Isolation and characterization of genomic DNA sequences that enhance the stability of plasmid DNA in mammalian cells /

The ease of production and manipulation has made plasmid DNA a prime target for its use in gene transfer technologies such as gene therapy and DNA vaccines. The major drawback of plasmid however is its stability within mammalian cells. Plasmid DNA is usually lost by cellular mechanisms or as a result of mitosis by simple dilution. This study set out to search for mammalian genomic DNA sequences that would enhance the stability of plasmid DNA in mammalian cells.Creating a plasmid based genomic DNA library, we were able to screen the human genome by transfecting the library into Human Embryonic Kidney (HEK 293) Cells. Cells that contained plasmid DNA were selected, using G418 for 14 days. The resulting population was then screened for the presence of biologically active plasmid DNA using the process of transformation as a detector.A commercially available plasmid DNA isolation kit was modified to extract plasmid DNA from mammalian cells. The standardized protocol had a detection limit of -0.6 plasmids per cell in one million cells. This allowed for the detection of 45 plasmids that were maintained for 32 days in the HEK 293 cells. Sequencing of selected inserts revealed a significantly higher thymine content in comparison to the human genome. Sequences with high A/T content have been associated with Scaffold/Matrix Attachment Region (S/MAR) sequences in mammalian cells. Therefore, association with the nuclear matrix might be required for the stability of plasmids in mammalian cells.

Benjamin Woodruff Price Papers, 1851, 1853-1860, 1862-1863, 1874, n.d.

The fonds includes sixty two items of correspondence between Benjamin Woodruff Price, aka Woodruff, Ben or Uncle, and various family members, both immediate and distant cousins. Also included is business correspondence related to Price’s activities as a watchmaker and/or jeweler. Benjamin Woodruff Price was born in Thorold Township ca. 1831, the son of Joseph Price and Mary Smith. B.W. Price married Ella or Ellen McGlashan (1851-1906) ca. 1868. Price died between 1891 and 1901, his burial location is unknown at present. A watchmaker and jeweler, Price lived most of his life in Fonthill, Ont. He also included auctioneer, undertaker and photographer as some of his other professional activities. His siblings included David Smith Price (wife Isabella Ann), John Smith Price (wife Elizabeth Jane), and sisters Susan Page (husband Edward Rice Page), Jerusha Price, Mary Price and Martha W. Stone (husband Dudley Ward Stone). John Smith Price died 18 April 1860, leaving no descendents. It is likely that G.W. Stone was a nephew to B.W. Price, the son of his sister Martha W. Stone and her husband Dudley Ward Stone. Susan Page was a sister of Benjamin Woodruff Price. She was married to Edward Rice Page and they had at least two children, Joseph and Clayton. At the time of this correspondence they lived in Suspension Bridge, NY, now part of Niagara Falls, New York. Edward Rice Page’s occupation was listed as saloon keeper. The Price family appears to have had a very large extended family. This information was gleaned from the contents of letters of Maggie Tisdale, daughter of Ephraim and Hannah (Price) Tisdale, P.A. or Ann Morgan, [may also be Phebe Ann] of Newark, NY? and Marietta House of Bayham Township. DeWitt Higgins of Suspension Bridge, NY aka Niagara Falls, NY was an auctioneer, specialized in buying jewellery, watches, clocks, from individuals and reselling his product to others like B.W. Price.

Welland Canal Survey of Lands David Price, 1834

Survey map and description of David Price's land created by The Welland Canal Company. Included is a written description of the land along with a drawing of the land. Noteable features include; Hellem's Creek, road, bridge, Chippewa, Griffith's land, canal. The deed for the land is dated October 14, 1834. The land totals 5 acres, 3 roads and 7 perches. Surveyor notes are seen in pencil and red pen on the map.

The Impact of a Corporate Name Change on Stock Price and Trading Volume of Canadian Companies

This thesis examines the impact of a corporate name change on stock price and trading volume of Canadian companies around the announcement date, the approval date, and the adoption date over the time period from 1997 to 2011. Name changes are classified into six categories: major and minor, structural and pure, diversified and focused, accompanied with a change in ticker symbol and without a change in ticker symbol, “Gold” name addition and deletion, and different reasons for name changes (e.g., merger and acquisition, change of structure, change of strategy, and better image). The thesis uses the standard event study methodology to perform abnormal return and trading volume analyses. In addition, regression analysis is employed to examine which type of a name change has the largest impact on cumulative abnormal returns. Sample stocks exhibit a significant positive abnormal return one-day prior to the approval day and one day after the adoption date. Around the approval date we observe significant abnormal returns for stocks with a structural name change. On the day after the adoption date we document abnormal returns for stocks with major, minor, structural, pure, focused, and ticker symbol name changes. If a merger or acquisition is the reason for a name change, companies tend to experience a significant positive abnormal return one-day before the approval date and on the adoption date. If a change of structure is the reason for a name change, companies exhibit a significant positive abnormal return on the approval date and a significant negative abnormal return on the adoption date. In case of a change of strategy as the reason for a name change, companies show a significant negative abnormal return around the approval date and a significant positive abnormal return around the adoption date.

Investigating the role of apoptosis regulator EndoG on exogenous DNA uptake, stability, replication and recombination

Endonuclease G (EndoG) is a well conserved mitochondrial nuclease with dual lethal and vital roles in the cell. It non-specifically cleaves endogenous DNA following apoptosis induction, but is also active in non-apoptotic cells for mitochondrial DNA (mtDNA) replication and may also be important for replication, repair and recombination of genomic DNA. The aim of our study was to examine whether EndoG exerts similar activities on exogenous DNA substrates such as plasmid DNA (pDNA) and viral DNA vectors, considering their importance in gene therapy applications. The effects of EndoG knockdown on pDNA stability and levels of encoded reporter gene expression were evaluated in the cervical carcinoma HeLa cells. Transfection of pDNA vectors encoding short-hairpin RNAs (shRNAs) reduced levels of EndoG mRNA and nuclease activity in HeLa cells. In physiological circumstances, EndoG knockdown did not have an effect on the stability of pDNA or the levels of encoded transgene expression as measured over a four day time-course. However, when endogenous expression of EndoG was induced by an extrinsic stimulus (a cationic liposome transfection reagent), targeting of EndoG by shRNA improved the perceived stability and transgene expression of pDNA vectors. Therefore, EndoG is not a mediator of exogenous DNA clearance, but in non-physiological circumstances it may non-specifically cleave intracellular DNA regardless of its origin. To investigate possible effects of EndoG on viral DNA vectors, we constructed and evaluated AdsiEndoG, a first generation adenovirus (Ad5 ΔE1) vector encoding a shRNA directed against EndoG mRNA, along with appropriate Ad5 ΔE1 controls. Infection of HeLa cells with AdsiEndoG at a multiplicity of infection (MOI) of 10 p.f.u./cell resulted in an early cell proliferation defect, absent from cells infected at equivalent MOI with control Ad5 ΔE1 vectors. Replication of Ad5 ΔE1 DNA was detected for all vectors, but AdsiEndoG DNA accumulated to levels that were 50 fold higher than initially, four days after infection, compared to 14 fold for the next highest control Ad5 ΔE1 vector. Deregulation of the cell cycle by EndoG depletion, which is characterized by an accumulation of cells in the G2/M transition, is the most likely reason for the observed cell proliferation defect. The enhanced replication of AdsiEndoG is consistent with this conclusion, as Ad5 ΔE1 DNA replication is intimately related to cell cycling and prolongation or delay in G2/M greatly enhances this process. Furthermore, infection of HeLa with AdsiEndoG at MOI of 50 p.f.u./cell resulted in an almost complete disappearance of viable, adherent tumour cells from culture, whereas almost a third of the cells were still adherent after infection with control Ad5 ΔE1 vectors, relative to the non-infected control. Therefore, targeting of EndoG by RNAi is a viable strategy for improving the oncolytic properties of first generation adenovirus vectors. In addition, AdsiEndoG-mediated knockdown of EndoG reduced homologous recombination between pDNA substrates in HeLa cells. The effect was modest but, nevertheless demonstrated that the proposed role of EndoG in homologous recombination of cellular DNA also extends to exogenous DNA substrates.

Alberta Price List No. 3., January 1st, 1925

A price list form for the Alberta Liquor Control Board from 1 January 1925. The price list includes "General Information" in regards to permits for individuals and special permits. The alcohol is then listed by category for pricing: Scotch Whisky, Irish Whisky, Rye Whisky, Bourbon Whisky, Rum, Gin, Brandy, Port, Native Wine, Italian (Type) Wines, Sherry, Claret, Burgundy, White Wine, Sparkling Wine, Vermouth, Cocktails, Liqueurs, Champagne, Bitters, Ale and Stout, Chinese Liquors.

General Price List, Ontario, 1 January 1924

A general price list from the year 1924 for the province of Ontario. The cover of the list reads: "Ontario Government Dispensaries Conducted Under Direction of Board of License Commissioners for Ontario By Authority of The Ontario Temperance Act. General Price List, Dispensaries sell liquor for medicinal, sacramental, scientific and manufacturing purposes only. The sale of liquor for beverage purposes in the Province of Ontario is prohibited by The Ontario Temperance Act. Dispensaries: No.1-154 Wellington Street West, Toronto; No.2-1271 Dundas Street West, Toronto; No.3-29 Charles Street, Hamilton; No.4-425 Talbot Street, London; No.5-30 Sandwich Street West, Windsor; No.6-Golden Lion Block, Kingston; No.7-92 Kent Street, Ottawa; No.8-109 Simpson Street, Fort William."

Newfoundland Price List, 1 June 1928

A price list for Newfoundland by the Board of Liquor Control, St. John's. The list is one page in length and has a few handwritten changes to prices.

Price List - Lawrence A. Wilson Co. Limited, 1916

A price list for Lawrence A. Wilson Co. Limited, 87 James St., Montreal Quebec. It is addressed to The Toronto Hunt, 52 Bay Street, Toronto. There are additional handwritten notes. One of the handwritten notes reads "ck to W. Stephen Haas"

Wiehl & Widmann Price List, August 1903

A price list for Wiehl & Widmann (Wholesale Importers of and Dealers in Wines, Liquors and Mineral Waters). The price list is dated August 1903 and the location of the dealer is New York.

The "Bodega" Company Price List

A price list for The "Bodega" Company Limited. The company has branches throughout the United Kingdom, including: London, Birmingham, Manchester, Liverpool, Glasgow, Edinburgh, Dundee, Brighton, and Ryde. The final pages of the price list also include a reprinted article from "The Irish Times" concerning a case against the use of the Bodega Company name.

Artists Rendering of Isobel Price

Artist’s rendering in colour of Isobel Price in an oval frame with a hook at the top and a section at the back which contains a lock of hair. This painting was done by Gerald Sinclair Hayward who was a renowned artist whose work was displayed at an exhibition in New York in 1899. He painted Theodore Roosevelt, William K. Vanderbilt and members of the ruling families of Britain, Germany and Russia. The frame is enclosed in a folding case lined with velvet and silk. The silk is quite worn. The outside of the case appears to be leather and has a stand for setting it upright. It closes with a metal latch. This is accompanied by a note by R. Band.

Indenture between Samuel Street and Eunice Price

Indenture (vellum) of bargain and sale between Samuel Street of Stamford and Eunice Price widow of the late David Price of Willoughby regarding a part of Lot no. 15 in the 5th Concession in Humberstone - instrument no. 1141, June 14, 1842.

Letter to Isabel from Phil [Phil Ingram Price]

Letter to Isabel from Phil [Phil Ingram Price] in which he expresses a desire to divide the estate with as little delay as possible. He has divided the books into 4 lots, but he asks if Isabel has some of the books as they are missing, March 28, 1900.

Letter to Isabel from someone whose last name is Price [the first name is illegible]

Letter to Isabel from someone whose last name is Price [the first name is illegible] in which the writer says that the row with Phil regarding the bonds is settled (1 ½ pages). This person does not anticipate any more trouble with Phil. They are sending the personal effects to Isabel as well as a cheque for $3000 as her share of the estate, and more next month, July 15, 1901.