John Andrews : Architecture a Performing Art

The distinguished Australian architect surveys his career and examines how his architectural theories are expressed in his designs.

Large-scale interlaboratory study to develop, analytically validate and apply highly multiplexed, quantitative peptide assays to measure cancer-relevant proteins in plasma

There is an increasing need in biology and clinical medicine to robustly and reliably measure tens-to-hundreds of peptides and proteins in clinical and biological samples with high sensitivity, specificity, reproducibility and repeatability. Previously, we demonstrated that LC-MRM-MS with isotope dilution has suitable performance for quantitative measurements of small numbers of relatively abundant proteins in human plasma, and that the resulting assays can be transferred across laboratories while maintaining high reproducibility and quantitative precision. Here we significantly extend that earlier work, demonstrating that 11 laboratories using 14 LC-MS systems can develop, determine analytical figures of merit, and apply highly multiplexed MRM-MS assays targeting 125 peptides derived from 27 cancer-relevant proteins and 7 control proteins to precisely and reproducibly measure the analytes in human plasma. To ensure consistent generation of high quality data we incorporated a system suitability protocol (SSP) into our experimental design. The SSP enabled real-time monitoring of LC-MRM-MS performance during assay development and implementation, facilitating early detection and correction of chromatographic and instrumental problems. Low to sub-nanogram/mL sensitivity for proteins in plasma was achieved by one-step immunoaffinity depletion of 14 abundant plasma proteins prior to analysis. Median intra- and inter-laboratory reproducibility was <20%, sufficient for most biological studies and candidate protein biomarker verification. Digestion recovery of peptides was assessed and quantitative accuracy improved using heavy isotope labeled versions of the proteins as internal standards. Using the highly multiplexed assay, participating laboratories were able to precisely and reproducibly determine the levels of a series of analytes in blinded samples used to simulate an inter-laboratory clinical study of patient samples. Our study further establishes that LC-MRM-MS using stable isotope dilution, with appropriate attention to analytical validation and appropriate quality c`ontrol measures, enables sensitive, specific, reproducible and quantitative measurements of proteins and peptides in complex biological matrices such as plasma.

Genome-wide association study identifies eight loci associated with blood pressure

Elevated blood pressure is a common, heritable cause of cardiovascular disease worldwide. To date, identification of common genetic variants influencing blood pressure has proven challenging. We tested 2.5 million genotyped and imputed SNPs for association with systolic and diastolic blood pressure in 34,433 subjects of European ancestry from the Global BPgen consortium and followed up findings with direct genotyping (N 71,225 European ancestry, N 12,889 Indian Asian ancestry) and in silico comparison (CHARGE consortium, N = 29,136). We identified association between systolic or diastolic blood pressure and common variants in eight regions near the CYP17A1 (P = 7 × 10 24), CYP1A2 (P = 1 × 10 23), FGF5 (P = 1 × 10 21), SH2B3 (P = 3 × 10 18), MTHFR (P = 2 × 10 13), c10orf107 (P = 1 × 10 9), ZNF652 (P = 5 × 10 9) and PLCD3 (P = 1 × 10 8) genes. All variants associated with continuous blood pressure were associated with dichotomous hypertension. These associations between common variants and blood pressure and hypertension offer mechanistic insights into the regulation of blood pressure and may point to novel targets for interventions to prevent cardiovascular disease.

Association scan of 14,500 nonsynonymous SNPs in four diseases identifies autoimmunity variants

We have genotyped 14,436 nonsynonymous SNPs (nsSNPs) and 897 major histocompatibility complex (MHC) tag SNPs from 1,000 independent cases of ankylosing spondylitis (AS), autoimmune thyroid disease (AITD), multiple sclerosis (MS) and breast cancer (BC). Comparing these data against a common control dataset derived from 1,500 randomly selected healthy British individuals, we report initial association and independent replication in a North American sample of two new loci related to ankylosing spondylitis, ARTS1 and IL23R, and confirmation of the previously reported association of AITD with TSHR and FCRL3. These findings, enabled in part by increased statistical power resulting from the expansion of the control reference group to include individuals from the other disease groups, highlight notable new possibilities for autoimmune regulation and suggest that IL23R may be a common susceptibility factor for the major 'seronegative' diseases.

Localization of type 1 diabetes susceptibility to the MHC class I genes HLA-B and HLA-A

The major histocompatibility complex (MHC) on chromosome 6 is associated with susceptibility to more common diseases than any other region of the human genome, including almost all disorders classified as autoimmune. In type 1 diabetes the major genetic susceptibility determinants have been mapped to the MHC class II genes HLA-DQB1 and HLA-DRB1 (refs 1–3), but these genes cannot completely explain the association between type 1 diabetes and the MHC region4, 5, 6, 7, 8, 9, 10, 11. Owing to the region's extreme gene density, the multiplicity of disease-associated alleles, strong associations between alleles, limited genotyping capability, and inadequate statistical approaches and sample sizes, which, and how many, loci within the MHC determine susceptibility remains unclear. Here, in several large type 1 diabetes data sets, we analyse a combined total of 1,729 polymorphisms, and apply statistical methods—recursive partitioning and regression...

Konstruktiivisen linjakkuuden perusteita etsimässä : Systemaattinen analyysi John Biggsin esittämistä yliopisto-opetuksen lähtökohdista

The aim of this study is to explore by systematic textual analysis the crucial conceptions of constructive alignment and to reconstruct the concept of constructive alignment and examine the relation between conceptual relationships in John Biggs’s texts. In this study, I have also analyzed the presuppositions of the concept of constructive alignment and its possible implications. The research material includes Biggs’s (1996b; 2003) article entitled Enhancing Teaching through Constructive Alignment and book entitled Teaching for Quality Learning at University. The primary purpose of the systematic textual analysis is to reconstruct concepts and gain access to a new or more profound understanding of the concepts. The main purpose of the constructive alignment is to design a teaching system that supports and encourages students to adopt a deep approach learning. At the center of the constructive alignment are two concepts: constructivism in learning and alignment in teaching. A tension was detected between these concepts. Biggs assumes that students’ learning activities are primed by the teaching. Because of this it is not important what the teacher does. At the same time he emphasizes that teaching interacts with learning. The teacher’s task is to support student’s appropriate learning activities. On the basis of the analysis, I conclude these conceptions are not mutually exclusive. Interaction between teaching and learning has an effect on student’s learning activities. The most essential benefit of the model of constructive alignment is that Biggs brings together and considers teaching at the same level with learning. A weakness of Biggs’s model relates to the theoretical basis and positions of the concept of constructive alignment. There are some conflicts between conceptions of epistemology in Biggs’s texts. In addition, Biggs writes about constructivism also as conceptions of epistemology, but doesn’t consider implications of that position or what follows or doesn’t follow from that commitment. On the basis of the analysis, I suggest that constructivism refers in Biggs’s texts rather to constructivism in learning than philosophical constructivism. In light of this study, constructive alignment doesn´t lead to philosophical constructivism. That’s why constructive alignment stays out of idealism. Biggs’s way of thinking about teachers possibility to confronting students’ misconceptions and evaluate and assess students’ constructions support a realist purpose in terms of philosophical stance. Realism does not drift toward general problems of relativism, like lack of criteria for assessing or evaluate these constructions.

Estimating cell diffusivity and cell proliferation rate by interpreting IncuCyte ZOOM™ assay data using the Fisher-Kolmogorov model

Background: Standard methods for quantifying IncuCyte ZOOM™ assays involve measurements that quantify how rapidly the initially-vacant area becomes re-colonised with cells as a function of time. Unfortunately, these measurements give no insight into the details of the cellular-level mechanisms acting to close the initially-vacant area. We provide an alternative method enabling us to quantify the role of cell motility and cell proliferation separately. To achieve this we calibrate standard data available from IncuCyte ZOOM™ images to the solution of the Fisher-Kolmogorov model. Results: The Fisher-Kolmogorov model is a reaction-diffusion equation that has been used to describe collective cell spreading driven by cell migration, characterised by a cell diffusivity, D, and carrying capacity limited proliferation with proliferation rate, λ, and carrying capacity density, K. By analysing temporal changes in cell density in several subregions located well-behind the initial position of the leading edge we estimate λ and K. Given these estimates, we then apply automatic leading edge detection algorithms to the images produced by the IncuCyte ZOOM™ assay and match this data with a numerical solution of the Fisher-Kolmogorov equation to provide an estimate of D. We demonstrate this method by applying it to interpret a suite of IncuCyte ZOOM™ assays using PC-3 prostate cancer cells and obtain estimates of D, λ and K. Comparing estimates of D, λ and K for a control assay with estimates of D, λ and K for assays where epidermal growth factor (EGF) is applied in varying concentrations confirms that EGF enhances the rate of scratch closure and that this stimulation is driven by an increase in D and λ, whereas K is relatively unaffected by EGF. Conclusions: Our approach for estimating D, λ and K from an IncuCyte ZOOM™ assay provides more detail about cellular-level behaviour than standard methods for analysing these assays. In particular, our approach can be used to quantify the balance of cell migration and cell proliferation and, as we demonstrate, allow us to quantify how the addition of growth factors affects these processes individually.

John and Ursula Schlosstein nee Gottschalk with their infant son Ralph. portraits family

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John Ettinger Collection 1941-1946

John Ettinger, 1987.

Association analyses of 249,796 individuals reveal 18 new loci associated with body mass index

Obesity is globally prevalent and highly heritable, but its underlying genetic factors remain largely elusive. To identify genetic loci for obesity susceptibility, we examined associations between body mass index and approximately 2.8 million SNPs in up to 123,865 individuals with targeted follow up of 42 SNPs in up to 125,931 additional individuals. We confirmed 14 known obesity susceptibility loci and identified 18 new loci associated with body mass index (P < 5 x 10(-)(8)), one of which includes a copy number variant near GPRC5B. Some loci (at MC4R, POMC, SH2B1 and BDNF) map near key hypothalamic regulators of energy balance, and one of these loci is near GIPR, an incretin receptor. Furthermore, genes in other newly associated loci may provide new insights into human body weight regulation.

John and Ursula Schlosstein nee Gottschalk with their infant son Ralph. portraits family

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Meta-analysis identifies 13 new loci associated with waist-hip ratio and reveals sexual dimorphism in the genetic basis of fat distribution

Waist-hip ratio (WHR) is a measure of body fat distribution and a predictor of metabolic consequences independent of overall adiposity. WHR is heritable, but few genetic variants influencing this trait have been identified. We conducted a meta-analysis of 32 genome-wide association studies for WHR adjusted for body mass index (comprising up to 77,167 participants), following up 16 loci in an additional 29 studies (comprising up to 113,636 subjects). We identified 13 new loci in or near RSPO3, VEGFA, TBX15-WARS2, NFE2L3, GRB14, DNM3-PIGC, ITPR2-SSPN, LY86, HOXC13, ADAMTS9, ZNRF3-KREMEN1, NISCH-STAB1 and CPEB4 (P = 1.9 × 10−9 to P = 1.8 × 10−40) and the known signal at LYPLAL1. Seven of these loci exhibited marked sexual dimorphism, all with a stronger effect on WHR in women than men (P for sex difference = 1.9 × 10−3 to P = 1.2 × 10−13). These findings provide evidence for multiple loci that modulate body fat distribution independent of overall adiposity and reveal strong gene-by-sex interactions.

Nineteenth century narratives reveal historic catch rates for Australian snapper (Pagrus auratus)

Snapper (Pagrus auratus) is widely distributed throughout subtropical and temperate southern oceans and forms a significant recreational and commercial fishery in Queensland, Australia. Using data from government reports, media sources, popular publications and a government fisheries survey carried out in 1910, we compiled information on individual snapper fishing trips that took place prior to the commencement of fisherywide organized data collection, from 1871 to 1939. In addition to extracting all available quantitative data, we translated qualitative information into bounded estimates and used multiple imputation to handle missing values, forming 287 records for which catch rate (snapper fisher−1 h−1) could be derived. Uncertainty was handled through a parametric maximum likelihood framework (a transformed trivariate Gaussian), which facilitated statistical comparisons between data sources. No statistically significant differences in catch rates were found among media sources and the government fisheries survey. Catch rates remained stable throughout the time series, averaging 3.75 snapper fisher−1 h−1 (95% confidence interval, 3.42–4.09) as the fishery expanded into new grounds. In comparison, a contemporary (1993–2002) south-east Queensland charter fishery produced an average catch rate of 0.4 snapper fisher−1 h−1 (95% confidence interval, 0.31–0.58). These data illustrate the productivity of a fishery during its earliest years of development and represent the earliest catch rate data globally for this species. By adopting a formalized approach to address issues common to many historical records – missing data, a lack of quantitative information and reporting bias – our analysis demonstrates the potential for historical narratives to contribute to contemporary fisheries management.