Transfection of newt blastema mesenchyme using the techniques of lipofection and direct gene transfer

The regenerating amphibian limb provides a useful system for studying genes involved in the establishment of positional information. While a number of candidate genes that may playa role in pattern formation have been identified, their function in vivo is unknown in this system. To better ascertain the role of these genes, it would be useful to be able to alter their normal patterns of expression in vivo and to assess the effects of this misexpression on limb pattern. In order to achieve this, a method of introducing a plasmid containing the eDNA of a gene of interest into a newt blastema (a growth zone of mesenchymal progenitor cells) is needed. Unfortunately, most commonly used transfection techniques cannot be used with newt blastema cells. In this study, I have used the techniques of lipofection and direct gene transfer to introduce plasmid DNA containing reporter genes into the cells of a regenerating newt limb. The technique of lipofection was most effective when the blastema cells were transfected in vitro. The optimal ratio for transfection was shown to be 1:3 DNA:Lipofectin (W/w) , and an increase in the amount of DNA present in the mixture (1:3 ratio maintained) resulted in a corresponding increase in gene expression. The technique of direct gene transfer was used to transfect newt blastema cells with and without prior complex formation with Lipofectin. Injection of plasmid DNA alone provided the most 3 promising results. It was possible to introduce plasmid DNA containing the reporter gene ~-galactosidase and achieve significant gene expression in cells associated with the injection site. In the future, it would be interesting to use this technique to inject plasmid DNA containing a gene which may have a role in pattern formation into specific areas of the newt blastema and to analyze the resulting limb pattern that emerges.

Examining the inclusion experiences of students with intellectual disabilities : a study focusing on the transition from elementary to secondary school

Transitioning from elementary to secondary school is a major event in adolescents' lives and can be associated with academic, social, and emotional challenges (Shaffer, 2005; Sirsch, 2003). Considerably less research has focused on the transitional experiences of students with intellectual disabilities (lD) as they enter secondary school and the role of educational inclusion in this process (Noland, Cason, & Lincoln, 2007). Conceivably, students with ID who leave inclusive elementary schools, where they have been educated alongside their peers without ID, and who enter segregated secondary educational placements may experience unique social and emotional challenges (Farmer, Pearl, & Van Acker, 1996; Fryxell & Kennedy, 1995; Shaffer, 2005). This study examined the transitional experiences of 6 students with ID and the role of educational inclusion, with a focus on elementary to secondary school transitions from inclusive to segregated settings and vice versa. This study included the collection of multiple sources of data. Semi-structured interviews with 6 caregivers and students with ID were conducted. Students' Individual Education Transitional Plans were discussed in caregivers' interviews to determine how they shaped students' educational inclusion experiences (Ontario Ministry of Education & Training, 1999/2000/2004). Parts ofthe following questionnaires were "qualitized" (Tashakkori & Teddlie, 1998) and administered orally: "Youth Self-Report" (YSR; Achenbach, 2001 c) and "Child Behaviour Checklist Caregivers Form" (CBLC/6-18; Achenbach, 200la). The findings of this study contribute to the literature on educational inclusion by highlighting the positive/negative social and emotional impact of congruent and incongruent transitional experiences of students with ID and the role of educational inclusion.

Rittermere Farm Craft Studio Fonds, 1951-2006

During the 1950’s, the Rittenhouse family of Vineland in the Niagara Peninsula opened a craft store and studio. Within a short period of time, they realized that resources for the craft of rug hooking were in demand and they began to build their business around this niche. Edna Rittenhouse, the mother, was the wool dyer; Margaret Rowan, the daughter, was the pattern designer; Ted Rowan, the son-in-law, changed careers and became the manager of the family business. The 1960’s were a prosperous time, not only in the Niagara Peninsula, but also for the Rittenhouse business. Edna Rittenhouse had been hooking rugs for decades but she and her family worked at developing and sharing newer techniques with newer materials. Shading manuals were authored and published; students became teachers; creativity abounded in the demand for and the creation of new designs. Instead of using woolen yarn, they were using pure woolen fabric; instead of using a standard cutter, they began using a uniquely designed cutter; instead of using frames, they employed a table top method. The new material and technique resulted in a rug with a smooth, uniform texture and a soft nap. Since many crafters belonged to crafters guilds, Margaret and Ted Rowan began promoting the idea of a guild for rug hookers and in time the Ontario Hooking Craft Guild was also a reality. A joint project between Chatelaine magazine and the Rittermere studio for Canada’s centennial year of 1967 was extremely well received within the circle of hooking crafters and the Rittermere Farm Craft Studio became a North American landmark for crafters. From this point onward the studio had a large customer base not only in North America but also overseas. The studio remained popular until 1984 when Margaret and Ted Rowan decided to retire. The Rittermere name has been preserved in the name of Rittermere-Hurst-Field which is a similar business located in Aurora which is just north of Toronto.

Rick Bell Family fonds, 1874-1875, 1879, 1884, 1886, 1888-1889, 1896, 1898, 1909-1910, 1939, 1945, n.d.

Lewis Tyrell married Jane Gains on August 31, 1849 in Culpeper Court House, Virginia. Jane Gains was a spinster. Lewis Tyrell died September 25, 1908 at his late residence, Vine St. and Welland Ave., St. Catharines, Ont. at the age of 81 years, 5 months. Jane Tyrell died March 1, 1886, age 64 years. Their son? William C. Tyrell died January 15, 1898, by accident in Albany, NY, age 33 years, 3 months. John William Taylor married Susan Jones were married in St. Catharines, Ont. on August 10, 1851 by William Wilkinson, a Baptist minister. On August 9, 1894 Charles Henry Bell (1871-1916), son of Stephen (1835?-1876) and Susan Bell, married Mary E. Tyrell (b. 1869?) daughter of Lewis and Alice Tyrell, in St. Catharines Ontario. By 1895 the Bell’s were living in Erie, Pennsylvania where children Delbert Otto (b. 1895) and Edna Beatrice (b. 1897) were born. By 1897 the family was back in St. Catharines where children Lewis Tyrell (b. 1899), Gertrude Cora (b. 1901), Bessie Jane (b. 1902), Charles Henry (b. 1906), Richard Nelson (b. 1911) and William Willoughby (b. 1912) were born. Charles Henry Bell operated a coal and ice business on Geneva Street. In the 1901 Census for St. Catharines, the Bell family includes the lodger Charles Henry Hall. Charles Henry Hall was born ca. 1824 in Maryland, he died in St. Catharines on November 11, 1916 at the age of 92. On October 24, 1889 Charles Hall married Susan Bell (1829-1898). The 1911 Census of Canada records Charles Henry Hall residing in the same household as Charles Henry and Mary Bell. The relationship to the householder is step-father. It is likely that after Stephen Bell’s death in 1876, his widow, Susan Bell married Hall. In 1939, Richard Nelson Bell, son of Charles Henry and Mary Tyrell Bell, married Iris Sloman. Iris (b. 22 May 1912 in Biddulph Township, Middlesex, Ontario) was the daughter of Albert (son of Joseph b. 1870 and Elizabeth Sloman, b. 1872) and Josie (Josephine Ellen) Butler Sloman of London, Ont. Josie (b. 1891) was the daughter of Everett Richard and Elizabeth McCarthy (or McCarty) Butler, of Lucan Village, Middlesex North. According to the 1911 Census of Canada, Albert, a Methodist, was a porter on the railroad. His wife, Josephine, was a Roman Catholic. Residing with Albert and Josie were Sanford and Sadie Butler and Sidney Sloman, likely siblings of Albert and Josephine. The Butler family is descended from Peter Butler, a former slave, who had settled in the Wilberforce Colony in the 1830s. Rick Bell b. 1949 in Niagara Falls, Ont. is the son of Richard Nelson Bell. In 1979, after working seven years as an orderly at the St. Catharines General Hospital while also attending night school at Niagara College, Rick Bell was hired by the Thorold Fire Dept. He became the first Black professional firefighter in Niagara. He is a founding member of the St. Catharines Junior Symphony; attended the Banff School of Fine Arts in 1966 and also performed with the Lincoln & Welland Regimental Band and several other popular local groups. Upon the discovery of this rich archive in his mothers’ attic he became passionate about sharing his Black ancestry and the contributions of fugitive slaves to the heritage Niagara with local school children. He currently resides in London, Ont.

Bell Family Bible, Miscellaneous, 1895 - 1918

A page from the Bell Family Bible entitled "Miscellaneous" recording the dates and places of birth of the children of Charles and Mary Bell. The birth dates for the eight children listed range from 1895 to 1918. There is also a single entry under the heading "Age of Children of Delbert Bell". This Bible was in the possession of the Rick Bell of St. Catharines. The Bell family is descended from former Black slaves from the United States who settled in Canada.The handwritten entries appear to be as follows: "Ages of the children of Charles and Mary Bell are Wilbert Otto Bell born November 7th 1895 Erie Pa. Edna Beatrice Bell born May 25th 1897 Erie Pa. Lewis Terrell Bell born April 8th 1899 St. Catharines Ont. Gertrude Bell born November 26th 1902 St. Catharines Ont. Charles Henry Bell born June 6th 1906 St. Catharines Ont. Richard Wilson Bell born March 19th 1911 William Willoughby Bell born May 2nd 1912 both in St. Catharines Ontario Age of Children of Delbert Bell March 12th 1918 Delbert Charlie Bell"

Niagara Postcards, 1902, 1908, 1910, undated.

Consists of 33 postcards featuring various scenes and landmarks in the Niagara region. The postcards feature the Great Gorge Route, the Niagara River and Falls, Queenston Heights, Ridley College, military camps in Niagara-on-the-Lake, and a Catholic Church in Port Colborne. Most of the postcards are blank, but four have been postmarked. One of these is addressed to Miss Ada Misner, Espanola, Ont., from M.G. and is postmarked August 18, 1908. Another postcard is addressed to Mr. Alfred H. Smith, Stamford Hill, London N., from Hedley Smith, France and is postmarked 1902. Another is addressed to Miss Annabel Bishop, Buffalo, N.Y. from Irene Lalour(?), and is postmarked April 30, 1908. The last postcard is addressed to Miss Edna Lackie, Toronto, from R.M. and is postmarked August 1, 1910.

Identification of novel retinoid receptors and their roles in vertebrate and invertebrate nervous systems

In vertebrates, signaling by retinoic acid (RA) is known to play an important role in embryonic development, as well as organ homeostasis in the adult. In organisms such as adult axolotls and newts, RA is also important for regeneration of the CNS, limb, tail, and many other organ systems. RA mediates many of its effects in development and regeneration through nuclear receptors, known as retinoic acid receptors (RARs) and retinoid X receptors (RXRs). This study provides evidence for an important role of the RA receptor, RAR~2, in ,( '. regeneration ofthe spinal cord and tail of the adult newt. It has previously been proposed that the ability of the nervous system to regenerate might depend on the presence or absence of this RAR~2 isoform. Here, I show for the very first time, that the regenerating spinal cord of the adult newt expresses this ~2 receptor isoform, and inhibition of retinoid signaling through this specific receptor with a selective antagonist inhibits tail and spinal cord regeneration. This provides the first evidence for a role of this receptor in this process. Another species capable of CNS ~~generation in the adult is the invertebrate, " Lymnaea stagnalis. Although RA has been detected in a small number of invertebrates (including Lymnaea), the existence and functional roles of the retinoid receptors in most invertebrate non-chordates, have not been previously studied. It has been widely believed, however, that invertebrate non-chordates only possess the RXR class of retinoid receptors, but not the RARs. In this study, a full-length RXR cDNA has been cloned, which was the first retinoid receptor to be discovered in Lymnaea. I then went on to clone the very first full-length RAR eDNA from any non-chordate, invertebrate species. The functional role of these receptors was examined, and it was shown that normal molluscan development was altered, to varying degrees, by the presence of various RXR and RAR agonists or antagonists. The resulting disruptions in embryogenesis ranged from eye and shell defects, to complete lysis of the early embryo. These studies strongly suggest an important role for both the RXR and RAR in non-chordate development. The molluscan RXR and RAR were also shown to be expressed in the adult, nonregenerating eNS, as well as in individual motor neurons regenerating in culture. More specifically, their expression displayed a non-nuclear distfibution, suggesting a possible non-genomic role for these 'nuclear' receptors. It was shown that immunoreactivity for the RXR was present in almost all regenerating growth cones, and (together with N. Farrar) it was shown that this RXR played a novel, non-genomic role in mediating growth cone turning toward retinoic acid. Immunoreactivity for the novel invertebrate RAR was also found in the regenerating growth cones, but future work will be required to determine its functional role in nerve cell regeneration. Taken together, these data provide evidence for the importance of these novel '. retinoid receptors in development and regeneration, particularly in the adult nervous system, and the conservation of their effects in mediating RA signaling from invertebrates to vertebrates.

Functional genomics of BAHD acyltransferases in different varieties of the wine grape, Vilis vinifera

The characteristic "foxy" aroma of Vilis labrusca Concord grapes is due in large part to methyl anthranilate, a volatile ester formed by the enzyme anthraniloyl- CoA:methanol anthraniloyltransferase (VIAMAT) of the superfamily of BARD acyltransferases. The publication of the genome ofthe closely related wine grape Vilis vinifera, which does not accumulate methyl anthranilate, permitted the searching for any putative VlAU4T-like genes, with the result of 5 highly homologous candidates being found, with one candidate sharing 95% identity to VlAU4T. Probing the gene expression of 18 different cultivars of V. vinifora ripe berries by RT -PCR showed that many varieties do indeed express VlAU4T-like genes. Subsequent cloning of the full-length open reading frame of one of these genes from eDNA prepared from the cultivar Sauvignon Blanc permitted preliminary biochemical characterization of the enzyme after heterologous expression in E. coli. It was determined that this alcohol acyltransferase (named VvsbAATl) catalyzes the formation of cis-3-hexenyl acetate, a "green-leaf' volatile. Although the cloned gene from Sauvignon Blanc had 95% identity at the amino acid level to VIAMAT, it displayed an altered substrate specificity and expression pattern. These results highlight the difficulty in predicting substrate specificity and function of enzymes through the basis of sequence homology, which is a common finding in the study of BARD acyltransferases. Also, the determination of function of VvsbAATl and other BARD acyltransferases in V. vinifera could be used as a genetic marker for certain aroma characteristics in grape breeding programs.