Partial characterization of the cloned dihydrofolate reductase gene of Saccharomyces cerevisiae

The cloned dihydrofolate reductase gene of Saccharomyces cerevisiae (DFR 1) is expressed in Escherichia coli. Bacterial strain JF1754 transformed with plasmids containing DFR 1 is at least 5X more resistant to inhibition by the folate antagonist trimethoprim. Expression of yeast DFR 1 in E. coli suggests it is likely that the gene lacks intervening sequences. The 1.8 kbp DNA fragment encoding yeast dhfr activity probably has its own promotor, as the gene is expressed in both orientations in E. coli. Expression of the yeast dhfr gene cloned into M13 viral vectors allowed positive selection of DFR 1 - M13 bacterial transfectants in medium supplemented with trimethoprim. A series of nested deletions generated by nuclease Bal 31 digestion and by restriction endonuclease cleavage of plasmids containing DFR 1 physically mapped the gene to a 930 bp region between the Pst 1 and Sal 1 cut sites. This is consistent with the 21,000 molecular weight attributed to yeast dhfr in previous reports. From preliminary DNA sequence analysis of the dhfr DNA fragment the 3' terminus of DFR 1 was assigned to a position 27 nucleotides from the Eco Rl cut site on the Bam Hi - Eco Rl DNA segment. Several putative yeast transcription termination consensus sequences were identified 3' to the opal stop codon. DFR 1 is expressed in yeast and it confers resistance to the antifolate methotrexate when the gene is present in 2 - 10 copies per cell. Plasmid-dependent resistance to methotrexate is also observed in a rad 6 background although the effect is somewhat less than that conferred to wild-type or rad 18 cells. Integration of DFR 1 into the yeast genome showed an intermediate sensitivity to folate antagonists. This may suggest a gene dosage effect. No change in petite induction in these yeast strains was observed in transformed cells containing yeast dhfr plasmids. The sensitivity of rad 6 , rad 18 and wild-type cell populations to trimethoprim were unaffected by the presence of DFR 1 in transformants. Moreover, trimethoprim did not induce petites in any strain tested, which normally results if dhfr is inhibited by other antifolates such as methotrexate. This may suggest that the dhfr enzyme is not the only possible target of trimethoprim in yeast. rad 6 mutants showed a very low level of spontaneous petite formation. Methotrexate failed to induce respiratory deficient mutants in this strain which suggested that rad 6 might be an obligate grande. However, ethidium bromide induced petites to a level approximately 50% of that exhibited by wild-type and rad 18 strains.

We need to have a conversation about OpenURL, a close look at a corpus of error reports from the 2013 academic year. Or, How I learned to stop worrying and give up on clean metadata.

Presented at Access 2014, winner of poster contest.

Evaluating three self-help smoking cessation interventions for post-secondary smokers : a randomized controlled trial on six Ontario campuses

Objective. Smoking prevalence is highest among the young adult cohort. Postsecondary students are no exception. Although many students intend to quit smoking, no research has established what methods best promote reductions in, or complete abstinence from smoking. This randomized controlled trial examined the effectiveness of three self-help smoking cessation interventions. Method. On six post-secondary campuses, 483 smokers who voluntarily accessed Leave The Pack Behind (a tobacco control initiative) were randomly assigned to one of three smoking cessation interventions: One Step At A Time (a 2-booklet, *gold standard' program for adults); Smoke|Quit (a newly-developed 2-booklet program for young adult students); and usual care (a 'Quit Kit' containing a booklet on stress management, information about pharmacological quitting aides and novelty items). All participants also received one proactive telephone support call from a peer counsellor. During the study, 85 participants withdrew. The final sample of 216 students who completed baseline questionnaires and 12-week follow-up telephone interviews was representative of the initial sample in terms of demographic characteristics, and smokingquitting- related variables. Results. Whether participants quit smoking depended upon treatment condition, ^(2, N=2\6) = 6.34, p = .04, with Smoke|Quit producing more successfijl quitters (18.4%) than One Step At A Time (4.5%) or the Quit Kit (1 1.4%). On average, participants had quit 53.46 days, with no significant difference across treatments. Selfefficacy also increased. Use of the intervention or other quitting aides was not associated with treatment condition. Among the 191 participants who did not quit smoking, treatment condition did not influence outcomes. Overall, 46.2% had made a quit attempt. Significant decreases in weekly tobacco consumption and increases in self-efficacy to resist smoking were observed from baseline to follow-up. Conclusion. Post-secondary institutions represent a potentially final opportunity for age-targeted interventions. Self-help resources tailored to students' social and contextual characteristics will have considerable more impact than stage-only tailored interventions. Both reduction and abstinence outcomes should be emphasized to positively support students to stop smoking.

Six-month outcomes of a multi-campus smoking cessation contest

Background: Up to 40% of North American post-secondary students smoke at least occasionally, and most want to quit. Given students' preferences for free, easy-to-access, self-directed, convenient cessation methods, a motivational, incentive-based cessation contest may be an effective way to assist students to quit. The current study describes 3- and 6-month outcomes experienced by post-secondary student smokers who entered the 'Let's Make A Deal!' contest. Methodology: Contestants from five university campuses who chose to quit completely ('Quit For Good') or reduce their tobacco consumption by 50% ('Keep The Count') were invited to participate in a study of the contest. Three and six months after registration, participants were contacted by phone to assess their smoking and quitting behaviours. Qualitative and quantitative measures were collected, including weekly tobacco consumption, efficacy to resist temptations to smoke, use of quitting aids, and strategies to cope with withdrawal. Quitting was assessed using 7-day point prevalence and continuous abstinence. Results: Seventy-four (64.9%) of the 114 participants recruited for the study completed the follow-ups. Over 31 % of participants who entered Quit For Good and 23.5% of participants who entered Keep The Count were identified as quitters at the 6-month follow-up. Among the quitters, 45.5% experienced sustained abstinence from smoking for the 6-month duration of the study. Keep The Count contestants reduced their tobacco consumption by 57.2% at 3-month follow-up and sustained some of this reduction through to the 6-month follow-up. Qualitative data provides insights into how quitters coped with withdrawal and what hampered continuing smokers' efforts to quit. Significance: A motivational, incentive-based contest for post-secondary students can facilitate both smoking cessation and harm reduction. The contest environment, incentives, resources, and "buddies" provide positive structural and social supports to help smokers overcome potential barriers to quitting, successfully stop smoking, and manage potential triggers to relapse. The contest cessation rates are higher than the typical 5-7% associated with unassisted quitting.

Adolescent girls and mathematics: classroom performance, attitudes toward mathematics and perceptions of parental attitudes /

The purpose of this study was to determine the extent to which gender differences exist in student attitudes toward mathematics and in their performance in mathematics at the Grade Seven and Eight level. The study also questioned how parents influence the attitudes of this grade level of male and female students toward mathematics. Historically, the literature has demonstrated gender differences in the attitudes of students toward mathematics, and in parental support for classroom performance in mathematics. This study was an attempt to examine these differences at one senior public school in the Peel Board of Education. One hundred three Grade Seven and Eight students at a middle school in the Peel Board of Education volunteered to take part in a survey that examined their attitudes toward mathematics, their perceptions of their parents' attitudes toward mathematics and support for good performance in the mathematics classroom, parental expectations for education and future career choices. Gender differences related to performance levels in the mathematics classroom were examined using Pearson contingency analyses. Items from the survey that showed significant differences involved confidence in mathematics and confidence in writing mathematics tests, as well as a belief in the ability to work on mathematics problems. Male students in both the high and low performance groups demonstrated higher levels of confidence than the females in those groups. Female students, however, indicated interest in careers that would require training and knowledge of higher mathematics. Some of the reasons given to explain the gender differences in confidence levels included socialization pressures on females, peer acceptance, and attribution of success. Perceived parental support showed no significant differences across gender groups or performance levels. Possible explanations dealt with the family structure of the participants in the study. Studies that, in the past, have demonstrated gender differences in confidence levels were supported by this study, and discussed in detail. Studies that reported on differences in parental support for student performance, based on the gender of the parent, were not confirmed by this study, and reasons for this were also discussed. The implications for the classroom include: 1) build on the female students' strengths that will allow them to enjoy their experiences in mathematics; 2) stop using the boys as a comparison group; and 3) make students more aware of the need to continue studying mathematics to ensure a wider choice of future careers.

Analytical modelling of bacterial migration and accumulation with rate-limited sorption in discrete fractures

An analytical model for bacterial accumulation in a discrete fractllre has been developed. The transport and accumlllation processes incorporate into the model include advection, dispersion, rate-limited adsorption, rate-limited desorption, irreversible adsorption, attachment, detachment, growth and first order decay botl1 in sorbed and aqueous phases. An analytical solution in Laplace space is derived and nlln1erically inverted. The model is implemented in the code BIOFRAC vvhich is written in Fortran 99. The model is derived for two phases, Phase I, where adsorption-desorption are dominant, and Phase II, where attachment-detachment are dominant. Phase I ends yvhen enollgh bacteria to fully cover the substratllm have accllillulated. The model for Phase I vvas verified by comparing to the Ogata-Banks solution and the model for Phase II was verified by comparing to a nonHomogenous version of the Ogata-Banks solution. After verification, a sensitiv"ity analysis on the inpllt parameters was performed. The sensitivity analysis was condllcted by varying one inpllt parameter vvhile all others were fixed and observing the impact on the shape of the clirve describing bacterial concentration verSllS time. Increasing fracture apertllre allovvs more transport and thus more accllffilliation, "Vvhich diminishes the dllration of Phase I. The larger the bacteria size, the faster the sllbstratum will be covered. Increasing adsorption rate, was observed to increase the dllration of Phase I. Contrary to the aSSllmption ofllniform biofilm thickness, the accllffilliation starts frOll1 the inlet, and the bacterial concentration in aqlleous phase moving towards the olitiet declines, sloyving the accumulation at the outlet. Increasing the desorption rate, redllces the dliration of Phase I, speeding IIp the accllmlilation. It was also observed that Phase II is of longer duration than Phase I. Increasing the attachment rate lengthens the accliffililation period. High rates of detachment speeds up the transport. The grovvth and decay rates have no significant effect on transport, althollgh increases the concentrations in both aqueous and sorbed phases are observed. Irreversible adsorption can stop accllillulation completely if the vallIes are high.

A school-based cognitive behavior modification intervention strategy for learned helpless mentally retarded students

This study assessed the usefulness of a cognitive behavior modification (CBM) intervention package with mentally retarded students in overcoming learned helplessness and improving learning strategies. It also examined the feasibility of instructing teachers in the use of such a training program for a classroom setting. A modified single subject design across individuals was employed using two groups of three subjects. Three students from each of two segregated schools for the mentally retarded were selected using a teacher questionnaire and pupil checklist of the most learned helpless students enrolled there. Three additional learned helplessness assessments were conducted on each subject before and after the intervention in order to evaluate the usefulness of the program in alleviating learned helplessness. A classroom environment was created with the three students from each school engaged in three twenty minute work sessions a week with the experimenter and a tutor experimenter (TE) as instructors. Baseline measurements were established on seven targeted behaviors for each subject: task-relevant speech, task-irrelevant speech, speech denoting a positive evaluation of performance, speech denoting a negative evaluation of performance, proportion of time on task, non-verbal positive evaluation of performance and non-verbal negative evaluation of performance. The intervention package combined a variety of CBM techniques such as Meichenbaum's (1977) Stop, Look and Listen approach, role rehearsal and feedback. During the intervention each subject met with his TE twice a week for an individual half-hour session and one joint twenty minute session with all three students, the experimentor and one TE. Five weeks after the end of this experiment one follow up probe was conducted. All baseline, post-intervention and probe sessions were videotaped. The seven targeted behaviors were coded and comparisons of baseline, post intervention, and probe testing were presented in graph form. Results showed a reduction in learned helplessness in all subjects. Improvement was noted in each of the seven targeted behaviors for each of the six subjects. This study indicated that mentally retarded children can be taught to reduce learned helplessness with the aid of a CBM intervention package. It also showed that CBM is a viable approach in helping mentally retarded students acquire more effective learning strategies. Because the TEs (Tutor experimenters) had no trouble learning and implementing this program, it was considered feasible for teachers to use similar methods in the classroom.

Manpower development and nation-building : Singapore's experience

Manpower is a basic resource. It is the indispensable means of converting other resources to mankind '.s use and benefit. As a process· of increasing the knowledge, skills, and dexterity of the people of a society, manpower development is the most fundamental means of enabling a nation to acquire the capacities to bring about its desired future state of affairs -- a more mighty and wealthier nation. Singapore's brief nation-building history justifies the emphasis accorded to the importance of good quality human resources and manpower development in economic and socio-political developments. As a tiny island-state with a poor natural resource base, Singapore's long-term survival and development depend ultimately upon the quality and the creative energy of her people. In line with the nation-building goals and strategies of the Republic, as conditioned by her objective setting, Singapore's basic manpower development premise has been one of "quality and not quantity". While implementing the "stop-at-two" family planning and population control programs and the relevant immigration measures to guard against the prospect of a "population explosion", the Government has energetically fostered various educational programs, including vocational training schemes, adult education programs, the youth movement, and the national service scheme to improve the quality of Singaporeans. There is no denying that some of the manpower development measures taken by the Government have imposed sacrifice and hardship on the Singapore citizens. Nevertheless, they are the basic conditions for the island-Republic's long-term survival and development. It is essential iii to note that Singapore's continuing existence and phenomenal-success are largely attributable to the will, capacities and efforts of her leaders and people. In the final analysis, the wealth and the strength of a nation are based upon its ability to conserve, develop and utilize effectively the innate capacities of its people. This is true not only of Singapore but necessarily of other developing nations. It can be safely presumed that since most developing states' concerns about the quality of their human resources and the progress of their nation-building work are inextricably bound to those about the quantity of their population, the "quality and not quantity" motto of Singapore's manpower development programs can also be their guiding principle.

Primary structure and expression studies of the dihydrofolate reductase gene (DFRI) of Saccharomyces cerevisiae

The nucleotide sequence of a genomic DNA fragment thought previously to contain the dihydrofolate reductase gene (DFR1) of Saccharomyces cerevisiae by genetic criteria was determined. This DNA fragment of 1784' basepairs contains a large open reading frame from position 800 to 1432, which encodes a enzyme with a predicted molecular weight of 24,229.8 Daltons. Analysis of the amino acid sequence of this protein revealed that the yeast polypep·tide contained 211 amino acids, compared to the 186 residues commonly found in the polypeptides of other eukaryotes. The difference in size of the gene product can be attributed mainly to an insert in the yeast gene. Within this region, several consensus sequences required for processing of yeast nuclear and class II mitochondrial introns were identified, but appear not sufficient for the RNA splicing. The primary structure of the yeast DHFR protein has considerable sequence homology with analogous polypeptides from other organisms, especially in the consensus residues involved in cofactor and/or inhibitor binding. Analysis of the nucleotide sequence also revealed the presence of a number of canonical sequences identified in yeast as having some function in the regulation of gene expression. These include UAS elements (TGACTC) required for tIle amino acid general control response, and "TATA H boxes as well as several consensus sequences thought to be required for transcriptional termination and polyadenylation. Analysis of the codon usage of the yeast DFRl coding region revealed a codon bias index of 0.0083. this valve very close to zero suggestes 3 that the gene is expressed at a relatively low level under normal physiological conditions. The information concerning the organization of the DFRl were used to construct a variety of fusions of its 5' regulatory region with the coding region of the lacZ gene of E. coli. Some of such fused genes encoded a fusion product that expressed in E.coli and/or in yeast under the control of the 5' regulatory elements of the DFR1. Further studies with these fusion constructions revealed that the beta-galactosidase activity encoded on multicopy plasmids was stimulated transiently by prior exposure of yeast host cells to UV light. This suggests that the yeast PFRl gene is indu.ced by UV light and nlay in1ply a novel function of DHFR protein in the cellular responses to DNA damage. Another novel f~ature of yeast DHFR was revealed during preliminary studies of a diploid strain containing a heterozygous DFRl null allele. The strain was constructed by insertion of a URA3 gene within the coding region of DFR1. Sporulation of this diploid revealed that meiotic products segregated 2:0 for uracil prototrophy when spore clones were germinated on medium supplemented with 5-formyltetrahydrofolate (folinic acid). This finding suggests that, in addition to its catalytic activity, the DFRl gene product nlay play some role in the anabolisln of folinic acid. Alternatively, this result may indicate that Ura+ haploid segregants were inviable and suggest that the enzyme has an essential cellular function in this species.

Participation factors in optional high school health and physical education

Student enrolment rates in optional health and physical education (HPE) classes have been steadily declining, to the point where most Ontario students stop taking HPE after completion of their one required credit, typically taken in grade nine. This study looked at factors that could contribute to HPE enrolment, sampling 227 grade ten students from five schools. These factors included selfefficacy (SE), perceived autonomy support (PAS), task value (TV), motivational regulation (autonomous, AR; controlled, CR), HPE grade average and body size discrepancy (BSD). Qualitative information was also gathered from students regarding likes and dislikes ofHPE, as well as reasons for their HPE enrolment choice. Cronbach Alpha values of each scale fell within acceptable values. ANOVA analysis revealed differences between enrolment groups in SE, TV, AR, HPE grade average, and BSD (p < .05). Reasons students reported for not taking HPE included a dislike of health classes, scheduling challenges, not needing HPE for future endeavors, concerns about social self-presentation, and a dislike of sports and/or competition. This research shows important differences between students and their HPE class choices and calls for a re-evaluation of how HPE classes are structured, advertised and scheduled by high school practitioners. Future works should look toward what other factors could be at play in students' decisions for or against optional HPE and how those factors interact with the constructs that were found to be of significance in this study. Keywords: Health and physical education, high school students, participation.

John W. Fisher Fonds, 1937-1961, 1964-1989, 1993, n.d.

Norah and Fred Fisher welcomed John Fisher into the world on November 29, 1912, not knowing what an influential role he would play in shaping Canada's history. John Fisher grew up as the middle child of five brothers and sisters in Frosty Hollow, New Brunswick, close to today’s town of Sackville. Sackville’s main industry was the Enterprise Foundry which the Fisher family owned and operated; however, Fisher had no plans of going into the family business. He was more inspired by his maternal grandfather, Dr. Cecil Wiggins, who lived with the family after retiring from the Anglican ministry. Wiggins encouraged all his grandchildren to be well read and to take part in discussions on current events. There were often visitors in the Fisher household taking part in discussions about politics, religion, and daily life. Fisher forced himself to take part in these conversations to help overcome his shyness in social settings. These conversations did help with his shyness and also in forming many opinions and observations about Canada. It put Fisher on the road to becoming Mr. Canada and delivering the many eloquent speeches for which he was known. Fisher did not venture far from home to complete his first degree. In 1934 he graduated from Mount Allison University in Sackville, NB with an Arts degree. The same year Fisher enrolled in Dalhousie’s law school. During his time at Dalhousie, Fisher discovered radio through Hugh Mills. Mills or “Uncle Mel” was on CHNS, Halifax’s only radio station at the time. Fisher began by making appearences on the radio drama show. By 1941 he had begun writing and broadcasting his own works and joined the staff as an announcer and continuity writer. In 1936 the Canadian Broadcasting Corporation was formed, the first National radio station. Fisher joined the CBC shortly after it’s beginning and remained with them, as well as the Halifax Herald newspaper, even after his law school graduation in 1937. By 1943 Fisher’s talks became a part of the CBC’s programming for a group of maritime radio stations. Fisher once described his talks as follows “my talks weren’t meant to be objective. . . they were meant to be favourable. They were ‘pride builders’” He began his famed John Fisher Reports at CBC Toronto when he transfered there shortly after the war. This program brought emmence pride to the fellow Canadians he spoke about leading to approximately 3500 requests per year to speak at banquets and meeting throughout Canada and the United States. Fisher was a well travelled indivdual who would draw on personal experiences to connect with his audience. His stories were told in simple, straight forward language for anyone to enjoy. He became a smooth, dynamic and passionate speaker who sold Canada to Canadians. He became a renowned journalist, folk historian, writer and broadcaster. Fisher was able to reach a vast array of people through his radio work and build Canadian pride, but he did not stop there. Other ways Fisher has contributed to Canada and the Canadian people include: Honoured by five Canadian Universities. 1956, became the Director of the Canadian Tourist Association. 1961, was appointed Special Assistant to the Prime Minister of Canada. 1963, Commissioner of the Centennial Commission (the Federal Agency Responsible for Canada’s 100th birthday) 1968, received the Service Medal , a coveted Order of Canada. President of John Fisher Enterprises Ltd., private consultant work, specializing in Centennial planning, broadcasts, lectures and promotion. John Fisher continued recording radio broadcasts even after his diagnosis with cancer. He would record 3 or 4 at a time so he was free to travel across Canada, the U.S., Europe and Mexico in search of treatments. Fisher passed away from the disease on February 15, 1981 and he is buried at Mount Pleasant Cemetery in Toronto.

Letter - Wilfrid Laurier to A. Munro Grier, 18 September 1901

A letter from Prime Minister Wilfrid Laurier to A. Munro Grier regarding an invitation to His Royal Highness. This is possibly referencing the Royal visit of the Duke of Cornwall in the year 1901. The Royals were due to tour Canada September and October of that year, with a stop in the Niagara area October 13th.

Letter - E.C. Schmon to Arthur A. Schmon, 10 April 1920

The letter discusses Arthur's "dandy cruise in the woods. Two hundred miles-" She also mentions that the couple has been married for eight months, but have only lived together two and half months. She talks about a train strike and states "the Pennsylvania trains stopped running yesterday and this morning all ferry boats." She mentions that the trolleys may stop running too. The price of food has increased as a result.

Letter - Julian Porter to R.A. Brown 12 July 1967

A letter from Mr. Julian Porter to R.A. Brown of Longmans Green Publishing Company. The letter indicates that Mr. Porter was introduced to Sean O'Sullivan during a political function. Mr. O'Sullivan mentioned that he had written a manuscript about Mr. J. Diefenbaker. Mr. Porter suggests that Longmans Green Publishing Company take interest in the manuscript. The company eventually rejects the book due to it being "so highly personal". The publishing company also remarks "I am personally most impressed with Mr. O'Sullivan's sincerity. I can't imagine anything will stop him from eventually going into politics."

Letter with the letterhead “H.H. Warner and Co.” of Rochester, N.Y. from the H.H. Warner Co

Letter with the letterhead “H.H. Warner and Co.” of Rochester, N.Y. from the H.H. Warner Co. The writer asks why Mr. Woodruff did not stop there on his way home from Pennsylvania regarding his shooting share at Point Mouillee [a game area in Monroe County, Michigan]. This letter is accompanied by an envelope, May 19, 1886.