Loading fluorescent Ca²⁺ indicators into living cells

Small-molecule fluorescent Ca2+ reporters are the most widely used tools in the field of Ca2+ signaling. The excellent spatial and temporal resolution afforded by fluorescent reporters has driven the understanding of Ca2+ as a messenger in many different cell types. In many situations, the cellular loading and monitoring of fluorescent Ca2+ indicators is quite trivial. However, there are numerous pitfalls that require consideration to ensure that optimal data are recorded. Fluorescent Ca2+ indicators have carboxylic acid groups for binding of Ca2+. Because these “free-acid” forms of the indicators are hydrophilic they cannot readily cross cell membranes and need to be introduced into cells using techniques such as microinjection, pinocytosis, or diffusion from a patch pipette. However, the most convenient and widely used method for loading indicators into cells is as hydrophobic compounds in which the carboxylic acid groups are esterified (commonly as acetoxymethyl [AM] or acetate esters). The ester versions of the indicators permeate the plasma membrane. The Ca2+-sensitive, free-acid form of the indicator is liberated following hydrolysis of the ester groups by intracellular esterases

Saving lives in the time of COVID-19. Case study of harm reduction, homelessness and drug use in Dublin, Ireland.

Dublin has outperformed even best-case scenarios for COVID-19 mortality among homeless and drug using populations. The experience provides important lessons for policy discussions on the pandemic, as well as broader lessons about pragmatic responses to these key client groups irrespective of COVID-19. The overarching lessons is that when government policy is well coordinated and underpinned by a science-driven and fundamentally pragmatic approach, lives are saved. Within this, the importance of strategic clarity and delivery, housing, lowered thresholds to methadone provision, Benzodiazepine (BZD) provision and Naloxone availability were key determinants of policy success. Further, given the rapid collapse in policy barriers to these interventions that COVID-19 produced, it is important to secure and protect these improved policy responses into the post-COVID-19 era

A milestone in drug policy: saving the lives of people who use drugs and were homeless in Dublin during the covid-19 pandemic

Dublin has outperformed even best-case scenarios for COVID-19 mortality among homeless and drug-using populations, write Austin O’Carroll, Tony Duffin and John Collins (LSE). When government policy is well coordinated and underpinned by a science-driven and fundamentally pragmatic approach, lives are saved

Local and global spontaneous calcium events regulate neurite outgrowth and onset of GABAergic phenotype during neural precursor differentiation

Neural stem cells can generate in vitro progenitors of the three main cell lineages found in the CNS. The signaling pathways underlying the acquisition of differentiated phenotypes in these cells are poorly understood. Here we tested the hypothesis that Ca2+ signaling controls differentiation of neural precursors. We found low-frequency global and local Ca2+ transients occurring predominantly during early stages of differentiation. Spontaneous Ca2+ signals in individual precursors were not synchronized with Ca2+ transients in surrounding cells. Experimentally induced changes in the frequency of local Ca2+signals and global Ca2+ rises correlated positively with neurite outgrowth and the onset of GABAergic neurotransmitter phenotype, respectively. NMDA receptor activity was critical for alterations in neuronal morphology but not for the timing of the acquisition of the neurotransmitter phenotype. Thus, spontaneous Ca2+ signals are an intrinsic property of differentiating neurosphere-derived precursors. Their frequency may specify neuronal morphology and acquisition of neurotransmitter phenotype

Sonification of the Riemann Zeta function.

The Riemann zeta function is one of the great wonders of mathematics, with a deep and still not fully solved connection to the prime numbers. It is defined via an infinite sum analogous to Fourier additive synthesis, and can be calculated in various ways. It was Riemann who extended the consideration of the series to complex number arguments, and the famous Riemann hypothesis states that the non-trivial zeroes of the function all occur on the critical line 0:5 + ti, and what is more, hold a deep correspondence with the prime numbers. For the purposes of sonification, the rich set of mathematical ideas to analyse the zeta function provide strong resources for sonic experimentation. The positions of the zeroes on the critical line can be directly sonified, as can values of the zeta function in the complex plane, approximations to the prime spectrum of prime powers and the Riemann spectrum of the zeroes rendered; more abstract ideas concerning the function also provide interesting scope

Class, commercialism and community in the origins and development of the Northern Rugby Football Union 1857-1910.

This thesis examines the role of class, regional, commercial and other social and economic factors in the origins and growth of Rugby football in Yorkshire and Lancashire during the period 1857 to 1910, focusing on the formation of the Northern Rugby Football Union (later to become the Rugby Football League) in 1895 and its subsequent development into a "new" sport of rugby league. Its main sources are documents of clubs and leading bodies of the sport and the sporting press of the period.Starting from an analysis of the spread of rugby from the public schools to the north of England, it links the rapid growth of the sport in the 1870s and 1880s to the sense of civic pride which prevailed among the industrial towns of the North and Midlands. In particular, it examines the means by which working class men and women became involved in the sport and looks at the nature and activities of rugby players and spectators. Its key focus is on the ways in which working class cultural practices became part of the fabric of the sport and the counter-development of the ideology of amateurism as a method of suppressing this, culminating in the Rugby Football Union's introduction of its first set of regulations intended to stamp out incipient professionalism, which were supported by both the northern and southern leaderships of the sport. It argues that the demands of working class players for payment and the growing commercialism of the sport in the North undermined amateurism and made its implementation, despite the vigorous efforts of its partisans, impossible.The breakdown in the consensus among rugby's leaders about how to deal with mass working class participation led directly to the 1895 split and the formation of the Northern Rugby Football Union, based on the payment of "broken-time" allowances to players. However, despite its initial successes, the Northern Union was marginalised by a combination of national and class-based forces, and, by the turn of the century, rendered impotent when face by the overwhelming popularity of soccer in the early 1900s. The necessity of establishing its own identity and holding back the soccer threat saw the Northern Union move away from being merely a professional version of rugby union and initiate the rule changes which created a new sport, expand to other countries and develop a distinct ideology. This allowed the sport to become almost exclusively identified with the working classes and opened the door to the predominance of working class cultural norms, both on the field and in the crowds which watched the game.Although class is viewed as the motor force which ultimately drove rugby to schism, the role of civic pride, of both the working and middle classes, the relationship between rugby and masculinity, links between sport and nationalism, the north-south divide in English society, and the rise of commercialism in the form of the "entertainment industry" are also examined in the changing contexts of the period for the impact they had on the sport and for their importance to its eventual schism

Ariel - Volume 12 Number 2

Executive Editors David G. Polin Larry H. Pastor Business Manager Alex Macones Jean Lien Editorial Page Editor Deepak Kapoor Sports Editor Todd Hoover Photography Editors Lois Leach Ken Yonemur

Stronger Together: Tiny-housers’ Views of Community

Over the last decade, tiny homes (generally described as homes smaller than 500 ft2) have grown in popularity thanks to recent representation in the media (Ford and Gomez‐Lanier 2017). An increasing number of people have begun to adopt this reimagined way of living, and some tiny-housers have sought to live near other tiny-housers, forming tiny house communities (Kilman 2016, Mangold et al. Forthcoming). Little systematic research focusing on the tiny-housers views of community exists. To provide initial insights, 30 interviews were conducted with people at various stages in their tiny house journeys. The 30 interviews were transcribed and analyzed in Nvivo 11. Preliminary findings suggest that many of these tiny-housers want to redefine the normative American concept of community by rejecting sub-urban frameworks and borrowing elements from both strong place and interest-based community models. They seek to accomplish this vision through one or more of the following: (i) increased community participation, (ii) shared spaces and resources, and (iii) development of significant relationships with neighbors rooted in mutual support and frequent interaction. While participants strive for this type of community, they also insist upon a clear recognition of personal space and boundaries. Participants also express an interest in having diverse communities, yet desire to maintain many common interests and goals. These community elements are not entirely unique to the tiny house movement. The small space, however, both encourages and facilitates this kind of community participation. Key words: Tiny house, lifestyle, downsizing, good life, alternative lifestyle, minimalism Ford, Jasmine and Lilia Gomez‐Lanier. 2017. Are Tiny Homes Here to Stay? A Review of Literature on the Tiny House Movement. Family and Consumer Sciences Research Journal 45(4):394-405. Kilman, Charlie. 2016. Small House, Big Impact: The Effect of Tiny Houses on Community and Environment. Undergraduate Journal of Humanistic Studies 2. Mangold, Severin, Chelsey Willoughby, Devin Hing, Codey Collins and Toralf Zschau. Forthcoming. Why Live Tiny? A New Multi-Dimensional Model. Sociological Spectrum