Effect of acyl chain length on transfection efficiency and toxicity of polyethylenimine

Polyethylenimine (PEI) is an efficient nonviral gene delivery vector because of its high buffering capacity and DNA condensation ability. In our study, the amino groups on the polymeric backbone were acylated using acetic or propionic anhydride to alter the protonation behaviour and the hydrophilic/hydrophobic balance of the polymer. The concentration of acylated primary amines was determined using trinitrobenzene sulphonic acid assay. Results showed that our modified polymers had lower buffering capacities in solutions compared to PEI. The polymers were complexed with plasmid encoding enhanced green fluorescent protein at three different ratios (1:1, 1:2 and 1:10 w/w DNA to polymer) to form polyplexes and their toxicities and transfection efficiencies were evaluated in HEK 293 cells. Acylation reduced the number of primary amines on the polymer and the surface charge, improving haemocompatibility and reducing cytotoxicity. The reduction in the concentration of amino groups helped to optimise DNA compaction and facilitated polyplex dissociation in the cell, which increased transfection efficiency of the modified polymers compared to the parent polymer. Polymers with buffering capacities greater than 50% and less than 80% relative to PEI, showed higher transfection efficiencies than PEI. The propionic anhydride modified polymers had appropriate interactions with DNA which provided both DNA compaction and polyplex dissociation. These systems interacted better with the cell membrane because of their slightly higher lipophilicity and formed polyplexes which were less cytotoxic than polyplexes of acetic anhydride modified polymers. Among the vectors tested, 1:0.3 mol/mol PEI:propionic anhydride in a 1:2 w/w DNA:polymer composition provided the best transfection system with improved transfection efficiency and reduced cytotoxicity.

Long-term change, interannual and intra-seasonal variability in climate and glacier mass balance in the Central Greater Caucasus, Russia

Long-term trends, interannual and intra-seasonal variability in the mass-balance record from Djankuat glacier, central Greater Caucasus, Russia, are related to local climate change, synoptic and large-scale anomalies in atmospheric circulation. A clear warming signal emerged in the central Greater Caucasus in the early 1990s, leading to a strong increase in ablation. In the absence of a compensating change in winter accumulation, the net mass balance of Djankuat has declined. The highest value of seasonal ablation on record was registered in the summer of 2000. At the beginning of the 21st century these trends reversed. Ablation was below average even in the summer of 2003, which was unusually warm in western Europe. Precipitation and winter accumulation were high, allowing for a partial recovery of net mass balance. The interannual variability in the components of mass balance is weakly related to the North Atlantic Oscillation (NAO) and the Scandinavian teleconnection patterns, but there is a clear link with the large-scale circulation anomalies represented by the Rossby pattern. Five synoptic categories have been identified for the ablation season of 2005, revealing a strong separation between components of radiation budget, air temperature and daily melt. Air temperature is the main control over melt. The highest values of daily ablation are related to the strongly positive NAO which forces high net radiation, and to the warm and moist advection from the Black Sea.

Quantifying uncertainty in critical loads: (B) acidity mass balance critical loads on a sensitive site

This paper reports an uncertainty analysis of critical loads for acid deposition for a site in southern England, using the Steady State Mass Balance Model. The uncertainty bounds, distribution type and correlation structure for each of the 18 input parameters was considered explicitly, and overall uncertainty estimated by Monte Carlo methods. Estimates of deposition uncertainty were made from measured data and an atmospheric dispersion model, and hence the uncertainty in exceedance could also be calculated. The uncertainties of the calculated critical loads were generally much lower than those of the input parameters due to a "compensation of errors" mechanism - coefficients of variation ranged from 13% for CLmaxN to 37% for CL(A). With 1990 deposition, the probability that the critical load was exceeded was > 0.99; to reduce this probability to 0.50, a 63% reduction in deposition is required; to 0.05, an 82% reduction. With 1997 deposition, which was lower than that in 1990, exceedance probabilities declined and uncertainties in exceedance narrowed as deposition uncertainty had less effect. The parameters contributing most to the uncertainty in critical loads were weathering rates, base cation uptake rates, and choice of critical chemical value, indicating possible research priorities. However, the different critical load parameters were to some extent sensitive to different input parameters. The application of such probabilistic results to environmental regulation is discussed.

Too much information? : Drama teachers’ perceptions of the value of subject knowledge

This paper reports on research into what drama teachers consider they really need to know as drama specialists. In the first instance the very concept of knowledge is discussed as it pertains to education in the arts as is the current situation in England regarding the extent to which new drama teachers’ subject specialist knowledge has been formally accredited and what the implications of this may be to an evolving curriculum. The research itself initially involved using a questionnaire to investigate the way in which drama teachers prioritised different aspects of professional knowledge. Results of this survey were deemed surprising enough to warrant further investigation through the use of interviews and a multiple-sorting exercise which revealed why the participants prioritised in the way they did. Informed by the work of Bourdieu, Foucault and Kelly, a model is proposed which may help explain the tensions experienced by drama teachers as they try to balance and prioritise different aspects of professional knowledge.

Carbon balance of UK peatlands: current state of knowledge and future research challenges

The retention of peatland carbon (C) and the ability to continue to draw down and store C from the atmosphere is not only important for the UK terrestrial carbon inventory, but also for a range of ecosystem services, the landscape value and the ecology and hydrology of ~15% of the land area of the UK. Here we review the current state of knowledge on the C balance of UK peatlands using several studies which highlight not only the importance of making good flux measurements, but also the spatial and temporal variability of different flux terms that characterise a landscape affected by a range of natural and anthropogenic processes and threats. Our data emphasise the importance of measuring (or accurately estimating) all components of the peatland C budget. We highlight the role of the aquatic pathway and suggest that fluxes are higher than previously thought. We also compare the contemporary C balance of several UK peatlands with historical rates of C accumulation measured using peat cores, thus providing a long-term context for present-day measurements and their natural year-on-year variability. Contemporary measurements from 2 sites suggest that current accumulation rates (–56 to –72 g C m–2 yr–1) are at the lower end of those seen over the last 150 yr in peat cores (–35 to –209 g C m–2 yr–1). Finally, we highlight significant current gaps in knowledge and identify where levels of uncertainty are high, as well as emphasise the research challenges that need to be addressed if we are to improve the measurement and prediction of change in the peatland C balance over future decades.

The Greenland Ice Sheet's surface mass balance in a seasonally sea-ice free Arctic

General circulation models predict a rapid decrease in sea ice extent with concurrent increases in near surface air temperature and precipitation in the Arctic over the 21st century. This has led to suggestions that some Arctic land ice masses may experience an increase in accumulation due to enhanced evaporation from a seasonally sea ice free Arctic Ocean. To investigate the impact of this phenomenon on Greenland ice sheet climate and surface mass balance (SMB) a regional climate model, HadRM3, was used to force an insolation-temperature melt SMB model. A set of experiments designed to investigate the role of sea ice independently from sea surface temperature (SST) forcing are described. In the warmer and wetter SI + SST simulation Greenland experiences a 23% increase in winter SMB but 65% reduced summer SMB, resulting in a net decrease in the annual value. This study shows that sea ice decline contributes to the increased winter balance, causing 25% of the increase in winter accumulation; this is largest in eastern Greenland as the result of increased evaporation in the Greenland Sea. These results indicate that the seasonal cycle of Greenland's SMB will increase dramatically as global temperatures increase, with the largest changes in temperature and precipitation occurring in winter. This demonstrates that the accurate prediction of changes in sea ice cover is important for predicting Greenland SMB and ice sheet evolution.

Agricultural commodity markets : reference point for the real value of a currency

Using monthly time-series data 1999-2013, the paper shows that markets for agricultural commodities provide a yardstick for real purchasing power, and thus a reference point for the real value of fiat currencies. The daily need for each adult to consume about 2800 food calories is universal; data from FAO food balance sheets confirm that the world basket of food consumed daily is non-volatile in comparison to the volatility of currency exchange rates, and so the replacement cost of food consumed provides a consistent indicator of economic value. Food commodities are storable for short periods, but ultimately perishable, and this exerts continual pressure for markets to clear in the short term; moreover, food calories can be obtained from a very large range of foodstuffs, and so most households are able to use arbitrage to select a near optimal weighting of quantities purchased. The paper proposes an original method to enable a standard of value to be established, definable in physical units on the basis of actual worldwide consumption of food goods, with an illustration of the method.

Scoring rules and competitive behavior in best value construction auctions

This paper examines the extent to which engineers can influence the competitive behavior of bidders in Best Value or multi-attribute construction auctions, where both the (dollar) bid and technical non-price criteria are scored according to a scoring rule. From a sample of Spanish construction auctions with a variety of bid scoring rules, it is found that bidders are influenced by the auction rules in significant and predictable ways. The bid score weighting, bid scoring formula and abnormally low bid criterion are variables likely to influence the competitiveness of bidders in terms of both their aggressive/conservative bidding and concentration/dispersion of bids. Revealing the influence of the bid scoring rules and their magnitude on bidders’ competitive behavior opens the door for the engineer to condition bidder competitive behavior in such a way as to provide the balance needed to achieve the owner’s desired strategic outcomes.