Gene expression and splicing alterations analyzed by high throughput RNA sequencing of chronic lymphocytic leukemia specimens.

BackgroundTo determine differentially expressed and spliced RNA transcripts in chronic lymphocytic leukemia specimens a high throughput RNA-sequencing (HTS RNA-seq) analysis was performed.MethodsTen CLL specimens and five normal peripheral blood CD19+ B cells were analyzed by HTS RNA-seq. The library preparation was performed with Illumina TrueSeq RNA kit and analyzed by Illumina HiSeq 2000 sequencing system.ResultsAn average of 48.5 million reads for B cells, and 50.6 million reads for CLL specimens were obtained with 10396 and 10448 assembled transcripts for normal B cells and primary CLL specimens respectively. With the Cuffdiff analysis, 2091 differentially expressed genes (DEG) between B cells and CLL specimens based on FPKM (fragments per kilobase of transcript per million reads and false discovery rate, FDR q < 0.05, fold change >2) were identified. Expression of selected DEGs (n = 32) with up regulated and down regulated expression in CLL from RNA-seq data were also analyzed by qRT-PCR in a test cohort of CLL specimens. Even though there was a variation in fold expression of DEG genes between RNA-seq and qRT-PCR; more than 90 % of analyzed genes were validated by qRT-PCR analysis. Analysis of RNA-seq data for splicing alterations in CLL and B cells was performed by Multivariate Analysis of Transcript Splicing (MATS analysis). Skipped exon was the most frequent splicing alteration in CLL specimens with 128 significant events (P-value <0.05, minimum inclusion level difference >0.1).ConclusionThe RNA-seq analysis of CLL specimens identifies novel DEG and alternatively spliced genes that are potential prognostic markers and therapeutic targets. High level of validation by qRT-PCR for a number of DEG genes supports the accuracy of this analysis. Global comparison of transcriptomes of B cells, IGVH non-mutated CLL (U-CLL) and mutated CLL specimens (M-CLL) with multidimensional scaling analysis was able to segregate CLL and B cell transcriptomes but the M-CLL and U-CLL transcriptomes were indistinguishable. The analysis of HTS RNA-seq data to identify alternative splicing events and other genetic abnormalities specific to CLL is an added advantage of RNA-seq that is not feasible with other genome wide analysis

Specialized dynamical properties of promiscuous residues revealed by simulated conformational ensembles

The ability to interact with different partners is one of the most important features in proteins. Proteins that bind a large number of partners (hubs) have been often associated with intrinsic disorder. However, many examples exist of hubs with an ordered structure, and evidence of a general mechanism promoting promiscuity in ordered proteins is still elusive. An intriguing hypothesis is that promiscuous binding sites have specific dynamical properties, distinct from the rest of the interface and pre-existing in the protein isolated state. Here, we present the first comprehensive study of the intrinsic dynamics of promiscuous residues in a large protein data set. Different computational methods, from coarse-grained elastic models to geometry-based sampling methods and to full-atom Molecular Dynamics simulations, were used to generate conformational ensembles for the isolated proteins. The flexibility and dynamic correlations of interface residues with a different degree of binding promiscuity were calculated and compared considering side chain and backbone motions, the latter both on a local and on a global scale. The study revealed that (a) promiscuous residues tend to be more flexible than nonpromiscuous ones, (b) this additional flexibility has a higher degree of organization, and (c) evolutionary conservation and binding promiscuity have opposite effects on intrinsic dynamics. Findings on simulated ensembles were also validated on ensembles of experimental structures extracted from the Protein Data Bank (PDB). Additionally, the low occurrence of single nucleotide polymorphisms observed for promiscuous residues indicated a tendency to preserve binding diversity at these positions. A case study on two ubiquitin-like proteins exemplifies how binding promiscuity in evolutionary related proteins can be modulated by the fine-tuning of the interface dynamics. The interplay between promiscuity and flexibility highlighted here can inspire new directions in protein-protein interaction prediction and design methods. © 2013 American Chemical Society

The Genomic Analysis of Erythrocyte microRNA Expression in Sickle Cell Diseases

BACKGROUND: Since mature erythrocytes are terminally differentiated cells without nuclei and organelles, it is commonly thought that they do not contain nucleic acids. In this study, we have re-examined this issue by analyzing the transcriptome of a purified population of human mature erythrocytes from individuals with normal hemoglobin (HbAA) and homozygous sickle cell disease (HbSS). METHODS AND FINDINGS: Using a combination of microarray analysis, real-time RT-PCR and Northern blots, we found that mature erythrocytes, while lacking ribosomal and large-sized RNAs, contain abundant and diverse microRNAs. MicroRNA expression of erythrocytes was different from that of reticulocytes and leukocytes, and contributed the majority of the microRNA expression in whole blood. When we used microRNA microarrays to analyze erythrocytes from HbAA and HbSS individuals, we noted a dramatic difference in their microRNA expression pattern. We found that miR-320 played an important role for the down-regulation of its target gene, CD71 during reticulocyte terminal differentiation. Further investigation revealed that poor expression of miR-320 in HbSS cells was associated with their defective downregulation CD71 during terminal differentiation. CONCLUSIONS: In summary, we have discovered significant microRNA expression in human mature erythrocytes, which is dramatically altered in HbSS erythrocytes and their defect in terminal differentiation. Thus, the global analysis of microRNA expression in circulating erythrocytes can provide mechanistic insights into the disease phenotypes of erythrocyte diseases

Materials and Molecular Modelling at the Exascale

Progression of computational resources towards exascale computing makes possible simulations of unprecedented accuracy and complexity in the fields of materials and molecular modelling (MMM), allowing high fidelity in silico experiments on complex materials of real technological interest. However, this presents demanding challenges for the software used, especially the exploitation of the huge degree of parallelism available on exascale hardware, and the associated problems of developing effective workflows and data management on such platforms. As part of the UKs ExCALIBUR exascale computing initiative, the UK-led MMM Design and Development Working Group has worked with the broad MMM community to identify a set of high priority application case studies which will drive future exascale software developments. We present an overview of these case studies, categorized by the methodological challenges which will be required to realize them on exascale platforms, and discuss the exascale requirements, software challenges and impact of each application area

Crop Updates 2000 - Pulses

This session covers fifty nine papers from different authors: 1.1999 PULSE INDUSTRY HIGHLIGHTS 2. CONTRIBUTORS 3. BACKGROUND 4. SUMMARY OF PREVIOUS RESULTS 5. 1999 REGIONAL ROUNDUP 6. Northern Agricultural Region, W. O’Neill, AGWEST 7. Central Agricultural Region J. Russell and R.J. French AGWEST 8. Great Southern and Lakes N. Brandon, C. Gaskin and N. Runciman, AGWEST 9. Esperance Mallee M. Seymour, AGWEST PULSE PRODUCTION AGRONOMY AND GENETIC IMPROVEMENT 10. Faba Bean 11. Desi chickpea Traits associated with drought resistance in chickpea, J. Berger, N.C. Turner, CLIMA and CSIRO Plant Industry, R.J. French, AGWEST, R. Carpenter, C. Ludwig and R. Kenney, CSIRO Plant Industry 12. Genotype x environment analysis of chickpea adaptation, J. Berger and N. Turner, CLIMA and CSIRO Plant Industry, and K.H.M. Siddique, AGWEST 13. Carbon fixation by chickpea pods under terminal drought, Q. Ma, CLIMA, M.H. Behboudian, Massey University, New Zealand, N.C. Turner and J.A. Palta, CLIMA, and CSIRO Plant Industry 14. Influence of terminal drought on growth and seed quality, M.H. Behboudian, Massey University, New Zealand, Q. Ma, CLIMA, N.C. Turner and J.A. Palta, CSIRO Plant Industry 15. Resistance to chilling at flowering and to budworm, H. Clarke, CLIMA Chickpea nodulation survey, J. Stott and J. Howieson, Centre for Rhizobium Studies, Murdoch University 16. Kabuli chickpea 17. Premium quality kabuli chickpea development in the ORIA, K.H.M. Siddique CLIMA and AGWEST, K.L. Regan, AGWEST, R. Shackles, AGWEST 18. International screening for Ascochyta blight resistance, K.H.M. Siddique CLIMA and AGWEST, C. Francis, CLIMA, K.L. Regan, AGWEST, N. Acikgoz and N. Atikyilmaz, AARI, Turkey and R.S. Malholtra, ICARDA, Syria 19. Agronomic evaluation of Ascochyta resistant kabuli germplasm in WA, K.H.M. Siddique CLIMA and AGWESTC. Francis, CLIMA, K.L. Regan and M. Baker, AGWEST 20. Field Pea 21. Lentil 22. ACIAR project J. Clements, K.H.M. Siddique CLIMA and AGWEST and C. Francis CLIMA 23. Vetch 24. Rust, M. Seymour, AGWEST 25. Narbon bean 26. Agronomy, M. Seymour, AGWEST 27. Lupinus species 28. Screening lupins for tolerance to alkaline/calcareous soils, C. Tang, CLIMA andUniversity of WAand J.D. Brand, WAITE, University of Adelaide 29. Lathyrus development, C. Hanbury and K.H.M. Siddique, CLIMA and AGWEST 30. Sheep feeding studies, C. White, CSIRO, Perth, C. Hanbury, CLIMA and K.H.M. Siddique, CLIMA and AGWEST 31. Lathyrus: a potential new ingredient in pig diets, B.P. Mullan, C.D. Hanbury and K.H.M. Siddique, AGWEST 32. Species comparison 33. Species for horticultural rotations, K.H.M. Siddique, AGWEST, R. Lancaster and I. Guthridge AGWEST 34. Marrow fat field pea shows promise in the southwest, K.H.M. Siddique, AGWEST, N. Runciman, AGWEST, and I. Pritchard, AGWEST, 35. Pulses on grey clay soils, P. Fisher, M. Braimbridge, J. Bignell, N. Brandon, R. Beermier, W. Bowden, AGWEST 36. Nutrient management of pulses 37. Summary of pulse nutrition studies in WA, M.D.A. Bolland, K.H.M. Siddique, G.P. Riethmuller, and R.F. Brennan, AGWEST 38. Pulse species response to phosphorus and zinc, S. Lawrence, Zed Rengel, University of WA, S.P. Loss, CSBP futurefarm, M.D.A. Bolland, .H.M. Siddique, W. Bowden, AGWEST 39. Gypsum 40. Antitranspirants seed priming DEMONSTRATION OF PULSES IN THE FARMING SYSTEM 41. Foliar and soil applied nutrients for field peas in the south coast mallee,M. Seymour, AGWEST, and P. Vedeniapine, Phosyn Ltd 42. Demonstration of pulse species at Kendenup, C. Kirkwood, Farmer, Katanning, R. Beermier, N. Runciman and N. Brandon, AGWEST 43. Kabuli chickpea demonstration at Gnowangerup, R. Beermier and N. Brandon, AGWEST 44. Lathyrus sativus demonstration at Mindarabin, N. Brandon and R. Beermier, AGWEST 45. New field pea varieties in the central eastern region, J. Russell, AGWEST DISEASE AND PEST MANAGEMENT 46. Ascochyta blight of chickpea 47. Botrytis grey mould (BGM) of chickpea 48. Fungal disease diagnostics, Pulse disease diagnostics, D. Wright, AGWEST Plant Laboratories 49. Viruses in pulses, Luteovirus infection in field pea and faba bean crops, and viruses in seed, L. Latham, CLIMA and AGWEST, R. Jones, AGWEST 50. Screening of pulse species for pea seed-borne mosaic virus, L. Latham, CLIMAand AGWEST, and R. Jones, AGWEST 51. CMV in chickpea: effect of seed-borne sources on virus spread and seed yield, R. Jones, AGWEST and L. Latham, CLIMA and AGWEST 52. Insect pests 53. Evaluation of transgenic field pea against the pea weevil,M.J. de Sousa Majer, School of Environmental Biology, Curtin University of Technology,, D. Hardie, and N.C. Turner, CSIRO Division of Plant Industry 54. Development of a molecular marker for pea weevil resistance in field pea, Oonagh Byrne, CLIMA, Darryl Hardie, AGWEST and Penny Smith, UWA 55. Aphid feeding damage to faba bean and lentil crops, Françoise Berlandier, AGWEST 56. Taxonomy and control of bruchids in pulses, N. Keals, CLIMA, D. Hardie and R. Emery, AGWEST, 57. ACKNOWLEDGMENTS 58. PUBLICATIONS BY PULSE PRODUCTIVITY PROJECT STAFF 59. VARIETIES PRODUCED AND COMMERCIALLY RELEASE

Epithelial cell polarity: a major gatekeeper against cancer?

The correct establishment and maintenance of cell polarity are crucial for normal cell physiology and tissue homeostasis. Conversely, loss of cell polarity, tissue disorganisation and excessive cell growth are hallmarks of cancer. In this review, we focus on identifying the stages of tumoural development that are affected by the loss or deregulation of epithelial cell polarity. Asymmetric division has recently emerged as a major regulatory mechanism that controls stem cell numbers and differentiation. Links between cell polarity and asymmetric cell division in the context of cancer will be examined. Apical–basal polarity and cell–cell adhesion are tightly interconnected. Hence, how loss of cell polarity in epithelial cells may promote epithelial mesenchymal transition and metastasis will also be discussed. Altogether, we present the argument that loss of epithelial cell polarity may have an important role in both the initiation of tumourigenesis and in later stages of tumour development, favouring the progression of tumours from benign to malignancy