Swi6 protein interactions and the regulation of the yeast cell cycle

In budding yeast, the cell cycle is initiated at a point in G1 called Start, which marks the irrevocable commitment to a round of mitotic cell division. At Start there is an increase in the expression of over 200 cell cycle-regulated genes depending in part, on the Start-specific transcription factor, Swi6p. The aim of this work was to identify factors that interact with Swi6p and thereby regulate its activity. The search for novel Swi6p protein-protein interactions was performed using the new and relatively untested Sos Recruitment System (SRS). This allowed not only the investigation of Swi6p interactions, but also the assessment of the effectiveness of the SRS system in yeast. The results of the screening ultimately proved inconclusive, as putative interactions could not be verified further by alternative m vivo or in vitro systems. The type of proteins identified from the screening, suggested a bias in the SRS system towards detecting conditional interactions. Nevertheless, during the investigation of the results from the SRS screen, a novel Swi6p interaction was discovered serendipitously by other means. In vitro assays with affinity-purified proteins demonstrated that Swi6p was a substrate of the major yeast cyclin dependent kinase, Cdc28p resulting in the specific phosphorylation of Swi6p at serine 160. Earlier work had identified phosphorylation at serine 160 as the signal for nuclear export of Swi6p but concluded also that this was Cdc28p independent. However, phosphorylation of serine 160 is at a canonical Cdc28p-phosphorylation site and is mainly catalysed by Clb5p/6p- Cdc28p complexes in vitro. In vivo, peak activity of Clb5p and Clb6p corresponds to the time of Swi6 phosphorylation and nuclear export. Furthermore, there is a delay in Swi6p nuclear export in clb5 clb6 mutants as judged by cellular observations of Swi6p-GFP fusion protein throughout the cell cycle. The fact the expression of CLB5 and CLB6 is regulated by Swi6p at Start suggests a negative feedback loop promoting cell cycle progression by removing Swi6p from the nucleus after Gl. Interestingly, phosphorylated serine 160 is a specific in vitro substrate of the essential mitotic phosphatase, Cdcl4p that is active at the M to G1 transition. The ability of Cdcl4p to dephosphorylate Swi6p suggests a feed forward mechanism whereby completion of anaphase directly promotes passage through Start by stimulating import of Swi6p back into the nucleus

The demand side of farmers\u27 markets in Kitchener-Waterloo: An examination of the Waterloo, St. Jacobs and Kitchener farmers\u27 markets (Ontario)

Farmers\u27 markets have traditionally been outlets for a variety of everyday food items, many sold by local farmers. Today, many markets have changed from their original function, and now offer a variety of goods to a wider variety of people both demographically and geographically. In the Kitchener-Waterloo area, three farmers’ markets exist, the Kitchener Farmers’ Market, the Waterloo County Farmers’ Market and the St. Jacobs\u27 Farmers Market. These markets, identified as recognisable farmers’ markets (Tunbridge 1992), differ from other market typologies including traditional markets, public markets and festival market places. Though broad in nature, farmers’ markets can be broken down into further groupings based on social-economic indicators as exemplified by the market patrons. This thesis examines market differences using a customer survey administered at these three markets in November 1993. From this investigation the following market characteristics were found. The Kitchener Farmers’ Market has evolved from a traditional public market to a redeveloped, regular downtown farmers’ market, attracting a primarily localized customer base who patronize the market as part of a regular shopping trip. The Waterloo Farmers’ Market, a contemporary, urban fringe farmers’ market, serves a similar function to the Kitchener Market but draws a regular customer base from Waterloo and the surrounding rural areas. Lastly the St. Jacobs’ Farmers’ Market, a day-trip recreational market, attracts a customer base from a wide area, offering goods of both a farmer and flea market nature, including a weekly livestock auction. The typicality of these inter-market differences are vis-a-vis other market locations

The demand side of farmers\u27 markets in Kitchener-Waterloo: An examination of the Waterloo, St. Jacobs and Kitchener farmers\u27 markets (Ontario)

Farmers\u27 markets have traditionally been outlets for a variety of everyday food items, many sold by local farmers. Today, many markets have changed from their original function, and now offer a variety of goods to a wider variety of people both demographically and geographically. In the Kitchener-Waterloo area, three farmers’ markets exist, the Kitchener Farmers’ Market, the Waterloo County Farmers’ Market and the St. Jacobs\u27 Farmers Market. These markets, identified as recognisable farmers’ markets (Tunbridge 1992), differ from other market typologies including traditional markets, public markets and festival market places. Though broad in nature, farmers’ markets can be broken down into further groupings based on social-economic indicators as exemplified by the market patrons. This thesis examines market differences using a customer survey administered at these three markets in November 1993. From this investigation the following market characteristics were found. The Kitchener Farmers’ Market has evolved from a traditional public market to a redeveloped, regular downtown farmers’ market, attracting a primarily localized customer base who patronize the market as part of a regular shopping trip. The Waterloo Farmers’ Market, a contemporary, urban fringe farmers’ market, serves a similar function to the Kitchener Market but draws a regular customer base from Waterloo and the surrounding rural areas. Lastly the St. Jacobs’ Farmers’ Market, a day-trip recreational market, attracts a customer base from a wide area, offering goods of both a farmer and flea market nature, including a weekly livestock auction. The typicality of these inter-market differences are vis-a-vis other market locations

Strain System for the Motion Base Shuttle Mission Simulator

The Motion Base Shuttle Mission Simulator (MBSMS) Strain System is an innovative engineering tool used to monitor the stresses applied to the MBSMS motion platform tilt pivot frames during motion simulations in real time. The Strain System comprises hardware and software produced by several different companies. The system utilizes a series of strain gages, accelerometers, orientation sensor, rotational meter, scanners, computer, and software packages working in unison. By monitoring and recording the inputs applied to the simulator, data can be analyzed if weld cracks or other problems are found during routine simulator inspections. This will help engineers diagnose problems as well as aid in repair solutions for both current as well as potential problems

Using Facebook Advertising to Connect with Extension Audiences

There is considerable interest in using social media to reach Extension audiences. The study\u27s main objective was to assess the effectiveness of Facebook promotion and event advertising on creating new client contacts as measured by Likes. The results show the fan base for each county increased slowly prior to and following the Facebook ad, while it increased more rapidly during the advertisement period. Thus, Facebook advertising appears to be an effective tool to increase awareness of Extension Facebook pages. Extension professionals should consider investing in Facebook advertising to expand their fan base

Prolonged treatment with pimelic o-aminobenzamide HDAC inhibitors ameliorates the disease phenotype of a Friedreich ataxia mouse model

NOTICE: this is the author’s version of a work that was accepted for publication in Neurobiology of Disease. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication.Friedreich ataxia (FRDA) is an inherited neurodegenerative disorder caused by GAA repeat expansion within the FXN gene, leading to epigenetic changes and heterochromatin-mediated gene silencing that result in a frataxin protein deficit. Histone deacetylase (HDAC) inhibitors, including pimelic o-aminobenzamide compounds 106, 109 and 136, have previously been shown to reverse FXN gene silencing in short-term studies of FRDA patient cells and a knock-in mouse model, but the functional consequences of such therapeutic intervention have thus far not been described. We have now investigated the long-term therapeutic effects of 106, 109 and 136 in our GAA repeat expansion mutation-containing YG8R FRDA mouse model. We show that there is no overt toxicity up to 5 months of treatment and there is amelioration of the FRDA-like disease phenotype. Thus, while the neurological deficits of this model are mild, 109 and 106 both produced an improvement of motor coordination, whereas 109 and 136 produced increased locomotor activity. All three compounds increased global histone H3 and H4 acetylation of brain tissue, but only 109 significantly increased acetylation of specific histone residues at the FXN locus. Effects on FXN mRNA expression in CNS tissues were modest, but 109 significantly increased frataxin protein expression in brain tissue. 109 also produced significant increases in brain aconitase enzyme activity, together with reduction of neuronal pathology of the dorsal root ganglia (DRG). Overall, these results support further assessment of HDAC inhibitors for treatment of Friedreich ataxia.This work was supported by Repligen Corporation; Muscular Dystrophy Association (MDA) USA; Ataxia UK; Friedreich's Ataxia Research Alliance (FARA); GoFAR; and the Wellcome Trust [089757]

Addressing cancer survivors\u27 cardiovascular health using the Automated Heart Health Assessment (AH-HA) EHR tool: Initial protocol and modifications to address COVID-19 challenges

BACKGROUND: The purpose of this paper is to describe the Automated Heart-Health Assessment (AH-HA) study protocol, which demonstrates an agile approach to cancer care delivery research. This study aims to assess the effect of a clinical decision support tool for cancer survivors on cardiovascular health (CVH) discussions, referrals, completed visits with primary care providers and cardiologists, and control of modifiable CVH factors and behaviors. The COVID-19 pandemic has caused widespread disruption to clinical trial accrual and operations. Studies conducted with potentially vulnerable populations, including cancer survivors, must shift towards virtual consent, data collection, and study visits to reduce risk for participants and study staff. Studies examining cancer care delivery innovations may also need to accommodate the increased use of virtual visits. METHODS/DESIGN: This group-randomized, mixed methods study will recruit 600 cancer survivors from 12 National Cancer Institute Community Oncology Research Program (NCORP) practices. Survivors at intervention sites will use the AH-HA tool with their oncology provider; survivors at usual care sites will complete routine survivorship visits. Outcomes will be measured immediately after the study visit, with follow-up at 6 and 12 months. The study was amended during the COVID-19 pandemic to allow for virtual consent, data collection, and intervention options, with the goal of minimizing participant-staff in-person contact and accommodating virtual survivorship visits. CONCLUSIONS: Changes to the study protocol and procedures allow important cancer care delivery research to continue safely during the COVID-19 pandemic and give sites and survivors flexibility to conduct study activities in-person or remotely

Recent ASA presidents and ‘top’ journals: observed publication patterns, alleged cartels and varying careers

It has been common for studies presented as about American sociology as a whole to rely on data compiled from leading journals (American Sociological Review [ASR] and American Journal of Sociology [AJS]), or about presidents of the American Sociological Association [ASA], to represent it. Clearly those are important, but neither can be regarded as providing a representative sample of American sociology. Recently, Stephen Turner has suggested that dominance in the ASA rests with a ‘cartel’ initially formed in graduate school, and that it favors work in a style associated with the leading journals. The adequacy of these ideas is examined in the light of available data on the last 20 years, which show that very few of the presidents were in the same graduate schools at the same time. All presidents have had distinguished academic records, but it is shown that their publication strategies have varied considerably. Some have had no ASR publications except their presidential addresses, while books and large numbers of other journals not normally mentioned in this context have figured in their contributions, as well as being more prominent in citations. It seems clear that articles in the leading journals have not been as closely tied to prestigious careers as has sometimes been suggested, and that if there is a cartel it has not included all the presidents