Quantitation of transgenic plant DNA in leachate water: real-time polymerase chain reaction analysis

Roundup Ready (RR) genetically modified (GM) corn and soybean comprise a large portion of the annual planted acreage of GM crops. Plant growth and subsequent plant decomposition introduce the recombinant DNA (rDNA) into the soil environment, where its fate has not been completely researched. Little is known of the temporal and spatial distribution of plant-derived rDNA in the soil environment and in situ transport of plant DNA by leachate water has not been studied before. The objectives of this study were to determine whether sufficient quantities of plant rDNA were released by roots during growth and early decomposition to be detected in water collected after percolating through a soil profile and to determine the influence of temperature on DNA persistence in the leachate water. Individual plants of RR corn and RR soybean were grown in modified cylinders in a growth room, and the cylinders were flushed with rain water weekly. Immediately after collection, the leachate was subjected to DNA purification followed by rDNA quantification using real-time Polymerase Chain Reaction (PCR) analysis. To test the effects of temperature on plant DNA persistence in leachate water, water samples were spiked with known quantities of RR soybean or RR corn genomic DNA and DNA persistence was examined at 5, 15, and 25 degrees C. Differences in the amounts and temporal distributions of root-derived rDNA were observed between corn and soybean plants. The results suggest that rainfall events may distribute plant DNA throughout the soil and into leachate water. Half-lives of plant DNA in leachate water ranged from 1.2 to 26.7 h, and persistence was greater at colder temperatures (5 and 15 degrees C).     

Composition of suberin extracted upon gradual alkaline methanolysis of Quercus suber L. cork

The monomeric composition of suberin extracts obtained by gradual alkaline methanolysis of Quercus suber cork was determined by gas chromatography-mass spectrometry (GC-MS). Results show that 1-alkanols and alkanoic and alpha,omega-alkanedioic acids are preferentially removed upon mild alkaline conditions, whereas mid-chain-modified omega-hydroxyalkanoic acids are preferentially removed under stronger alkaline conditions. Saturated omega-hydroxyalkanoic acids are found to be abundant in all suberin extracts. These results are consistent with two distinct suberin fractions with different locations in cork cell walls and/or esterification degrees. It is proposed that these fractions correlate with the two main suberin peaks in the solid state (13)C NMR spectra of cork and suberin extracts. Quantitative GC-MS analysis showed that suberin monomers comprise approximately 30% (w/w) of the suberin extracts, the remaining comprising nonvolatile structures with high M(n) values, as measured by vapor pressure osmometry. The presence of a large fraction of high molecular weight aliphatic structures in suberin extracts is supported by the corresponding NMR spectra.     

The self-incompatible RNase S-alleles of Brazilian apple cultivars

Apple plants are self-incompatible because a genetic mechanism allows the female reproductive organ to recognize and reject self-pollen or pollen from genetic related individuals and allows non-self pollen to effect fertilization. Thus, there are implications to both breeding strategies and orchard management for fruit production. The purpose of this study was to identify and to characterize the S-RNase alleles of the gametophytic incompatibility among apple cultivars developed in Brazil, seeking to give support for choosing right combinations of parent in the apple breeding programs. It also sought to identify correct combinations of scion/pollinator cultivars of commercial apple orchards. A total of 16 specific S-RNase alleles primers were tested against DNA extracted from 12 Brazilian cultivars and their parents. The molecular analysis confronted to the reference cultivars, showed that the cultivars Daiane, Imperatriz and Princesa have the same incompatibility S3 and S5 alleles, while ‘Lisgala’ showed the alleles S2 and S5; ‘Suprema’, S1 and S9; ‘Catarina’, S1 and S19; ‘Joaquina’ and ‘Fred Hough’, S5 and S19; ‘Baronesa’, S3 and S9; ‘Duquesa’, S2 and S3. For ‘Primícia’ and ‘Condessa’ it was only possible to identify one of the S-alleles, namely S24 and S2, respectively, with the second allele remaining to be identified. Progeny test indicated the Mendelian inheritance for RNase alleles. Results of this study will be helpful to judiciously choose parents in apple breeding programs to improve compatibility.     

Human muscle phosphoglycerate mutase deficiency: newly discovered metabolic myopathy

Muscle phosphoglycerate mutase activity was decreased (5.7 percent of the lowest control value) in a 52-year-old man with intolerance for strenuous exercise and recurrent pigmenturia since adolescence. All of the other enzymes of glycolysis had normal activities, and glycogen concentration was normal. Electrophoretic, heat lability, and mercury inhibition studies showed that the small residual activity in the patient's muscle was represented by the brain (BB) isoenzyme of phosphoglycerate mutase, suggesting a genetic defect of the M subunit which predominates in normal muscle. The prevalence of the BB isoenzyme in other tissues, including muscle culture, may explain why symptoms were confined to muscle.     

Io volcanism seen by new horizons: a major eruption of the Tvashtar volcano

Jupiter's moon Io is known to host active volcanoes. In February and March 2007, the New Horizons spacecraft obtained a global snapshot of Io's volcanism. A 350-kilometer-high volcanic plume was seen to emanate from the Tvashtar volcano (62 degrees N, 122 degrees W), and its motion was observed. The plume's morphology and dynamics support nonballistic models of large Io plumes and also suggest that most visible plume particles condensed within the plume rather than being ejected from the source. In images taken in Jupiter eclipse, nonthermal visible-wavelength emission was seen from individual volcanoes near Io's sub-Jupiter and anti-Jupiter points. Near-infrared emission from the brightest volcanoes indicates minimum magma temperatures in the 1150- to 1335-kelvin range, consistent with basaltic composition.     

Design and chemical synthesis of a homogeneous polymer-modified erythropoiesis protein

We report the design and total chemical synthesis of "synthetic erythropoiesis protein" (SEP), a 51-kilodalton protein-polymer construct consisting of a 166-amino-acid polypeptide chain and two covalently attached, branched, and monodisperse polymer moieties that are negatively charged. The ability to control the chemistry allowed us to synthesize a macromolecule of precisely defined covalent structure. SEP was homogeneous as shown by high-resolution analytical techniques, with a mass of 50,825 +/-10 daltons by electrospray mass spectrometry, and with a pI of 5.0. In cell and animal assays for erythropoiesis, SEP displayed potent biological activity and had significantly prolonged duration of action in vivo. These chemical methods are a powerful tool in the rational design of protein constructs with potential therapeutic applications.     

Using BCG,MPT-51 and Ag85 as antigens in an indirect ELISA for the diagnosis of bovine tuberculosis

This study evaluated the Mycobacterium tuberculosis protein antigen MPT-51, the trimeric antigen 85 (Ag85) complex, and Bacillus Calmette-Guérin (BCG) in an indirect ELISA to diagnose bovine tuberculosis (TB) from serum samples. Serum was collected from 208 intra-dermal tuberculin test (ITT)-positive and 54 ITT-negative animals from a region where bovine TB is endemic. Using the Ag85 and BCG antigens, the indirect ELISA was able to discriminate ITT-positive from ITT-negative animals. This level of discrimination was not achieved when using the MPT-51 antigen. The highest sensitivity (Se) and specificity (Sp) of the test was found when BCG was used (Se, 82%; Sp, 91%). Further work in different epidemiological settings and with larger numbers of animals will be required to validate these findings.     

A semi-empirical model to assess uncertainty of spatial patterns of erosion

Distributed erosion models are potentially good tools for locating soil sediment sources and guiding efficient Soil and Water Conservation (SWC) planning, but the uncertainty of model predictions may be high. In this study, the distribution of erosion within a catchment was predicted with a semi-empirical erosion model that combined a semi-distributed hydrological model with the Morgan, Morgan and Finney (MMF) empirical erosion model. The model was tested in a small catchment of the West Usambara Mountains (Kwalei catchment, Tanzania). Soil detachability rates measured in splash cups (0.48–1.16 g J^− 1) were close to model simulations (0.30–0.35 g J^− 1). Net erosion rates measured in Gerlach troughs (0.01–1.05 kg m^− 2 per event) were used to calibrate the sediment transport capacity of overland flow. Uncertainties of model simulations due to parameterisation of overland flow sediment transport capacity were assessed with the Generalized Likelihood Uncertainty Estimation (GLUE) methodology. The quality of the spatial predictions was assessed by comparing the simulated erosion pattern with the field-observed erosion pattern, measuring the agreement with the weighted Kappa coefficient of the contingency table. Behavioural parameter sets (weighted Kappa > 0.50) were those with short reinfiltration length (< 1.5 m) and ratio of overland flow power α to local topography power γ close to 0.5. In the dynamic Hortonian hydrologic regime and the dissected terrain of Kwalei catchment, topography controlled the distribution of erosion more than overland flow. Simulated erosion rates varied from − 4 to + 2 kg m^− 2 per season. The model simulated correctly around 75% of erosion pattern. The uncertainty of model predictions due to sediment transport capacity was high; around 10% of the fields were attributed to either slight or severe erosion. The difficult characterisation of catchment-scale effective sediment transport capacity parameters poses a major limit to distributed erosion modelling predicting capabilities.