The analysis of gene transcripts associated with conidiation in the insect pathogenic fungus, Metarhizium anisopliae /

Conidia of the insect pathogenic fungus, Metarhizium anisopliae play an important role in pathogenicity because they are the infective propagules that adhere to the surface of the insect, then germinate and give rise to hyphal penetration of the insect cuticle. Conidia are produced in the final stages of insect infection as the mycelia emerge from the insect cadaver. The genes associated with conidiation have not yet been studied in this fiingus. hi this study we used the PCR-based technique, suppression subtractive hybridization (SSH) to selectively amplify conidial-associated genes in M. anisopliae. We then identified the presence of these differentially expressed genes using the National Center for Biotechnology Information database. One of the transcripts encoded an extracellular subtilisin-like protease, Prl, which plays a fundamental role in cuticular protein degradation. Analysis of the patterns of gene expression of the transcripts using RT-PCR indicated that conidial-associated cDNAs are expressed during the development of the mature conidium. RT-PCR analysis was also performed to examine in vivo expression of Prl during infection of waxworm larvae {Galleria mellonelld). Results showed expression of Prl as mycelia emerge and produce conidia on the surface of the cadaver. It is well documented that Prl is produced during the initial stages of transcuticular penetration by M. anisopliae. We suggest that upregulation of Prl is part of the mechanism by which reverse (from inside to the outside of the host) transcuticular penetration of the insect cuticle allows subsequent conidiation on the cadaver.

Collaborative learning in a Japanese as a foreign language classroom /

This study examined the influence of training on Asian learners' beliefs, interaction, and attitudes during collaborative learning (CL) and explored the processes of their CL in pairs. The literature contains few studies on the effect of collaborative training in language learning. In addition, it shows gaps between SLA theory and practice resulting from learners' cultural differences. Although second/subsequent language acquisition (SLA) theory assumes that CL contributes to language learning, implementing CL in a multicultural classroom is often considered to be unsuccessful by teachers. The research questions designed to address this gap explore: (a) the extent to which tra~ng affects Asian learners' attitudes towards and interaction during CL; (b) how Asian learners accomplish collaborative tasks in pairs. In the quasi-experimental research design, the learners in the treatment group received special training in CL for 5 weeks while the learners in the comparison group did not receive similar training. Data were collected from 45 McMaster University students through pre- and posttests, pre- and postintervention questionnaires, student information, and informal classroom observations. To detennine the influence of training, the frequency of communication units (c-units), Language Related Episodes (LREs), Collaborative Dialogue (CD) from audio-taped data, and the fmal draft scores were compared between pre- and posttests. The learners' pre- and postintervention questionnaires were also compared. Transcripts from audio-taped data, students' information, their responses and comments from questionnaires, and informal observations served to investigate the processes of Asian learners' CL. Overall, this study found that training had significant influence on the frequency of c-units and CD, and considerable impact on the draft scores, although little influence on the frequency of LREs was observed. The results from the questionnaires in the treatment group showed positive changes in the learners' beliefs on pair work after training. On the other hand, analyses of the transcription data showed that the learners did not conduct enough discussion for a resolution of problems with peers. In conclusion, results suggested the need for teacher intervention, a longer period of collaborative training, and an implementation of self-evaluation into the course grade to encourage the learners to succeed in collaborative learning.

The essence of a tacit intersection : a heuristic inquiry into nature-based leisure and personal transformation

There are a considerable number of programs and agencies that count on the existence of a unique relationship between nature and human development. In addition, there are significant bodies of literature dedicated to understanding developmentally focused nature-based experiences. This research project was designed to flirther the understanding of this phenomenon. Consequently, the purpose of this research endeavour was to discover the essence ofthe intersection ofpersonal transformation and nature-based leisure, culminating in a rich and detailed account of this otherwise tacit phenomenon. As such, this research built on the assumption of this beneficial intersection of nature and personal transformation and contributes to the understanding ofhow this context is supporting or generating of selfactualization and positive development. Heuristic methods were employed because heuristics is concerned with the quality and essence of an experience, not causal relationships (Moustakas, 1990). Heuristic inquiry begins with the primary researcher and her personal experience and knowledge of the phenomenon. This study also involved four other coresearchers who had also experienced this phenomenon intensely. Co-researchers were found through purposeful and snowball sampling. Rich narrative descriptions of their experiences were gathered through in-depth, semi-structured interviews, and artifact elicitation was employed as a means to get at co-researchers' tacit knowledge. Each coresearcher was interviewed twice (the first interview focused on personal transformation, the second on nature) for approximately four and a half hours in total. Transcripts were read repeatedly to discern patterns that emerged from the study of the narratives and were coded accordingly. Individual narratives were consolidated to create a composite narrative of the experience. Finally, a creative synthesis was developed to represent the essence of this tacit experience. In conclusion the essence of the intersection of nature-based leisure and personal transformation was found to lie in the convergence of the lived experience of authenticity. The physical environment of nature was perceived and experienced to be a space and context of authenticity, leisure experiences were experienced as an engagement of authenticity, and individuals themselves encountered a true or authentic self that emanated from within. The implications of these findings are many, offering suggestions, considerations and implications from reconsidered approaches to environmental education to support for selfdirected human development.

Spiritual journeys in the classroom: elementary public school teachers' perspectives /

This thesis explored 5 public elementary school teachers ' perceptions of spirituality and its implications for classroom practice. A generic qualitative study was conducted where each teacher shared her experiences and perceptions in one audiotaped semistructured interview. Transcripts were generated and coded for themes which emerged, resulting in the findings of the study. Following this process, the participants verified the accuracy of the transcripts and findings through a member-checking system. The research found that each teacher has her own definition of spirituality. Furthermore, one's personal connection with spirituality can involve a relationship with religion, the self, a higher being, others, and nature. These spiritual relationships were nurtured through a variety of methods outlined by the teachers. This resulted in the creation of a personal spiritual profile for each teacher which contained each teacher's spiritual connections or facets and the methods used to develop these facets. The teachers identified spiritual needs in their students warranting the need for and importance of spiritual education. Given this, a number of classroom practices were identified with the intention of meeting the spiritual needs of students. Among these practices, the teacher as role model was identified as a significant practice for students' spiritual development in the classroom. The teachers further outlined a number of professional development initiatives with the intention to promote greater awareness for spiritual education and to provide resources for educators.

Creativity and the working artist/teacher : the relationships

The purpose of this qualitative study was to understand the relationships between creativity and the working artist/teacher employed by an art college. The topic emerged from my job as an instructor at The Ontario College of Art which was used as the primary data resource and provided the highest caliber of professionals to chose from. Existent data were used to facilitate the study generated by the research of Cawelti, Rappaport, and Wood (1992). The data were generated by a group of 5 faculty members from The University of Northern Iowa, recognized for their expertise in the arts (a painter, a poet, a sculptor, a novelist, and a photographer). They were asked to respond to the following statement: "In as much detail as you like, list the things that you did, thought, or felt the last time you created an artistic product. II Cawelti, Rappaport, and Wood (1992) produced three models of the creative process, each building on the previous, with the resultant third,being in my opinion, an excellent illustration (text/visual) of the creative process. Model three (Appendix D) presented a "multi-dimensional view of the creative process: time, space, observatility, and consciousnessll (p. 90). Model three utilized a visual mapping device along the bottom of the page linked to text segments above. Both the visual and the text were interrelated so that they harmonized into a comprehensive "picture." The parti'cipants of this qualitative study were asked to consider model three from their professional perspective as artist/teachers. The interpretive sciences directed the methodology. The hermeneutic circle of continuous reflection from the whole to the part and back to the whole was an important aspect of the data analyses. Four members of the Foundation Department at The Ontario College of Art were the key participants. A series of conversational interviews was the primary source of data collection, this was augmented by observation, fie,ldnotes, and follow up telephone interviews. Transcripts of interviews were returned to participants for reflection and the telephone was used to discuss any additional -points raised. Analysis consisted of coding and organizing data according to emerging themes. These themes formed the basis for the narrative stories. The text of the narrative stories were given back to each participant for further comment. Revisions were made until both the researcher and the participants felt that the stories reflected reality. The resultant whole was critiqued from the researcher's perspective. The significance of this study was discussed as it pertains to the working artist/teacher and areas in need of further study are pointed out.

Facilitating self-directed learning: the nurse educators' perspective

The concept of self-directed learning was examined from the nurse educators' perspective. One structured interview, lasting between one and one-and-a half hours, was conducted with each of 14 nurse educators in two community college continuing education programs in nursing. One community college program encourages selfdirected learning; the other encourages self-study and active participation in the teaching/learning process. All 14 interviews were tape-recorded and transcribed verbatim. Verbatim transcripts were analyzed for themes, patterns, and relationships utilizing analyst-constructed typologies. six prerequisites or necessary conditions for facilitating self-directed learning in a community college continuing education program in nursing were identified. ~he crucial issue in facilitating self-directed learning was found to be the issue of teacher-control.

The establishment of two novel bovine cell lines by transfection with the putative transforming region of bovine adenovirus type 3

Human adenoviruses (Ads), members of the family adenoviridae, are medium-sized DNA viruses which have been used as valuable research tools for the study of RNA processing, oncogenic transformation, and for the development of viral vectors for use in gene delivery and immunization technology. The left 12% of the linear Ad genollle codes for products which are necessary for the efficient replication of the virus, as well as being responsible for the forlllation of tumors in animallllodels. The establishlllent of the 293 cell line, by immortalization of human embryonic kidney cells with th~ E1 region of Ad type S (AdS), has facilitated extensive manipulation of the Ads and the development of recombinant Ad vectors. The study of bovine adenoviruses (BAVs), which cause mild respiratory and gastrointestinal infections in cattle has, on the other hand, been limited primarily to that of infectivity, immunology and clinicallllanifestations. As a result, any potential as gene delivery vehicles has not yet been realized. Continued research into the molecular biolo~gy of BAVs and the development of recolllbinant vectors would benefit from the development of a cell line analogous to that of the 293 cells. In an attelllpt to establish such a cell line, the recombinant plaslllid pKC-neo was constructed, containing the left 0-19.7% of the BAV type 3 (BAV3) genome, and the selectable marker for resistance to the aminoglycoside G418, a neomycin derivative. The plasmid construct was then used to transfect both the Madin-Darby bovine kidney (MDBK) -iicell line and primary bovine lung cells, after which G418-resistant foci were selected for analysis. Two cell lines, E61 (MDBK) and E24 (primary lung), were subsequently selected and analysed for DNA content, revealing the presence of the pKC-neo sequences in their respective genomes. In addition, BAV3 RNA transcripts were detected in the E61 cells. Although the presence of E1 products has yet to be confirmed in both cell lines, the E24 cells exhibit a phenotype characteristic of partial transformation by E1. The apparent immortalization of the primary lung cells will permit exploitation of their ability to take up exogenous DNA at high efficiency.

Teachers' perspectives on the implementation of expectations-based curricula /

This qualitative study examines teachers' experiences implementing new standardized curricula in Ontario schools. This new curricula contained several policy changes and an expectations based format which directed what knowledge and skills students were to demonstrate in each subject. This level of specificity of subject-content served to control teachers in relation to curricula; however, data suggested that at the same time, teachers had enormous flexibility in terms of pedagogy. Four secondary teachers who were implementing a Grade 10 course in the 2000-2001 school year participated in the study. The qualitative framework supported the researcher's emphasis on examining the participants' perspectives on the implementation of expectation-based curricula. Data collected included transcripts from interviews conducted with teacher participants and a representative of the Ontario Ministry of Education and Training, field notes, and a research journal. Many of the factors often cited in the literature as influencing implementation practices were found to have affected the participants' experiences of curriculum implementation: time, professional development, and teachers' beliefs, particularly concerning students. In addition, the format of the policy documents proved to both control and free teachers during the implementation process. Participants believed that the number of specific expectations did not provide them an opportunity to add content to the curriculum; at the same time, teachers also noted that the general format of the policy document allowed them to direct instruction to match students' needs and their own teaching preferences. Alignment between teachers' beliefs about education and their understanding of the new curriculum affected the ways in which many participants adapted during the implementation process.

Moving from values inaction to values-in-action : an exploration of how values can be managed intentionally by national sports organizations

The study examined the intentional use of National Sport Organizations' (NSOs) stated values. Positive Organizational Scholarship (POS) was applied to an Appreciative Inquiry (AI) approach of interviewing NSO senior leaders. One intention of this research was to foster a connection between academia and practitioners, and in so doing highlight the gap between values inaction and values-in-action. Data were collected from nine NSOs through multiple-case studies analysis of interview transcripts, websites, and constitutional statements. Results indicated that while the NSOs operated from a Management by Objectives (MBO) approach they were interested in exploring how Management by Values (MBV) might improve their organization's performance. Eleven themes from the case studies analysis contributed to the development of a framework. The 4-1 framework described how an NSO can progress through different stages by becoming more intentional in how they use their values. Another finding included deepening our understanding of how values are experienced within the NSO and then transferred across the entire sport. Participants also spoke about the tension that arises among their NSO' s values as well as the dominant values held by funding agents. This clash of values needs to be addressed before the tension escalates. Finally, participants expressed a desire to learn more about how values can be used more intentionally to further their organization's purpose. As such, strategies for intentionally leveraging values are also suggested. Further research should explore how helpful the 4-1 framework can be to NSOs leaders who are in the process of identifying or renewing their organization's values.

Alternative splicing of DNA polymerase beta in vertebrates

Alternative splicing (AS) is the predominant mechanism responsible for increasing eukaryotic transcriptome and proteome complexity. In this phenomenon, numerous mRNA transcripts are produced from a single pre-mRNA sequence. AS is reported to occur in 95% of human multi-exon genes; one specific gene that undergoes AS is DNA polymerase beta (POLB). POLB is the main DNA repair gene which performs short patch base excision repair (BER). In primate untransformed primary fibroblast cell lines, it was determined that the splice variant (SV) frequency of POLB correlates positively with species lifespan. To date, AS patterns of POLB have only been examined in mammals primarily through the use of cell lines. However, little attention has been devoted to investigating if such a relationship exists in non-mammals and whether cell lines reflect what is observed in vertebrate tissues. This idea was explored through cloning and characterization of 1,214 POLB transcripts from four non-mammalian species (Gallus gallus domesticus, Larus glaucescens, Xenopus laevis, and Pogona vitticeps) and two mammalian species (Sylvilagus floridanus and Homo sapiens) in two tissue types, liver and brain. POLB SV frequency occurred at low frequencies, < 3.2%, in non-mammalian tissues relative to mammalian (>20%). The highest POLB SV frequency was found in H. sapiens liver and brain tissues, occurring at 65.4% and 91.7%, respectively. Tissue specific AS of POLB was observed in L. glaucescens, P. vitticeps, and H. sapiens, but not G. gallus domesticus, X. laevis and S. floridanus.The AS patterns of a second gene, transient receptor potential cation channel subfamily V member 1 (TRPV1), were compared to those of POLB in liver and brain tissues of G. gallus domesticus, X. laevis and H. sapiens. This comparison was performed to investigate if any changes (either increase or decrease) observed in the AS of POLB were gene specific or if they were tissue specific, in which case similar changes in AS would be seen in POLB and TRPV1. Analysis did not reveal an increase or decrease in both the AS of POLB and TRPV1 in either the liver or brain tissues of G. gallus domesticus and H. sapiens. This result suggested that the AS patterns of POLB were not influenced by tissue specific rates of AS. Interestingly, an increase in the AS of both genes was only observed in X. laevis brain tissue. This result suggests that AS in general may be increased in the X. laevis brain as compared to liver tissue. No positive correlation between POLB SV frequency and species lifespan was found in non-mammalian tissues. The AS patterns of POLB in human primary untransformed fibroblast cell lines were representative of those seen in human liver tissue but not in brain tissue. Altogether, the AS patterns of POLB from vertebrate tissues and primate cell lines revealed a positive correlation between POLB SV frequency and lifespan in mammals, but not in non-mammals. It appears that this positive correlation does not exist in vertebrate species as a whole.

The Meaning of the Virtual YMCA After School Program for Former Participants: An Exploratory Study

Research has shown that after school programs can provide children with opportunities that help foster positive development and adaptation. Current research meets the need for identifying short term outcomes, program standards, and short term evaluation techniques, however less understood is the lasting meaning of participation for previous participants after their participation. The purpose of this study was to explore the meaning and perceived impacts of participation in a preventative, skill-building after school program for former participants. Using an exploratory case study approach, interviews were conducted with six previous participants of the Virtual YMCA and a former school principal. Reconstructed narratives and analyzed transcripts show that participants do still derive meaning and significance from their participation in the program, although highly individualized. Significant aspects of the program are identified, as well as the participant’s perceived impacts which are still present today. Discussion for practitioners, researchers, and funders is provided.

Exploring Perceptions of Accessibility, Necessity and Use of Social Support for Wilderness Therapy Field Instructors

Organizations offering therapeutic wilderness programming have a responsibility to ensure the well-being of their front line employees. A system of social support that is formed through communication with others, either personally or professionally, can assist field instructors in effectively managing the demands arising from their work. Phenomenological analysis of semi-structured interview transcripts from seven participants provided insight on perceptions of necessity, accessibility and use of social support. Fourteen main themes and thirteen subthemes emerged from the data. Findings are presented using the six components of Parsons’ (1980) staff development model and strongly suggest program managers consider and apply specific measures aimed at increasing the social support for front line field instructors in a wilderness therapy work context.

Bullying in the Same-Sex Friendships of Young Adolescent Girls: A Qualitative Study

This qualitative, narrative study explored the bullying experiences of young adolescent girls within their same-sex dyadic and group friendships. The participants were 5 female students, ages 11 and 12 years old, from 1 private, religious school in southern Ontario. Each girl participated in an audiotaped, 30-minute, personal interview based on an unstructured interview protocol. Interview transcripts were analyzed for bullying behaviors using Marini and Dane's (2008) subtypes of bullying, including the form, function, and involvement in bullying. Interview transcripts were also analyzed for common and emerging themes using aspects of L. M., Brown and Gilligan's (1992) "Listener's Guide." The findings of this study suggested that within their same-sex friendships girls assume the roles of all participants in bullying, including bullies, victims, bystanders, and bully-victims. The findings also suggested that bullying behaviors within young adolescent girls' same-sex friendships are mainly indirect in their mode of attack and that they are both proactive and reactive. The bully behaviors identified in this study were used to inform the major themes or salient features within the dynamics of girls' same-sex friendships also identified. These themes included acceptance, intimacy, negotiation, inclusion/exclusion, moral character judgements, and power. The findings of this study will be used to inform current theory, personal and professional practice, as well as future research.

A Survey of hMLH1 mRNA Splicing Profiles in Human Cell Lines: Comparing Primary Cultured Fibroblasts Before and After Oxidative Stress and Transiently versus Stably Demethylated Colon Cancer Derived Cell Lines

Most human genes undergo alternative splicing and loss of splicing fidelity is associated with disease. Epigenetic silencing of hMLH 1 via promoter cytosine methylation is causally linked to a subset of sporadic non-polyposis colon cancer and is reversible by 5-aza-2' -deoxycytidine treatment. Here I investigated changes in hMLHI mRNA splicing profiles in normal fibroblasts and colon cancer-derived human cell lines. I established the types and frequencies of hMLHI mRNA transcripts generated under baseline conditions, after hydrogen peroxide induced oxidative stress, and in acutely 5-aza-2' -deoxycytidine-treated and stably derepressed cancer cell lines. I found that hMLHI is extensively spliced under all conditions including baseline (50% splice variants), the splice variant distribution changes in response to oxidative stress, and certain splice variants are sensitive to 5- aza-2' -deoxycytidine treatment: Splice variant diversity and frequency of exon 17 skipping correlates with the level of hMLHI promoter methylation suggesting a link between promoter methylation and mRNA splicing.

Integrating Children with Emotional and Behavioural Disabilities into Community Recreation Programs: A Handbook for Staff

This study examined the process of integrating children with Emotional Behavioural Disorders (EBDs) with their peers into recreation programs. The purpose was to develop a set of recommendations for the development of a handbook to help workers in recreation with the integration process. To this end, a needs assessment was conducted with experienced recreation workers in the form of semistructured interviews. Participants were recruited from two community centers in a large southern Ontario city. Themes were drawn from the analysis of the interview transcripts and combined with findings from the research literature. The results were a set of recommendations on the content and format of a handbook for integrating children with EBDs into recreation programs.