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.

A Temporal Analysis of Emotions in Girls' Secondary School Physical Education Games and Fitness Classes

Female enthusiasm towards engaging in physical education (PE) significantly decreases with age as it provides females with positive and negative emotional experiences. This study examined emotions within four grade nine female PE soccer and fitness classes (N = 67). Emotional patterns were studied over time and across two units of instruction and in relation to student grades. A mixed-method approach was utilized assessing the state emotions of shame, enjoyment, anxiety, and social physique anxiety (SPA). Results revealed unsatisfactory internal consistency for shame and thus it was removed. Statistical analysis revealed no significant changes in emotions over time, whereas qualitative analysis found that state emotions were inconsistent. Statistical analysis indicated that students in the fitness classes reported significantly higher levels of anxiety and SPA on the final class (p < .01). Qualitative analysis signaled different origins and themes of students‟ emotions. No predictive relationship between emotion and students‟ grade was found.

Effects of computer-mediated communication and face-to-face communication on the quantity and quality of discourse produced by English as a second language students /

The effects oftwo types of small-group communication, synchronous computer-mediated and face-to-face, on the quantity and quality of verbal output were con^ared. Quantity was deiSned as the number of turns taken per minute, the number of Analysis-of-Speech units (AS-units) produced per minute, and the number ofwords produced per minute. Quality was defined as the number of words produced per AS-unit. In addition, the interaction of gender and type of communication was explored for any differences that existed in the output produced. Questionnaires were also given to participants to determine attitudes toward computer-mediated and face-to-face communication. Thirty intermediate-level students fi-om the Intensive English Language Program (lELP) at Brock University participated in the study, including 15 females and 15 males. Nonparametric tests, including the Wilcoxon matched-pairs test, Mann-Whitney U test, and Friedman test were used to test for significance at the p < .05 level. No significant differences were found in the effects of computer-mediated and face-to-face communication on the output produced during follow-up speaking sessions. However, the quantity and quality of interaction was significantly higher during face-to-face sessions than computer-mediated sessions. No significant differences were found in the output produced by males and females in these 2 conditions. While participants felt that the use of computer-mediated communication may aid in the development of certain language skills, they generally preferred face-to-face communication. These results differed fi-om previous studies that found a greater quantity and quality of output in addition to a greater equality of interaction produced during computer-mediated sessions in comparison to face-to-face sessions (Kern, 1995; Warschauer, 1996).