Recreating Bioenergetic Elements In Protein Maquettes

In all organisms, protein-mediated electron transfers underlie energy metabolism and countless other critical metabolic pathways. Deciphering the factors governing electron transfers offers enormous practical value, including finding reliable guidelines for metabolic engineering to produce clean solar fuels. Protein maquettes provide simple, flexible scaffolds to study biological electron transfer processes. The maquette approach to protein design builds man-made oxidoreductases from first principles with minimal reference to natural protein sequences. This scheme has produced impressive in vitro and in vivo results to replicate the functions of natural oxidoreductases using simple, straightforward α-helical bundles. The redox function of maquettes has so far been limited to electron transfers within a single water-soluble molecule. This work extends the maquette project to examine electron transfers A) between diffusing redox partners and B) in amphiphilic proteins. Nature employs complimentary electrostatic surfaces to bring diffusing redox proteins together, and a series of maquettes of varying surface charge demonstrate that the same principles apply to flexible manmade proteins. A heme-binding maquette of complimentary surface charge is shown to reduce natural cytochrome c at physiological rates, while a maquette variant similar in charge to cyt c shows a far weaker interaction. The ionic strength dependence of these interactions is shown to be broadly similar to that in natural proteins. This work also presents the first intra-protein redox function in a de novo amphiphilic protein and describes progress toward transmembrane electron transfer analogous to that in the cytochrome bc1 complex. These functional achievements are remarkable given that the proteins are designed from first principles without atomically resolved structures, and they hold promise for future efforts in applying artificial proteins to new metabolic pathways

Patients' preferences for the management of non-metastatic prostate cancer: discrete choice experiment

Objective To establish which attributes of conservative treatments for prostate cancer are most important to men. Design Discrete choice experiment. Setting Two London hospitals. Participants 129 men with non-metastatic prostate cancer, mean age 70 years; 69 of 118 (58%) with T stage 1 or 2 cancer at diagnosis. Main outcome measures Men's preferences for, and trade-offs between, the attributes of diarrhoea, hot flushes, ability to maintain an erection, breast swelling or tenderness, physical energy, sex drive, life expectancy, and out of pocket expenses. Results The men's responses to changes in attributes were all statistically significant. When asked to assume a starting life expectancy of five years, the men were willing to make trade-offs between life expectancy and side effects. On average, they were most willing to give up life expectancy to avoid limitations in physical energy (mean three months) and least willing to trade life expectancy to avoid hot flushes (mean 0.6 months to move from a moderate to mild level or from mild to none). Conclusions Men with prostate cancer are willing to participate in a relatively complex exercise that weighs up the advantages and disadvantages of various conservative treatments for their condition. They were willing to trade off some life expectancy to be relieved of the burden of troublesome side effects such as limitations in physical energy

Transverse field-induced nucleation pad switching modes during domain wall injection

We have used magnetic transmission soft X-ray microscopy (M-TXM) to image in-field magnetization configurations of patterned Ni80F20 domain wall "nucleation pads" with attached planar nanowires. Comparison with micromagnetic simulations suggests that the evolution of magnetic domains in rectangular injection pads depends on the relative orientation of closure domains in the remanent state. The magnetization reversal pathway is altered by the inclusion of transverse magnetic fields. These different modes explain previous results of domain wall injection into nanowires

Direct imaging of domain-wall interactions in Ni80Fe20 planar nanowires

We have investigated magnetostatic interactions between domain walls in Ni80Fe20 planar nanowires using magnetic soft x-ray microscopy and micromagnetic simulations. In addition to significant monopole-like attraction and repulsion effects we observe that there is coupling of the magnetization configurations of the walls. This is explained in terms of an interaction energy that depends not only on the distance between the walls, but also upon their internal magnetization structure

A systematic review of the effectiveness of adalimumab, etanercept and infliximab for the treatment of rheumatoid arthritis in adults and an economic evaluation of their cost-effectiveness

Objectives: This report reviews the clinical effectiveness and cost-effectiveness of adalimumab, etanercept and infliximab, agents that inhibit tumour necrosis factor-a (TNF-a), when used in the treatment of rheumatoid arthritis (RA) in adults. \ud \ud Data sources: Electronic databases were searched up to February 2005. \ud \ud Review methods: Systematic reviews of the literature on effectiveness and cost-effectiveness were undertaken and industry submissions to the National Institute for Health and Clinical Excellence (NICE) were reviewed. Meta-analyses of effectiveness data were also undertaken for each agent. The Birmingham Rheumatoid Arthritis Model (BRAM), a simulation model, was further developed and used to produce an incremental cost-effectiveness analysis. \ud \ud Results: Twenty-nine randomised controlled trials (RCTs), most of high quality, were included. The only head-to-head comparisons were against methotrexate. For patients with short disease duration (≤3 years) who were naïve to methotrexate, adalimumab was marginally less and etanercept was marginally more effective than methotrexate in reducing symptoms of RA. Etanercept was better tolerated than methotrexate. Both adalimumab and etanercept were more effective than methotrexate in slowing radiographic joint damage. Etanercept was also marginally more effective and better tolerated than methotrexate in patients with longer disease durations who had not failed methotrexate treatment. Infliximab is only licensed for use with methotrexate. All three agents, either alone (where so licensed) or in combination with ongoing disease-modifying antirheumatic drugs (DMARDs), were effective in reducing the symptoms and signs of RA in patients with established disease. At the licensed dose, the numbers needed to treat (NNTs) (95% CI) required to produce an American College for Rheumatology (ACR) response compared with placebo were: ACR20: adalimumab 3.6 (3.1 to 4.2), etanercept 2.1 (1.9 to 2.4), infliximab 3.2 (2.7 to 4.0); ACR50: adalimumab 4.2 (3.7 to 5.0), etanercept 3.1 (2.7 to 3.6), infliximab 5.0 (3.8 to 6.7); and ACR70: adalimumab 7.7 (5.9 to 11.1), etanercept 7.7 (6.3 to 10.0), infliximab 11.1 (7.7 to 20.0). In patients who were naïve to methotrexate, or who had not previously failed methotrexate treatment, a TNF inhibitor combined with methotrexate was significantly more effective than methotrexate alone. Infliximab combined with methotrexate had an increased risk of serious infections. All ten published economic evaluations met standard criteria for quality, but the incremental cost-effectiveness ratios (ICERs) ranged from being within established thresholds to being very high because of varying assumptions and parameters. All three sponsors who submitted economic models made assumptions favourable to their product. BRAM incorporates improvements in quality of life and mortality, but assumes no effect of TNF inhibitors on joint replacement. For use in accordance with current NICE guidance as the third DMARD in a sequence of DMARDs, the base-case ICER was around £30,000 per quality-adjusted life-year (QALY) in early RA and £50,000 per QALY in late RA. Sensitivity analyses showed that the results were sensitive to the estimates of Health Assessment Questionnaire (HAQ) progression while on TNF inhibitors and the effectiveness of DMARDs, but not to changes in mortality ratios per unit HAQ. TNF inhibitors are most cost-effective when used last. The ICER for etanercept used last is £24,000 per QALY, substantially lower than for adalimumab (£30,000 per QALY) or infliximab (£38,000 per QALY). First line use as monotherapy generates ICERs around £50,000 per QALY for adalimumab and etanercept. Using the combination of methotrexate and a TNF inhibitor as first line treatment generates much higher ICERs, as it precludes subsequent use of methotrexate, which is cheap. The ICERs for sequential use are of the same order as using the TNF inhibitor alone. \ud \ud Conclusions: Adalimumab, etanercept and infliximab are effective treatments compared with placebo for RA patients who are not well controlled by conventional DMARDs, improving control of symptoms, improving physical function, and slowing radiographic changes in joints. The combination of a TNF inhibitor with methotrexate was more effective than methotrexate alone in early RA, although the clinical relevance of this additional benefit is yet to be established, particularly in view of the well-established effectiveness of MTX alone. An increased risk of serious infection cannot be ruled out for the combination of methotrexate with adalimumab or infliximab. The results of the economic evaluation based on BRAM are consistent with the observations from the review of clinical effectiveness, including the ranking of treatments. TNF inhibitors are most cost-effective when used as last active therapy. In this analysis, other things being equal, etanercept may be the TNF inhibitor of choice, although this may also depend on patient preference as to route of administration. The next most cost-effective use of TNF inhibitors is third line, as recommended in the 2002 NICE guidance. Direct comparative RCTs of TNF inhibitors against each other and against other DMARDs, and sequential use in patients who have failed a previous TNF inhibitor, are needed. Longer term studies of the quality of life in patients with RA and the impact of DMARDs on this are needed, as are longer studies that directly assess effects on joint replacement, other morbidity and mortality

Observation of field-induced domain wall propagation in magnetic nanowires by magnetic transmission X-ray microscopy

Magnetic transmission X-ray microscopy (M-TXM) is used to image domain walls in magnetic ring structures formed by a 300 nm wide, 24 nm thick Ni{sub 81}Fe{sub 19} nanowire. Both transverse and vortex type domain walls are observed after application of different field sequences. Domain walls can be observed by comparing images obtained from opposite field sequences, or else domain wall propagation observed by comparing successive images in a particular field sequence. This demonstrates the potential use of M-TXM in developing and understanding planar magnetic nanowire behavior

Observation of field-induced domain wall propagation in magnetic nanowires by magnetic transmission X-ray microscopy

Abstract Magnetic transmission X-ray microscopy (M-TXM) is used to image domain walls in magneti

The Australian public's preferences for emergency care alternatives and the influence of the presenting context: a discrete choice experiment

Objectives The current study seeks to quantify the Australian public's preferences for emergency care alternatives and determine if preferences differ depending on presenting circumstances. Setting Increasing presentations to emergency departments have led to overcrowding, long waiting times and suboptimal health system performance. Accordingly, new service models involving the provision of care in alternative settings and delivered by other practitioners continue to be developed. Participants A stratified sample of Australian adults (n=1838), 1382 from Queensland and 456 from South Australia, completed the survey. This included 951 females and 887 males from the 2045 people who met the screening criteria out of the 4354 people who accepted the survey invitation. Interventions A discrete choice experiment was used to elicit preferences in the context of one of four hypothetical scenarios: a possible concussion, a rash/asthma-related problem involving oneself or one's child and an anxiety-related presentation. Mixed logit regression was used to analyse the dependent variable choice and identify the relative importance of care attributes and the propensity to access care in each context. Results Results indicated a preference for treatment by an emergency physician in hospital for possible concussion and treatment by a doctor in ambulatory settings for rash/asthma-related and anxiety-related problems. Participants were consistently willing to wait longer before making trade-offs in the context of the rash/asthma-related scenario compared with when the same problem affected their child. Results suggest a clear preference for lower costs, shorter wait times and strong emphasis on quality care; however, significant preference heterogeneity was observed. Conclusions This study has increased awareness that the public's emergency care choices will differ depending on the presenting context. It has further demonstrated the importance of service quality as a determinant of healthcare choices. The findings have also provided insights into the Australian public's reactions to emergency care reforms

A proteomics and transcriptomics investigation of the venom from the Barychelid spider Trittame loki (brush-foot trapdoor)

Although known for their potent venom and ability to prey upon both invertebrate and vertebrate species, the Barychelidae spider family has been entirely neglected by toxinologists. In striking contrast, the sister family Theraphosidae (commonly known as tarantulas), which last shared a most recent common ancestor with Barychelidae over 200 million years ago, has received much attention, accounting for 25% of all the described spider toxins while representing only 2% of all spider species. In this study, we evaluated for the first time the venom arsenal of a barychelid spider, Trittame loki, using transcriptomic, proteomic, and bioinformatic methods. The venom was revealed to be dominated by extremely diverse inhibitor cystine knot (ICK)/knottin peptides, accounting for 42 of the 46 full-length toxin precursors recovered in the transcriptomic sequencing. In addition to documenting differential rates of evolution adopted by different ICK/knottin toxin lineages, we discovered homologues with completely novel cysteine skeletal architecture. Moreover, acetylcholinesterase and neprilysin were revealed for the first time as part of the spider-venom arsenal and CAP (CRiSP/Allergen/PR-1) were identified for the first time in mygalomorph spider venoms. These results not only highlight the extent of venom diversification in this neglected ancient spider lineage, but also reinforce the idea that unique venomous lineages are rich pools of novel biomolecules that may have significant applied uses as therapeutics and/or insecticides