Genetic and Biochemical Approaches for <em>In Vivo</em> and <em>In Vitro</em> Assessment of Protein Oligomerization: The Ryanodine Receptor Case Study

Oligomerization is often a structural requirement for proteins to accomplish their specific cellular function. For instance, tetramerization of the ryanodine receptor (RyR) is necessary for the formation of a functional Ca2+ release channel pore. Here, we describe detailed protocols for the assessment of protein self-association, including yeast two-hybrid (Y2H), co-immunoprecipitation (co-IP) and chemical cross-linking assays. In the Y2H system, protein self-interaction is detected by β-galactosidase assay in yeast co-expressing GAL4 bait and target fusions of the test protein. Protein self-interaction is further assessed by co-IP using HA- and cMyc-tagged fusions of the test protein co-expressed in mammalian HEK293 cells. The precise stoichiometry of the protein homo-oligomer is examined by cross-linking and SDS-PAGE analysis following expression in HEK293 cells. Using these different but complementary techniques, we have consistently observed the self-association of the RyR N-terminal domain and demonstrated its intrinsic ability to form tetramers. These methods can be applied to protein-protein interaction and homo-oligomerization studies of other mammalian integral membrane proteins

What Explains the Gender Gap in Financial Literacy? The Role of Household Decision-Making

Research has shown that financial illiteracy is widespread among women, and that many women are unfamiliar with even the most basic economic concepts needed to make saving and investment decisions. This gender gap in financial literacy may contribute to the differential levels of retirement preparedness between women and men. However, little is known about the determinants of the gender gap in financial literacy. Using data from the RAND American Life Panel, we examined potential explanations for the gender gap including the role of marriage and division of financial decision-making among couples. We found that differences in the demographic characteristics of women and men did not explain much of the financial literacy gap, whereas education, income and current and past marital status reduced the observed gap by around 25%. Oaxaca decomposition revealed the great majority of the gender gap in financial literacy is not explained by differences in covariates - characteristics of men and women - but due to coefficients, or how literacy is produced. We did not find strong support for specialization in financial decision-making within couples by gender. Instead, we found that decision-making within couples was sensitive to the relative education level of spouses for both women and men

Marital Histories and Economic Well-Being

Compared to unmarried individuals married individuals report greater average wealth. A restricted focus on current marital status risks misrepresenting the effects of marriage on wealth, as an increasing proportion of older adults have been divorced and remarried, having lived through the dramatic upheavals in family structure from the 1960s through the 1980s. To shed light on the associations between a lifetime of marriage events and wealth near retirement, we used panel data from the Health and Retirement Study and developed categories of marital experiences that acknowledged current status, type, number and date of past marital disruptions and total duration of time spent married across the lifespan. We found that the route individuals took to get to their current marital status were important predictors of wealth levels near retirement and were different for males and females. Observable differences in lifetime earnings, mortality risk, risk aversion, other characteristics such as education and number of children, explained much of the wealth difference between married and remarried individuals however neither observable characteristics nor sources of other wealth from pensions and Social Security were enough to explain the large differences in wealth accumulation between single and married women and individuals experiencing more than one marital disruption. Given the higher divorce rate, prevalence of multiple divorces and earlier age of divorce of the Baby Boomer cohort compared to earlier cohorts, an understanding of how marriage disruptions over the lifecycle impact savings is increasingly important for understanding the economic security of retirees.Social Security Administrationhttp://deepblue.lib.umich.edu/bitstream/2027.42/61806/1/wp180.pd

Analyzing an Abbreviated Dynamics Concept Inventory and Its Role as an Instrument for Assessing Emergent Learning Pedagogies

The Dynamics Concept Inventory (DCI) is a validated assessment tool commonly used to evaluate student growth within core, gateway-level mechanics courses. This research explored the evaluative use of this tool within the context of Freeform – an emergent course system that buttresses active class meetings with blended and collaborative virtual learning environments, themselves founded upon extensive multimedia content and interactive forums – at Purdue University. The paper specifically considers a number of related issues including: (i) the thoughtful development (via expert content validation) and statistical reliability of an abbreviated DCI instrument, which is more amenable to in-class implementation than the much longer full DCI; (ii) the correlation of abbreviated-DCI performance with exam scores and final course grades for a dynamics course using the Freeform framework with an emphasis on both conceptual understanding and traditional problem-solving skills; and (iii) various inter-section performance metrics in a preliminary study on how an implementation of the abbreviated-DCI may help elucidate the impact of the instructor within the Freeform framework. The results of these analyses supported the validity and reliability of the abbreviated DCI tool, and demonstrated its usefulness in a formal research setting. The preliminary study suggested that the Freeform framework might normalize differences in instructor pedagogical choices and student performance across class sections. These findings indicate that the abbreviated DCI holds promise as a research instrument and lay the groundwork for future inquiry into the impact of the Freeform instructional framework on students and instructors alike

Ageing and entrepreneurial preferences

Previous research on age and entrepreneurship assumed homogeneity and downplayed age-related differences in the motives and aims underlying enterprising behaviour. We argue that the heterogeneity of entrepreneurship influences how the level of entrepreneurial activity varies with age. Using a sample of 2566 respondents from 27 European countries we show that entrepreneurial activity increases almost linearly with age for individuals who prefer to only employ themselves (self-employers), whereas it increases up to a critical threshold age (late 40s) and decreases thereafter for those who aspire to hire workers (owner-managers). Age has a considerably smaller effect on entrepreneurial behaviour for those who do not prefer self-employment but are pushed into it by lack of alternative employment opportunities (reluctant entrepreneurs). Our results question the conventional wisdom that entrepreneurial activity declines with age and suggest that effective responses to demographic changes require policy makers to pay close attention to the heterogeneity of entrepreneurial preferences

Public health insurance and entry into self-employment

We estimate the impact of a differential treatment of paid employees versus self-employed workers in a public health insurance system on the entry rate into entrepreneurship. In Germany, the public health insurance system is mandatory for most paid employees, but not for the selfemployed, who usually buy private health insurance. Private health insurance contributions are relatively low for the young and healthy, and until 2013 also for males, but less attractive at the other ends of these dimensions and if membership in the public health insurance allows other family members to be covered by contribution-free family insurance. Therefore, the health insurance system can create incentives or disincentives to starting up a business depending on the family’s situation and health. We estimate a discrete time hazard rate model of entrepreneurial entry based on representative household panel data for Germany, which include personal health information, and we account for non- random sample selection. We estimate that an increase in the health insurance cost differential between self-employed workers and paid employees by 100 euro per month decreases the annual probability of entry into selfemployment by 0.38 percentage points, i.e. about a third of the average annual entry rate. The results show that the phenomenon of entrepreneurship lock, which an emerging literature describes for the system of employer provided health insurance in the USA, can also occur in a public health insurance system. Therefore, entrepreneurial activity should be taken into account when discussing potential health care reforms, not only in the USA and in Germany

Cholesterol catalyses Aβ42 aggregation through a heterogeneous nucleation pathway in the presence of lipid membranes.

Alzheimer's disease is a neurodegenerative disorder associated with the aberrant aggregation of the amyloid-β peptide. Although increasing evidence implicates cholesterol in the pathogenesis of Alzheimer's disease, the detailed mechanistic link between this lipid molecule and the disease process remains to be fully established. To address this problem, we adopt a kinetics-based strategy that reveals a specific catalytic role of cholesterol in the aggregation of Aβ42 (the 42-residue form of the amyloid-β peptide). More specifically, we demonstrate that lipid membranes containing cholesterol promote Aβ42 aggregation by enhancing its primary nucleation rate by up to 20-fold through a heterogeneous nucleation pathway. We further show that this process occurs as a result of cooperativity in the interaction of multiple cholesterol molecules with Aβ42. These results identify a specific microscopic pathway by which cholesterol dramatically enhances the onset of Aβ42 aggregation, thereby helping rationalize the link between Alzheimer's disease and the impairment of cholesterol homeostasis

Entrepreneurs' exit and paths to retirement : theoretical and empirical considerations

The number of ageing entrepreneurs in micro- and small-sized companies is rapidly increasing in Finland and other European Union countries. Over half a million jobs, in over one hundred thousand companies within the EU, are lost annually due to unsuccessful, predominantly retirement-related transfers of businesses. This challenge coincides with EU Grand Challenges and has been highlighted in the Entrepreneurship 2020 Action Plan (European Commission 2013). It has been estimated that in Finland, some 8000 jobs are lost yearly due to the ageing of entrepreneurs. Therefore, entrepreneur ageing has implications not only for the ageing individual but also for the company and the society at large. As entrepreneurs age it becomes more essential for them to start planning when and how they transition into retirement. While they may experience several exits and subsequent re-entries into working life via buying or starting new companies, exiting ones entrepreneurial career due to old age retirement differs from exits that occur earlier during the career. In this chapter, we provide a short overview of the entrepreneur retirement and exit literature from an age perspective. Furthermore, we present a theoretical conceptualization which combines entrepreneur retirement process with exit theories. This will enable scholars to better understand the retirement process, including decision-making, transitioning, and adjustment to retirement. We also provide empirical evidence using data collected among Finnish entrepreneurs in 2012 and 2015, where we outline the types of exits and assess several factors, including age, in association with exit intentions.fi=vertaisarvioitu|en=peerReviewed