Structure determination of the O-specific polysaccharides from Citrobacter O4- and O27-lipopolysaccharides by methylation analysis and one- and two-dimensional 1H-NMR spectroscopy

Using sugar and methylation analyses, and one- and two-dimensional 1H-NMR spectroscopy at 500 MHz it was established that poly-beta-1,2-4-deoxy-D-arabinohexopyranose occurs as O-specific chains of lipopolysaccharides in Citrobacter serotypes O4, O27 and O36. Strong serological cross-reactivity between these serotypes is in full agreement with the chemical identity of their O-specific polysaccharides.     

Increased Summer Temperatures Reduce the Growth and Regeneration of Larix sibirica in Southern Boreal Forests of Eastern Kazakhstan

The larch forests at the southern limit of the Siberian boreal forest in Central Asia have repeatedly experienced strong recent growth declines attributed to decreasing summer precipitation in the course of climate warming. Here, we present evidence from the southernmost Larix sibirica forests in eastern Kazakhstan that these declines are primarily caused by a decrease in effective moisture due to increasing summer temperatures, despite constant annual, and summer precipitation. Tree-ring chronologies (>800 trees) showed a reduction by 50–80% in mean ring width and an increase in the frequency of missing rings since the 1970s. Climate-response analysis revealed a stronger (negative) effect of summer temperature (in particular of the previous year’s June and July temperature) on radial growth than summer precipitation (positive effect). It is assumed that a rise in the atmospheric vapor pressure deficit, which typically increases with temperature, is negatively affecting tree water status and radial growth, either directly or indirectly through reduced soil moisture. Larch rejuvenation ceased in the 1950s, which is partly explained by increasing topsoil desiccation in a warmer climate and a high drought susceptibility of larch germination, as was demonstrated by a germination experiment with variable soil moisture levels. The lack of regeneration and the reduced annual stem increment suggest that sustainable forest management aiming at timber harvesting is no longer feasible in these southern boreal forests. Progressive climate warming is likely to cause a future northward shift of the southern limit of the boreal forest.     

Iron and phosphate uptake explains the calcifuge–calcicole behavior of the terricolous lichens Cladonia furcata subsp. furcata and C. rangiformis

Mechanisms causing the calcifuge–calcicole behavior of lichens are largely unexplored. Studying the case examples of two closely related terricolous lichens, the calcifuge Cladonia furcata subsp. furcata and the calcicole C. rangiformis, we found that preference for acidic or calcareous soils in these lichens is related to iron and phosphate uptake as in vascular plants. In laboratory studies, the calcicole species was more efficient in the intracellular uptake of Fe³⁺ and phosphate at pH 8 than the calcifuge species. At pH 3, intracellular uptake of Fe²⁺ in the calcicole species significantly exceeded that in the calcifuge species suggesting that calcicole lichens suffer from toxicity symptoms by excess Fe²⁺ at acidic sites. Though these observations parallel findings from calcifuge and calcicole vascular plants, mechanisms leading to the different iron and phosphate uptake characteristics in the studied calcifuge and calcicole lichens may differ from those in vascular plants and should be the topic of future research. A role of the depside atranorin in facilitating iron uptake by reducing Fe³⁺ in the apoplast is hypothesized.     

Reconstitution of the glucose transporter from bovine heart

Reconstitution of the glucose transporter from heart should be useful as an assay in its purification and in the study of its regulation. We have prepared plasma membranes from bovine heart which display D-glucose reversible binding of cytochalasin B (33 pmol sites/mg protein; Kd = 0.2 muM). The membrane proteins were reconstituted into liposomes by the freeze-thaw procedure. Reconstituted liposomes showed D-glucose transport activity which was stereospecific, saturable and inhibited by cytochalasin B, phloretin, and mercuric chloride. Compared to membrane proteins reconstituted directly, proteins obtained by dispersal of the membranes with low concentrations of cholate or by cholate solubilization showed 1.2- or 2.3-fold higher specific activities for reconstituted transport, respectively. SDS-polyacrylamide gel electrophoresis followed by electrophoretic protein transfer and labeling with antisera prepared against the human erythrocyte transporter identified a single band of about 45 kDa in membranes from both dog and bovine hearts, a size similar to that reported for a number of other glucose transporters in various animals and tissues.     

Biomass Stock and Productivity of Primeval and Production Beech Forests: Greater Canopy Structural Diversity Promotes Productivity

Our knowledge of temperate broadleaf forest ecology is based mostly on the study of production forests, which lack the terminal stage of forest development and have a simpler stand structure than old-growth and primeval forests. How primeval and production forests differ in net primary production (NPP) is not well known. In three primeval and three nearby production forests of European beech (Fagus sylvatica) in the Slovakian Carpathians, we measured aboveground biomass stocks (live and dead), aboveground NPP (ANPP) and parameters characterizing canopy structural diversity (leaf area index and its spatial variation). Our study aims were (1) to explore the role of canopy structural diversity for ANPP and (2) to assess evidence of a productivity decline in the terminal stage. While aboveground live biomass stocks were on average 20% greater in the primeval forests (386 vs. 320 Mg ha^?1; insignificant difference at two sites), deadwood mass stocks were on average four times larger than in the production forests (86 vs. 19 Mg ha^?1). ANPP was similarly high in the primeval and production forests (10.0 vs. 9.9 Mg ha^?1 y^?1) and did not decrease towards the terminal stage. Production models indicate that, in the primeval forests, about 10% of ANPP (ca. 1 Mg ha^?1 y^?1) was generated by effects related to leaf area heterogeneity, evidencing a positive effect of structural diversity on forest productivity, even though species diversity was low. This study helps to better understand the impact of forest management on the productivity and carbon storage in temperate woodlands.     

Noncomplementing Diploids from Bacillus Subtilis Protoplast Fusion: Relationship between Maintenance of Chromosomal Inactivation and Segregation Capacity

Fusions of Bacillus subtilis protoplasts from two genetically marked strains produce noncomplementing heterodiploid bacteria. These noncomplementing diploids (Ncds) carry both parental chromosomes, but only one is expressed. Fusion products of strains polymorphic for NotI restriction sites provide new physical evidence to support the conclusion that Ncds are not an artifact of cross feeding or cell adhesion. We show that reversible chromosomal inactivation can only account for the biparental trait of unstable Ncds. Two types of cells were recovered from the late progeny of unstable Ncds: Ncds with irreversible chromosome silencing (stable Ncds) and secondary recombinants that displayed a genomic mosaic NotI profile. Segregants from an unstable Ncd population gave rise to two viable haploid cell types. By contrast, stable Ncds segregated into a population of viable and inviable haploid cells. We propose that the latter are derived from irreversible chromosome silencing. Our results indicate that clonal populations of stable Ncds are heterogenous and suggest that segregation and inactivation are independent parameters.     

Common ragweed (Ambrosia artemisiifolia L.): allergenicity and molecular characterization of pollen after plant exposure to elevated NO2

Ragweed pollen is the main cause of allergenic diseases in Northern America, and the weed has become a spreading neophyte in Europe. Climate change and air pollution are speculated to affect the allergenic potential of pollen. The objective of this study was to investigate the effects of NO₂, a major air pollutant, under controlled conditions, on the allergenicity of ragweed pollen. Ragweed was exposed to different levels of NO₂ throughout the entire growing season, and its pollen further analysed. Spectroscopic analysis showed increased outer cell wall polymers and decreased amounts of pectin. Proteome studies using two‐dimensional difference gel electrophoresis and liquid chromatography–tandem mass spectrometry indicated increased amounts of several Amb a 1 isoforms and of another allergen with great homology to enolase Hev b 9 from rubber tree. Analysis of protein S‐nitrosylation identified nitrosylated proteins in pollen from both conditions, including Amb a 1 isoforms. However, elevated NO₂ significantly enhanced the overall nitrosylation. Finally, we demonstrated increased overall pollen allergenicity by immunoblotting using ragweed antisera, showing a significantly higher allergenicity for Amb a 1. The data highlight a direct influence of elevated NO₂ on the increased allergenicity of ragweed pollen and a direct correlation with an increased risk for human health.     

Product quality research institute evaluation of cascade impactor profiles of pharmaceutical aerosols, part 3: Final report on a statistical procedure for determining equivalence

The purpose of this article is to report final results of the evaluation of a chi-square ratio test proposed by the US Food and Drug Administration (FDA) for demonstrating equivalence of aerodynamic particle size distribution (APSD) profiles of nasal and orally inhaled drug products. A working group of the Product Quality Research Institute previously published results demonstrating some limitations of the proposed test. In an effort to overcome the test's limited discrimination, the group proposed a supplemental test, a population bioequivalence (PBE) test for impactor-sized mass (ISM). In this final report the group compares the chi-square ratio test to the ISM-PBE test and to the combination of both tests. The basis for comparison is a set of 55 realistic scenarios of cascade impactor data, which were evaluated for equivalence by the statistical tests and independently by the group members. In many instances, the combined application of these 2 tests appeared to increase the discriminating ability of the statistical procedure compared with the chi-square ratio test alone. In certain situations the chi-square ratio test alone was sufficient to determine equivalence of APSD profiles, while in other situations neither of the tests alone nor their combination was adequate. This report describes all of these scenarios and results. In the end, the group did not recommend a statistical test for APSD profile equivalence. The group did not investigate other in vitro tests, in vivo issues, or other statistical tests for APSD profile comparisons. The studied tests are not intended for routine quality control of APSD.