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.

The effects of three oral dosages (0.4 mg, 0.8 mg, and 1.6 mg) of quercetin on Colon-25 tumors implanted in Balb-c mice were studied. The data in this study show that: (1) certain dosages of quercetin in alcohol solutions, reduces the weight, and size of implanted Colon-25 tumors in Balb-c mice, (2) these same dosages of quercetin all produce a profound neutrophilia combined with a significant lymphopenia at day 20 post-implantation, and (3) there was relatively little evidence of histological changes in the quercetin-treated tumor section which would indicate that the action(s) of quercetin is primarily at the subcellular level probably within the nuclei of the tumor cells.

Spontaneously active monolayer neuronal networks cultured on photoetched multimicroelectrode plates (MMEPs) offer great potential for use in studying neuronal networks. However, there are many problems associated with frequent, long-term use of MMEPs. The major problems include (1) polysiloxane insulation deterioration and breakdown, (2) and loss of gold at the gold electroplated indium-tin oxide (ITO) electrodes. The objective of this investigation was to correct these major problems. Quality control measures were employed to monitor MMEP fabrication variables. The phenotypes of polysiloxane degradation were identified and classified. Factors that were found to contribute most to insulation deterioration were (1) moisture contamination during MMEP insulation, (2) loss of the quartz barrier layer from excessive exposure to basic solutions, and (3) repetitive use in culture. As a result, the insulation equipment and methods were modified to control moisture-dependent insulation deterioration, and the KOH reprocessing solution was replaced with tetramethylguanidine to prevent damage to the quartz. The problems associated with gold electroplating were solved via the addition of a pulsed-DC application of gold in a new citrate buffered electroplating solution.

Seventy-one aquatic species including the copepodids and nauplii were identified from Lake Texoma from August 1996 to September 1997. Zooplankton community structure, abundance and spatial and temporal distributions were compared among five lake zones delineated a priori based on chloride concentration. The zones, in order of decreasing chloride concentration, are the Red River zone (RRZ), Red river Transition zone (RRTZ), Main Lake zone (MLZ), Washita River Transition zone (WRTZ) and Washita River zone (WRZ). Bray Curtis Similarity Index showed community structure was most similar in the two Red River arm zones, the two Washita River arm zones and the MLZ. Zooplankton abundance was greatest in the Red River arm (312 org/L), intermediate in the Washita River arm (217 org/L) and least in the Main Lake body (103 org/L). A significant increase in the abundance of a deformed rotifer, Keratella cochlearis, was observed mainly in the Red River arm during a second study from March 1999 to June 1999. Seasonal dynamics, rather than spatial dynamics, were more important in structuring the zooplankton community, especially in the two river arms. Spatial variance was solely attributed to station and zone effects independent of time for a few crustacean species and many of the …

The proposed study examined the role of anxiety and risk-taking in driving performance in adolescents. In addition to examining the sample as a whole, gender differences were assessed given earlier reports from our laboratory and others indicating that males and females differ with respect to risky behaviors to driving performance and anxiety. Adolescents' subjective and physiological responses to a driving simulator task were assessed. Anxiety was measured via self report and salivary cortisol. Participants provided a baseline saliva sample and 3 post-task samples for cortisol analysis. Subjective anxiety scores were obtained at both baseline and following the driving stressor. Information concerning impulsivity, as well as other psychological constructs was also collected at baseline. Unlike the pilot study, there were no relationships (with or without respect to gender) between salivary cortisol and both self-reported anxiety (state and trait) or impulsively measures for this sample. These results suggest that this group of adolescents may not have been anxious about the driving task. This discrepancy may stem from error introduced by the smaller sample size obtained from the initial findings or to other factors remaining outside the parameters of the current study. The task did, however, induce a slight hypothalamic pituitary adrenal (HPA) …

Earthworms, Lumbricus terrestris, were challenged orally and intracoelomically with two bacterial species, Aeromonas hydrophila and Pseudomonas aeruginosa, and mortality rates were observed. Neither were found to be particularly pathogenic at injected doses of up to 108 bacteria per earthworm. The influence of Cu++ (as CuSO4) on the earthworm's response to bacterial challenge was investigated by exposing earthworms to sublethal levels of Cu++ prior to bacterial challenge. Exposure at sublethal concentrations up to 3 m g/cm2 did not have a pronounced influence on host resistance to challenge as measured by earthworm mortality. Cu++ increased the earthworm's ability to agglutinate rabbit erythrocytes, indicating that Cu++ exposure caused coelomocyte death, autolysis and release of agglutinins into the coelom, possibly explaining resistance to bacterial challenge.

Four populations of nine-banded armadillos, Dasypus novemcinctus, were investigated in the south-central United States in order to assess genetic variation in an isolated population (Electric Island, Lake Hamilton, Garland County, Arkansas); a semi-isolated population (Arkansas Post, Arkansas County, Arkansas), and two free ranging populations (southern Arkansas and central Texas). A 233 basepair sequence of the D-loop region of mitochondrial DNA was sequenced in individuals from each population. Individuals and populations were compared to assess relatedness among populations and individuals. Higher sequence diversity was detected in the semi-isolated population, while lower sequence diversity was observed in the isolated and free ranging populations. Overall, all populations exhibited low genetic variation when compared to genetic variation for other mammals. The results support the hypothesis that rapid range expansion combined with the organism's unique reproductive strategies have promulgated low genetic variation in the North American populations of nine-banded armadillos.

This study focused on the enzyme, aspartate transcarbamoylase (ATCase) from A. hydrophila, a Gram-negative bacterium found in fresh water. The molecular mass of the ATCase holoenzyme from A. hydrophila is 310 kDa. The enzyme is likely composed of 6 catalytic polypeptides of 34 kDa each and 6 regulatory polypeptides of 17 kDa each. The velocity-substrate curve for A. hydrophila ATCase is sigmoidal for both aspartate and carbamoylphosphate. The Km for aspartate was the highest to date for an enteric bacterium at 97.18 mM. The Km for carbamoylphosphate was 1.18 mM. When heated to 60 ºC, the specific activity of the enzyme dropped by more than 50 %. When heated to 100 ºC, the enzyme showed no activity. The enzyme's activity was inhibited by ATP, CTP or UTP.

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.

GABAA receptor binding is transiently increased in rat whisker barrels during the second postnatal week, at a time when neurons in the developing rat cortex are vulnerable to excitotoxic effects. To test whether these GABAA receptors might serve to protect neurons from excessive excitatory input, polymer implants containing the GABAA receptor antagonist bicuculline were placed over barrel cortex for a 4-day period in young (postnatal days 8 - 12) and adult rats. In the cortex of young, but not adult rats, the chronic blockade of GABAA receptors resulted in substantial tissue loss and neuron loss. The greater loss of neurons in young rats supports the hypothesis that a high density of GABAA receptors protects neurons from excessive excitatory input during a sensitive period in development.

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 …

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.

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 …

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 …

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.

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.

Like its enteric counterpart, aspartate transcarbamoylase (ATCase) from Pseudomonas putida is a dodecamer of two different polypeptides. Unlike the enterics, the Pseudomonas ATCase lacks regulatory polypeptides but employs instead inactive dihydroorotases for an active dodecamer. Previous work showed that PyrB contains not only the active site but also the effector binding sites for ATP, UTP and CTP at its N-terminus. In this work, 11 amino acids were deleted from the N-terminus of PyrB and the ATCase with the truncated protein was expressed in E. coli pyrB- and purified. The wild type enzyme was similarly treated. Velocity-substrate plots without effectors gave Michaelis-Menten kinetics in all cases. Deleting 11 amino acids did not affect dodecameric assembly but altered effector responses. When carbamoylphosphate was varied, the mutant enzyme was inhibited by UTP while the wild type enzyme was activated 2-fold. When the aspartate was varied, CTP had no effect on the mutant enzyme but strongly inhibited the wild type enzyme.

A life history and productivity study of Enallagma civile was conducted in a playa that was located in the southern High Plains of Texas. Other odonates were also studies to identify their contributions to the habitat.

An anti-G"11 antibody was used to label neuronal cilia throughout the rat central nervous system. Immunoreactive cilia were observed in every examined region of the rat CNS, but not in monkey or mouse tissue. Antibodies to G"q and G"q/11 failed to label cilia. Immunoreactive cilia were observed as early as postnatal day 0 in spinal tissue, and postnatal day 3 in hypothalamic tissue. There was a statistically significant negative correlation between a region's mean cilium length and that region's distance to the nearest ventricle; regions nearest ventricles were those with the longest cilia. This correlation suggests neuronal cilia may function as chemosensors, detecting substances as they move out from the cerebrospinal fluid and into the extracellular space of the brain.

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.

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.

The pyrimidine salvage enzyme, nucleoside hydrolase, is catalyzes the irreversible hydrolysis of nucleosides into the free nucleic acid base and D-ribose. Nucleoside hydrolases have varying degrees of specificity towards purine and pyrimidine nucleosides. In E. coli, three genes were found that encode homologues of several known nucleoside hydrolases in protozoa. All three genes (designated yaaF, yeiK, and ybeK) were amplified by PCR and cloned. Two of the gene products (yeiK and ybeK) encode pyrimidine-specific nucleoside hydrolases, while the third (yaaF) encodes a nonspecific nucleoside hydrolase. All three were expressed at low levels and had different modes of regulation. As a comparative analysis, the homologous genes of Pseudomonas aeruginosa and P. fluorescens (designated nuh) were cloned. Both were determined to encode nonspecific nucleoside hydrolases. The nucleoside hydrolases of the pseudomonads exhibited markedly different modes of regulation. Both have unique promoter structures and genetic organization. Furthermore, both pseudomonad nucleoside hydrolase were found to contain an N-terminal extension of 30-35 amino acids that is shown to act as a periplasmic-signaling sequence. These are the first two nucleoside hydrolases, to date,that have been conclusively demonstrated to be exported to the periplasmic space. The physiological relevance of this is explained.

How temperature affects physiological development in the chicken embryo is unknown. Embryos incubated at 38°C or 35°C showed no difference in growth or survival. The time to hatching was longer in 35°C than 38°C embryos (23.7 vs. 20.6 days), but unaffected was the relative timing of appearance of developmental landmarks (internal, external pipping). At stage 43-44, 38°C embryos maintained oxygen consumption around 1 mL/g/h despite acute temperature reduction (suggesting thermoregulatory maturation), unlike 35°C embryos. In 35°C embryos the lower oxygen-carrying capacity and temperature insensitive blood O2 affinity (P50 about 30 mmHg) may restrict O2 delivery to tissues, limiting metabolism during decreased ambient temperature. Reduced incubation temperature retards normal hematological and thermoregulatory development.

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 …