This syllabus includes both lecture notes and laboratory activities for a two-semester anatomy and physiology community college course. The syllabus is based on a 16-week semester that is comprised of a three-hour lecture section and a one-hour laboratory class each week. Both the lecture course and laboratory are necessary to fulfill the requirement for anatomy and physiology. Laboratory activities coincide with lectures to enhance understanding of each topic by providing visual and hands-on experiments for the concepts learned in the lecture. Laboratory quizzes will be given each week to help students maintain a working knowledge of the material learned in the laboratory. This course is appropriate for the typical anatomy and physiology student and should benefit both students who plan to major in biology and also those who are non-biology majors. Because subject matter in anatomy and physiology is quite difficult, the importance of attendance and good study skills is stressed.
Aspartate transcarbamoylase (ATCase) catalyzes the first committed step in the pyrimidine biosynthetic pathway. Bacterial ATCases have been divided into three classes, class A, B, and C, based on their molecular weight, holoenzyme architecture, and enzyme kinetics. Moraxella bovis is a fastidious organism, the etiologic agent of infectious bovine keratoconjunctivitis (IBK). The M. bovis ATCase was purified and characterized for the first time. It is a class A enzyme with a molecular mass of 480 to 520 kDa. It has a pH optimum of 9.5 and is stable at high temperatures. The ATCase holoenzyme is inhibited by CTP > ATP > UTP. The Km for aspartate is 1.8 mM and the Vmax 1.04 µmol per min, where the Km for carbamoylphosphate is 1.05 mM and the Vmax 1.74 µmol per min.
An aspartate transcarbamoylase (ATCase) gene from Bordetella pertussis was amplified by PCR and ligated into pT-ADV for expression in Escherichia coli. This particular ATCase (pyrB) was an inactive gene found adjacent to an inactive dihydroorotase (DHOase) gene (pyrC'). This experiment was undertaken to determine whether this pyrB gene was capable of expression alone or if it was capable of expression only when cotransformed with a functional pyrC'. When transformed into E. coli TB2 pyrB-, the gene did not produce any ATCase activity. The gene was then co-transformed into E. coli TB2 pyrB- along with a plasmid containing the pyrC' gene from Pseudomonas aeruginosa and assayed for ATCase activity. Negative results were again recorded.
The ability to synthesize pyrimidine nucleotides is essential for most organisms. Pyrimidines are required for RNA and DNA synthesis, as well as cell wall synthesis and the metabolism of certain carbohydrates. Recent findings, however, indicate that the pyrimidine biosynthetic pathway and its intermediates maybe more important for bacterial metabolism than originally thought. Maksimova et al., 1994, reported that a P. putida M, pyrimidine auxotroph in the third step of the pathway, dihydroorotase (DHOase), failed to produce the siderophore pyoverdin. We created a PAO1 DHOase pyrimidine auxotroph to determine if this was also true for P. aeruginosa. Creation of this mutant was a two-step process, as P. aeruginosa has two pyrC genes (pyrC and pyrC2), both of which encode active DHOase enzymes. The pyrC gene was inactivated by gene replacement with a truncated form of the gene. Next, the pyrC2 gene was insertionally inactivated with the aacC1 gentamicin resistance gene, isolated from pCGMW. The resulting pyrimidine auxotroph produced significantly less pyoverdin than did the wild type. In addition, the mutant produced 40% less of the phenazine antibiotic, pyocyanin, than did the wild type. As both of these compounds have been reported to be vital to the virulence response of P. aeruginosa, we decided to test the ability of the DHOase mutant strain to produce other virulence factors as well. Here we report that a block in the conversion of carbamoyl aspartate (CAA) to dihydroorotate significantly impairs the ability of P. aeruginosa to affect virulence. We believe that the accumulation of CAA in the cell is the root cause of this observed defect. This research demonstrates a potential role for pyrimidine intermediates in the virulence response of P. aeruginosa and may lead to novel targets for chemotherapy against P. aeruginosa infections.
The University of North Texas Stream Research Facility (UNTSRF) was designed to examine contaminant impacts on effluent-dominated stream ecosystems. Stream mesocosms, fed municipal effluent from the City of Denton, TX, Pecan Creek Water Reclamation Plant (PCWRP), were treated with 0, 15 or 140 µg/L cadmium for a 10-day study in August 2000. Laboratory toxicity test and stream macroinvertebrate responses indicated that cadmium bioavailability was reduced by constituents of effluent-dominated streams. The Biotic Ligand Model (BLM) for Cd was used to predict a 48 hour Cd EC50 for Ceriodaphnia dubia of 280 µg/L in these effluent-dominated streams. This value is higher that an EC50 of 38.3 µg/L Cd and a 7-day reproduction effect level of 3.3 µg/L Cd generated for C. dubia in reconstituted laboratory hard water. These results support use of a cadmium BLM for establishing site-specific acute water quality criteria in effluent-dominated streams. Although not affected by 15 µg/L treatments, organisms accumulated Cd in 15 µg/L treated streams. Hence, over longer exposure periods, Cd accumulation may increase and a no effect level may be lower than the observed 10-day no effect level of 15 µg/L. A toxicity identification evaluation procedure was utilized with in vitro and in vivo bioassays to identify estrogenic compounds in PCWRP effluent, previously identified to seasonally induce vitellogenin (VTG) in male fathead minnows. Steroids, nonylphenol ethoxylate metabolites, and other unidentified compounds were identified as causative effluent estrogens. These findings suggest that in vivo VTG bioassays should be used to confirm in vitro Yeast Estrogen Screening assay activity when effluents are fractionated or screened for estrogenicity. A subsequent 90-day cadmium study was initiated to assess long-term effluent and cadmium effects on fish endocrine function. Juvenile fathead minnows were placed in UNTSRF pool sections of replicate streams treated with 0, 5, 20 or 80 µg/L Cd. Male …
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.
Wildland fires in the United States are not always confined to wilderness areas. The growth of population centers and housing developments in wilderness areas has blurred the boundaries between rural and urban. This merger of human development and natural landscape is known in the wildland fire community as the wildland urban interface or WUI, and it is within this interface that many wildland fires increasingly occur. As wildland fire intrusions in the WUI increase so too does the need for tools to assess potential impact to valuable assets contained within the interface. This study presents a methodology that combines real-time weather data, a wildland fire behavior model, satellite remote sensing and geospatial data in a geographic information system to assess potential risk to human developments and natural resources within the Austin metropolitan area and surrounding ten counties of central, Texas. The methodology uses readily available digital databases and satellite images within Texas, in combination with an industry standard fire behavior model to assist emergency and natural resource managers assess potential impacts from wildland fire. Results of the study will promote prevention of WUI fire disasters, facilitate watershed and habitat protection, and help direct efforts in post wildland fire mitigation and restoration.
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.
This study focused on the effectiveness of integrating traditional plant community analyses with landscape ecological analyses based on remotely sensed data. A temporal analysis of plant community diversity was conducted for major plant communities of the Ray Roberts Lake area using transect monitoring data collected between 1987 and 1997. Landscape analyses were performed with FRAGSTATS*ARC using classified SPOT satellite imagery for 1987 and 1997. Although the methodology developed in this work was exploratory, it was found that characterizing the dynamics of major plant communities in the study area produced a more effective and insightful analysis of Ray Roberts Lake area landscape dynamics.
This dissertation comprises three parts and is presented in two chapters. Chapter 1 concerns Arthrobacter, a bacterium with an intriguing growth cycle. Whereas most bacteria exist as either a rod or coccus, this bacterium shares the rod/coccus lifestyle. It therefore seemed important to examine the growth regulatory pathways from the rod and coccus. The committed step, that catalyzed by aspartate transcarbamoylase (ATCase), in the pyrimidine biosynthetic pathway was chosen. The ATCase in Arthrobacter is like the well known Pseudomonas enzyme except that it has an active dihydroorotase (DHOase) associated. Included in Chapter 1 is the description of a microorganism, Burkholderia cepacia, whose ATCase has characteristics that are at once reminiscent of bacteria, mammals, and fungi. It differs in size or aggregation based on environmental conditions. In addition, it has an active DHOase associated with the ATCase, like Arthrobacter. B. cepacia is important both medically and for bioremediation. Since B. cepacia is resistant to most antibiotics, its unique ATCase is a prime target for inhibition. Whereas the first chapter deals with the de novo pathway to making pyrimidines, which is found mainly in the lag and log phase, Chapter 2 addresses the salvage pathway, which comes more into play during the stationary phase. This section focuses on the isolation, identification, and grouping of a number of natural soil bacteria from various soil locations. These organisms are important agriculturally, medically, and industrially. Addition of these soil isolates to poor soils has been found to improve the soil. In a previous study by D.A. Beck, the salvage schemes for a number of laboratory strains of microorganisms were determined. Nine separate classes of salvage were designated by determining the salvage enzymes present. In this study emphasis has been placed on soil bacteria, which had not previously been analyzed. A number of species of soil …
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 …
Laboratory single-species toxicity tests are used to assess the effects of contaminants on aquatic biota. Questions remain as to how accurately these controlled toxicity tests predict sitespecific bioavailability and effects of metals. Concurrent 42-day Hyalella azteca exposures were performed with cadmium and final treated municipal effluent in the laboratory and at the University of North Texas Stream Research Facility. Further laboratory testing in reconstituted hard water was also conducted. Endpoints evaluated include survival, growth, reproduction, and Cd body burden. My results demonstrate that laboratory toxicity tests may overestimate toxicity responses to cadmium when compared to effluent dominated stream exposures. Discrepancies between endpoints in the three tests likely resulted from increased food sources and decreased cadmium bioavailability in stream mesocosms
Nutrient availability, water depth, competition, and soil management effects on cattail (Typha latifolia) growth in wetland systems were examined. Soluble reactive phosphorous (SRP), nitrate-nitrogen (NO3-N), and ammonia-nitrogen (NH3-N) removals were tested at a constructed wetland receiving municipal wastewater effluent. Over all, no significant differences in nutrients occurred between diverse planted and cattail areas. T. latifolia seeds, under the canopy of Eleochoris macrostachya, had low seed germination. Established stands of emergent vegetation can prevent cattail colonization and spread. Germination of T. latifolia at various water depths was tested, and depth impacts on cattail seedling growth and survival were ascertained using various moist soil management techniques in three ponds. Water levels at 0cm and >40cm can adversely impact cattail establishment.
On-road remote sensing has the ability to operate in real-time, and under real world conditions, making it an ideal candidate for detecting gross polluters on major freeways and thoroughfares. In this study, remote sensing was employed to detect carbon monoxide (CO), hydrocarbons (HC), and nitrogen oxide (NO). On-road remote sensing data taken from measurements performed in six states, (Arizona, California, Colorado, Illinois, Texas, and Utah) were cleaned and analyzed. Data mining and exploration were first undertaken in order to search for relationships among variables such as make, year, engine type, vehicle weight, and location. Descriptive statistics were obtained for the three pollutants of interest. The data were found to have non-normal distributions. Applied transformations were ineffective, and nonparametric tests were applied. Due to the extremely large sample size of the dataset (508,617 records), nonparametric tests resulted in "p" values that demonstrated "significance." The general linear model was selected due to its ability to handle data with non-normal distributions. The general linear model was run on each pollutant with output producing descriptive statistics, profile plots, between-subjects effects, and estimated marginal means. Due to insufficient data within certain cells, results were not obtained for gross vehicle weight and engine type. The "year" variable was not directly analyzed in the GLM because "year" was employed in a weighted least squares transformation. "Year" was found to be a source of heteroscedasticity; and therefore, the basis of a least-squares transformation. Grouped-years were analyzed using medians, and the results were displayed graphically. Based on the GLM results and descriptives, Japanese vehicles typically had the lowest CO, HC, and NO emissions, while American vehicles ranked high for the three. Illinois, ranked lowest for CO, while Texas ranked highest. Illinois and Colorado were lowest for HC emissions, while Utah and California were highest. For NO, Colorado ranked highest …
Constructed wetlands are considered an inefficient technology for long-term phosphorus (P) removal. The P retention effectiveness of subsurface wetlands can be improved by using appropriate substrates. The objectives of this study were to: (i) use sorption isotherms to estimate the P sorption capacity of the two materials, masonry sand and light weight expanded shale; (ii) describe dissolved P removal in small (2.7 m3) subsurface flow wetlands; (iii) quantify the forms of P retained by the substrates in the pilot cells; and (iv) use resulting data to assess the technical and economic feasibility of the most promising system to remove P. The P sorption capacity of masonry sand and expanded shale, as determined with Langmuir isotherms, was 60 mg/kg and 971 mg/kg respectively. In the pilot cells receiving secondarily treated wastewater, cells containing expanded shale retained a greater proportion of the incoming P (50.8 percent) than cells containing masonry sand (14.5 percent). After a year of operation, samples were analyzed for total P (TP) and total inorganic P (TIP). Subsamples were fractionated into labile-P, Fe+Al-bound P, humic-P, Ca+Mg-bound P, and residual-P. Means and standard deviations of TP retained by the expanded shale and masonry sand were 349 + 169 and 11.9 + 18.6 mg/kg respectively. The largest forms of P retained by the expanded shale pilot cells were Fe+Al- bound P (108 mg/kg), followed by labile-P (46.7 mg/kg) and humic-P (39.8). Increases in the P forms of masonry sand were greatest in labile-P (7.5 mg/kg). The cost of an expanded shale wetland is within the range of costs conventional technologies for P removal. Accurate cost comparisons are dependent upon expansion capacity of the system under consideration. Materials with a high P sorption capacity also have potential for enhancing P removal in other constructed wetland applications such as stormwater wetlands and wetlands …
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.
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.
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