Biology section. (Collegiate Division 2002).

Aubrey, Douglas P., and D. Alexander Wait. NEED SCHOOL@@@ FIRE EFFECTS ON SEEDLING AND SAPLING COMPOSITION IN AN OZARK FOREST: A DEMOGRAPHIC AND PHYSIOLOGICAL APPROACH. Understanding species and community level responses of seedlings and saplings to fire management is important in determining the effectiveness of habitat restoration. We have initiated a long-term demographic and physiological investigation into the effects of fire on the composition of seedlings and saplings of the dominant overstory trees, Oak (Quercus), Hickory (Carya), and Cedar (Juniperus) within the Drury-Mincy Conservation Area in Taney County, Missouri. The Missouri Department of Conservation has implemented controlled bums for the purpose of savanna restoration in this area. We are sampling in three different habitat types; unburned closed forest, recently burned closed forest, and regularly burned glade/savanna. All target species within a 308m plot/habitat were tagged and height, basal area, and dominance determined. Leaf photosynthetic rates and nitrogen (N) were collected, and habitat leaf area index (LAD was determined. Demographic results indicated that total density was greatest in the glade/s avanna habitats compared to either the burned or unburned closed forest habitats, with White Oaks dominating the glade/savanna habitat and Red Oaks dominating both closed forest habitats. Total density was similar in the burned closed forest and the unburned closed forest. Photosynthetic rates for all species were highest in glade/savanna habitats, which also had the lowest LAI and highest total leaf N. White Oak had the highest photosynthetic rates, followed by Hickory and Red Oak; however, Red Oak had the highest leaf N. In total, these results indicate only very subtle effects of recent burning on saplings and seedlings in closed Ozark forests.

Beale, A. William Jewell College. Caedibacter taeniospiralis, an endosymbiotic bacterium that lives inside Paramecia tetraurelia, produces an R body and a toxin. The R body encoding locus, or reb locus, has been cloned into E. coli. Although the R body protein is expressed, no toxic activity has been detected. The reb locus contains 4 open reading frames: reb A, reb B, reb C, and reb D. However, protein expression assays did not detect expression of the reb D protein, even though the reb D open reading frame appears to encode a functional gene. The analysis of the putative amino acid sequences showed a close homology between the four polypeptides. To determine why the reb D gene was not being expressed, sequence analysis upstream of the open reading frame was perforrned, and a potential catabolite repressor sequence was identified. This has led to the hypothesis that reb D is a highly regulated gene and may encode the toxin. We constrncted two deletion derivatives of the reb locus to determine the function of reb D. Using over-lap extension PCR we have generated pBQ88[DELTA]CRS, that lacks the catabolite regulatory sequence, and pBQ88ArebD, that is missing the entire rebD open reading frame. We will now use RT-FCR to measure gene expression in these clones prior to the performance of killing tests.

Coleman, M. and L. Wetmore William Jewell College EXPRESSION OF CYCLIN Bi IN U373 GLIOBLASTOMA MULTIFORME CELLS. Cyclin B 1 is an intracellular protein that is involved in the [G.sub.2]/M cell cycle transition, controlling cell division. This [G.sub.2]/M transition is inhibited by DNA damage. Glioblastoma multiforme is one of the most invasive and aggressive forms of brain tumor, and makes up 45% of all brain tumors. In this experiment the expression of cyclin B 1 is examined in U373 glioblastoma multiforme cells by treating the cells with one or more of the following chemotherapeutic agents: etoposide (VP-16), genistein, and caffeine. VP-16 and genistein, both topoisomerase II inhibitors, activate apoptosis (programmed cell death) pathways. Caffeine overrides the [G.sub.2]/M checkpoint of the cell cycle and forces the cells with DNA damage to continue into mitosis (cell division). A testable hypothesis is in the presence of VP- 16, genistein, and caffeine, cydin B1 expression in U373 cells will increase. Imm unohistochemistry, an antibody reaction, is used to determine the expression of cyclin B1 within the cell. A pink-colored product appears in the cell, where cyclin B1 is located. Cells are then examined for the intensity of cyclin B1 expression using a microscopy.

Cooper, S., (1) R. Kupfer (1), B. Neil (1), L. Shimkets (2) and K. Lee1 (1.) Division of Science (Biology), Truman State. (2.) Department of Microbiology, University of Georgia University. CHARACTERIZATION OF THE PUTATIVE ZINC METALLOPROTEASE, prtl, IN MYXOCOCCUS XANTHUS BY GENE DISRUPTION. Zinc metalloproteases have been implicated in the production of extracellular matrix that plays a significant role in biofilm formation. Recently finished M. xanthus genome sequencing data revealed presence of five putative zinc metalloproteases. The internal fragment of one of them, prt1, was amplified, cloned into a vector, and introduced into the wild type and two different motility mutants to disrupt prtl through a single homologous recombination event. The effect of prtl disruption on the extracellular matrix production, cohesion, and development is under investigation with final data to be presented at the research symposium. Cohesion assays have demonstrated a fifty percent increase within prtl mutants with respect to the wild type. Development is also affected with current data pointing toward a fifty percent reduction in spore formation.

Davis, H.K. and D.P. Heruth. Department of Biology, William Jewell College. ISOLATION AND CHARACTERIZATION OF A DRUG RESISTANT COLON CARCINOMA CELL LINE. Previous studies by our research group determined that a de-differentiated cell line, SW837-DH2 exhibited minimal decreased c-myc levels when exposed to sodium butyrate. However, when treated with genistein, a tyrosine kinase inhibitor versus a histone deacetylase inhibitor, a dramatic reduction in myc levels resulted. This suggests an alternate form of drug resistance than the "Multidrug resistant" phenotype found in many types of metastatic cancers. With treatment of 2 mM Sodium Butyrate daily, a de-differentiated SW837 cell line has been developed similar to the SW837-DH2 cell line. This dedifferentiated cell line exhibits an altered morphology with pseudopod extensions. In addition, under microscopy an increase in flourescent granules is noted indicating a rise in mitochondrial activity. Currently, experiments are underway to produce a pure clone of the de-differentiated cell line and to test whether this resistance is synonymous to histone deacetylase inhibitors as expected or follows a more common "multidrug resistant" phenotype.

Dunn, L., M.F., and S. Vortriede. Haskins Department of Biology, Rockhurst University. A TWO-YEAR STUDY OF BLISTER BEETLE (MELOIDAE) OCCURRENCE AND BEHAVIOR IN WESTERN MISSOURI AND EASTERN KANSAS ALFALFA FIELDS. Eleven fields, totaling approximately 57 hectares (140 acres), of alfalfa in Jackson and Johnson Counties, MO and Franldin County, KS were sampled weekly between June-September 2000 and 2001. Walking a randomly-generated grid, investigators alternated sweeping with visual inspections. No correlations were found between field size and the following variables: number of blister beetle swarms, population size of the swarms, and total number of beetles observed in each field. Mark-recapture studies, which produced only a few recaptured individuals, indicated some beetles tended to remain for several weeks in the initial area of capture. Global positioning system data were used to record locations of swarms and their dispersal patterns within fields. Other data recorded included: weather, status of alfalfa development, and types of plant damage, if any. In four of the alfalfa fields, general insect samples were obtained weekly by collecting all insects from five randomly-selected sweep samples and preserved for later analysis.

Englert, V.D., (1) R.L. Maser,. (2) J.A. Dilts, (1) and D.P. Heruth Daniel P.1. (1.) Department of Biology, William Jewell College. (2.) University of Kansas Medical Center. ISOLATION AND CHARACTERIZATION OF THE REB LOCUS FROM PSEUDOMONAS TAENIOSPIRALIS. Pseudomonas taeniospiralis, a hydrogen-oxidizing, gram-negative soil bacterium, produce large protein ribbons, termed R bodies. R bodies are coiled into cylindrical structures within bacterial cells and have been found in both free-living bacteria within the Pseudomonas genus and endosymbiotic bacteria within the Caedibacter genus. Five types of R-bodies have been characterized (types 51, 7, Cc, Pt, and Pa) based on physical dimensions, morphology, and behavior in chemical environments. The genetic determinants, reb locus, for type 51 R-body production in C. taeniospiralis have been determined previously and we have demonstrated that the rebB genes from both C. taeniospiralis and P. taeniospiralis are identical. In this study we utilized the known similaritie s between the two rebB genes to clone and sequence the complete P. taeniospiralis type Pt reb locus. Comparison of the two reb loci allows for a better understanding of the evolutionary relationship between these two types of R bodies.

Fairchild, S.N. and T.J. Allen. Department of Biology. William Jewell College. COMPARISON OF TWO FILTRATION SYSTEMS IN THE REMOVAL OF CRYPTOSPORIDIUM PARVUM AND GIARDIA LABLIA. Only about one-third of the people in the world's least developed countries have dependable access to a safe water supply and adequate sanitary facilities. There are two solutions to this problem. The first is education and the other is and effective water filtration. The latter solution should have a low construction cost, can be easily maintained, and that utilizes native materials. David Manz created a filtration system that uses sand to filter out the impurities in water, however, this system is used primarily for towns and cities; therefore, it is difficult for people to make multiple trips a day just for clean water. The people avoid making the trips and sacrifice themselves by using contaminated water. Smaller scaled versions of Manz's filter have been constructed. The purpose of the study is to compare Manz's system set up (gra vel and sand only) to another system with additional sand layering. The efficacy of the two filters' set up will be determined quantitatively and by the ability to remove Cryptosporidium and Giardia lamblia from sampled water. My hypothesis is that the new set up will not only induce better filtration capabilities but also enable a wide distribution of the system to families rather than to only communities. Water samples will be taken from Rush Creek in Liberty, Missouri, two times a day (A.M. and P.M.). Twelve samples per week will be analyzed. Temperatures and counts of Cryptosporidium and Giardia lamblia before and after will be recorded.

Fongue, V., G. O'Conner (1), and D. Gibbs.2 (1.) Biology Department and (2.) Chemistry Department, Rockhurst University. THE FEEDING DETERRENT EFFECT OF PHLOXINE-B AND OTHER LIGHT ACTIVATED COMPOUNDS ON LARVAE OF TENEBRIO MOLITOR. Previous bioassays showed that plant extracts from the class of terpenes are good feeding deterrents for insects that normally feed on stored grain products or live plants. The present bioassays were conducted on the class of compound called Chromosomes. We attempted to ascertain the effects of anthraquinone and its sodium salt, Phloxine-B, erythrocin-B, fluorencein, 4-phenylazofluorescein, juglone and Phloxine-B associated to an azo group. Each chemical was tested at concentrations varying between 200 ppm and 800 ppm. Insects were placed in glass bowls with 50 g of Carolina Biological Bran Meal, for culturing mealworms. One group was exposed to the sunlight and the other was continuously kept in the dark. At concentrations as low as 200 ppm, Phloxine-B showed some killing activity, which increased with its concentration. The other Photosensitizers showed some killing activity, but with a less intensity. The addition of an azo group to anthraquinone increases its photo-activity. In the group exposed to light, a maximum of 90% of the insects died while in the group kept in the dark a maximum of 10% died. In each control group, no more than 10% died. Results suggest that photoactivated compounds such as Phloxine-B or anthraquinone may be used as feeding deterrents at very low concentrations.

Graham, J.M. and T.J. Allen, Department of Biology, William Jewell College and CVOR St. John's Regional Hospital. ENDOVASCULAR VERSUS OPEN REPAIR OF ABDOMINAL AORTIC ANEURYSMS: A SINGLE-CENTER STUDY. Aortic abdominal aneurysms (AAAs) represent a degenerative process and are characterized by a bulge in the arteriole wall. Given their propensity to rupture, AAAs pose a significant health care problem. The conventional method of repairing AAAs involves a large abdominal incision. However, there is a new method that removes the need of this incision and the resulting complications. This new method has made it possible to replace the defective artery with an artificial one called an endograft via two small incisions in the groin. Much controversy exists as to the feasibility of this new method. To address this question, a matched-pair analysis on the outcomes of the two methods was conducted. In the first year of an endografting program, 62 patients underwent endovascular therapy for the treatment of AAA. Data col lected was then retrospectively compared to a matched group of 55 patients undergoing open repair. Patients treated by the conventional method had significantly higher blood loss, mean operative time, and hospital stay. However, complication rates and incidence of reoperation were significantly higher in the patients treated with an endograft. While there are obvious advantages to endovascular treatment of AAAs, individual risk factors should be considered in deciding which procedure to employ.

Hirschman, M., S. Addison, T. Randall, S. Recker, K. McConkey, S. Tye, and R. Enochs, Department of Science, Central Methodist College. EFFECT OF STRESS AND HOUSING ON IgM RESPONSE TO IMMUNE CHALLENGE BY LIPOPOLYSACCHARIDE (LPS) IN RATS. Numerous studies have indicated that immune function is influenced by stress as well as social support. The effect may vary under acute versus chronic stress conditions. This study was designed to investigate both the effects of an acute stress, as well as group versus individual housing on production of IgM following injection of LPS. Two groups of rats were housed individually and two groups were in groups of four. Each rat was injected with 100 ug/kg LPS at 8 am. Fifteen minutes later stressed rats were subjected to an eight-minute forced swim at 15[degrees]C. Tail vein blood was drawn every 24 hrs for 4 days and then every other day until 20 days after injection. Blood was refrigerated for 24 hrs then the serum collected and frozen until assay. Total serum IgM was determi ned by performing an Enzyme-Linked Immunosorbent Assay (ELISA). Preliminary results indicated that both individually housed and group housed unstressed rats showed elevated levels of total IgM between three and twelve days after injection of LPS. This increase was greater in the group which was housed individually. Rats which were subjected to cold swim stress after LPS injection failed to show a consistent increase in IgM response whether housed individually or in groups. While these results are preliminary they do suggest that acute stress has a negative impact on the ability to respond to immune challenge. The results suggest that housing differences may also have some effect. Additional studies of LPS specific IgM level should help elucidate the nature of this response.

Johnson, A.L., (1) R. L. Maser (2), J. A. Dilts (1), and D.P. Heruth (1), (1.) Department of Biology, William Jewell College. (2.) University of Kansas Medical Center. ISOLATION AND CHARACTERIZATION OF THE CAEDIBACTER TAENIOSPIRALIS PLASMID, PKAP116. Paramecium tetraurelia 116 contain Caedibacter taeniospiralis, bacterial endosymbionts that demonstrate killing activity towards Paramecium without endosy bionts. Caedibacter maintain a 43 kb plasmid (pKAP1 16) and produce rolled proteins called refractile bodies or R bodies. R bodies are associated with killing activity and are coded for by the reb locus, a 1931 bp region on the plasmid. Of the four closely related reb locus genes, two encode for R bodies, one allows for post-translational modification of these genes, and the fourth is thought to encode for the killing toxin. pKAP116 has been isolated from Caedibacter, modified by insertion of a kan (R) Mu transposon in vitro, and transformed by electroporation into E. coli. The E. ccli transformants produce R b odies. DNA sequence analysis of the plasmid is possible by using sequencing primers located at both ends of the transposon. Here we report the characterization of the potential open reading frames (ORFs), consisting of both ORFs with homologies to known genes and newly identified ORFS. Analysis of these sequences provides an understanding of (1) the evolution of the plasmid, (2) the genes necessary for killing, and (3) the endosymbiotic relationship between C. taeniospiralis and P. tetraurelia 116.

* Kupfer, R., B. Hughes, and K. Lee. Department of Biology, Truman State University. EFFECTS OF LIGHT EXPOSURE ON MYXOCOCCUS XANTHUS DEVELOPMENT. Previous studies have demonstrated that exposure to light during development impedes M. xanthus fruiting body formation by an unknown mechanism. This study was conducted to determine the effects of varying intensities and durations of light exposure in order to gain more knowledge of the inhibition process. Cells at mid-exponential phase of growth were harvested and spotted on TPM non-nutrient agar to induce sporulation. To examine the effects of light exposure time, samples were incubated at 32[degrees]C under light conditions and placed in subsequent dark conditions at increments of 0 (control), 3, 6, 12, 24, 48, 96, and 144 hours. Fruiting body formation under natural light conditions was studied in a greenhouse by incubating samples in complete darkness (control), semi-shade, and direct sunlight. In both phases of the experiment, inhibitive effects were quantifi ed by conducting a spore count using a Petroff-Hausser counting chamber under a phase contrast microscope. M. xanthus spore development exhibited a significant quantitative decline as light exposure time was increased over the entire 144-hour period (p<0.0001), particularly between the 24-hour and 48-hour exposure times. Development was also inhibited in cultures incubated under direct sunlight (p=0.013) as well as in semi-shade (p=0.011) compared to the control. The results of this study suggest that inhibition of development is directly proportional to duration and intensity of light exposure, and that the critical stage of development most afflicted by light occurs 24 to 48 hours after starvation.

Marquardt, S. R. and C.A. Schmidt. Department of Biology, Central Missouri State University. CORRELATES OF URBANIZATION AND USE OF URBAN HABITAT BY BATS. Urban development often results in a reduction of natural areas, an increase in the number of artificial light sources, an increase in human population density that results in more manmade structures, and expansion in total area as well as changes in the general shape of the city. It is important to determine bow urbanization and human activity impact bat communities because bats are sensitive indicators of ecosystem health. The goal of this study is to examine the relationship between urbanization factors and bat activity within urban areas. Seven cities in Missouri were chosen as sampling locations based on population size and general shape. A total of eight monitoring sites (outer zone: sites 14; inner zone: sites 5-8) were selected in a circular pattern within each city. Using broad-band ultrasound detectors two research teams recorded the number of bat passes observed at each site. The total number of bat passes and the number of passes in each sampling zone was regressed against six urbanization factors. Of the factors considered in this study, the strongest associations were between the number of passes in the outer zone and both percentage of agricultural land (positive association) and percentage of forested land (negative association). These associations suggest that even though streetlights may provide concentrations of prey species, bats still use forested areas extensively to forage. Under this scenario, as agricultural land replaces suitable forest habitat, bats would shift some of their foraging activity to the clusters of insects attracted to the city lights.

Meler J. William Jewell College FATTY ACID INDUCED APOPTOSIS IN VASCULAR SMOOTH MUSCLE CELLS (VSMCS). Palmitate (C16:O) and oleate (C18.1) are the most concentrated fatty acids in blood plasma and serve in such cellular functions as metabolism, intracellular signaling molecules, and cell membrane formation. In addition, certain saturated fatty acids, such as palmitate, have been implicated in cellular apoptosis (programmed cell death) of many cell types including cardiac myocytes and granulosa cells. This data suggests the possibility that other cell types may apoptose along similar pathways. The purpose of this study is to observe VSMCs exposed to palmitate and oleate to determine whether saturated fatty acids induce apoptosis in VSMCs. VSMCs have a contractile role in vivo, and are the primary component of blood vessel walls where they mediate changes in and maintenance of blood pressure. In addition, VSMCs in a synthetic state are found in atherosclerotic plaque, and therefore may be implicated in plaque d estabilization, rupture, and thrombosis once apoptosed. To determine whether fatty acids induce apoptosis in VSMCs, cultured VSMCs (which resemble synthetic VSMCs in vivo) will be exposed to palmitate and oleate (0.5 mM each) for 24 hours, after which the extent of apoptosis will be determined by flow cytometry. It is hypothesized that palmitate will induce apoptosis while oleate will not.

Mompati, F., and M. F. Haskins, Department of Biology, Rockhurst University. OCCURRENCE OF PENTATOMIDAE IN ALFALFA FROM WESTERN MISSOURI AND EASTERN KANSAS. During the summers of 2000 and 2001 insects were collected from alfalfa fields located in Johnson and Jackson Counties in Missouri, and Franklin County, Kansas. Sweep nets were used to sample the fields on a weekly basis between June and September of each year. Captured stink bugs were classified into two genera. Although stink bugs have piercing-sucking mouthparts which enable them to feed on plants, the population sizes of stink bugs in alfalfa is typically low, as supported by our data, and the insects are not considered a major threat to alfalfa.

Myers, J.L., and M.C. Barnhart. Department of Biology, Southwest Missouri State University. EFFECTS OF HYPOXIA ON SURVIVORSHIP AND GROWTH OF JUVENILE MUSSELS. Native freshwater mussels (Unionidae) are the most threatened family of animals in North America. This investigation explores low oxygen (hypoxia) as a possible threat to the survivorship of juvenile mussels less than one month old. Juveniles of Lampsilis rafinesqueana, L. reeve iana, and L. cardium were subjected to dissolved oxygen (DO) levels between 4% and 85% of air saturation (AS). In each experiment, 4 replicate groups of 5-6 juveniles were placed into each of 6 DO levels at 20[degrees]C. The number of surviving juveniles was recorded at 2-day intervals. Time to 50% mortality ([LT.sub.50]) was determined for each treatment group. [LT.sub.50] ranged from 7-33 days and varied among species and DO treatments. [LT.sub.50] declined significantly below about 10% AS (1 mg/L) in 3 of 4 species. In contrast, [LT.sub.50] of L. cardium increased below 10% A S. Growth was not affected by DO. The results indicate that juvenile Lampsilis are highly tolerant of low levels of dissolved oxygen. However, this study did not consider other factors, such as [CO.sub.2] and pH, which vary with DO in field conditions. This study was supported by funding from the U.S. Fish and Wildlife Service and the Missouri Department of Conservation.

Pittman, A., K. Baumgartner, L.A. Wetmore, and D.P. Heruth. Departments of Biology and Chemistry, William Jewell College. GENISTEIN-INDUCED APOPTOSIS IN HCT8 COLON CARCINOMA CELLS Our research has been directed toward an understanding of how the c-myc oncogene is regulated in cells of the colorectal mucosa. In previous work we have found that genistein, a tyrosine kinase inhibitor, caused a dramatic decrease in the abundance of c-myc RNA and an inhibition of proliferation in the HCT8 colon carcinoma cells. In the present work we demonstrate that genistein induces apoptosis in HCT8 cells. Apoptosis was determined by flow cytometry to measure the sub G1 population, microscopy with ethidium bromide/acridine orange staining to visualize nuclear condensation, and identification of DNA fragmentation by gel electrophoresis. Treatment of HCT8 cells with 60 ug/ml resulted in a 3-, 11-, and 4.5-fold increase at 24, 48, and 72 hours, respectively, compared with untreated cells. To understand further the role of the c-my c pathway and to potentially identify additional genes involved in the apoptotic response to genistein, we performed DNA microarray assays to measure differential gene expression between treated and untreated HCT8 cells. We report here a profile of genes with at least a two-fold gene difference in gene expression.

Schumacher, S.A., (1) K.A. McCullough, (1) R. Maser (2), J.A. Dilts, (1) and D.P. Heruth. (1) Department of Biology, William Jewell College. ISOLATION AND CHARACTERIZATION OF A RECQ DNA HELICASE FROM TETRAHYMENA THERMOPHILA. Aging occurs in nearly all organisms, yet the exact cause for aging remains unknown. However, the discovery of genes that, when mutated, cause premature aging conditions in humans (WRN, BLM) and yeast (Sgs1) has provided model systems for understanding the aging process. These genes encode a specific class of proteins, RecQ DNA helicases, that are involved in the maintenance of DNA. Due to the complexity of higher organisms, less complex organisms provide useful model systems to try to identify key molecular components and their functions within the aging process. We have chosen to study the aging process in Tetrahymena thermophila, a ciliated protozoan, because, unlike most organisms, it has an unlimited clonal lifespan. Therefore, we hypothesized that the RecQ helicase from T. thermophi la may provide insight into mechanisms that control clonal lifespan. We report here that we have cloned a RecQ DNA helicase from T. thermophila that is similar to yeast Sgs1 and human WRN and BLM. We will now be able to complete functional assays on the Tetrahymena protein that will help us to understand further the aging process in all organisms.

Sesser, L.M. and A. R. Oller, Ph.D. Department of Biology and Earth Science, Central Missouri State University. MANGANESE II AND ULTRAVIOLET RADIATION EFFECTS ON PROTEUS MIRABILIS ATCC 7002. Deinococcus radiodurans is a bacterium that has the capability of resisting high levels of ultraviolet (UV) radiation. Proteus mirabilis is also considered highly resistant to UV radiation. According to Chou and Tan (1990), manganese II can cause abnormal spurts of cell growth after the normal log phase, referred to as Mn(II)-induced cell division (Mn-CD) in D. radiodurans. Thus far, other bacterial species tested for the Mn-CD growth do not exhibit a resistance to UV radiation. This study tested P. mirabilis ATCC 7002 to determine if Mn-CD growth phase can be induced in another bacterial genus upon exposure to manganese, as well as if managanse causes susceptibility changes to UV or antibiotics. Manganese II was added to 250 mL flasks and samples were taken every two hours to perform plate counts, to prepare slides for m icroscopy, and to determine growth spectrophotometrically. Rifampicin and norfloxcincin-laden plates were subjected to UV irradiation and growth was measured. Control groups were similarly performed and all experiments were performed in duplicate. Results suggest that manganese has a significant effect on the number of P. mirabilis cells in relation to the control group. Manganese also appeared to cause P. mirabilis to become more sensitive to UV radiation. This study was supported by the McNair Central Achievers Program.

Stroup, D., and D. Wetmore. Department of Biology and Chemistry, William Jewell College. EXPRESSION OF CYCLIN B1 IN P53-MUTANT U138 MG GLIOBLASTOMA CELLS AFTER EXPOSURE TO ETOPOSIDE (VP16). Glioblastoma multiforme is the most aggressive of the glioma cancers. [1] Gross pathology displays regions of necrosis and hemorrhage. [1] Microscopically, regions of pseudopalisading necrosis, pleomorphic nuclei and cells, and microvascular proliferation can be observed. [1] Genetically deletions, amplifications and point mutations have been identified. [1] If damaged DNA, in a normal cell, is discovered then p53 regulates the level of cyclin B1 so that the initiation of mitosis cannot begin and the cell either arrests or goes into apoptosis. [3] The hypotheses in this experiment were after dosing U138 MG glioblastoma cells with VP16, cyclin B1 expression will increase in the presence of caffeine, and cyclin B1 expression will decrease in the presence of genistein. The cells were rehydrated, and the primary monoclonal cyc lin B1 antibody was applied to the cells. After incubation with the primary antibody, the cells were reacted with a streptovidis-linked secondary antibody that will allow time for chromagin formation at the site of cyclin B1 expression. The cells were then counterstained with hematoxylin and sealed under a coverslip with aqueous mounting medium. A cellular histogram of cyclin B1 will then be analyzed by microscopy.

Walters, D.S. and L.A. Wetmore. Department of Biology and Chemistry, William Jewell College. A DYSFUNCTIONAL [G.sub.2] CELL CYCLE CHECKPOINT PATHWAY IN p53-MUTANT GLIOMA CELLS. Glioblastoma tumors are aggressive brain tumors which are inoperable and resistant to chemotherapy and radiation. Cell cycle checkpoints monitor DNA integrity and control cell cycle division. Because these tumors are resistant to traditional cancer treatments that target DNA, investigation of the cell cycle checkpoints in glioblastoma may reveal more effective methods of treatment. In this research, the [G.sub.2] cell cycle checkpoint in two p53 -mutant glioma cell lines, U138MG and U373MG, was analyzed by treatment with one or more of the following reagents: etoposide, a topoisomerase II inhibitor; genistein, a topoisomerase II and tyrosine kinase inhibitor known to activate a [G.sub.2] blockade via the ATM pathway; and caffeine, a [G.sub.2] cell cycle checkpoint ATM pathway inhibitor. Some glioblastoma cells are hyperploidy, meaning they contain excessive amounts of DNA. When dosed with a combination of the above reagents, U138 display a decreased hyperploidy population of cells, while U373 display an increase. These results confirm a dysfunction in the [G.sub.2]/M cell cycle checkpoint. Based on these results, cyclin B levels, the cyclin which regulates entrance into mitosis, requires further study in U138 and U373.

Wieberg, C.G., (1) R.E.Kissell Jr., (1) L. Hansen, (2) and J. Beringer. (2) (1.) Division of Agriculture, Natural Sciences and Mathematics, Lincoln University. (2.) Missouri Department of Conservation. SPATIAL AND TEMPORAL RELATIONSHIPS OF WHITE-TAILED DEER ANTLER CHARACTERISTICS IN MISSOURI. How antler size varies across Missouri is unknown. In addition, the effect of the environment on antler size has received limited attention and has not been examined in the Midwest. Our objectives were to 1) determine geographic patterns of antler size across Missouri and 2) determine spatio-temporal changes of antler characteristics in Missouri between the periods 1951-1970 and 1997-2001. Antler size, as indicated by beam circumference (mm) and the number of points, was collected at county check-stations by MDC personnel and volunteers during 2 time periods, 1951 to 1970 and 1997 to 2001. Mean circumference and mean number of points by county and physiographic region were determined for 1.5-year-old deer. Regression of antler characteristics and latitude/longitude indicated latitude was important (p < 0.0001). Geographic distribution of antler characteristics has changed over time. Variation of antler size became less over time and indicated a northwest to southeast trend. We will discuss possible explanations of our findings.