Role of the electrochemical proton gradient in genetic transformation of Haemophilus influenzae.

The uptake of homologous DNA by Haemophilus influenzae was studied as a function of the proton motive force in completely competent cultures in the pH range of 6 to 8. The composition and magnitude of the proton motive force were varied by using the ionophores valinomycin and nigericin (in the presence of various potassium ion concentrations) and by using protonophores. No interaction of the ionophores with the DNA transformation system itself was observed. Either component of the proton motive force, the electrical potential or the pH gradient, can drive the uptake of DNA, and the extent of the uptake of DNA is ultimately determined by the total proton motive force. The transformation frequency increases with the proton motive force, which reaches a maximum value at around -130 mV. These results are consistent with an electrogenic proton-DNA symport mechanism, but direct evidence for such a system is not available. The proton motive force was followed during competence development of H. influenzae at pH 8. In the initial phase (up to 50 min), the proton motive force remained constant at about -90 mV, whereas the transformation frequency rose steeply. In the second phase, the proton motive force increased. The transformation frequency in this phase increased with the proton motive force, as in completely competent cultures. These observations and the observed inhibition by NAD of both the proton motive force and the transformation frequency indicate that structural components of the competent state are formed in the initial phase of competence development, whereas the second phase is characterized by an increase of the proton motive force.     

Root-soil contact of maize,as measured by a thin-section technique

In models of oxygen, water and nutrient uptake by plant roots, the degree of root-soil contact is an important parameter. An observation technique is required to evaluate to what extent root-soil contact depends on plant species, soil texture and structure. Thin sections for studying soil structure may be used for this purpose, provided that roots do not shrink during section preparation, and that all root cross sections are recognized.Maize was grown in pots with soil aggregates obtained by sieving and compacting to three bulk densities. Thin sections were made by freeze-drying samples before impregnating the soil with resin. Two checks were made on the validity of the method. Firstly, visual appearance of roots with intact epidermis, cortex and other tissues did not show signs of shrinkage. Secondly, the agreement was checked between root lengths obtained by washing duplicate soil samples and the number of root cross sections counted on horizonal and vertical thin sections. For the latter, the angle at which roots intersected the thin-section plane was determined from the shape of the cross sections. The frequency distribution of calculated angles was in agreement with the frequency distribution expected for a randomly oriented set of cylinders when an error term was included in the simulated measurements.Some results are presented for a field test of the thin-section method with barley on a calcareous marine sandy loam. Root hairs, apparently undamaged by sample preparation, are important for bridging the gap between roots and soil in this situation. According to the experience presented, the thin-section technique is suitable to derive the degree of root-soil contact, as influenced by species, soil texture and structure, in samples obtained from pot or field experiments.Communication No. 43 of the Dutch Programme on Soil Ecology of Arable Farming Systems.Communication No. 43 of the Dutch Programme on Soil Ecology of Arable Farming Systems.     

A conserved helicase motif of the AddA subunit of the Bacillus subtilis ATP-dependent nuclease (AddAB) is essential for DNA repair and recombination

Various mutations were introduced in a conserved helicase domain (motif VI) of the AddA subunit of the Bacillus subtilis ATP-dependent nuclease (AddAB) by site-directed mutagenesis. These mutations affected the helicase activity and the ATP-dependent exonuclease activity on double-stranded DNA (dsDNA) as the substrate to various degrees, but had hardly any effect on the exonuclease activity on single-stranded DNA (ssDNA), suggesting that exonuclease activity on dsDNA of the enzyme requires unwinding of the DNA. This idea was supported by the finding that, initially, the rate and extent of unwinding of the DNA were higher than those of its degradation to acid-soluble products by the exonucleolytic activity. The effects of the mutations on DNA repair and recombination correlated strongly with their effects on helicase activity. Taken together, these results suggest that motif VI is essential for the helicase activity, and that this activity is required for DNA repair and recombination.     

Characterizing Forest Change Using Community-Based Monitoring Data and Landsat Time Series

Increasing awareness of the issue of deforestation and degradation in the tropics has resulted in efforts to monitor forest resources in tropical countries. Advances in satellite-based remote sensing and ground-based technologies have allowed for monitoring of forests with high spatial, temporal and thematic detail. Despite these advances, there is a need to engage communities in monitoring activities and include these stakeholders in national forest monitoring systems. In this study, we analyzed activity data (deforestation and forest degradation) collected by local forest experts over a 3-year period in an Afro-montane forest area in southwestern Ethiopia and corresponding Landsat Time Series (LTS). Local expert data included forest change attributes, geo-location and photo evidence recorded using mobile phones with integrated GPS and photo capabilities. We also assembled LTS using all available data from all spectral bands and a suite of additional indices and temporal metrics based on time series trajectory analysis. We predicted deforestation, degradation or stable forests using random forest models trained with data from local experts and LTS spectral-temporal metrics as model covariates. Resulting models predicted deforestation and degradation with an out of bag (OOB) error estimate of 29% overall, and 26% and 31% for the deforestation and degradation classes, respectively. By dividing the local expert data into training and operational phases corresponding to local monitoring activities, we found that forest change models improved as more local expert data were used. Finally, we produced maps of deforestation and degradation using the most important spectral bands. The results in this study represent some of the first to combine local expert based forest change data and dense LTS, demonstrating the complementary value of both continuous data streams. Our results underpin the utility of both datasets and provide a useful foundation for integrated forest monitoring systems relying on data streams from diverse sources.     

The Receptor Concept in 3D: From Hypothesis and Metaphor to GPCR–Ligand Structures

The first mentioning of the word “receptor” for the structure with which a bioactive compound should react for obtaining its specific influence on a physiological system goes back to the years around 1900. The receptor concept was adapted from the lock and key theory for the enzyme substrate and blockers interactions. Through the years the concept, in the beginning rather being a metaphor, not a model, was refined and became reality in recent years. Not only the structures of receptors were elucidated, also the receptor machineries were unraveled. Following a brief historical review we will describe how the recent breakthroughs in the experimental determination of G protein-coupled receptor (GPCR) crystal structures can be complemented by computational modeling, medicinal chemistry, biochemical, and molecular pharmacological studies to obtain new insights into the molecular determinants of GPCR–ligand binding and activation. We will furthermore discuss how this information can be used for structure-based discovery of novel GPCR ligands that bind specific (allosteric) binding sites with desired effects on GPCR functional activity.     

Internal transcribed spacer polymorphism in Pseudo-nitzschia multistriata (Bacillariophyceae) in the Gulf of Naples: recent divergence or intraspecific hybridization?

The planktonic diatom Pseudo-nitzschia multistriata is a potentially toxic species recorded during late summer-autumn in the Gulf of Naples (Tyrrhenian Sea, Italy). We analysed the genetic structure by amplifying the internal transcribed spacer (ITS-1-5.8S-ITS-2) region of the ribosomal DNA of 44 strains isolated along 2 years. Polymorphism in the ITS region was detected by direct sequencing and the PCR-products from selected strains were thus cloned to assess intra-strain ITS variability. Strains clustered into three main types: type A and B - differing by 0.6% sequence divergence - and type A/B, showing both A and B variants within the same genome. The three types showed no differences in the large subunit sequences (LSU) of the rDNA, ultrastructure of the valve, secondary structure of ITS-1 and ITS-2, ploidy level and they were sexually compatible. Based on the results of these multiple approaches, we can state that the three ITS-types belong to the same reproductive unit (or "species" sensu Mayr [(1942). Systematics and the Origin of Species. Columbia University Press, New York]). We suggest that ITS polymorphism in P. multistriata may be related to the contemporary occurrence of different but still inbreeding populations which either diverged recently or originated in different geographic areas and became sympatric in the studied area.     

Improving the analysis of designed studies by combining statistical modelling with study design information

Background  

In the fields of life sciences, so-called designed studies are used for studying complex biological systems. The data derived from these studies comply with a study design aimed at generating relevant information while diminishing unwanted variation (noise). Knowledge about the study design can be used to decompose the total data into data blocks that are associated with specific effects. Subsequent statistical analysis can be improved by this decomposition if these are applied on selected combinations of effects.