University. Libraries, 2004) Werner, Sara B., author; Sandau, Ursula S., author; Hinds, Laura R., author; Handa, Robert J., authorThe Sarco/Endoplasmic Reticulum Ca2+-ATPase Pump (serca2) plays a critical role in intracellular Ca2+ dynamics. One function is to store intracellular Ca2+ in the endoplasmic reticulum. By sequestering Ca2+ from the cytoplasm, serca pumps can act to prevent Ca2+ induced excitotoxicity. Recent studies have shown that androgens can also prevent excitotoxic cell death. Therefore, in this study, we investigated if androgens can regulate serca2. Primary hippocampal neurons were prepared from E18 rat fetuses and after 14 days in vitro, were treated with either dihydrotestosterone (DHT) or vehicle (control) for 48 hours. Following treatment, cells were harvested, total RNA was isolated, and serca2 mRNA was measured using quantitative real time RT-PCR. GAPDH mRNA was measured as an internal control. Levels of serca2 mRNA were compared between the two treatment groups. A dose response curve for DHT was generated (1nM and 10nM). A 1nM concentration of DHT significantly upregulated serca2 mRNA as opposed to that of 10nM where a down-regulation was seen. The upregulation of serca2 mRNA following DHT treatment could be responsible for the neuroprotective actions attributed to androgen receptors.Item Open Access Study of the factors that affect attitudes towards reporting sexual assault of females at Colorado State University(Colorado State University. Libraries, 2004) Cordova, Caitlyn L., author; Downey, Eleanor, authorThe purpose of this study was to determine the factors discourage female students at Colorado State University from reporting sexual assault incidences to the police or other law enforcement agencies. For the purpose of this study sexual assault was defined as any sexual contact without consent, this includes but is not limited to: touching of intimate body areas, intercourse, or penetration. This study was done using a questionnaire to survey 100 females on the CSU campus. The survey consisted of 27 questions that assessed the respondents 19 attitudes about reporting the assault based on individual feelings about sexual assault, circumstances related to the assault, and individual characteristics. The questionnaire also contained 2 questions related to the respondents 19 knowledge of and willingness to use resources on the CSU campus and/or in the Fort Collins community and 4 questions that collected demographic information about the respondents.Item Open Access The emb proteins in Mycobacterium smegmatis(Colorado State University. Libraries, 2004) Nagy, Toni A., author; Vissa, Varalakshmi, author; Chatterjee, Delphi, authorMycobacterium tuberculosis, the causative agent of tuberculosis, was once thought to be a ghost of the past. However, the emergence of drug resistant strains has brought this bacterium back into the spot light. Ethambutol, a compound with structural similarity to D-arabinose, has been shown to have many inhibitory effects on M. tuberculosis and other mycobacterial species. Some of these activities include: inhibition of synthesis of the arabinan constituent of the cell wall arabinogalactan (AG), inhibition of RNA metabolism, and phospholipid synthesis. Likewise, this inhibition of arabinan synthesis extends to that of lipoarabinomannan (LAM). One of the most significant components of all mycobacterial cell walls is LAM; it has been implicated as a central molecule involved in the virulence and immunopathogenesis of tuberculosis. The arabinan of LAM is attached to a mannan backbone which extends from a phosphatidylinositol mannoside anchor at the reducing end. Recent research efforts have been directed towards showing that the Emb proteins are involved in arabinan synthesis. These proteins are conserved throughout all mycobacterial species, and through the use of computer program algorithms (such as SOSUI, TMPRED), it has been predicted that there are 11-13 transmembrane domains in the N-terminal region of the proteins. This correlates to approximately 670 amino acids. Also, these programs showed a soluble globular C-terminal domain accounting for approximately 430 amino acids. Specifically, it was found that EmbC targets the arabinan of LAM. This was made possible by the inactivation of the embA, embB, and embC genes individually, and subsequently looking at the resulting structural alterations on the arabinan component of cell wall AG and LAM. With this knowledge, the establishment of the catalytic site of the embC gene that specifically controls the arabinosylation of lipomannan to give mature lipoarabinomannan was necessary. Generation of hybrids with variation in the N-terminus of the EmbC protein is the current strategy being used to pursue this information. An EmbC/B hybrid was created by the fusion of the N-terminus of EmbC and the C-terminal domain of EmbB. This included 668 amino acids from the N-terminal of EmbC, fused with the last 407 amino acids of EmbB. This gene fusion was cloned into the pVV16 shuttle vector and electroporated into M. smegmatis ΔembC. LAM was then extracted from these hybrid cells, and analysis showed that the EmbC/B hybrid resulted in a shortened form of LAM. After further biochemical studies on this truncated LAM such as glycosyl compositional analysis, endoarabinase digestion followed by HPAEC (High-pH anion-exchange chromatography) profiling, and mass spectrometry analysis, it was shown that LAM was not only truncated, but had a structural alteration where the nonreducing end resembled that of the arabinan of AG. After such exciting results, current work continues with the generation of hybrids. The formation of the 50:50 EmbC/B hybrid containing approximately 580 amino acids of each of the EmbC and EmbB genes has been completed. This final construct has been transformed into the ΔembC mutant by electroporation and biochemical analysis is being performed. Results pertaining to the hybrids capacity to complement the LAM defect will be discussed in this poster presentation. This new 50:50 fusion will be essential in giving us the knowledge on the contribution of the first eight transmembrane domains of the EmbC protein in LAM biosynthesis.Item Open Access The physics of cutmarks(Colorado State University. Libraries, 2004) Potter, Sheridan L., author; Todd, Larry C., authorCutmarks are the most direct evidence of faunal butchery by humans ; however, understanding the physical properties associated with their creation is critical when interpreting the archaeological record. By quantifying the minimum amount of force required to cut through soft tissue and the minimum amount of force required to produce a visible cutmark on the surface of bone, and then correlating those values with the maximum amount of force exerted by a human butchering with a stone tool, archaeologists will better understand the conditions conducive to creating cutmarks. A porcine metatarsal served as the specimen for the cutting experiment, while obsidian and chert flakes, and a scalpel blade were used as the cutting tools. Axial cutting force was measured with a dynamic loading cell, accurate to the nearest Newton. Cutmarks were replicated with rubber latex and were analyzed using a scanning electron microscope at varying degrees of magnification, and depth was measured to the nearest micrometer. Twenty adults (10 male and 10 female) volunteered to perform an experiment measuring the maximum amount of force that could be exerted in a kneeling position while holding a small flake and a large biface. Force was measured using a digital scale accurate to the nearest tenth of a kilogram. Results have shown that less force is required to cut through soft tissue using obsidian as opposed to chert flakes, the amount of force required to produce a visible cutmark on a bone is constant, and that on average males can exert a greater maximum force using both large and small stone tools than females.Item Open Access The Sea Gecko at Pirate Cove Aquarium and Resort(Colorado State University. Libraries, 2004) Shepard, Charles B., author; Leigh, Katharine E., authorMy senior Capstone project dives into the waters of Costa Rica where design has taken a new step toward embracing the future of sustainable living. This special resort yacht offers scuba diving, dining and bar facilities, glass bottom boat viewing, hot tub, and accommodates sailing. The concept that separates this boat from the competition is its sustainable design. The boat runs completely off clean energy, through solar and wind power. The electric engines use solar and wind power as a backup system to giant sails that provide the main source of transport across ocean waters. The boat type is trimaran, with three hulls, a large center cabin, and two smaller cabins on port and starboard sides. This hull design makes the boat virtually impossible to capsize, and allows a larger deck expanse. The boat is almost as wide as it is tall yet remains very streamlined allowing the hull to cut through the water, leaving minimal wake. The 'glass bottom boat' feature is unique in the sense that it acts as an inverse aquarium. Large aquarium windows on the sides and bottom give the space an open ocean-viewing experience to all who come aboard. My objective for this project was to demonstrate the capabilities that this aquatic facility brings to sustainable study.Item Open Access Third metacarpal condyle bone mineral density in relation to equine condylar fractures(Colorado State University. Libraries, 2004) Zimmerman, Chelsea A., author; Shearin, M. G., author; Kawcak, Chris E., authorFractures of the third metacarpal condyle in metacarpophalangeal joints frequently occur in young racehorses during high-speed training or racing, and can be career or life ending. Prior research suggests that equine condylar fractures may occur from periods of continued loading, as opposed to a single traumatic event. Also, repeated trauma may lead to higher subchondral bone density, possibly reducing bone quality making it more susceptible to microfractures. The coalescence of these microfractures can lead to gross fracture. The objectives of this study were to evaluate density patterns within the distal third metacarpal (MC3) bone of forelimbs in racing and non-racing horses, and to determine how density patterns might relate to areas of condylar fracture commonly seen in racehorses. Computed Tomographic (CT) scans of the metacarpophalangeal joints were taken bilaterally on eight racehorses and eight control horses. Using OsteoApp, a program that utilizes CT data to create three-dimensional volumes, bone mineral density was measured from fifteen slices, taken in the frontal plane, radiating at 2mm increments from the palmar aspect to the dorsal aspect of the distal third metacarpus. A specific color scheme, based on CT pixel values, was established to identify visual patterns of the slices. Density data were then calibrated and analyzed using SAS software (Cary, NC). There was a statistically significant difference (p<.0005) between the mean densities of racehorses vs. control horses. Although there was no evidence that the limb (L, R) played a significant role, mean slice density of racehorses was significantly different between slice locations (p<.0001). In conclusion, racing and high speed training leads to a significant increase in density, and a density pattern that appears to create a sharp density gradient. Large changes in bone density are thought to increase the chances of fracture, therefore predisposing racehorses to injury. Further research is needed to identify the changes preceding a condylar fracture in order to prevent them.