De-novo design of complementary (antisense) peptide mini-receptor inhibitor of interleukin 18 (IL-18).

Complementary (antisense) peptide mini-receptor inhibitors are complementary peptides designed to be receptor-surrogates that act by binding to selected surface features of biologically important proteins thereby inhibiting protein-cognate receptor interactions and subsequent biological effects. Previously, we described a complementary peptide mini-receptor inhibitor of interleukin-1beta (IL-1beta) that was designed to bind to an external surface loop (beta-bulge) of IL-1beta (Boraschi loop) clearly identified in the X-ray crystal structure of this cytokine. Here, we report the de-novo design and rational development of a complementary peptide mini-receptor inhibitor of cytokine interleukin-18 (IL-18), a protein for which there is no known X-ray crystal structure. Using sequence homology comparisons with IL-1beta, putative IL-18 surface loops are identified and used as a starting point for design, including a loop region 1 thought to be equivalent with the Boraschi loop of IL-1beta. Only loop region 1 complementary peptides are found to be promising leads as mini-receptor inhibitors of IL-18 but these are prevented from being properly successful owing to solubility problems. The application of "M-I pair mutagenesis" and inclusion of a C-terminal arginine residue are then sufficient to solve this problem and convert one lead peptide into a functional complementary peptide mini-receptor inhibitor of IL-18. This suggests that the biophysical and biological properties of complementary peptides can be improved in a rational and logical manner where appropriate, further strengthening the potential importance of complementary peptides as inhibitors of protein-protein interactions, even when X-ray crystal structural information is not readily available

Influence of wind-speed on short-duration NO2 measurements using Palmes and Ogawa passive diffusion samplers

We assessed the precision and accuracy of nitrogen dioxide (NO2) concentrations over 2-day, 3-day and 7-day exposure periods measured with the following types of passive diffusion samplers: standard (open) Palmes tubes; standard Ogawa samplers with commercially-prepared Ogawa absorbent pads (Ogawa[S]); and modified Ogawa samplers with absorbent-impregnated stainless steel meshes normally used in Palmes tubes (Ogawa[P]). We deployed these passive samplers close to the inlet of a chemiluminescence NO2 analyser at an urban background site in Glasgow, UK over 32 discrete measurement periods. Duplicate relative standard deviation was < 7% for all passive samplers. The Ogawa[P], Ogawa[S] and Palmes samplers explained 93%, 87% and 58% of temporal variation in analyser concentrations respectively. Uptake rates for Palmes and Ogawa[S] samplers were positively and linearly associated with wind-speed (P < 0.01 and P < 0.05 respectively). Computation of adjusted uptake rates using average wind-speed observed during each sampling period increased the variation in analyser concentrations explained by Palmes and Ogawa[S] estimates to 90% and 92% respectively, suggesting that measurements can be corrected for shortening of diffusion path lengths due to wind-speed to improve the accuracy of estimates of short-term NO2 exposure. Monitoring situations where it is difficult to reliably estimate wind-speed variations, e.g. across multiple sites with different unknown exposures to local winds, and personal exposure monitoring, are likely to benefit from protection of these sampling devices from the effects of wind, for example by use of a mesh or membrane across the open end. The uptake rate of Ogawa[P] samplers was not associated with wind-speed resulting in a high correlation between estimated concentrations and observed analyser concentrations. The use of Palmes meshes in Ogawa[P] samplers reduced the cost of sampler preparation and removed uncertainty associated with the unknown manufacturing process for the commercially-prepared collection pads

Consistency of Urban Background Black Carbon Concentration Measurements by Portable AE51 and Reference AE22 Aethalometers: Effect of Corrections for Filter Loading

Monitoring exposure to black carbon with portable devices is an important part of researching the health impacts of combustion-related air pollutants. We collected 786 hourly averaged equivalent black carbon (eBC) measurements from co-located duplicate portable AE51 Aethalometers and a UK Government reference AE22 Aethalometer (the data for the latter were corrected for filter darkening effects using a standard procedure), at an urban background site in Glasgow, UK. The AE51 and the reference concentrations were highly correlated (R 2 ≥ 0.87) for the combined deployment periods. The application of a previously reported method for correcting the AE51’s underestimation of concentrations, associated with filter loading, generally led to an overestimation of values (specifically, the normalised mean bias values for the two AE51s increased from –2% and +3% to +14% and +25% across the full range of measurements after correction). We found only limited and inconsistent evidence that the AE51 Aethalometers (attenuation [AE51_ATN] ≤ ~52) underestimated the eBC concentrations compared to the reference measurements. Thus, our observations indicate that the AE51 can achieve close agreement with the reference AE22 monitor without applying corrections for filter loading at relatively low AE51_ATN values in environments with low eBC concentrations

Estimation of spatial patterns of urban air pollution over a 4-week period from repeated 5-min measurements

Determination of intra-urban spatial variations in air pollutant concentrations for exposure assessment requires substantial time and monitoring equipment. The objective of this study was to establish if short-duration measurements of air pollutants can be used to estimate longer-term pollutant concentrations. We compared 5-min measurements of black carbon (BC) and particle number (PN) concentrations made once per week on 5 occasions, with 4 consecutive 1-week average nitrogen dioxide (NO2) concentrations at 18 locations at a range of distances from busy roads in Glasgow, UK. 5-min BC and PN measurements (averaged over the two 5-min periods at the start and end of a week) explained 40 - 80%, and 7 - 64% respectively, of spatial variation in the intervening 1-week NO2 concentrations for individual weeks. Adjustment for variations in background concentrations increased the percentage of explained variation in the bivariate relationship between the full set of NO2 and BC measurements over the 4-week period from 28% to 50% prior to averaging of repeat measurements. The averages of five 5-min BC and PN measurements made over 5 weeks explained 75% and 33% respectively of the variation in average 1-week NO2 concentrations over the same period. The relatively high explained variation observed between BC and NO2 measured on different time scales suggests that, with appropriate steps to correct or average out temporal variations, repeated short-term measurements can be used to provide useful information on longer-term spatial patterns for these traffic-related pollutants

Temporal changes in field calibration relationships for Aeroqual S500 O3 and NO2 sensor-based monitors

Sensor-based monitors are increasingly used to measure air pollutant concentrations, but require calibration under field conditions. We made intermittent comparisons (6 times over 6-month period) between ozone and nitrogen dioxide concentrations measured by Aeroqual gas-sensitive semiconductor (O3) and electrochemical (NO2) sensors (two of each) and reference analysers in the UK Automatic Urban and Rural Network. Each deployment period was split into equal (n = 48 x1-hour) training and test datasets, to derive and test calibration equations respectively. We observed significant bivariate linear relationships between Aeroqual O3 and Reference O3 concentrations, and significant multiple linear relationships between Aeroqual NO2 and both Reference NO2 and Aeroqual O3 concentrations. Changes in monitor responses over time (including apparent baseline drift in O3 sensor output, and discrepancies between the 2 Aeroqual NO2 sensors) resulted in relatively inaccurate concentrations estimates (cf. reference concentrations) from calibration equations derived in the first training period and applied to subsequent test deployments (e.g. NO2 RMSE = 47.2 μg m-3 (n = 286) for a dataset of all test periods combined, for one of the two monitor pairs). Substantial improvements in accuracy of estimated concentrations were achieved by combination of repeated intermittent training data into a single calibration dataset (NO2 RMSE = 8.5 μg m-3 for same test dataset described above). This latter approach to field calibration is recommended