Recognition of the lipoyl domain is the ultimate determinant of substrate channelling in the pyruvate dehydrogenase multienzyme complex

Reductive acetylation of the lipoyl domain (E2plip) of the dihydrolipoyl acetyltransferase component of the pyruvate dehydrogenase multienzyme complex of Escherichia coli is catalysed specifically by its partner pyruvate decarboxylase (E1p), and no productive interaction occurs with the analogous 2-oxoglutarate decarboxylase (E1o) of the 2-oxoglutarate dehydrogenase complex. Residues in the lipoyl-lysine beta-turn region of the unlipoylated E2plip domain (E2plip(apo)) undergo significant changes in both chemical shift and transverse relaxation time (T(2)) in the presence of E1p but not E1o. Residue Gly11, in a prominent surface loop between beta-strands 1 and 2 in the E2plip domain, was also observed to undergo a significant change in chemical shift. Addition of pyruvate to the mixture of E2plip(apo) and E1p caused larger changes in chemical shift and the appearance of multiple cross-peaks for certain residues, suggesting that the domain was experiencing more than one type of interaction. Residues in both beta-strands 4 and 5, together with those in the prominent surface loop and the following beta-strand 2, appeared to be interacting with E1p, as did a small patch of residues centred around Glu31. The values of T(2) across the polypeptide chain backbone were also lower than in the presence of E1p alone, suggesting that E2plip(apo) binds more tightly after the addition of pyruvate. The lipoylated domain (E2plip(holo)) also exhibited significant changes in chemical shift and decreases in the overall T(2) relaxation times in the presence of E1p, the residues principally affected being restricted to the half of the domain that contains the lipoyl-lysine (Lys41) residue. In addition, small chemical shift changes and a general drop in T(2) times in the presence of E1o were observed, indicating that E2plip(holo) can interact, weakly but non-productively, with E1o. It is evident that recognition of the protein domain is the ultimate determinant of whether reductive acetylation of the lipoyl group occurs, and that this is ensured by a mosaic of interactions with the Elp.     

BMP-2-modulated chondrogenic differentiation in vitro involves down-regulation of membrane-bound beta-catenin

Bone morphogenetic proteins (BMPs) are important regulators of cellular differentiation and embryonic development. Beta catenin mediated nuclear signaling has been implicated in BMP-2-modulated chondrogenic differentiation in the pluripotential stem cell line C3H10T1/2. However, there is little information on the functional role of beta catenin in BMP-2-modulated differentiation of primary nontransformed mesenchymal cells. Here, we present evidence to show that BMP-2-induced chondrogenic differentiation in high-density primary mesenchymal culture is associated with a significant decrease in membrane-bound beta catenin by 72 hours compared to controls. Nuclear localization of beta catenin is not detectable by immunofluorescence and the TCF-responsive reporter vector TOPFLASH shows only background activity during chondrogenic differentiation. BMP-2-treated cultures show reduced cell-cell adhesion by 72 hours, which correlates with the changes in levels of membrane-bound beta catenin. Up-regulation of membrane-bound beta catenin blocks the effect of BMP-2 on both chondrogenic differentiation and cell-cell adhesiveness. These findings suggest that BMP-2 can modulate the adhesivity of adherens junctions through regulation of membrane bound beta catenin.     

Solution studies of chymotrypsin inhibitor-2 glutamine insertion mutants show no interglutamine interactions

Mutants of chymotrypsin inhibitor protein 2 have previously been studied in which 4 or 10 glutamine residues were inserted into the inhibitory loop of the protein between residues 59 and 60, as potential models for the behaviour of glutamine tracts in proteins associated with polyglutamine-expansion neurodegenarative diseases. These mutants form very stable monomers, dimers and trimers. Although the cause of oligomerisation was found to be domain-swapping, it was thought that the glutamine insertions might nevertheless show evidence of weak interglutamine interactions in solution that could mimic those occurring in disease-associated proteins. In the present NMR study, we used steady-state (15)N[(1)H] NOE measurements and chemical shift comparisons to characterise the motional properties of the inserted glutamines in these CI2 mutants. We found the glutamines to be highly mobile, with no evidence of interactions amongst them in either monomers or dimers.     

Management and outcome of winter upper respiratory tract infections in children aged 0-9 years.

Age-specific incidences for upper respiratory tract infections in children from a new-town population during 1975-7 were studied, and 965 consecutive upper respiratory tract infections in children aged under 10 during two winters were analysed in detail. Significantly different management plans made by seven doctors did not correlate with the clinical outcome as judged by complications, recall rates, and demand for treatment for similar episodes in the future. Two hundred and thirty-two children (24%) returned for another consultation for the same episode of upper respiratory tract infection. The main reason for these repeat consultations seemed to be that parental expectations about the natural history of the illness were not fulfilled. More realistic parental expectations might be set and safer clinical standards maintained if doctors warned parents about symptoms such as cough and occasional diarrhoea or vomiting that are commonly associated with upper respiratory tract infections in children.     

Humboldt's spa: microbial diversity is controlled by temperature in geothermal environments

Over 200 years ago Alexander von Humboldt (1808) observed that plant and animal diversity peaks at tropical latitudes and decreases toward the poles, a trend he attributed to more favorable temperatures in the tropics. Studies to date suggest that this temperature–diversity gradient is weak or nonexistent for Bacteria and Archaea. To test the impacts of temperature as well as pH on bacterial and archaeal diversity, we performed pyrotag sequencing of 16S rRNA genes retrieved from 165 soil, sediment and biomat samples of 36 geothermal areas in Canada and New Zealand, covering a temperature range of 7.5–99 °C and a pH range of 1.8–9.0. This represents the widest ranges of temperature and pH yet examined in a single microbial diversity study. Species richness and diversity indices were strongly correlated to temperature, with R² values up to 0.62 for neutral–alkaline springs. The distributions were unimodal, with peak diversity at 24 °C and decreasing diversity at higher and lower temperature extremes. There was also a significant pH effect on diversity; however, in contrast to previous studies of soil microbial diversity, pH explained less of the variability (13–20%) than temperature in the geothermal samples. No correlation was observed between diversity values and latitude from the equator, and we therefore infer a direct temperature effect in our data set. These results demonstrate that temperature exerts a strong control on microbial diversity when considered over most of the temperature range within which life is possible.     

A framework for studying transient dynamics of population projection matrix models

Empirical models are central to effective conservation and population management, and should be predictive of real-world dynamics. Available modelling methods are diverse, but analysis usually focuses on long-term dynamics that are unable to describe the complicated short-term time series that can arise even from simple models following ecological disturbances or perturbations. Recent interest in such transient dynamics has led to diverse methodologies for their quantification in density-independent, time-invariant population projection matrix (PPM) models, but the fragmented nature of this literature has stifled the widespread analysis of transients. We review the literature on transient analyses of linear PPM models and synthesise a coherent framework. We promote the use of standardised indices, and categorise indices according to their focus on either convergence times or transient population density, and on either transient bounds or case-specific transient dynamics. We use a large database of empirical PPM models to explore relationships between indices of transient dynamics. This analysis promotes the use of population inertia as a simple, versatile and informative predictor of transient population density, but criticises the utility of established indices of convergence times. Our findings should guide further development of analyses of transient population dynamics using PPMs or other empirical modelling techniques.     

Changes to soil organic N dynamics with leguminous woody plant encroachment into grasslands

Encroachment of nitrogen-fixing trees and shrubs into grasslands and savannas is a well-documented land cover change that occurs worldwide. In the Rio Grande Plains region of southern Texas, previous studies have shown woody encroachment by leguminous Prosopis glandulosa (mesquite) trees increases soil C and N, decreases microbial biomass N relative to soil N, and accelerates N mineralization and nitrification. We examined responses of the dominant organic N components in soil (amino acids and amino sugars) and two soil-bound protein-N acquiring enzymes (arylamidase and β-N-acetylglucosaminidase) along a grassland-to-woodland successional chronosequence to determine changes to soil N chemistry and extractability. The proportion of total N held within amino compounds was significantly lower in the woodlands (47 %) relative to the grassland soils (62 %). This increase in non-hydrolysable N was accompanied by increases in plant cell wall derived amino acids (e.g. hydroxyproline, serine) and losses of microbial amino sugars, indicating the woodland organic N pool was altered in composition and potentially in quality, either because it was more structurally protected or difficult to degrade due to polymerization/condensation reactions. Soil carbon-normalized activities of both soil-bound N-acquiring enzymes were significantly higher in woodland soils, consistent with changes in the biochemical composition of organic N. Although soil total N increases following woody encroachment, this additional organic N appears to be less extractable by chemical hydrolysis and thus potentially in more refractory forms, which may limit microbial N accessibility, slow the cycling of soil organic carbon, and contribute to observed soil C and N accrual in these systems.