Biolistic introduction of a synthetic Bt gene into elite maize

Summary A synthetic Bt gene encoding a truncated version of the CryIA(b) protein derived from Bacillus thuringiensis was successfully introduced into elite maize using microprojectile bombardment of immature embryos. The method used to initiate and identify transformation events is described. We describe the detailed parameters used for the Biolistics device as well as the plasmids used for the transformations. The plasmids contained the synthetic Bt gene driven by either the 35S CaMV promoter or a combination of two tissue-specific promoters, leaf and pollen, derived from maize. Specific conditions for the culture of Type I callus from immature embryos, the phosphinothricin (PPT) selection protocol, and the regeneration of plants are discussed. T0 and T1 plants were initially identified using the pH-dependent chlorophenol red test and/or the histochemical -glucuronidase (GUS) assay. PCR and Southern data confirm the presence of the 35S CaMV promoter and the synthetic Bt gene.     

Feline herpesvirus

Feline herpesvirus (FHV-1; felid herpesvirus 1 (FeHV-1)) is an alphaherpesvirus of cats closely related to canine herpesvirus-1 and phocine herpesvirus-1. There is only one serotype of the virus and it is relatively homogenous genetically. FeHV-1 is an important cause of acute upper respiratory tract and ocular disease in cats. In addition, its role in more chronic ocular disease and skin lesions is increasingly being recognised. Epidemiologically, FeHV-1 behaves as a typical alphaherpesvirus whereby clinically recovered cats become latently infected carriers which undergo periodic episodes of virus reactivation, particularly after a stress. The primary site of latency is the trigeminal ganglion. Conventional inactivated and modified-live vaccines are available and protect reasonably well against disease but not infection, although viral shedding may be reduced. Genetically engineered vaccines have also been developed, both for FeHV-1 and as vector vaccines for other pathogens, but none is as yet marketed.     

Water-table management in lowland UK peat soils and its potential impact on CO2 emission

The rate of oxidation of peat soils is highly seasonal and varies with temperature and soil moisture content. Large variations in soil moisture content result in wet–dry cycles that can enhance peat degradation. Water‐table management plays a crucial role in controlling and damping the effect of these environmental factors. However, maintaining high ditch water levels in fields bounded by ditches does not guarantee a high field groundwater level. The effect of installing subsurface irrigation at different spacings on water table elevation was studied in a low‐lying peat grassland. The water table elevation data were compared against values predicted with a water balance model. In addition, greenhouse experiments were carried out on undisturbed soil core samples collected from the peat grassland as well as a low‐lying peatland under intensive arable faming to measure CO₂ evolution under different water regimes. The field data from the peat grassland suggest that sub‐irrigation spacing as low as 10 m is necessary during summer periods to maintain groundwater levels similar to those in the ditches. Over the same period of observation, the difference in water level between the ditches and the non‐irrigated fields is as high as 0.7 m. Modelled outputs are in good correlation with the field observations, and demonstrate that simple water balance models can provide an effective tool to study the effect of water management practices and potential changes in subsurface conditions, climate and land use on water‐table levels. The measurement of CO₂ emission from undisturbed peat soil columns shows that the rate of oxidation of soil organic matter from peat soils is highly seasonal and that drainage exacerbates the rate of peat mineralization.     

Green tea polyphenol extract regulates the expression of genes involved in glucose uptake and insulin signaling in rats fed a high fructose diet

Green tea has antidiabetic, antiobesity, and anti-inflammatory activities in animal models, but the molecular mechanisms of these effects have not been fully understood. Quantitative real-time polymerase chain reaction (PCR) was used to investigate the relative expression levels and the effects of green tea (1 and 2 g solid extract/kg diet) on the expression of glucose transporter family genes (Glut1/Slc2a1, Glut2/Slc2a2, Glut3/Slc2a3, and Glut4/Slc2a4) and insulin signaling pathway genes (Ins1, Ins2, Insr, Irs1, Irs2, Akt1, Grb2, Igf1, Igf2, Igf1r, Igf2r, Gsk3b, Gys1, Pik3cb, Pik3r1, Shc1, and Sos1) in liver and muscle of rats fed a high-fructose diet known to induce insulin resistance and oxidative stress. Glut2 and Glut4 were the major Glut mRNAs in rat liver and muscle, respectively. Green tea extract (1 g) increased Glut1, Glut4, Gsk3b, and Irs2 mRNA levels by 110, 160, 30, and 60% in the liver, respectively, and increased Irs1 by 80% in the muscle. Green tea extract (2 g) increased Glut4, Gsk3b, and Pik3cb mRNA levels by 90, 30, and 30% but decreased Shc1 by 60% in the liver and increased Glut2, Glut4, Shc1, and Sos1 by 80, 40, 60, and 50% in the muscle. This study shows that green tea extract at 1 or 2 g/kg diet regulates gene expression in the glucose uptake and insulin signaling pathway in rats fed a fructose-rich diet.     

Effect of processing conditions on trace elements in fish roe from six commercial new zealand fish species

The concentrations of trace elements in fish roes and the effect of processing conditions (karasumi-like or karashi mentaiko) were investigated in six commercial fish species from New Zealand. The studied elements were As, Cd, Cr, Cu, Hg, Pb, and Zn, and the roes were from the following species: chinook salmon ( Oncorhynchus tshawytscha), hoki ( Macruronus novaezelandiae), southern blue whiting ( Micromesistius australis), hake ( Merluccius australis), blue warehou ( Seriolella brama), and barracouta ( Thyrsites atun). The concentrations of As, Cd, Cr, Hg, and Pb in the roes were lower than literature values for fish muscles. Only Zn in barracouta roe and Cu in salmon roe and their products were relatively higher than the generally accepted levels in fish muscles and could be of safety concern. Hence, the consumption of barracouta and salmon roes among certain parts of the population needs to be monitored and assessed. Dry salting (karasumi-like) processing increased ( P < 0.001) the concentrations of the studied trace elements while salting fermentation (karashi mentaiko) processing tended to decrease the levels of trace elements. Fermentation may be a useful process to decrease the level of toxic trace elements.     

Effects of microbial supplements containing yeast and lactobacilli on roughage-fed ruminal microbial activities

Effects of two microbial feed supplements on microbial activities in rumen-stimulating cultures and the rumens of steers fed a fescue hay-based roughage diet were evaluated. The yeast culture supplement contained Saccharomyces cerevisiae (1.4 to 4.2 x 10(9) colony-forming units [cfu]/g), whereas the mixed microbial supplement contained yeast, lactobacilli and enterococci (1.4 to 2.7 x 10(9) cfu/g, 1.2 to 2.3 x 10(9) cfu/g, and 1.5 to 2.6 x 10(10) cfu/g, respectively). Concentrations of viable yeast cells were increased consistently in continuous cultures and rumens of steers receiving either supplement (1 g/kg of feed). However, neither supplement consistently altered the relative concentrations of volatile fatty acids or ammonia in continuous cultures and rumens of steers. The pH tended to be greater (P = .13) in continuous cultures receiving yeast culture supplement than in cultures receiving the unsupplemented diet (6.50 vs 6.36), but pH in the rumens of steers was not affected by the supplements. Concentrations of cellulolytic microorganisms in cultures and the rumens of steers receiving supplements containing only yeast were from 5 to 40 times greater than those observed in cultures or steers receiving the unsupplemented diet. Supplements that had been treated with heat (121 degrees C for 15 min) to inactive yeast cells did not alter the concentrations of cellulolytic bacteria in rumen-stimulating cultures. These results suggest that live yeast culture supplements stimulate growth of cellulolytic microorganisms in the rumen.     

Glycolipid and mucopolysaccharide abnormality in fibroblasts of fabry's disease

Cultures of skin fiibroblasts from a patient with Fabry's disease showed an accumulation of the glycolipid, galactosyl-galactosyl-glucosyl ceramide. Such cells also showed metachromasia on staining with toluidine blue and a markedly elevated acid mucopolysaccharide content.