Robust 3D DNA FISH using directly labeled probes.

3D DNA FISH has become a major tool for analyzing three-dimensional organization of the nucleus, and several variations of the technique have been published. In this article we describe a protocol which has been optimized for robustness, reproducibility, and ease of use. Brightly fluorescent directly labeled probes are generated by nick-translation with amino-allyldUTP followed by chemical coupling of the dye. 3D DNA FISH is performed using a freeze-thaw step for cell permeabilization and a heating step for simultaneous denaturation of probe and nuclear DNA. The protocol is applicable to a range of cell types and a variety of probes (BACs, plasmids, fosmids, or Whole Chromosome Paints) and allows for high-throughput automated imaging. With this method we routinely investigate nuclear localization of up to three chromosomal regions

RNA-binding proteins ZFP36L1 and ZFP36L2 promote cell quiescence.

Progression through the stages of lymphocyte development requires coordination of the cell cycle. Such coordination ensures genomic integrity while cells somatically rearrange their antigen receptor genes [in a process called variable-diversity-joining (VDJ) recombination] and, upon successful rearrangement, expands the pools of progenitor lymphocytes. Here we show that in developing B lymphocytes, the RNA-binding proteins (RBPs) ZFP36L1 and ZFP36L2 are critical for maintaining quiescence before precursor B cell receptor (pre-BCR) expression and for reestablishing quiescence after pre-BCR-induced expansion. These RBPs suppress an evolutionarily conserved posttranscriptional regulon consisting of messenger RNAs whose protein products cooperatively promote transition into the S phase of the cell cycle. This mechanism promotes VDJ recombination and effective selection of cells expressing immunoglobulin-μ at the pre-BCR checkpoint.This work was funded by the Biotechnology and Biological Sciences Research Council, a Medical Research Council CASE studentship with GSK, an MRC centenary award (A.G) and project grants from Bloodwise. DJH was supported by a Medical Research Council Clinician Scientist FellowshipThis is the author accepted manuscript. The final version is available from the American Association for the Advancement of Science via http://dx.doi.org/10.1126/science.aad597

Myc dynamically and preferentially relocates to a transcription factory occupied by Igh.

Transcription in mammalian nuclei is highly compartmentalized in RNA polymerase II-enriched nuclear foci known as transcription factories. Genes in cis and trans can share the same factory, suggesting that genes migrate to preassembled transcription sites. We used fluorescent in situ hybridization to investigate the dynamics of gene association with transcription factories during immediate early (IE) gene induction in mouse B lymphocytes. Here, we show that induction involves rapid gene relocation to transcription factories. Importantly, we find that the Myc proto-oncogene on Chromosome 15 is preferentially recruited to the same transcription factory as the highly transcribed Igh gene located on Chromosome 12. Myc and Igh are the most frequent translocation partners in plasmacytoma and Burkitt lymphoma. Our results show that transcriptional activation of IE genes involves rapid relocation to preassembled transcription factories. Furthermore, the data imply a direct link between the nonrandom interchromosomal organization of transcribed genes at transcription factories and the incidence of specific chromosomal translocations

Crop Updates 2011 - Nutrition, Precision Agriculture & Climate and Forecasting

This session covers sixteen papers from different authors: Nutrition 1. Balance® used in conventional cropping practice with half of the upfront fertiliser rate can sustain crop yield and build soil biological fertility, Deb Archdeacon1, Andrew Gulliver2 and David Cullen2, 1Agronomica, Wellington Mill, WA, 2Custom Composts, Nambeelup, WA 2. Effects of potassium (K) supply on plant growth, potassium uptake and grain Yield in wheat grown in grey sand, Qifu Ma1, Richard Bell1, Ross Brennan2 and Craig Scanlan2, 1School of Environmental Science, Murdoch University, 2Department of Agriculture and Food 3. Improving fertiliser management: redefining the relationship between soil tests and crop responses for wheat in WA, Wen Chen1, 2, Ross Brennan2, Geoff Anderson2, Richard Bell1 and Mike Bolland2, 1School of Environmental Science, Murdoch University, 2Department of Agriculture and Food 4. Improved phosphorus and potassium management: redefining the soil test and lupin response relationships in WA, Wen Chen1, 2, Ross Brennan2, Geoff Anderson2, Richard Bell1 and Mike Bolland2, 1School of Environmental Science, Murdoch University, Western Australia, 2Department of Agriculture and Food 5. Converting phosphorus retention index (PRI) to phosphorus buffering index (PBI) for Western Australian soils, Peter Rees and Sandy Alexander, Summit Fertilizers 6. Variability of radiometric potassium and Colwell potassium relationships across the Great Southern, Frank D’Emden, Precision Agronomics Australia 7. Rotary spading and mouldboard ploughing of water-repellent sandplain soils fulfils promise, Stephen Davies, Craig Scanlan and Breanne Best, Department of Agriculture and Food 8. Soil nitrous oxide (N2O) fluxes are low from a grain legume crop grown in a semi-arid climate Louise Barton1, Klaus Butterbach-Bahl2, Ralph Kiese2 and Daniel Murphy1, 1 School of Earth & Environment, University of Western Australia, 2 Karlsruhe Institute of Technology, Institute for Meteorology & Climate Research, Garmisch-Partenkirchen, Germany, 9. Mouldboard ploughing of sandplain soils – more grain, fewer weeds, Peter Newman Department of Agriculture and Food Precision Agriculture 10.What’s preventing growers from implementing precision agriculture (PA)? Roger Mandel1, Roger Lawes2 and Michael Robertson2, 1Curtin University, 2CSIRO 11. On how many paddocks does precision agriculture (PA) deliver a return? Roger Lawes1, Michael Robertson1 and Roger Mandel2, 1CSIRO Ecosystem Sciences, Floreat, WA, 2Curtin University 12. Demonstration pf precision agriculture (PA) principles in the Great Southern, Western Australia, Derk Bakker1, Jeremy Lemon1, Alison Lacey1, John Paul Collins1, Roger Mandel2, Frank D’Emden3, Glen Riethmuller1, 1Department of Agriculture and Food, 2Curtin University, 3Precision Agronomics Australia Climate and Forecasting 13. Statistical seasonal rainfall forecasts for south west Australia, Fiona H Evans Department of Agriculture of Food 14. How has changing climate recently affected Western Australia’s capacity to increase crop productivity and water use efficiency? David Stephens, Department of Agriculture and Food 15. Is Yield Prophet® a useful tool in Western Australia? — an agribusiness perspective, Caroline Peek, Department of Agriculture and Food 16. A season of Yield Prophet® — how it saw the dry, Tim Scanlon and Caroline Peek Department of Agriculture of Foo

The adjuvant GLA-SE promotes human Tfh cell expansion and emergence of public TCRβ clonotypes

The generation of protective humoral immunity after vaccination relies on the productive interaction between antigen-specific B cells and T follicular helper (Tfh) cells. Despite the central role of Tfh cells in vaccine responses, there is currently no validated way to enhance their differentiation in humans. From paired human lymph node and blood samples, we identify a population of circulating Tfh cells that are transcriptionally and clonally similar to germinal center Tfh cells. In a clinical trial of vaccine formulations, circulating Tfh cells were expanded in Tanzanian volunteers when an experimental malaria vaccine was adjuvanted in GLA-SE but not when formulated in Alum. The GLA-SE–formulated peptide was associated with an increase in the extrafollicular antibody response, long-lived antibody production, and the emergence of public TCRβ clonotypes in circulating Tfh cells. We demonstrate that altering vaccine adjuvants is a rational approach for enhancing Tfh cells in humans, thereby supporting the long-lived humoral immunity that is required for effective vaccines.</jats:p

Diverse values of nature for sustainability

Twenty-five years since foundational publications on valuing ecosystem services for human well-being(1,2), addressing the global biodiversity crisis(3) still implies confronting barriers to incorporating nature's diverse values into decision-making. These barriers include powerful interests supported by current norms and legal rules such as property rights, which determine whose values and which values of nature are acted on. A better understanding of how and why nature is (under)valued is more urgent than ever(4). Notwithstanding agreements to incorporate nature's values into actions, including the Kunming-Montreal Global Biodiversity Framework (GBF)(5) and the UN Sustainable Development Goals(6), predominant environmental and development policies still prioritize a subset of values, particularly those linked to markets, and ignore other ways people relate to and benefit from nature(7). Arguably, a 'values crisis' underpins the intertwined crises of biodiversity loss and climate change(8), pandemic emergence(9) and socio-environmental injustices(10). On the basis of more than 50,000 scientific publications, policy documents and Indigenous and local knowledge sources, the Intergovernmental Platform on Biodiversity and Ecosystem Services (IPBES) assessed knowledge on nature's diverse values and valuation methods to gain insights into their role in policymaking and fuller integration into decisions(7,11). Applying this evidence, combinations of values-centred approaches are proposed to improve valuation and address barriers to uptake, ultimately leveraging transformative changes towards more just (that is, fair treatment of people and nature, including inter- and intragenerational equity) and sustainable futures

Diverse values of nature for sustainability

Twenty-five years since foundational publications on valuing ecosystem services for human well-being1,2, addressing the global biodiversity crisis3 still implies confronting barriers to incorporating nature’s diverse values into decision-making. These barriers include powerful interests supported by current norms and legal rules such as property rights, which determine whose values and which values of nature are acted on. A better understanding of how and why nature is (under)valued is more urgent than ever4. Notwithstanding agreements to incorporate nature’s values into actions, including the Kunming-Montreal Global Biodiversity Framework (GBF)5 and the UN Sustainable Development Goals6, predominant environmental and development policies still prioritize a subset of values, particularly those linked to markets, and ignore other ways people relate to and benefit from nature7. Arguably, a ‘values crisis’ underpins the intertwined crises of biodiversity loss and climate change8, pandemic emergence9 and socio-environmental injustices10. On the basis of more than 50,000 scientific publications, policy documents and Indigenous and local knowledge sources, the Intergovernmental Platform on Biodiversity and Ecosystem Services (IPBES) assessed knowledge on nature’s diverse values and valuation methods to gain insights into their role in policymaking and fuller integration into decisions7,11. Applying this evidence, combinations of values-centred approaches are proposed to improve valuation and address barriers to uptake, ultimately leveraging transformative changes towards more just (that is, fair treatment of people and nature, including inter- and intragenerational equity) and sustainable futures

Effect of simplifications of bone and components inclination on the elastohydrodynamic lubrication modeling of metal-on-metal hip resurfacing prosthesis

It is important to study the lubrication mechanism of metal-on-metal hip resurfacing prosthesis in order to understand its overall tribological performance, thereby minimize the wear particles. Previous elastohydrodynamic lubrication studies of metal-on-metal hip resurfacing prosthesis neglected the effects of the orientations of the cup and head. Simplified pelvic and femoral bone models were also adopted for the previous studies. These simplifications may lead to unrealistic predictions. For the first time, an elastohydrodynamic lubrication model was developed and solved for a full metal-onmetal hip resurfacing arthroplasty. The effects of the orientations of components and the realistic bones on the lubrication performance of metal-on-metal hip resurfacing prosthesis were investigated by comparing the full model with simplified models. It was found that the orientation of the head played a very important role in the prediction of pressure distributions and film profiles of the metal-on-metal hip resurfacing prosthesis. The inclination of the hemispherical cup up to 45 had no appreciable effect on the lubrication performance of the metal-on-metal hip resurfacing prosthesis. Moreover, the combined effect of material properties and structures of bones was negligible. Future studies should focus on higher inclination angles, smaller coverage angle and microseparation related to the occurrences of edge loading. © IMechE 2012