Hood College Student Works
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Item Assessing School-Related Anxiety Levels and Self-Efficacy Among First-Generation College Students(2024-12) Chamberlin, Casey; Gupta, Sangeeta; Kundey, Shannon; Gricus, Michelle; Hood College Department of Psychology & Counseling; Departmental HonorsCurrent research on student success suggests that first-generation college students (FGCS) experience worse academic outcomes, and greater barriers to success compared to their continuing-education peers (Dennis et al., 2005; Koh et al., 2022). There is also a body of literature on how various student factors, such as anxiety and self-efficacy, drive student success (Barrows et al., 2013; Kumari & Jain, 2014). When a student specifically experiences school-related anxiety and does not believe in their ability to perform well in the classroom, this could impact their academic outcomes. The present study aims to examine the extent to which FGCS struggle with school-related anxiety and self-efficacy versus their continuing-education peers. The present study assessed anxiety and self-efficacy in 170 undergraduate first-year students, 38% of whom were FGCS, using the anxiety subscale of the Learning and Study Skills Inventory (LASSI), the Revised Test Anxiety Scale, and an Academic Self-Efficacy Scale. FGCS students reported significantly higher anxiety levels based on their LASSI scores. However, FGCS were not significantly different than their non-FCGS peers in their test anxiety or self-efficacy, as revealed by independent samples t-tests. Additionally, there were significant negative correlations between self-efficacy and test anxiety as well as between self-efficacy and LASSI anxiety across all students. Implications for FGCS, school-related anxiety, and self-efficacy are discussed.Item Communities of Practice: How Teachers' Relationships Underscore Suburban Middle School Culture and Organizational Commitment(2024-12) Sutherin, Kristen G.; Locraft Cuddapah, Jennifer; Harris, Keith; Markoe, Michael; Hood College Organizational Leadership; Doctor of Organizational LeadershipTeacher attrition poses significant challenges in education systems, leading to inconsistent instruction, reliance on substitute teachers, and increased costs for hiring and training replacements (Carver-Thomas & Darling-Hammond, 2019). Despite considerable research on this issue in various educational settings, a gap exists in understanding the factors influencing teacher commitment specific to suburban middle school teachers. Drawing from theories on communities of practice (Lave & Wenger, 1991), organizational commitment (Allen & Meyer, 1990), and cultural frameworks (Schein & Schein, 2017), this multiple case study of middle school teachers explored how school culture impacted their commitment to their schools and districts. The central research questions focused on the influence of school culture on organizational commitment; the elements of culture manifested in artifacts, beliefs, and assumptions; and the effects of team, school, and district cultures on teacher commitment. Additionally, the study examined how relationships within these contexts shape teacher commitment and explored the costs and benefits associated with sustaining commitment. Data procedures involved distributing questionnaires to general education teachers across four suburban middle schools followed by in-depth, semi-structured interviews with selected participant volunteers (Creswell & Creswell, 2018). Key themes emerged, highlighting the significance of relationships and trust in fostering commitment, the overall culture of the school, the role of leadership support and stability, and the ultimate decision to stay or leave. Study findings underscored that neglecting culture and commitment issues could exacerbate teacher attrition, necessitating continual investments in new teachers and depriving students of the stability and expertise that experienced teachers provide. Teachers want to stay in schools where they feel a sense of belongingness which leads to organizational commitment. To address these challenges, recommendations include ensuring teachers begin to connect and build relationships within their schools, creating communities of practice for school leaders to share best practices, minimizing principal attrition and movement year to year, establishing opportunities for teachers to engage with district leaders, and building trust and respect through clear communication. By prioritizing these strategies, educational stakeholders can work toward operational excellence and mitigate teacher turnover, ultimately ensuring a more stable and enriching learning environment for students in suburban middle schools.Item Conflict as the Accelerant of Social Change(2024-11) Rogers, Cameron; Verzosa, Noel; Dodman, Trevor; Campion, Corey; Hood College English & Communication Arts; Humanities (M.A.)Even as humanity has advanced and become “civilized” over its existence, conflict clings to it in varying scopes and impacts. The changing power of war, conflict, and bloodshed stretches far and wide, from benefits like medical advancements to its numerous detriments of killing, destruction, and crimes against humanity. While most conflicts are regarded as tragedies and needless bloodshed, they still spur change through the actions taken during or after their occurrence. Some small-scale conflicts can spark massive social changes, such as the Wounded Knee Occupation of 1973. Other conflicts can lead to gradual changes in perception about war and its necessity, such as the Wars in Vietnam and Iraq. In my portfolio, I will examine conflict’s ability to accelerate social change through specific engagements, arguing that it can serve as a harsh but necessary tool for societies to advance and to right past wrongs.Item AMYLOID PLAQUES IN CHRONIC WASTING DISEASE(1990-09) Guiroy, Don C.; Hood College Biology; Biomedical and Environmental ScienceChronic wasting disease (CWD), a progressive neurological disorder of captive mule deer and Rocky Mountain elk, is characterized neuropathologically by widespread spongiform change of the neuropil, intracytoplasmic vacuolation of the neuronal perikarya and astrocytic hypertrophy and hyperplasia. Histochemically demonstrable amyloid plaques and amyloid plaques reactive to antibodies prepared against scrapie amyloid in captive mule deer, mule deer hybrids and Rocky Mountain elk naturally affected with CWD are presented. Amyloid plaques in CWD-affected mule deer were congophilic, birefringent and periodic acid-Schiff (PAS) positive. In mule deer hybrids, only occasional PAS-positive plaques were observed. No histochemically demonstrable plaques were observed in CWD-affected Rocky Mountain elk. Antibody raised against scrapie amyloid showed robust immunoreactivity with amyloid plaques in CWD-affected captive mule deer and were found in the cerebral grey and white matter in clusters or in isolation, in deep subcortical nuclei, in all layers of the cerebellum, in areas of extensive vacuolation and in subpial and perivascular regions. In hybrid deer, plaques were rarely observed in cerebellum and subpia. A notable finding in brain sections of CWD-affected hybrid deer and Rocky Mountain elk was a circumscribed collection of scrapie amyloid-immunopositive nuclei with non-congophilic, non-birefringent amyloid deposits located at its center. In elk, plaques were not observed in cerebellum, subpia and subependyma. Furthermore, amyloid plaques in CWD-affected captive mule deer were alcianophilic at 0.3 M magnesium chloride indicating the presence of weakly to moderately sulfated glycosaminoglycans. Similar scrapie amyloid-immunoreactive plaques are also present in Creutzfeldt-Jakob disease, Gerstmann- Straussler syndrome and kuru in humans. The data presented here corroborate that CWD belongs to the subacute spongiform virus encephalopathies (transmissible cerebral amyloidoses).Item SIGNIFICANT FACTORS, ISSUES, AND THE ROLE OF POPULAR FORCES INVOLVED IN THE PASSAGE OF THE VOTING RIGHTS ACT OF 1965 IN THE UNITED STATES SENATE(1987-05) Guilfoyle, Jean Marie; Hood College Counseling, Care and Interdisciplinary Studies in Human Behavior; Human SciencesItem AMMONIA AND NITRATE UPTAKE KINETICS BY THREE SPECIES OF AQUATIC VASCULAR PLANTS(2007-12) Guedes, Marcia; Hood College Biology; Biomedical and Environmental ScienceItem THE DEVELOPMENT OF A WESTERN BLOT (IMMUNOBLOT) ASSAY FOR THE DETECTION OF IgG AND IgM ANTIBODIES TO TREPONEMA PALLIDUM IN HUMAN SERUM(1996-12) Gross, Robert W.; Hood College Biology; Biomedical and Environmental ScienceA Western Immunoblot assay was developed to detect IgG and IgM antibodies to Treponema pallidum, the causative agent of Syphilis, in human serum. A total of 220 human serum samples and 3 cerebral spinal fluid samples were run on the Syphilis Western ImmunoBlot (SWIB) IgG assay and the Fluorescent Treponemal Antibody Absorption (FTA-ABS) assay to determine relative sensitivity, specificity and accuracy. High specificity for the 14, 15 and 48 kDa antigens was demonstrated with FTA-ABS positive samples versus FTA-ABS negative samples. The development of only one of these three bands in the SWIB IgG assay was indicative of a positive sample. Sensitivity, specificity and accuracy were checked further by running 30 human serum samples from a well-defined CDC syphilis panel and 14 potentially cross- reactive autoimmune samples on the SWIB IgG assay. The sensitivity, specificity and accuracy for all 267 samples were 97.4% (111/114), 98% (150/153) and 97.8% (261/267), respectively. A total of 151 human serum samples were tested on the SWIB IgM assay and a commercially available Syphilis IgM Capture ELISA to determine relative sensitivity, specificity and accuracy. A lack of specificity with the 37 kDa antigen was demonstrated. Overall, very few bands were visualized on IgM blots and no definitive positive cut-off was evident. Three samples were equivocal by ELISA and were not included in the calculations. The sensitivity, specificity and accuracy of the SWIB IgM assay were checked further by running 14 potentially cross-reactive autoimmune samples. With the appearance of one band other than the 37 kDa antigen band as indicative of a positive, the sensitivity, specificity and accuracy for 162 samples tested were 46.7% (14/30), 84.8% (112/132) and 77.8% (126/162), respectively. Immunochemical analysis of separated Treponema pallidum antigens immobilized on nitrocellulose strips was conducted using mild periodate oxidation to identify any carbohydrate moieties. The 31, 34 and 41 kDa antigen bands lost reactivity with mild periodate oxidation compared with control strips. These antigens are possible glycoproteins of Treponema pallidum.Item THE DEVELOPMENT OF AN ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) FOR THE DETECTION OF IgG AND IgM ANTIBODIES TO LEGIONELLA PNEUMOPHILA SEROGROUPS ONE THROUGH SIX(1993-12) Gross, Catherine S.; Hood College Biology; Human SciencesAn enzyme linked immunosorbent assay (ELISA) was developed for the detection of IgG and IgM antibodies to Legionella pneumophila serogroups 1 through 6. The ELISA compared favorably with the L. pneumophila serogroup 1 through 6 Indirect Fluorescent Antibody (IFA), the current serological method of choice. Two clinical comparisons were conducted to evaluate the performance of the LEGIONELLA ELISA assay relative to the IFA method for detection of IgG and IgM antibodies to L. pneumophila. In the first comparison, a total of 203 clinical specimens were tested in the ELISA and resulted in specificity, sensitivity, and accuracy percentages of 93.0%, 99.4% and 98.0%, respectively, when qualitatively compared to the IFA method. The second clinical comparison was performed by personnel at Maryland Department of Health using the LEGIONELLA ELISA and was based on a collection of samples primarily from the Legionella outbreak in Western Maryland. LEGIONELLA ELISA results indicated 100% sensitivity, 94.115 specificity and an overall accuracy of 95% when qualitatively compared to the IFA method. The specificity of the LEGIONELLA ELISA was further challenged by evaluation of 94 potential cross reactive sera that included other Legionella app., mycoplasma, chlamydia and syphilis specimens. Cross reactivity was observed in the LEGIONELLA ELISA in one L. micdadei specimen with a confirmed negative L. pneumophila IFA titer. A selection of these antisera were additionally evaluated by western immuno-blot analysis in an effort to detect potential cross reactive proteins. Detection of significant increases in antibody against L. pneumophila is critical to diagnosis by the IFA method. Such detection was assessed in the LEGIONELLA ELISA using 131 paired acute and convalescent sera. The ELISA demonstrated a 100% sensitivity, 98.9% specificity and an overall accuracy of 99.2% in detection of significant increases in antibody titer. Paired acute and convalescent sera from four individuals with diagnosed legionellosis were evaluated by western immuno-blot analysis in an effort to define the immune response to L. pneumophila antigens. Differences in the IgG and IgM immune responses were apparent. An IgG only seroconversion was clearly demonstrated in one individual. Monoclonal antibodies specific to the 58 kDa heat shock protein, 24 kDa Mip and common lipopolysaccharide epitopes were used as markers in the western immuno-blots. Periodate oxidation was performed on L. pneumophila serogroup specific antigens to determine the participation of carbohydrate moieties in the serologic reactivity of selected L. pneumophila positive antisera. Monoclonal serogroup specific anti-lipopolysaccharide antibodies were used in the periodate oxidation experiments to verify the effectiveness of various concentrations of periodate on carbohydrate epitopes. Sensitivity to periodate was indicated in the ELISA for all serogroups by percent absorbance reductions in the binding of the monoclonal and polyclonal antisera when compared against the absorbance of the controls. Periodate oxidation was additionally performed on transblotted L. pneumophila serogroup specific antigens in an effort to characterize the effect of periodate on serogroup specific and common epitopes and to possibly identify periodate sensitive antigens. Some sensitivity to periodate was indicated in the western immuno-blot analysis by absence of banding. In overall performance, the LEGIONELLA ELISA compared favorably with the IFA method and will be a rapid and reliable objective alternative for detection of IgG and IgM antibodies to L. pneumophila.Item QUANTIFICATION OF PHOTODIMERS FROM ULTRAVIOLET RADIATION-INDUCED DNA DAMAGE IN THE SEA ANEMONE, AIPTASIA PALLIDA USING ENDONUCLEASE SENSITIVE SITES (ESS)(2011-05) Griebel, Jessica M.; Hood College Biology; Biomedical and Environmental ScienceTropical littoral zones offer biologically harmful environments to marine invertebrates due to high levels of exposure to ultraviolet radiation. I documented the extent of DNA damage in the sea anemone Aiptasia pallida under laboratory conditions by applying UVR to live animals as well as pooled DNA that had previously been extracted from A. pallida. Cultured A. pallida from Walsingham Pond, Bermuda were subjected to varying lengths of UVR exposure to quantify DNA damage in the form of number of cyclobutane pyrimidine dimers (CPD's). An endonuclease was applied to those treated with UVR to determine varying amounts of DNA damage. Samples without endonuclease were more significant compared to those treated with endonuclease. Overall, all treatments were not statistically significant at the 0.05 alpha level. Small sample sizes and the inability to extract DNA efficiently without causing damage to the controls, were not found to follow a specific pattern.Item Effects of Diazinon and Diazinon-HC1 Reaction Mixture On Acetylcholinesterase Activity In the Brain and Blood Plasma Of Rainbow Trout (Salmo gairdneri)(1978-07) Greene, Sefronia Ann; Hood College Biology; Human SciencesRainbow trout (Salmo gairdneri) were exposed to various concentrations of diazinon and diazinon-HC1 reaction mixture over a 64 hour period during which time the brain and blood plasma of the fish were analyzed for acetylcholinesterase (AChE) activity. The fish were placed into glass jars containing 14 liters of well water and were sampled at 4, 8, 16, 32, and 64 hours time periods; there were five treatments per time period and three replicates per treatment. The treatments consisted of a positive control, a negative control and three concentrations of diazinon or diazinon-HC1 reaction mixture. Test concentrations for diazinon were 0.1, 0.56, and 1.0 ppm., and 0.01, 0.032, and 0.056 70 volume for the diazinon-HC1 reaction mixture. The data collected indicated that the rate of absorption of diazinon and diazinon-HC1 reaction mixture increased rapidly during the first 16 hours and for the most part, remained at or near this level throughout the duration of the remaining test periods. However, the trout were more sensitive to the residual toxicity (due to an AChE inhibitor) in the diazinon-HC1 reaction mixture than to the diazinon. This was indicated by a 504 kill in the test population over a 64 hour period in the reaction mixture as opposed to 1870 mortality over the same period with diazinon.Item SOME LEGAL ASPECTS OF NON-MARITAL COHABITATION(1974-07) Graf, Catherine K.; Hood College Psychology; Human SciencesNon-marital cohabitation, on college campuses in particular, has been of professional interest recently, with ongoing research concentrated on the prevalence of cohabitation, the social factors leading to cohabitation, characteristics of the relationship, problems experienced, and the effect of cohabitation on the individuals involved, on their relationship to each other, and on society. At the Groves Conference on Marriage and the Family at Myrtle Beach, South Carolina, on May 10-13, 1973, persons who were directly involved in research on cohabitation reviewed what was known about the subject up to that date and discussed what needed to be done in the future to understand the phenomenon more completely and to evaluate its influence on the institutions of marriage and family. The prime recommendation for policy and position statements made by the group at the close of the Groves Conference was that consideration be given to rethinking existing laws prohibiting cohabitation, since, in the opinion of the group, the decision to cohabit should be an individual one and persons should have freedom of choice in the matter. In addition, persons who cohabit may encounter problems in regard to housing, insurance, employment, property settlement, financial proceedings (wills, trusts, Social Security payments, loans), restrictions imposed by military regulations, and establishment of paternity of offspring. All of these areas will be discussed in an effort to ascertain the cohabitor's legal position.Item Personality Traits and Personal History Variables Correlated with Decisions to Have Children or to be Childfree(1980) Gottlieb, Catherine Wilson; Hood College Psychology; Human SciencesThe purpose of this study was to assess the relationship between personal history variables, personality variables as measured by the Edwards Personal Preference Schedule, and a person's decision either to have children or to voluntarily remain childfree. The personality variables being considered were Murray's manifest needs associated with achievement, deference, order, exhibition, autonomy, affiliation, intraception, succorance, dominance, abasement, heterosexuality and aggression. Participants were selected by their response to a preliminary letter distributed to all Hood College graduate students; they and their spouses, if any, were invited to participate in the study and were asked to indicate whether or not they desire having children. The sample was heterogeneous in age, sex and personal background. Participants were asked to complete a background data sheet and the Edwards Personal Preference Schedule. Results obtained using the background questionnaire showed several significant differences: childfree persons more often had friends who were also childfree; the mothers of childfree persons more often had higher levels of education; and childfree persons more often live in urban settings. Results obtained using the Edwards Personal Preference Schedule indicate significantly higher scores for the group with children on the heterosexuality scale. The childfree group had higher scores approaching significance on the scales of abasement and endurance.Item THE ISOLATION AND TRANSMISSION OF Helicobacter hepaticus sp. nov., ISOLATED FROM THE LIVERS AND INTESTINAL CONTENTS OF MICE(1994-07) Gorelick, Peter L.; Hood College Biology; Biomedical and Environmental ScienceIn the fall of 1992 at the National Cancer Institute - Frederick Cancer Research and Development Center (NCI-FCRDC), A/JCr stock mice which showed suppurative skin lesions and treated and untreated A/JCr mice from a long-term toxicologic study were found to have a unique chronic active hepatitis of unknown etiology. No biological agent was known to cause such lesions. A major effort was undertaken to determine the cause of the hepatitis, since all these mice were obtained from the NCI-FCRDC Animal Production Area (NCI-FCRDC-APA). The initial approach was to examine potential environmental toxins. Upon failure to identify any specific environmental cause, a search for a possible biological agent was pursued. In the course of utilizing a Steiner's modification of the Warthin-Starry stain (Steiner's stain), pathologists found what appeared to be a helical organism in the hepatic parenchyma of affected mice. The hepatitis was successfully reproduced in A/J mice obtained from Jackson Laboratories (Jax), Bar Harbor, ME, which had been injected with liver suspensions from affected A/JCr mice. A histopathologic survey employing the Steiner's stain on livers from adult NCI-FCRDC-APA mice demonstrated hepatic lesions in numerous mouse strains (A/JCr, C3H/HeNCr, SJL/NCr, BALB/cAnNCr and SCID/NCr), while hepatic lesions were absent in others (C57BL/6NCr, B6C3F1 and nude [nu/nu]). Hepatic lesions were found more frequently in male mice than female mice. Isolation of the organism was accomplished by culturing the livers of affected A/JCr and SCID/NCr mice and incubating the suspensions on trypticase soy agar plates with 5% sheep blood and brucella blood agar with antibiotics at 37°C under microaerophilic conditions. The organism was motile, catalase positive, oxidase positive and rapidly hydrolyzed urea. The organism was tentatively designated Helicobacter hepaticus sp. nov. Subsequently, Helicobacter hepaticus sp. nov. has been cultured from the intestinal tract and feces of affected mice from the NCI-FCRDC-APA and research holding colonies. To demonstrate infectivity, the organism was injected into unaffected A/J mice from Jax. At several time points livers were aseptically collected, cultured, and a portion submitted for histopathological examination. The hepatitis and presence of the organism was confirmed by histopathology and culture. In a third transmission study, CB.17 SCID mice from Taconic Farms (Tac), Germantown, N.Y., were exposed by direct animal or fecal contact to affected SCID/NCr mice from NCI-FCRDC-APA. Over time the organism and the hepatitis were found to have been transmitted to the CB.17 SCID mice, thus demonstrating that the route of infection was fecal-oral. To date, there are no other organisms known to cause the same type of hepatitis as Helicobacter hepaticus sp. nov.. This novel organism has the potential to provide a much needed animal model for biocarcinogenesis and other human health problems.Item PURIFICATION AND CHARACTERIZATION OF RECOMBINANT TOBACCO ETCH VIRUS PROTEASE(1996-05) Goldstein, Adam Stuart; Hood College Biology; Biomedical and Environmental ScienceThe development of expression systems which permit the overproduction of large amounts of protein is a very important advancement in modern biotechnology. Recent developments in cloning have eased some of the traditional problems of obtaining high yields of pure product. New techniques enable the researcher to express and purify proteins of interest by fusing the expressed protein to a variety of commercially available affinity tags such as, f3-galactosidase, glutathione-S-transferase (GST) (Smith, D. 1988), and polyhistidine peptides (Guan, C. 1987). A new site specific protease, Tobacco Etch Virus protease (rTEV), has been cloned and overexpressed for the use in rapid cloning and purification procedures. In a purification scheme, the protein of interest will contain a TEV cleavage site flanked with six histidines. When the protein is cleaved only a single glycine residue at the amino terminus of the protein remains from the inserted rTEV site. With only one glycine residue left on the purified protein there is less of a chance for incorrect foldings, or for unexpected activities.Item DEVELOPMENT OF A HIV-1 NUCLEOCAPSID (p7) PROTEIN CAPTURE ASSAY(1994-04) Goebel, P. Bradley; Hood College Biology; Biomedical and Environmental ScienceThe p24 antigen capture assay, the current immunoassay used to detect and quantitate human immunodeficiency virus, is an undesirable assay when analyzing plasma or serum from an HIV-1 infected person because anti-p24 antibodies interfere with the assay. Before development of an antigen capture assay that may be useful in analyzing samples from HIV-1 infected people, sera from HIV-1 positive people were tested for antibodies to the nucleocapsid protein of HIV-1, p7. Of 801 HIV-1 antibody positive sera tested, 100 (12.5%) were positive for anti-p7, indicating low anti-p7 seroprevalence in infected persons. An antigen capture assay for p7 was developed and compared to the p24 antigen capture assay in reconstruction experiments. HIV-1 diluted in normal plasma was readily detected by both the p7 and p24 antigen capture assays. The p7 antigen capture assay detected virus diluted in the HIV-1 positive plasma as efficiently as from normal plasma. However, the p24 capture assay was ineffective in detecting virus diluted in HIV-1 positive plasma, even at p24 input concentrations as high as 460 ng/ml. Even though the p7 antigen capture assay currently lacks the sensitivity to detect virus in the plasma if HIV-1 infected people, the reconstruction experiments indicate that with increased sensitivity the assay may prove to be useful in assessing viral quantity in HIV-1 infected individuals. The p7 antigen capture assay will be useful in neutralization studies even at the current level of sensitivity.Item CLONING, SEQUENCING AND EXPRESSION OF THE MEDIUM GENOMIC RNA SEGMENT OF SANDFLY FEVER SICILIAN VIRUS(1995-07) Glass, Pamela J.; Hood College Biology; Biomedical and Environmental ScienceSandfly fever virus Sicilian strain (SFS) is a member of the Phlebovirus genus in the family Bunyaviridae. This virus is the causative agent of an acute, self-limiting flu-like illness lasting 2-5 days. Serologic studies have shown that SFS virus is endemic in southern Europe, the Mediterranean, North Africa, and central Asia which coincides with the distribution of its vector, Phlebotomus papatasi. Epidemics of sandfly fever occur when nonimmune adults, such as tourists or soldiers, enter an area where the virus is endemic. Sandfly fever is of military importance due to its short incubation period, capable of rendering large numbers of nonimmune troops ineffective. The development of a vaccine against SFS virus would allow vaccination of troops prior to entering endemic areas. The phlebovirus genome contains three single-stranded RNA segments designated L (large), M (medium), and S (small) of negative or ambisense polarity. The M segment encodes two viral glycoproteins, G1 and G2, and a nonstructural protein, NSм. Studies of other phleboviruses have demonstrated that the glycoproteins are important for viral infection, pathogenesis, and are the primary targets of the immune response. Analysis of the SFS virus M genomic segment was initiated to determine the sequence and genomic organization for use in the development of both nucleic acid-based diagnostic methods and recombinant vaccines. A series of cDNA clones representing the M genome segment were produced by conventional cloning methods and by polymerase chain reaction amplification. All clones utilized for sequence determination were found to be specific for the SFS virus M RNA segment when used as hybridization probes against SFS virus RNA in northern blot analysis. The M-segment cDNA clones were sequenced by the dideoxy method utilizing both manual and automated procedures. The SFS virus M RNA segment was found to be 4402 nucleotides in length. Computer analysis of this sequence predicted a single open reading frame (ORF) 1341 amino acids in length. Based on comparison of the predicted protein sequence of the SFS M segment with other reported phlebovirus M segments, PCR primers were designed to prepare expression cassettes for producing the viral glycoproteins in recombinant baculoviruses. The downstream glycoprotein region was cloned into Autographa califomica nuclear polyhedrosis virus (AcNPV) and expressed in Spodoptera frugiperda (Sf9) cells. The expressed protein was characterized by immunoprecipitation with SFS virus hyperimmune mouse ascitic fluid and was found to be indistinguishable, based on electrophoretic mobilities in denaturing SDS-PAGE, from the authentic SFS virus G2 envelope glycoprotein. These results led to the conclusion that the genomic organization of the SFS M RNA segment is 5'-NSм-G1-G2-3'.Item OVEREXPRESSION OF HUMAN P21ʷᵃᶠ¹⸍ᶜⁱᵖ¹ ARRESTS THE GROWTH OF CHICKEN EMBRYO FIBROBLASTS TRANSFORMED BY INDIVIDUAL ONCOGENES(1998-01) Givol, Iris; Hood College Biology; Biomedical and Environmental ScienceIn normal cells, cell growth and division are controlled by the interplay between proto-oncogenes and tumor suppressor genes. Cancer cells usually have both activated an oncogene and lost a functional tumor suppressor gene. This study addresses the interrelationship between these positive and negative growth regulators. The main focus is on p21ʷᵃᶠ¹⸍ᶜⁱᵖ¹, a cyclin/cdk inhibitor whose expression is regulated by p53. High level expression of the p53 tumor suppressor can block the growth of cancer cells. Wafl/cipl is transactivated by p53 and the p21ʷᵃᶠ¹⸍ᶜⁱᵖ¹ protein is itself a suppressor of cell growth. To test the growth suppression effect of p21ʷᵃᶠ¹⸍ᶜⁱᵖ¹ or p53 on the growth of normal cells and cells transformed by individual oncogenes, we used replication-competent retroviral vectors to induce high level expression of p53 and p21ʷᵃᶠ¹⸍ᶜⁱᵖ¹ in chicken embryo fibroblasts (CEF). These are primary cells and by using them we avoid the complications that arise when such experiments are done with established cell lines. This study shows that overexpression of p21ʷᵃᶠ¹⸍ᶜⁱᵖ¹ or p53 arrests the growth of CEF at the G 1 phase of the cell cycle by inhibiting DNA synthesis. We next asked whether the growth inhibition induced by p21ʷᵃᶠ¹⸍ᶜⁱᵖ¹ could be overcome by mitogenic signals delivered by a variety of oncogenes. We show that the growth of CEF transformed by v-Src, tf-Ras, c-Mos and c-Myc is inhibited by overexpression of p21ʷᵃᶠ¹⸍ᶜⁱᵖ¹. This suggests that p21ʷᵃᶠ¹⸍ᶜⁱᵖ¹ functions downstream of these oncogenes, suggesting that mitogenic signals converge at the cyclin/cdk complex. To address the interplay between positive and negative signals within the cell cycle machinery, we used this system to test the effects of overexpressing both p21ʷᵃᶠ¹⸍ᶜⁱᵖ¹ and E2F1, a subunit of the E2F transcription factor, on the growth of CEF. The E2F transcription factors are released from binding to hypophosphorylated pRb when pRb is phosphorylated by cyclin/cdks. This allows for transcription of S phase genes and progression from G₁ to S phase in the cell cycle. In this system very high levels of E2F1 overexpression cause considerable apoptosis, however the surviving cells still overexpress E2F1. These cells are transformed and their growth is blocked by overexpression of p21ʷᵃᶠ¹⸍ᶜⁱᵖ¹. These data suggest that p21ʷᵃᶠ¹⸍ᶜⁱᵖ¹ might be useful for gene therapy for human cancer. We also show that the apoptosis induced by high levels of E2F1 can be blocked by Bc1-2, an inhibitor of apoptosis.Item A PLATFORM FOR THE SCREENING OF BREAST CANCER-ASSOCIATED POLYMORPHISIMS USING A PRIMER EXTENSION MICROARRAY(2007-04) Gilbert, Joseph T.; Hood College Biology; Biomedical and Environmental ScienceIt is of great societal and scientific importance to be able to identify low penetrance polymorphic loci and the combinations of allelic variation that give the greatest risk to an individual for the development of breast cancer. Understanding about this variation will be beneficial both for diagnosis, prevention and in the development of drug targets for breast cancer. To this end I propose the creation of a diagnostic platform that will help to identify the risk factors associated with known or suspected polymorphic markers that may influence the progression of breast cancer. The platform that I propose would entail the creation of an oligonucleotide microarray consisting of oligonucleotide detection primers that would anneal to genomic DNA from a donor and detect polymorphic mutations among known and suspected breast cancer-associated alleles. Genomic DNA would be taken from a patient and then amplified. The loci of interest would be copied out of the genomic material by using a multiplexing PCR primer set. The amplified polymorphic loci would then be cleaned up and combined to create a hybridization probe for the microarray. Once the probe is hybridized to the microarray, a polymerase will then be added along with fluorescent-labeled dNTPs and the detection primer would be extended using the labeled dNTPs. This extension would only occur when there is a complete nucleotide match between the sample DNA from the patient and the detection primer's 3' end. A microarray scanner should be able to detect the presence of incorporated labeled nucleotides. The data from large scale studies using this platform coupled with the monitoring of an individual's medical history that has been genotyped using this method, could provide both scientists and clinicians with data available to make a better judgment regarding future risk and prevention of this disease.Item MOLECULAR ANALYSIS OF FACTORS AFFECTING LYMPHOMA SUSCEPTIBILITY OF AKXD RECOMBINANT INBRED MOUSE STRAINS(1987-06) Gilbert, Deborah J.; Hood College Biology; Biomedical and Environmental ScienceRecombinant inbred (RI) strains of mice result from the inbreeding of an F₂ generation which itself was produced by crosses between two existing inbred strains. The genomes of individual RI strains represent stable segregant populations resulting from the reassortment of the two progenitor genotypes. The AKXD RI strains have been derived from progenitor strains AKR/J and DBA/2J that differ significantly in lymphoma incidence. As a result of how the genes affecting lymphoma susceptibility have segregated and been fixed in the RI strains, expression of different levels of lymphoma susceptibility, different ages of onset of disease and involvement of different cell types in the tumors have been observed among the AKXD strains. Ten independently derived AKXD RI strains were analyzed for lymphoma susceptibility. Nine strains were found to be highly susceptible to lymphomas although the level of susceptibility varied greatly. Southern blot analysis of the ten different AKXD strains using molecular probes representing the joint regions of the immunoglobulin heavy chain, kappa light chain and T-cell receptor β-chain genes was performed to classify the strains according to predominant lymphoma type. Additionally a histological classification based on the cell morphology scheme of Pattengale and Taylor was used to verify the molecular classifications of stem, Pre-B, B, T and myeloid cell lineages. Of the nine highly lymphomatous strains analyzed, three died primarily of T cell tumors, two died primarily of B-cell tumors and four were susceptible to both B and T cell tumors. Using molecular probes specific for ecotropic proviruses or ecotropic and oncogenic class I mink cell focus forming (MCF) proviruses it was shown that 91% of the 135 tumors analyzed contained detectable somatically acquired proviruses. Generally MCF proviruses were associated with T-cell lymphomas whereas ecotropic proviruses were associated with B-cell lymphomas. The segregation of a dominant gene, Rmcf, which alters the sensitivity of cells to infection by MCF MuLVs, was recently identified by Hartley et al. on mouse chromosome 5. The allele of this gene carried by each strain also correlated with the predominant lymphoma types seen in these tumors. The three strains that died predominantly of MCF virus -associated T cell lymphomas inherited the Rmcfˢ (sensitive) allele from the parental AKR/J strain. AKXD-2 inherited the Rmcfʳ (resistant) allele from the DBA/2J parental strain and lymphomas from this strain contained no detectable MCF proviruses. Using probes homologous to Myc, Pvt-1, Pim-1 and Fis-1, rearrangements were detected in lymphomas and shown to result from viral integration in the respective regions. Proviral integration near Myc, Pvt-1, Fis-1 and Pi -1 were detected primarily in T-cell lymphomas. In three cases rearrangements near Fis-1 and Pim-1 were detected in the same tumor tissue as rearrangements near Myc. Proviral integration near Myb was not detected in any of the lymphomas analyzed. The significance of the genetic loci analyzed will be discussed with respect to their role in the molecular basis of the disease process.Item COMPARING THE COMPREHENSION OF PARENTAL INFORMED CONSENT AND THE CORRESPONDING MINOR ASSENT DOCUMENT(2010-03) Giambarresi, Leah; Hood College Biology; Biomedical and Environmental ScienceThe informed consent process has had a long history which led to the development of explicit requirements regarding the information disclosed to adult subjects in clinical trials. There are very few similar requirements for the information that is disclosed to a minor who participates in the same research. This study aims to examine the difference in the understanding of an adult consent document as compared to its respective child assent document from the same trial. Two document pairs were selected based upon the presence of the 14 required and additional elements of informed consent as described in 21 CFR 50.25. In the first pair ("consistent"), there was very little difference in the number of these elements presented; the second pair ("inconsistent") had a far greater difference. A total of 131 volunteers each read one of the documents and answered a series of written questions designed to analyze their understanding of the trial and their feelings about participation. Results were tabulated, and the difference in understanding between each consent and its respective assent was calculated. Then, the differences in understanding between the two pairs were compared. Analysis showed an approximate 20% difference in understanding between the pairs. Data was also analyzed to determine how the lack of elements in the assent documents would affect the volunteer s willingness to participate. Close to 40% of the individuals in both assent groups who declined to participate in the described trial attributed this decision to the lack of at least one element. This diminished understanding and difference in participation suggest that a reevaluation of the assent process may be useful in order to respect the autonomy of the minor clinical trial participant.