The cloning and reconstitution of a bovine adenovirus type 2 E3 deletion mutant and the sequencing and analysis of the early 4 region

Recombinant Adenoviruses (Ads) have been shown to have potential applications in three areas: gene therapy, high level protein expression and recombinant vaccines.' At least three different locations within the Ad genome can be deleted and subsequently used for the insertion of foreign sequences. These include the Early 3 (E3), Early 1 (E1) and Early 4 (E4) regions. Viral vectors of this type have been well studied in Human Ads 2 and 5, however one has not yet been constructed for Bovine Adenovirus Type 2 (BAV2). The E3 region is located between 76.6 and 86 m.u. on the r-strand and is transcribed in a rightward direction. The gene products of the Early 3 region (E3) have been shown to be non-essential for viral replication, in vitro, but are required for host immunosurveillance. This study represents the cloning and reconstitution of a BAV2 E3 deletion mutant. A deletion of 1800bp was made within the E3 region of BAV2 and the thymidine kinase gene was subsequently inserted in the deleted area . . The plasmid pdlE3-4tk1 (23.4Kbp) was constructed and used to to facilitate homologous recombination with the wild type BAV2 to produce a mutant. Southern Blotting and Hybridization results suggest the presence of a BAV2 E3 deletion mutant with thymidine kinase sequences present. The E4 region of Human Adenovirus types 2 and 5 is located at the extreme right end of the genome (91.3 map units - 99.1 map units) and is transcribed in a leftward direction giving rise to a complicated set of differentially spliced mRNAs. Essentially there are 7 open reading frames (ORFs) encoding for at least 7 polypeptides. The gene products encoded by the E4 region have been shown to be essential for the expression of late viral genes, host cell shutoff and normal viral growth. We have cloned and sequenced the right end segment between 90.5 map units and 100 map units of the BAV2 genome. The results show several open reading frames which encode polypeptides exhibiting homology to three polypeptides encoded by the E4 region of human adenovirus type 2. These include the 14kDa protein encoded by ORF1, the 34kDa protein encoded by ORF6 and the 13kDa protein encoded by ORF3. The nucleotide sequence, restriction enzyme map, and ORF map of the E4 region could be very useful in future molecular manipulation of this region and could possibly explain the slow growth rate of BAV2 in MDBK cells.

Therapeutic riding : learning and recovery for people with disabilities

The purpose ofthis study was to explore the process oftherapeutic riding as an experiential and holistic approach to learning and recovery for people with disabilities as perceived by the providers oftherapeutic riding. To enhance the connection between theory and practice and to suggest future research, the researcher endeavoured to develop a theory that contributed to the knowledge base oftherapeutic riding, animal-assisted therapy and education, experiential education, and experiential therapy in addition to contributing to connections among them. This topic was investigated because ofthe lack ofresearch about the process of therapeutic riding, particularly from learning and a recovery perspective. Few studies have addressed how therapeutic riding outcomes are achieved or how the therapeutic riding process actually works. This study was identified as grounded theory using qualitative data through interviews and narrative reflections with therapeutic riding providers, a researcher's journal, field notes, and written documents. Grounded theory analysis was used to analyze the qualitative data. This consisted ofdoing open, axial, and selective coding. This study provided detailed descriptions ofthe research approach, researcher's involvement, participant and site selection, data collection and analysis, methodological assumptions and limitations, credibility established, and ethical considerations. The findings ofthe data analysis revealed the theme ofrelationships as central to the learning and recovery process oftherapeutic riding for people with disabilities. The significance ofthe team relationships, the horse and rider relationship, and the providers and rider relationship was found. The essential components ofthe learning and recovery process were presented in a diagram in the selective coding phase. Goals oftherapeutic riding included psycho-education; behavioural and social; physical; and equestrian. Parts ofthe process ofhow outcomes were achieved included motivation; "opens new doors;" risk; task analysis; control; communication; and environmental factors. Outcomes of therapeutic riding included independence and mobility; confidence; and transfer abilities or skills. The implications ofthese findings for theory, practice, and further research were also. explored.

Isolation and analysis of a corprinus cinereus DNA fragment showing homology to a fimbrial cDNA from ustilago violacea

Surface proteinaceous fibrils, termed fimbriae, were first identified on gram negative bacteria in the 1940s. Fungal fimbriae, discovered some 25 years later, are found on members of all fungal classes. In the present study, polyclonal antiserum raised against the fimbrial proteins of U. vio/acea were used in order to identify antigenically related proteins from Coprinus cinereus and Schizophy//um commune. Two polypeptides with molecular masses of 37 and 39 kDa from C. cinereus were observed and confirm earlier results. A single previously unidentified 50 kDa polypeptide in S. commune crossreacted with the antiserum. The 50 kDa protein was found to consist of 3 isoforms with isoelectric points ranging from 5.6 to 5.8. A fimbrial cDNA derived from U. vio/acea was used to identify DNA restriction fragments from C. cinereus and S. commune showing homology to the fimbrial transcript of U. vio/acea. Heterologous hybridization with this cDNA was used in order to screen a C. cinereus genomic DNA library. A single clone, A2-3A, with a 14 kbp insert showed strong homology to the pfim3-1 cDNA. The region of homology, a 700 bp Xba I fragment, was subcloned into pUG19. This plasmid was refered to as pXX8. DNA sequence determinations of pXX8 and adjacent fragments from A2-3A suggested that the cloned DNA was a portion of the rONA repeat encoding the small subunit rRNA. DNA sequence analysis of pfim3-1 yielded an incomplete open reading frame. The predicted amino acid sequence codes for a 206 amino acid, 22 kDa polypeptide which contains a domain similar to a transmembrane domain from rat leukocyte antigen, GDS3. As well, an untranslated 576 nucleotide domain showed 81 % homology to pXX8 and 830/0 homology to the 188 rRNA sequence of Ustilago maydis. This sequence was found adjacent to a region of adenine-thymine base pairs presumed to represent the polyadenylation sequence of the fimbrial transcript. The size and extent of homology is sufficient to account for the hybridization of pfim3-1 to rDNA. It is suggested that this domain represents a completely novel regulatory domain within eukaryotes that may enable the observed rapid regeneration of fimbriae in U. violacea.

A multidimensional measurement approach and analysis of children's motivation for reading, attributional style, and reading achievement

The present study investigates the usefulness of a multi-method approach to the measurement of reading motivation and achievement. A sample of 127 elementary and middle-school children aged 10 to 14 responded to measures of motivation, attributions, and achievement both longitudinally and in a challenging reading context. Novel measures of motivation and attributions were constructed, validated, and utilized to examine the relationship between ~ motivation, attributions, and achievement over a one-year period (Study I). The impact of classroom contexts and instructional practices was also explored through a study of the influence of topic interest and challenge on motivation, attributions, and persistence (Study II), as well as through interviews with children regarding motivation and reading in the classroom (Study III). Creation and validation of novel measures of motivation and attributions supported the use of a self-report measure of motivation in situation-specific contexts, and confirmed a three-factor structure of attributions for reading performance in both hypothetical and situation-specific contexts. A one-year follow up study of children's motivation and reading achievement demonstrated declines in all components of motivation beginning at age 10 through 12, and particularly strong decreases in motivation with the transition to middle school. Past perceived competence for reading predicted current achievement after controlling for past achievement, and showed the strongest relationships with reading-related skills in both elementary and middle school. Motivation and attributions were strongly related, and children with higher motivation Fulmer III displayed more adaptive attributions for reading success and failure. In the context of a developmentally inappropriate challenging reading task, children's motivation for reading, especially in terms of perceived competence, was threatened. However, interest in the story buffered some ofthe negative impacts of challenge, sustaining children's motivation, adaptive attributions, and reading persistence. Finally, children's responses during interviews outlined several emotions, perceptions, and aspects of reading tasks and contexts that influence reading motivation and achievement. Findings revealed that children with comparable motivation and achievement profiles respond in a similar way to particular reading situations, such as excessive challenge, but also that motivation is dynamic and individualistic and can change over time and across contexts. Overall, the present study outlines the importance of motivation and adaptive attributions for reading success, and the necessity of integrating various methodologies to study the dynamic construct of achievement motivation.

The Implementation of Jenkins’s 21st-Century Skills in the Curriculum: A Cross-National Policy Analysis

This meta-analytic study sought to determine if cross-national curricula are aligned with burgeoning digital learning environments in order to help policy makers develop curriculum that incorporates 21st-century skills instruction. The study juxtaposed cross- national curricula in Ontario (Canada), Australia, and Finland against Jenkins’s (2009) framework of 11 crucial 21st-century skills that include: play, performance, simulation, appropriation, multitasking, distributed cognition, collective intelligence, judgment, transmedia navigation, networking, and negotiation. Results from qualitative data collection and analysis revealed that Finland implements all of Jenkins’s 21st-century skills. Recommendations are made to implement sound 21st-century skills in other jurisdictions.

How health organizations can apply scenario based planning and analytic technologies to make strategic consequential decisions in a time of uncertainty

The magnitude of the cervical cancer problem, coupled with the potential for prevention with recent technological advances, made it imperative to step back and reassess strategic options for dealing with cervical cancer screening in Kenya. The purpose of this qualitative study was: 1) to explore the extent to which the Participatory Action Research (PAR) methodology and the Scenario Based Planning (SBP) method, with the application of analytics, could enable strategic, consequential, informed decision making, and 2) to determine how influential Kenyan decision makers could apply SBP with analytic tools and techniques to make strategic, consequential decisions regarding the implementation of a Cervical Self Sampling Program (CSSP) in both urban and rural settings. The theoretical paradigm for this study was action research; it was experiential, practical, and action oriented, and resulted in co-created knowledge that influenced study participants’ decision making. Action Africa Help International (AAHI) and Brock University collaborated with Local Decision Influencing Participants (LDIP’s) to develop innovative strategies on how to implement the CSSP. SBP tools, along with traditional approaches to data collection and analysis, were applied to collect, visualize and analyze predominately qualitative data. Outputs from the study included: a) a generic implementation scenario for a CSSP (along with scenarios unique to urban and rural settings), and b) 10 strategic directions and 22 supporting implementation strategies that address the variables of: 1) technical viability, 2) political support, 3) affordability, 4) logistical feasibility, 5) social acceptability, and 6) transformation/sustainability. In addition, study participants’ capacity to effectively engage in predictive/prescriptive strategic decision making was strengthened.

Major Sport Event Operational Planning Issues and Strategies: A Multi-Case Delphi Study

This study examined the operational planning, implementation and execution issues of major sport events, as well as the mitigation and management strategies used to address these issues, with the aim of determining best practices in sport event operational planning. The three Research Questions were: 1) What can previous major sport events provide to guide the operational management of future events? 2) What are the operational issues that arise in the planning and execution of a major sport event, how are they mitigated and what are the strategies used to deal with these issues? 3) What are the best practices for sport event operational planning and how can these practices aid future events? Data collection involved a modified Delphi technique that consisted of one round of in-depth interviews followed by two rounds of questionnaires. Both data collection and analysis were guided by an adaptation of the work of Parent, Rouillard & Leopkey (2011) with a focus on previously established issue and strategy categories. The results provided a list of Top 26 Prominent Issues and Top 17 Prominent Strategies with additional issue-strategy links that can be used to aid event managers producing future major sport events. The following issue categories emerged as having had the highest impact on previous major sport events that participants had managed: timing, funding and knowledge management. In addition, participants used strategies from the following categories most frequently: other, formalized agreements and communication.