Rapid fabrication of glass/PDMS hybrid µIMER for high throughput membrane proteomics

Mass spectrometry (MS) based proteomics has brought a radical approach to systems biology, offering a platform to study complex biological functions. However, key proteomic technical challenges remain, mainly the inability to characterise the complete proteome of a cell due to the thousands of diverse, complex proteins expressed at an extremely wide concentration range. Currently, high throughput and efficient techniques to unambiguously identify and quantify proteins on a proteome-wide scale are in demand. Miniaturised analytical systems placed upstream of MS help us to attain these goals. One time-consuming step in traditional techniques is the in-solution digestion of proteins (4-20 h). This also has other drawbacks, including enzyme autoproteolysis, low efficiency, and manual operation. Furthermore, the identification of α-helical membrane proteins has remained a challenge due to their high hydrophobicity and lack of trypsin cleavage targets in transmembrane helices. We demonstrate a new rapidly produced glass/PDMS micro Immobilised Enzyme Reactor (μIMER) with enzymes covalently immobilised onto polyacrylic acid plasma-modified surfaces for the purpose of rapidly (as low as 30 s) generating peptides suitable for MS analysis. This μIMER also allows, for the first time, rapid digestion of insoluble proteins. Membrane protein identification through this method was achieved after just 4 min digestion time, up to 9-fold faster than either dual-stage in-solution digestion approaches or other commonly used bacterial membrane proteomic workflows.     

Parallel receivers and sparse sampling in multidimensional NMR

The recent introduction of NMR spectrometers with multiple receivers permits spectra from several different nuclear species to be recorded in parallel, and several standard pulse sequences to be combined into a single entity. It is shown how these improvements in the flow and quality of spectral information can be significantly augmented by compressive sensing techniques--controlled aliasing, Hadamard spectroscopy, single-point evaluation of evolution space (SPEED), random sampling, projection-reconstruction, and hyperdimensional NMR. Future developments of these techniques are confidently expected to mitigate one of the most serious limitations in multidimensional NMR--the excessive duration of the measurements.     

Evidence for shape coexistence at medium spin in Rb

Four previously known rotational bands in ⁷⁶Rb have been extended to moderate spins using the Gammasphere and Microball γ ray and charged particle detector arrays and the ⁴⁰Ca(⁴⁰Ca, 3pn) reaction at a beam energy of 165 MeV. The properties of two of the negative-parity bands can only readily be interpreted in terms of the highly successful Cranked Nilsson-Strutinsky model calculations if they have the same configuration in terms of the number of g9/2 particles, but they result from different nuclear shapes (one near-oblate and the other near-prolate). These data appear to constitute a unique example of shape coexisting structures at medium spins.     

Using phase-type models to cost stroke patient care across health,social and community services

Stroke disease places a heavy burden on society, incurring long periods of time in hospital and community care, and associated costs. Also stroke is a highly complex disease with diverse outcomes and multiple strategies for therapy and care. Previously a modeling framework has been developed which clusters patients into classes with respect to their length of stay (LOS) in hospital. Phase-type models were then used to describe patient flows for each cluster. Also multiple outcomes, such as discharge to normal residence, nursing home, or death can be permitted. We here add costs to this model and obtain the Moment Generating Function for the total cost of a system consisting of multiple transient phase-type classes with multiple absorbing states. This system represents different classes of patients in different hospital and community services states. Based on stroke patients’ data from the Belfast City Hospital, various scenarios are explored with a focus on comparing the cost of thrombolysis treatment under different regimes. The overall modeling framework characterizes the behavior of stroke patient populations, with a focus on integrated system-wide costing and planning, encompassing hospital and community services. Within this general framework we have developed models which take account of patient heterogeneity and multiple care options. Such complex strategies depend crucially on developing a deep engagement with the health care professionals and underpinning the models with detailed patient-specific data.     

On the role of reachability and observability in NMR experimentation

It is shown that the system theoretic concepts of reachability and observability are relevant to the analysis of NMR experiments. Moreover, the sets of reachable states are examined and Lie theoretic criteria are given for the reachability of the system. The question is investigated how the set of reachable states depends on the class of input functions that are allowed. Both one‐dimensional and multi‐dimensional NMR experiments are considered. This revised version was published online in July 2006 with corrections to the Cover Date.     

HOW DOES THE WAY IN WHICH INDIVIDUAL STUDENTS BEHAVE AFFECT THE SHARED CONSTRUCTION OF MEANING?

Audio taped discussions between three students have been examined to shed light on the way in which the behaviour of individual students may affect the shared construction of meaning with graphical calculators. These discussions revealed a complex pattern of interaction between the students. Each student was responsible for defining his or her own role within the discourse and these roles appeared to change as the discussion progressed. With reference to the framework offered by Winbourne and Watson (1998), it is proposed that local communities of practice have been established and that the individual student's positioning within the community of practice determines their success as a learner and contributes towards the creation of shared knowledge.