Quantitative proteomic approach to study subcellular localization of membrane proteins

As proteins within cells are spatially organized according to their role, knowledge about protein localization gives insight into protein function. Here, we describe the LOPIT technique (localization of organelle proteins by isotope tagging) developed for the simultaneous and confident determination of the steady-state distribution of hundreds of integral membrane proteins within organelles. The technique uses a partial membrane fractionation strategy in conjunction with quantitative proteomics. Localization of proteins is achieved by measuring their distribution pattern across the density gradient using amine-reactive isotope tagging and comparing these patterns with those of known organelle residents. LOPIT relies on the assumption that proteins belonging to the same organelle will co-fractionate. Multivariate statistical tools are then used to group proteins according to the similarities in their distributions, and hence localization without complete centrifugal separation is achieved. The protocol requires approximately 3 weeks to complete and can be applied in a high-throughput manner to material from many varied sources.

Twitter and society : an introduction

Since its launch in 2006, Twitter has turned from a niche service to a mass phenomenon. By the beginning of 2013, the platform claims to have more than 200 million active users, who “post over 400 million tweets per day” (Twitter, 2013). Its success is spreading globally; Twitter is now available in 33 different languages, and has significantly increased its support for languages that use non-Latin character sets. While Twitter, Inc. has occasionally changed the appearance of the service and added new features—often in reaction to users’ developing their own conventions, such as adding ‘#’ in front of important keywords to tag them—the basic idea behind the service has stayed the same: users may post short messages (tweets) of up to 140 characters and follow the updates posted by other users. This leads to the formation of complex follower networks with unidirectional as well as bidirectional connections between individuals, but also between media outlets, NGOs, and other organisations. While originally ‘microblogs’ were perceived as a new genre of online communication, of which Twitter was just one exemplar, the platform has become synonymous with microblogging in most countries. A notable exception is Sina Weibo, popular in China where Twitter is not available. Other similar platforms have been shut down (e.g., Jaiku), or are being used in slightly different ways (e.g., Tumblr), thus making Twitter a unique service within the social media landscape.

Structural layers of communication on Twitter

Twitter is used for a range of communicative purposes. These extend from personal tweets that address what used to be Twitter’s default question, “What’s happening?”, through one-on-one @reply conversations between close friends and attempts at getting the attention of celebrities and other public actors, to discussions in communities built around specific issues—and back again to broadcast-style statements from well-known individuals and brands to their potentially very large retinue of followers.

Metrics for Understanding Communication on Twitter

As the systematic investigation of Twitter as a communications platform continues, the question of developing reliable comparative metrics for the evaluation of public, communicative phenomena on Twitter becomes paramount. What is necessary here is the establishment of an accepted standard for the quantitative description of user activities on Twitter. This needs to be flexible enough in order to be applied to a wide range of communicative situations, such as the evaluation of individual users’ and groups of users’ Twitter communication strategies, the examination of communicative patterns within hashtags and other identifiable ad hoc publics on Twitter (Bruns & Burgess, 2011), and even the analysis of very large datasets of everyday interactions on the platform. By providing a framework for quantitative analysis on Twitter communication, researchers in different areas (e.g., communication studies, sociology, information systems) are enabled to adapt methodological approaches and to conduct analyses on their own. Besides general findings about communication structure on Twitter, large amounts of data might be used to better understand issues or events retrospectively, detect issues or events in an early stage, or even to predict certain real-world developments (e.g., election results; cf. Tumasjan, Sprenger, Sandner, & Welpe, 2010, for an early attempt to do so).

Twitter and Sports : Football Fandom in Emerging and Established Markets

Twitter and other social media have become increasingly important tools for maintaining the relationships between fans and their idols across a range of activities, from politics and the arts to celebrity and sports culture. Twitter, Inc. itself has initiated several strategic approaches, especially to entertainment and sporting organisations; late in 2012, for example, a Twitter, Inc. delegation toured Australia in order to develop formal relationships with a number of key sporting bodies covering popular sports such as Australian Rules Football, A-League football (soccer), and V8 touring car racing, as well as to strengthen its connections with key Australian broadcasters and news organisations (Jackson & Christensen, 2012). Similarly, there has been a concerted effort between Twitter Germany and the German Bundesliga clubs and football association to coordinate the presence of German football on Twitter ahead of the 2012–2013 season: the Twitter accounts of almost all first-division teams now bear the official Twitter verification mark, and a system of ‘official’ hashtags for tweeting about individual games (combining the abbreviations of the two teams, e.g. #H96FCB) has also been instituted (Twitter auf Deutsch, 2012).

Crisis Communication in Natural Disasters : The Queensland Floods and Christchurch Earthquakes

Over the past decade, social media have gone through a process of legitimation and official adoption, and they are now becoming embedded as part of the official communications apparatus of many commercial and public-sector organisations— in turn, providing platforms like Twitter with their own sources of legitimacy. Arguably, the demonstrated utility of social media platforms and tools in times of crisis—from civil unrest and violent crime through to natural disasters like bushfires, earthquakes, and floods—has been a crucial driver of this newfound legitimacy. In the mid-2000s, user-created content and ‘Web 2.0’ platforms were known to play a role in crisis communication; back then, the involvement of extra-institutional actors in providing and sharing information around such events involved distributed, ad hoc, or niche platforms (like Flickr), and was more likely to be framed as ‘citizen journalism’ or ‘crowdsourcing’ (see, for example, Liu, Palen, Sutton, Hughes, & Vieweg, 2008, on the then-emerging role of photo-sharing in disasters). Since then, the dramatically increased take-up of mainstream social media platforms like Facebook and Twitter means that the pool of potential participants in online crisis communication has broadened to include a much larger proportion of the general population, as well as traditional media and official emergency response organisations.

Epilogue : Why Study Twitter?

Each of the thirty-one contributions in this volume implicitly spells out its own answer to this question. Surprisingly perhaps even for such a highly interdisciplinary volume as this one, these answers vary considerably in their approaches, their objectives, and their underlying assumptions about the object of study. This diversity of scholarly perspectives on Twitter, barely half a decade since it first emerged as a popular platform, highlights its versatility. Beginning as a side project to a now-forgotten podcasting platform, rising to popularity as a social network service focussed around mundane communication and therefore widely lambasted as a cesspool of vanity and triviality by incredulous journalists (including technology journalists), it was later embraced by those same journalists, governments, and businesses as a crucial source of real-time information on everything from natural disasters to celebrity gossip, and from debates over sexual violence to Vatican politics.

The politics of Twitter data

Talk of Big Data seems to be everywhere. Indeed, the apparently value-free concept of ‘data’ has seen a spectacular broadening of popular interest, shifting from the dry terminology of labcoat-wearing scientists to the buzzword du jour of marketers. In the business world, data is increasingly framed as an economic asset of critical importance, a commodity on a par with scarce natural resources (Backaitis, 2012; Rotella, 2012). It is social media that has most visibly brought the Big Data moment to media and communication studies, and beyond it, to the social sciences and humanities. Social media data is one of the most important areas of the rapidly growing data market (Manovich, 2012; Steele, 2011). Massive valuations are attached to companies that directly collect and profit from social media data, such as Facebook and Twitter, as well as to resellers and analytics companies like Gnip and DataSift. The expectation attached to the business models of these companies is that their privileged access to data and the resulting valuable insights into the minds of consumers and voters will make them irreplaceable in the future. Analysts and consultants argue that advanced statistical techniques will allow the detection of ongoing communicative events (natural disasters, political uprisings) and the reliable prediction of future ones (electoral choices, consumption)...

Metaphors of big data

Metaphors are a common instrument of human cognition, activated when seeking to make sense of novel and abstract phenomena. In this article we assess some of the values and assumptions encoded in the framing of the term big data, drawing on the framework of conceptual metaphor. We first discuss the terms data and big data and the meanings historically attached to them by different usage communities and then proceed with a discourse analysis of Internet news items about big data. We conclude by characterizing two recurrent framings of the concept: as a natural force to be controlled and as a resource to be consumed.

Echolocation call production during aerial and terrestrial locomotion by New Zealand's enigmatic lesser short-tailed bat, Mystacina tuberculata

Linkage of echolocation call production with contraction of flight muscles has been suggested to reduce the energetic cost of flight with echolocation, such that the overall cost is approximately equal to that of flight alone. However, the pattern of call production with limb movement in terrestrially agile bats has never been investigated. We used synchronised high-speed video and audio recordings to determine patterns of association between echolocation call production and limb motion by Mystacina tuberculata Gray 1843 as individuals walked and flew, respectively. Results showed that there was no apparent linkage between call production and limb motion when bats walked. When in flight, two calls were produced per wingbeat, late in the downstroke and early in the upstroke. When bats walked, calls were produced at a higher rate, but at a slightly lower intensity, compared with bats in flight. These results suggest that M. tuberculata do not attempt to reduce the cost of terrestrial locomotion and call production through biomechanical linkage. They also suggest that the pattern of linkage seen when bats are in flight is not universal and that energetic savings cannot necessarily be explained by contraction of muscles associated with the downstroke alone.

The long and short of it: branch lengths and the problem of placing the New Zealand short-tailed bat.

The taxonomic position of the endemic New Zealand bat genus Mystacina has vexed systematists ever since its erection in 1843. Over the years the genus has been linked with many microchiropteran families and superfamilies. Most recent classifications place it in the Vespertilionoidea, although some immunological evidence links it with the Noctilionoidea (=Phyllostomoidea). We have sequenced 402 bp of the mitochondrial cytochrome b gene for M. tuberculata (Gray in Dieffenbach, 1843), and using both our own and published DNA sequences for taxa in both superfamilies, we applied different tree reconstruction methods to find the appropriate phylogeny and different methods of estimating confidence in the parts of the tree. All methods strongly support the classification of Mystacina in the Noctilionoidea. Spectral analysis suggests that parsimony analysis may be misleading for Mystacina's precise placement within the Noctilionoidea because of its long terminal branch. Analyses not susceptible to long-branch attraction suggest that the Mystacinidae is a sister family to the Phyllostomidae. Dating the divergence times between the different taxa suggests that the extant chiropteran families radiated around and shortly after the Cretaceous–Tertiary boundary. We discuss the biogeographical implications of classifying Mystacina within the Noctilionoidea and contrast our result with those classifications placing Mystacina in the Vespertilionoidea, concluding that evidence for the latter is weak.

The Long and Short of It: Branch Lengths and the Problem of Placing the New Zealand Short-Tailed Bat, Mystacina

The taxonomic position of the endemic New Zealand bat genus Mystacina has vexed systematists ever since its erection in 1843. Over the years the genus has been linked with many microchiropteran families and superfamilies. Most recent classifications place it in the Vespertilionoidea, although some immunological evidence links it with the Noctilionoidea (=Phyllostomoidea). We have sequenced 402 bp of the mitochondrial cytochrome b gene for M. tuberculata (Gray in Dieffenbach, 1843), and using both our own and published DNA sequences for taxa in both superfamilies, we applied different tree reconstruction methods to find the appropriate phylogeny and different methods of estimating confidence in the parts of the tree. All methods strongly support the classification of Mystacina in the Noctilionoidea. Spectral analysis suggests that parsimony analysis may be misleading for Mystacina's precise placement within the Noctilionoidea because of its long terminal branch. Analyses not susceptible to long-branch attraction suggest that the Mystacinidae is a sister family to the Phyllostomidae. Dating the divergence times between the different taxa suggests that the extant chiropteran families radiated around and shortly after the Cretaceous-Tertiary boundary. We discuss the biogeographical implications of classifying Mystacina within the Noctilionoidea and contrast our result with those classifications placing Mystacina in the Vespertilionoidea, concluding that evidence for the latter is weak.