Thesis (B.A.)--Haverford College, Dept. of History, 2003. / Includes bibliographical references.

My purpose with this thesis has been to find out how to choreograph a fight sequence in ananimated movie. With the help of professional stuntmen and dancers, I have tried to findmethods and techniques used on stage and in live-action film and apply those to animatedfilms. To successfully identify the techniques used, I conducted a number of interviews andanalyzed literature that deals with the subject of fight choreography. By the analysis of themethods provided by the choreographers and selected parts of the literature, I have compiled anumber of steps I recommend 3D animators should to take in their efforts to create their ownfight choreography. / Mitt syfte med den här uppsatsen har varit att ta reda på hur man koreograferar enslagsmålssekvens i en animerad film. Med hjälp av yrkesverksamma stuntmän och dansarehar jag försökt finna metoder och tillvägagångssätt som används på scen och film ochapplicera dessa på animerad film. För att lyckas urskilja de tekniker som används genomfördejag ett antal intervjuer samt analyserade litteratur som behandlade ämnet slagsmålskoreografi.Genom att analysera de metoder som koreograferna tillhandahöll samt utvaldadelar ur litteraturen sammanställde jag ett antal steg som en 3D-animatör bör vidta iskapandet av egna slagsmålskoreografier.

Tomato bushy stunt virus (TBSV) is a positive-sense single-stranded RNA virus. It encodes five open reading frames (ORFs), including two nested genes, expressing movement-associated proteins. One of these proteins, P22, interacts with a host transcription factor containing a homeodomain leucine-zipper motif, known as HFi22. Similar proteins of this type traffic their RNA from cell-to-cell, suggesting the possiblity that HFi22 is involved in the cell-to-cell movement of TBSV RNA. To further characterize the nature of the interaction between P22 and HFi22 on the cellular level, cellular fractionation experiments were conducted. To investigate the functional role of HFi22 in viral movement I attempted to inactivate its expression using a virus induced gene silencing system with a Tobacco rattle virus (TRV) vector. A final objective was based on the notion that different hosts can impact the stability of viruses used to express foreign genes of biotechnological interest. To compare virus stability in different hosts, TBSV expressing the green fluorescent protein (GFP) was inoculated onto various TBSV hosts, and infected leaf tissue was then used as inoculum to be rubbed onto a local lesion host. This technique made it possible to quantify the number of fluorescent foci versus total lesions. Results obtained for the first objective indicate that P22 and HFi22 co-fractionate in nucleus and membrane-enriched samples. In addition, it was found that HFi22 is largely conserved through a wide variety of plant species but not in lettuce, which was found to be compromised for effective virus spread. Control experiments for the second objective showed that plants were successfully silenced with TRV carrying the phytoene desaturase (PDS) gene resulting in photobleaching, however attempts to silence HFi22 have not yielded conclusive results. The results obtained for the third objective indicate there is a difference in how efficiently a foreign gene insert is maintained by TBSV in different host plants. In summary, the overall results of this research showed that host factors influence the host-virus interaction but their exact contributions remain elusive.

Tomato bushy stunt virus (TBSV) is a positive-sense single-stranded RNA virus. It encodes five open reading frames (ORFs), including two nested genes, expressing movement-associated proteins. One of these proteins, P22, interacts with a host transcription factor containing a homeodomain leucine-zipper motif, known as HFi22. Similar proteins of this type traffic their RNA from cell-to-cell, suggesting the possiblity that HFi22 is involved in the cell-to-cell movement of TBSV RNA. To further characterize the nature of the interaction between P22 and HFi22 on the cellular level, cellular fractionation experiments were conducted. To investigate the functional role of HFi22 in viral movement I attempted to inactivate its expression using a virus induced gene silencing system with a Tobacco rattle virus (TRV) vector. A final objective was based on the notion that different hosts can impact the stability of viruses used to express foreign genes of biotechnological interest. To compare virus stability in different hosts, TBSV expressing the green fluorescent protein (GFP) was inoculated onto various TBSV hosts, and infected leaf tissue was then used as inoculum to be rubbed onto a local lesion host. This technique made it possible to quantify the number of fluorescent foci versus total lesions. Results obtained for the first objective indicate that P22 and HFi22 co-fractionate in nucleus and membrane-enriched samples. In addition, it was found that HFi22 is largely conserved through a wide variety of plant species but not in lettuce, which was found to be compromised for effective virus spread. Control experiments for the second objective showed that plants were successfully silenced with TRV carrying the phytoene desaturase (PDS) gene resulting in photobleaching, however attempts to silence HFi22 have not yielded conclusive results. The results obtained for the third objective indicate there is a difference in how efficiently a foreign gene insert is maintained by TBSV in different host plants. In summary, the overall results of this research showed that host factors influence the host-virus interaction but their exact contributions remain elusive.

No description available.

Two Tomato bushy stunt virus (TBSV) proteins, P19 and P22, have been found to interact with the Nicotiana benthamiana host proteins Hin19 and HFi22 in yeast two,hybrid assays. To determine functional roles of these interacting host proteins, viral induced gene silencing (VIGS) was employed to knock,down their expression. TBSV has been demonstrated to activate a virus,specific antiviral response pathway in N. benthamiana. To characterize this pathway, the antiviral RNAi induced silencing complex (RISC) was isolated from TBSV-infected plants. Additionally, putative RISC-associated proteins were identified in silico and suggested roles for these have been identified through literature and database searches. A further aim was the identification of proteins that coimmunoprecipitate with the TBSV-induced RISC following RISC isolation. A primary aim of this investigation was to identify functional roles for host proteins that interact with the two TBSV 3-terminal encoded proteins, P22 and P19. Each of these has functional roles in viral movement and pathogenicity. In yeast two-hybrid assays, P22 has been shown to interact with HFi22 while P19 interacts with Hin19. VIGS was utilized in attempts to silence the expression of these two host proteins in order to determine their functional roles. VIGS-mediated suppression of the TBSV-interacting proteins Hin19 and HFi22 has not been accomplished. Despite multiple attempts and multiple approaches, these proteins have not been amenable to silencing. In light of this finding, it is proposed that rather than utilizing VIGS to down-regulate protein levels for Hin19 and HFi22, other approaches should be utilized. To characterize the TBSV-mediated RNAi pathway, functionally active antiviral RISC was purified from TBSV-infected N. benthamiana plants using ion-exchange chromatography. This RISC was found to be active only in the degradation of TBSV transcripts, indicating the specificity expected from a programmed RISC. Characterization and identification of proteins that copurify with RISC has not yet been accomplished, though in silico analysis has yielded over 150 putative RISC-associated proteins. Of these, a subset has been identified as highly likely candidates based upon function and/or homology to RISC-associated proteins in non-plant organisms, and a model for the TBSV-induced antiviral pathway has been proposed.

Thesis (Ph. D.)--University of Adelaide, Dept. of Plant Pathology, 1987. / Includes bibliographical references.

Stuntwork became a science when stuntman and technician Yakima Canutt left the rodeo to work in Hollywood westerns. Canutt perfected methods and designed mechanisms that made dangerous stunts safer and visually exciting. Many of Canutt's techniques are still used today by modern stuntmen like Hal Needham, Ronnie Rondell, and Paul Baxley. Directed by stuntman Hal Needham and starring "box office draw" Burt Reynolds, Hooper presented the stuntman as a rugged, fun-loving, almost suicidal superman. For the first time in film's short history, the stuntman and his craft became a topic of wide public interest. The stuntman had become "glamorous" almost rivaling his actor counterpart.

Tombusviruses are single, positive strand RNA viruses of plants, often associated with parasitic defective interfering (DI) RNAs. Two viral- coded gene products, namely p33 and p92, are required for tombusvirus replication. The overlapping domains of p33 and p92 contain an arginine/proline-rich (RPR) RNA binding motif. In this study, the role of RPR motif and viral RNA in tombusvirus replication and recombination, as well as involvement of viral RNA in tombusvirus replicase assembly was examined. Using site-directed mutagenesis I generated a series of RPR mutants of Cucumber necrosis tombusvirus (CNV). Analysis of RPR mutants defined that wild type RPR motif, especially two of the four arginines, were required for efficient RNA binding in vitro, for replication of tombusviruses, their associated DI RNAs, subgenomic (sg)RNA synthesis and DI RNA recombination in vivo. Experiments using a two-component tombusvirus replication system showed that RPR motif is critical for functions of both p33 and p92 in replication, but its role in these proteins might not be identical. Recombination studies using a novel tombusvirus three-component system revealed that mutations in RPR motif of p33 replicase protein resulted in an altered viral RNA recombination rate. Identified DI RNA recombinants were mostly imprecise, with recombination sites clustered around a replication enchancer and an additional putative cis-acting element that might facilitate the template switching events by the tombusvirus replicase. To study the role of RNA during the assembly of functional tombusvirus replicase, recombinant CNV replicase that showed similar properties to plant-derived CNV replicase was purified from Saccharomyces cerevisiae. When in addition to p33 and p92 proteins DI RNA was co-expressed in yeast cells, the isolated replicase activity was increased ~40 fold. Further studies defined RNA motifs within two short DI RNA regions that enhanced active CNV replicase formation. In summary, this study showed that the conserved RNA binding motif of the tombusvirus replicase proteins and viral RNA are involved in replicase assembly, viral RNA replication, subgenomic RNA synthesis and RNA recombination. This data shed new light on the complex roles of the viral elements in replication, and will help future studies aimed at interfering with viral infections.

Student Number : 0314429G - MSc research report - School of Molecular and Cell Biology - Faculty of Science / Tomato (Lycopersicon esculentum) is a horticultural commodity of great economic importance in many parts of the world, including South Africa. A previous study identified a new begomovirus, Tomato curly stunt virus (ToCSV), as the causative virus of a new and potentially devastating disease of tomatoes in South Africa. In this study, symptomatic plants, suspected of infection with an uncharacterized ToCSV isolate (01/2521) were collected for screening from Pietermaritzburg, South Africa. A host range study was conducted with the original ToCSV isolate (99/0631). Two small DNA molecules (1449 nts and 755 nts) were found associated with ToCSV [01/2521] using near-full length primers AL1c2745 and PAR1v32 specific for ToCSV. A single small DNA molecule (842 nts) was also found in association with the original ToCSV isolate. Nucleotide sequence analysis revealed that the two small DNA molecules (1449bp and 755bp) have no significant nucleotide sequence identity (less than 20%) with any known begomovirus. The 842bp molecule has the most significant nucleotide sequence identity (48%) to that of ToCSV (AF261885), while less than 20% nucleotide sequence identities were found when compared with other begomoviruses. Nucleotide sequence alignment of the 842bp DNA molecule to the ToCSV sequence, showed that this small DNA molecule is a chimeric molecule that could have arisen through recombination, partly from the coding regions of the ToCSV genome, but the rest of the molecule is of unknown origin. All three small DNA molecules identified in this study were compared to some known begomovirus associated subgenomic molecules and satellite molecules, and sequence identities of less than 20% were found. To our knowledge, this is the first report of a small DNA molecule found associated with the ToCSV genome. The complete genome sequence of ToCSV [01/2521] was not determined. Based on the results we obtained from the host range study, all the chosen test plants are not susceptible to ToCSV infection. The infectivity of all the small molecules identified in this study, is currently being investigated.