Construction of a "mutagenesis cartridge" for poliovirus genome-linked viral protein: Isolation and characterization of viable and nonviable mutants

By following a strategy of genetic analysis of poliovirus, we have constructed a synthetic "mutagenesis cartridge" spanning the genome-linked viral protein coding region and flanking cleavage sites in an infectious cDNA clone of the type 1 (Mahoney) genome. The insertion of new restriction sites within the infectious clone has allowed us to replace the wild-type sequences with short complementary pairs of synthetic oligonucleotides containing various mutations. A set of mutations have been made that create methionine codons within the genome-linked viral protein region. The resulting viruses have growth characteristics similar to wild type. Experiments that led to an alteration of the tyrosine residue responsible for the linkage to RNA have resulted in nonviable virus. In one mutant, proteolytic processing assayed in vitro appeared unimpaired by the mutation. We suggest that the position of the tyrosine residue is important for genome-linked viral protein function(s)

Putting Synthesis into Biology: A Viral View of Genetic Engineering through De Novo Gene and Genome Synthesis

SummaryThe rapid improvements in DNA synthesis technology hold the potential to revolutionize biosciences in the near future. Traditional genetic engineering methods are template dependent and make extensive but laborious use of site-directed mutagenesis to explore the impact of small variations on an existing sequence “theme.” De novo gene and genome synthesis frees the investigator from the restrictions of the pre-existing template and allows for the rational design of any conceivable new sequence theme.Viruses, being among the simplest replicating entities, have been at the forefront of the advancing biosciences since the dawn of molecular biology. Viral genomes, especially those of RNA viruses, are relatively short, often less than 10,000 bases long, making them amenable to whole genome synthesis with the currently available technology. For this reason viruses are once again poised to lead the way in the budding field of synthetic biology—for better or worse

Assessing Resilience of Pasture Production to Climatic Changes

Increasing temperatures and atmospheric carbon dioxide (CO2) concentrations, together with changes to rainfall patterns, will influence seasonal pasture production; however climate change projections for south eastern Australia are uncertain (CSIRO and BoM 2007). Despite this, climate change impact assessments generally rely on specific climate projections, but in this study an alternative approach was developed to test the resilience of production to incremental changes in climate

Comparison of Summer Forages and the Effect of Nitrogen Fertilizers on \u3ci\u3eBrassica\u3c/i\u3e Forages in Tasmania

Summer forage crops, and in particular Brassica spp., have become increasingly popular in dairy production systems in Tasmania. Field experiments were conducted for 3 years in northwestern Tasmania, in the spring/summers beginning in 1995. The study aimed to compare yield and quality of Brassica and Poaceae forages and the response of Brassica species to nitrogen (N) (50, 100 and 200 kg N/ha) and irrigation. The average total yields of dryland (rainfed) crops in 1995 to 1997 experiments, were turnip (Brassica rapa) 9.3 t/ha, rape (B. napus) 5.9 t/ha, oats (Aevena sativa) 5.2 t/ha, kale (B. oleracea) 5.1 t/ha, short-lived ryegrass (Lolium multiflorum) 5.1 t/ha, pasja (B. campestris × B. napus) 4.3 t/ha, perennial ryegrass (L. perenne) 4.2 t/ha, millet (Echinochloa utilis) 3.8 t/ha, and maize. (Zea mays) 2.9 t/ha. Irrigation increased the yield of turnips by 4.8 t/ha (mainly bulbs) and millet yields by 1.4 t/ha and reduced maize yield by 1.2 t/ha. Brassica species were higher in ME and lower in CP than the Poaceae forages. Nitrogen fertilizer increased the DM yield of tops of all Brassica crops in the 1997/98 experiments under irrigation, but it decreased the yield of turnips bulbs. The total yields with 50, 100 and 200 kg N/ha were 14, 15.2 and 15 t DM/ha for turnips, 7.5, 8.5 and 10 t for pasja and 10, 12 and 12.2 t DM/ha for rape, respectively. With 100 kg N/ha the average concentration of quality attributes of turnips, pasja and rape were CP 14, 22 and 19%, ME 12, 14.6 and 12.6 MJ/kg DM respectively. Nitrogen increased the CP, but had no effect on ME of any Brassica crops. Brassica forage are superior to Poaceae forages for summer feed production and as a part of pasture renovation process. They are higher in their yields, quality and water use efficiency and respond well to N fertilizer

The Use of Long-Term Modelling in Analysing N\u3csub\u3e2\u3c/sub\u3eO Abatement Strategies in Dairy Pastures

Concerns about the environmental impact of nitrogen (N) losses in Australia, in particular, nitrous oxide emissions are related to the rapid increase in N application on dairy pastures. Computer modelling is the most suitable method available to assess the potential of best management practices (BMP) to reduce field losses, as direct field measurements are frequently limited by the short term nature of many field trials

An evaluation of carbon offset supplementation options for beef production systems on coastal speargrass in central Queensland, Australia

In 2014, the Australian Government implemented the Emissions Reduction Fund to offer incentives for businesses to reduce greenhouse gas (GHG) emissions by following approved methods. Beef cattle businesses in northern Australia can participate by applying the 'reducing GHG emissions by feeding nitrates to beef cattle' methodology and the 'beef cattle herd management' methods. The nitrate (NO3) method requires that each baseline area must demonstrate a history of urea use. Projects earn Australian carbon credit units (ACCU) for reducing enteric methane emissions by substituting NO3 for urea at the same amount of fed nitrogen. NO3 must be fed in the form of a lick block because most operations do not have labour or equipment to manage daily supplementation. NO3 concentrations, after a 2-week adaptation period, must not exceed 50 g NO3/adult animal equivalent per day or 7 g NO3/kg dry matter intake per day to reduce the risk of NO3 toxicity. There is also a 'beef cattle herd management' method, approved in 2015, that covers activities that improve the herd emission intensity (emissions per unit of product sold) through change in the diet or management. The present study was conducted to compare the required ACCU or supplement prices for a 2% return on capital when feeding a low or high supplement concentration to breeding stock of either (1) urea, (2) three different forms of NO3 or (3) cottonseed meal (CSM), at N concentrations equivalent to 25 or 50 g urea/animal equivalent, to fasten steer entry to a feedlot (backgrounding), in a typical breeder herd on the coastal speargrass land types in central Queensland. Monte Carlo simulations were run using the software @risk, with probability functions used for (1) urea, NO3 and CSM prices, (2) GHG mitigation, (3) livestock prices and (4) carbon price. Increasing the weight of steers at a set turnoff month by feeding CSM was found to be the most cost-effective option, with or without including the offset income. The required ACCU prices for a 2% return on capital were an order of magnitude higher than were indicative carbon prices in 2015 for the three forms of NO3. The likely costs of participating in ERF projects would reduce the return on capital for all mitigation options. © CSIRO 2016

Asthma exacerbations and worsenings in patients aged 1-75 years with add-on tiotropium treatment

This review explores the effect of tiotropium Respimat® add-on therapy on asthma exacerbations and worsenings, adverse events (AEs) related to exacerbations and symptoms and any effects on seasonality across the 10 UniTinA-asthma® clinical trials comprising over 6000 patients. When added on to inhaled corticosteroids ± additional therapies, tiotropium significantly reduced the risk of exacerbations and worsenings in adults with symptomatic severe asthma and provided a non-significant improvement in worsenings in adults with symptomatic moderate and mild asthma, which was significant for patients with moderate asthma receiving tiotropium 2.5 µg once daily vs. placebo. Trials in paediatric patients were not powered to assess exacerbations or worsenings, but when AEs related to asthma exacerbations and symptoms were grouped into a composite endpoint and pooled, tiotropium improved outcomes vs. placebo (rate ratio 0.76; 95% confidence interval 0.63, 0.93). The reduction in exacerbations with tiotropium is apparent across all patients during the observed seasonal peaks of these events

Modelling Adaptation and Mitigation Strategies for Southern Livestock Industries of Australia

Climate change will impact on the Australian grazing industries both through mitigation policies and the impact of warmer temperatures, increased atmospheric CO2 and changed rainfall patterns (Cullen et al. 2009; Eckard et al. 2010). Mechanistic models are useful tools to inform our understanding of the complex interactions between future climates and the soil, plant, animal and management in livestock production systems. This paper summarises the results of a number of whole farm systems modelling studies investigating likely impacts of climate change, adaptation options and emissions implications for livestock production in southern Australia