Detecting variable (V), diversity (D) and joining (J) gene segment recombination using a two-colour fluorescence system

<p>Abstract</p> <p>Background</p> <p>Diversity of immunoglobulins and the T cell antigen receptors is achieved via the recombination activating gene (RAG)-mediated rearrangement of variable (V), diversity (D) and joining (J) gene segments, and this underpins the efficient recognition of a seemingly limitless array of antigens. Analysis of V(D)J recombination activity is typically performed using extrachromosomal recombination substrates that are recovered from transfected cells and selected using bacterial transformation. We have developed a two-colour fluorescence-based system that simplifies detection of both deletion and inversion joining events mediated by RAG proteins.</p> <p>Results</p> <p>This system employs two fluorescent reporter genes that differentially mark unrearranged substrates and those that have undergone RAG-mediated deletion or inversion events. The recombination products bear the hallmarks of true V(D)J recombination and activity can be detected using fluorescence microscopy or flow cytometry. Recombination events can be detected without the need for cytotoxic selection of recombination products and the system allows analysis of recombination activity using substrates integrated into the genome.</p> <p>Conclusions</p> <p>This system will be useful in the analysis and exploitation of the V(D)J recombination machinery and suggests that similar approaches could be used to replace expression of one gene with another during lymphocyte development.</p

Simple guide to starting a research group

Conducting cutting-edge research and scholarship becomes more complicated with each passing year; forming a collaborative research group offers a way to navigate this increasing complexity. Yet many individuals whose work might benefit from the formation of a collaborative team may feel overwhelmed by the prospect of attempting to build and maintain a research group. We propose this simple guide for starting and maintaining such an enterprise

The network structure of visited locations according to geotagged social media photos

Businesses, tourism attractions, public transportation hubs and other points of interest are not isolated but part of a collaborative system. Making such collaborative network surface is not always an easy task. The existence of data-rich environments can assist in the reconstruction of collaborative networks. They shed light into how their members operate and reveal a potential for value creation via collaborative approaches. Social media data are an example of a means to accomplish this task. In this paper, we reconstruct a network of tourist locations using fine-grained data from Flickr, an online community for photo sharing. We have used a publicly available set of Flickr data provided by Yahoo! Labs. To analyse the complex structure of tourism systems, we have reconstructed a network of visited locations in Europe, resulting in around 180,000 vertices and over 32 million edges. An analysis of the resulting network properties reveals its complex structure.Comment: 8 pages, 3 figure

Tumour Cell Generation of Inducible Regulatory T-Cells in Multiple Myeloma Is Contact-Dependent and Antigen-Presenting Cell-Independent

Regulatory T-cells (TReg cells) are increased in patients with multiple myeloma (MM). We investigated whether MM cells could generate and/or expand TReg cells as a method of immuno-surveillance avoidance. In an in vitro model, CD4+CD25-FoxP3- T-cells co-cultured with malignant plasma cells (primary MM cells and cell lines) induced a significant generation of CD4+CD25+FoxP3+ inducible TReg cells (tTReg cells; p<0.0001), in a contact-dependent manner. tTReg cells were polyclonal, demonstrated a suppressive phenotype and phenotypically, demonstrated increased FoxP3 (p = 0.0001), increased GITR (p<0.0001), increased PD1 (p = 0.003) and decreased CD62L (p = 0.007) expression compared with naturally occurring TReg cells. FACS-sorted tTReg cells differentiated into FoxP+IL-17+ and FoxP3-IL-17+ CD4+ cells upon TCR-mediated stimulation. Blocking experiments with anti-ICOS-L MoAb resulted in a significant inhibition of tTReg cell generation whereas both IL-10 & TGFβ blockade did not. MM tumour cells can directly generate functional TReg cells in a contact-dependent manner, mediated by ICOS/ICOS-L. These features suggest that tumour generation of TReg cells may contribute to evasion of immune surveillance by the host

Canopy nitrogen, carbon assimilation, and albedo in temperate and boreal forests: Functional relations and potential climate feedbacks

The availability of nitrogen represents a key constraint on carbon cycling in terrestrial ecosystems, and it is largely in this capacity that the role of N in the Earth\u27s climate system has been considered. Despite this, few studies have included continuous variation in plant N status as a driver of broad-scale carbon cycle analyses. This is partly because of uncertainties in how leaf-level physiological relationships scale to whole ecosystems and because methods for regional to continental detection of plant N concentrations have yet to be developed. Here, we show that ecosystem CO2 uptake capacity in temperate and boreal forests scales directly with whole-canopy N concentrations, mirroring a leaf-level trend that has been observed for woody plants worldwide. We further show that both CO2 uptake capacity and canopy N concentration are strongly and positively correlated with shortwave surface albedo. These results suggest that N plays an additional, and overlooked, role in the climate system via its influence on vegetation reflectivity and shortwave surface energy exchange. We also demonstrate that much of the spatial variation in canopy N can be detected by using broad-band satellite sensors, offering a means through which these findings can be applied toward improved application of coupled carbon cycle–climate models

The Co-Production of Sustainable Future Scenarios

Scenarios are a tool to develop plausible, coherent visions about the future and to foster anticipatory knowledge. We present the Sustainable Future Scenarios (SFS) framework and demonstrate its application through the Central Arizona-Phoenix Long-term Ecological Research (CAP LTER) urban site. The SFS approach emphasizes the co-development of positive and long-term alternative future visions. Through a collaboration of practitioner and academic stakeholders, this research integrates participatory scenario development, modeling, and qualitative scenario assessments. The SFS engagement process creates space to question the limits of what is normally considered possible, desirable, or inevitable in the face of future challenges. Comparative analyses among the future scenarios demonstrate trade-offs among regional and microscale temperature, water use, land-use change, and co-developed resilience and sustainability indices. SFS incorporate diverse perspectives in co-producing positive future visions, thereby expanding traditional future projections. The iterative, interactive process also creates opportunities to bridge science and policy by building anticipatory and systems-based decision-making and research capacity for long-term sustainability planning

Before the Pandemic Ends: Making Sure This Never Happens Again

Introduction On 30 January 2020, the World Health Organization (WHO) declared a Global Health Emergency of international concern attendant to the emergence and spread of SARS-CoV-2, nearly two months after the first reported emergence of human cases in Wuhan, China. In the subsequent two months, global, national and local health personnel and infrastructures have been overwhelmed, leading to suffering and death for infected people, and the threat of socio-economic instability and potential collapse for humanity as a whole. This shows that our current and traditional mode of coping, anchored in responses after the fact, is not capable of dealing with the crisis of emerging infectious disease. Given all of our technological expertise, why is there an emerging disease crisis, and why are we losing the battle to contain and diminish emerging diseases? Part of the reason is that the prevailing paradigm explaining the biology of pathogen-host associations (coevolution, evolutionary arms races) has assumed that pathogens must evolve new capacities - special mutations – in order to colonize new hosts and produce emergent disease (e.g. Parrish and Kawaoka, 2005). In this erroneous but broadly prevalent view, the evolution of new capacities creates new opportunities for pathogens. Further, given that mutations are both rare and undirected, the highly specialized nature of pathogen-host relationships should produce an evolutionary firewall limiting dissemination; by those definitions, emergences should be rare (for a historical review see Brooks et al., 2019). Pathogens, however, have become far better at finding us than our traditional understanding predicts. We face considerable risk space for pathogens and disease that directly threaten us, our crops and livestock – through expanding interfaces bringing pathogens and hosts into increasing proximity, exacerbated by environmental disruption and urban density, fueled by globalized trade and travel. We need a new paradigm that explains what we are seeing. Additional section headers: The Stockholm Paradigm The DAMA Protocol A Sense of Urgency and Long-Term Commitment Reference

Cultural and economic complementarities of spatial agglomeration in the British television broadcasting industry: Some explorations.

This paper considers the processes supporting agglomeration in the British television broadcasting industry. It compares and contrasts the insights offered by the cultural turn in geography and more conventionally economic approaches. It finds that culture and institutions are fundamental to the constitution of production and exchange relationships and also that they solve fundamental economic problems of coordinating resources under conditions of uncertainty and limited information. Processes at a range of spatial scales are important, from highly local to global, and conventional economics casts some light on which firms are most active and successful

The Bathypelagic Biome of the Atlantic Ocean: Character and Ecological Discreteness of the Fish Fauna

Recent global synthetic analyses have revealed that marine taxonomic inventories are far from complete, nowhere more so than in the deep-pelagic ocean. At over a billion km3, it is the largest biome on Earth, yet only a tiny fraction of the biogeographic records include the bathypelagic fauna. This data gap served as the impetus for recent deepwater surveys, many of which have altered our perceptions of pelagic ecosystems. Here we examine data from four deep-pelagic (0-5000+ m) sampling programs in the Atlantic (60°N-25°S) in order to assess the character of bathypelagic fish communities with respect to faunal distinctiveness and ecological connectivity. Regions studied include the Gulf of Mexico, Sargasso 702 Sea, eastern North/South Atlantic, and mid-North Atlantic. Quantitative analyses give contrasting pictures with respect to faunal composition and ecosystem operation. The discreteness of the bathypelagic zone is exhibited faunistically by the suite of ―holobathypelagic‖ species found only below 1000 m, most of which are highly modified morphologically. Geometric abundance class analyses reveal that the character of relative species abundance distributions between the meso- and bathypelagic zones is fundamentally dissimilar; the former exhibit a much higher proportion of common species, while the latter exhibit a much higher percentage of rarer species. From a community energetics perspective, however, the bathy- and mesopelagic zones are highly interconnected. Approximately 70% of fish species collected below 1000 m are also found in the mesopelagic zone, and in the far North Atlantic, are also found in the epipelagial. These species comprised 66 to \u3e90% of individuals collected below 1000 m in the regions sampled. In the mid-North Atlantic, these species contribute to the unexpected water-column biomass maximum observed between 1500-2300 m. Thus, the ―transient‖ taxa (primarily mesopelagic migrators and spanner taxa) add considerably to the ichthyofaunal diversity of the world ocean below 1000 m, and appear to be the vectors that support the diverse array of holobathypelagic fishes whose taxonomic composition is dominated by piscivores. Data from the four regions studied suggests that classic pelagic biogeographic boundaries do not apply to bathypelagic realm, as shared species are the rule rather than the exception. Last, cumulative species curves suggest we are far from understanding the true complexity of the bathypelagic zone