Homologue expression of a fungal endo-1,4-β-Dxylanase using submerged and solid substrate fermentations

The xyn5 gene, which encodes an endo-β-1,4-xylanase (Xyn5), in Aspergillus niger GS1 was cloned into an expression cassette under the control of constitutive glyceraldhehyde-3-phosphate dehydrogenase gene promoter. The expression system was designed to produce the recombinant enzyme containing a six-histidine peptide fused to the carboxyl end of the protein. The efficiency of Xyn5 production under submerged (SmF) and solid-state (SSF) fermentation was investigated using the homologous cotransformed A. niger AB4.1. A productivity of 17.1 U/(l·h) was estimated for SSF and 3.2 U/(l·h) for SmF calculated at peak value of enzyme titers. Recombinant Xyn5 obtained by SSF on polyurethane fiber, was purified 5.1-fold by anion exchange and immobilized metal affinity chromatography, with 35.7% recovery. The purified recombinant enzyme showed an apparent molecular weight of 30 kDa and optimal activity (522 U/mg protein) at pH 5.5 and 50°C.Key words: Aspergillus niger GS1, xylanolytic activity, solid-state fermentation, homologue expression, polyurethane fiber

Intrahepatic cholestasis of pregnancy

Intrahepatic cholestasis of pregnancy (ICP) is a cholestatic disorder characterized by (i) pruritus with onset in the second or third trimester of pregnancy, (ii) elevated serum aminotransferases and bile acid levels, and (iii) spontaneous relief of signs and symptoms within two to three weeks after delivery. ICP is observed in 0.4–1% of pregnancies in most areas of Central and Western Europe and North America, while in Chile and Bolivia as well as Scandinavia and the Baltic states roughly 5–15% and 1–2%, respectively, of pregnancies are associated with ICP. Genetic and hormonal factors, but also environmental factors may contribute to the pathogenesis of ICP. Intrahepatic cholestasis of pregnancy increases the risk of preterm delivery (19–60%), meconium staining of amniotic fluid (27%), fetal bradycardia (14%), fetal distress (22–41%), and fetal loss (0.4–4.1%), particularly when associated with fasting serum bile acid levels > 40 μmol/L. The hydrophilic bile acid ursodeoxycholic acid (10–20 mg/kg/d) is today regarded as the first line treatment for intrahepatic cholestasis of pregnancy. Delivery has been recommended in the 38th week when lung maturity has been established

Confirmation of general relativity on large scales from weak lensing and galaxy velocities

Although general relativity underlies modern cosmology, its applicability on cosmological length scales has yet to be stringently tested. Such a test has recently been proposed, using a quantity, EG, that combines measures of large-scale gravitational lensing, galaxy clustering and structure growth rate. The combination is insensitive to 'galaxy bias' (the difference between the clustering of visible galaxies and invisible dark matter) and is thus robust to the uncertainty in this parameter. Modified theories of gravity generally predict values of EG different from the general relativistic prediction because, in these theories, the 'gravitational slip' (the difference between the two potentials that describe perturbations in the gravitational metric) is non-zero, which leads to changes in the growth of structure and the strength of the gravitational lensing effect3. Here we report that EG = 0.39 +/- 0.06 on length scales of tens of megaparsecs, in agreement with the general relativistic prediction of EG $\approx$ 0.4. The measured value excludes a model within the tensor-vector-scalar gravity theory, which modifies both Newtonian and Einstein gravity. However, the relatively large uncertainty still permits models within f(R) theory, which is an extension of general relativity. A fivefold decrease in uncertainty is needed to rule out these models.Comment: Submitted version; 13 pages, 2 figures. Accepted version and supplementary material are available at: http://www.nature.com/nature/journal/v464/n7286/full/nature08857.html

Childhood and adult tuberculosis in a rural hospital in Southeast Ethiopia: a ten-year retrospective study

<p>Abstract</p> <p>Background</p> <p>Many DOTS experiences in developing countries have been reported. However, experience in a rural hospital and information on the differences between children and adults are limited. We described the epidemiology and treatment outcome of adult and childhood tuberculosis (TB) cases, and identified risk factors associated with defaulting and dying during TB treatment in a rural hospital over a 10-year period (1998 to 2007).</p> <p>Methods</p> <p>Retrospective data collection using TB registers and treatment cards in a rural private mission hospital. Information was collected on number of cases, type of TB and treatment outcomes using standardised definitions.</p> <p>Results</p> <p>2225 patients were registered, 46.3% of whom were children. A total of 646 patients had smear-positive pulmonary TB (PTB), [132 (20.4%) children]; 816 had smear-negative PTB [556 (68.2%) children], and 763 extra-PTB (EPTB) [341 (44.8%) children]. The percentage of treatment defaulters was higher in paediatric (13.9%) than in adult patients (9.3%) (p = 0.001). The default rate declined from 16.8% to 3.5%, and was independently positively associated with TB meningitis (AOR: 2.8; 95% CI: 1.2-6.6) and negatively associated with smear-positive PTB (AOR: 0.6; 95% CI: 0.4-0.8). The mortality rate was 5.3% and the greatest mortality was associated with adult TB (AOR: 1.7; 95% CI: 1.1-2.5), TB meningitis (AOR: 3.6; 95% CI:1.2-10.9), and HIV infection (AOR: 4.3; 95% CI: 1.9-9.4). Decreased mortality was associated with TB lymphadenitis (AOR: 0.24; 95% CI: 0.11-0.57).</p> <p>Conclusion</p> <p>(1) The registration of TB cases can be useful to understand the epidemiology of TB in local health facilities. (2) The defaulter and mortality rate of childhood TB is different to that of adult TB. (3) The rate of defaulting from treatment has declined over time.</p

Spike-Timing Precision and Neuronal Synchrony Are Enhanced by an Interaction between Synaptic Inhibition and Membrane Oscillations in the Amygdala

The basolateral complex of the amygdala (BLA) is a critical component of the neural circuit regulating fear learning. During fear learning and recall, the amygdala and other brain regions, including the hippocampus and prefrontal cortex, exhibit phase-locked oscillations in the high delta/low theta frequency band (∼2–6 Hz) that have been shown to contribute to the learning process. Network oscillations are commonly generated by inhibitory synaptic input that coordinates action potentials in groups of neurons. In the rat BLA, principal neurons spontaneously receive synchronized, inhibitory input in the form of compound, rhythmic, inhibitory postsynaptic potentials (IPSPs), likely originating from burst-firing parvalbumin interneurons. Here we investigated the role of compound IPSPs in the rat and rhesus macaque BLA in regulating action potential synchrony and spike-timing precision. Furthermore, because principal neurons exhibit intrinsic oscillatory properties and resonance between 4 and 5 Hz, in the same frequency band observed during fear, we investigated whether compound IPSPs and intrinsic oscillations interact to promote rhythmic activity in the BLA at this frequency. Using whole-cell patch clamp in brain slices, we demonstrate that compound IPSPs, which occur spontaneously and are synchronized across principal neurons in both the rat and primate BLA, significantly improve spike-timing precision in BLA principal neurons for a window of ∼300 ms following each IPSP. We also show that compound IPSPs coordinate the firing of pairs of BLA principal neurons, and significantly improve spike synchrony for a window of ∼130 ms. Compound IPSPs enhance a 5 Hz calcium-dependent membrane potential oscillation (MPO) in these neurons, likely contributing to the improvement in spike-timing precision and synchronization of spiking. Activation of the cAMP-PKA signaling cascade enhanced the MPO, and inhibition of this cascade blocked the MPO. We discuss these results in the context of spike-timing dependent plasticity and modulation by neurotransmitters important for fear learning, such as dopamine

Study of the reaction e^{+}e^{-} -->J/psi\pi^{+}\pi^{-} via initial-state radiation at BaBar

We study the process $e^+e^-\to J/\psi\pi^{+}\pi^{-}$ with initial-state-radiation events produced at the PEP-II asymmetric-energy collider. The data were recorded with the BaBar detector at center-of-mass energies 10.58 and 10.54 GeV, and correspond to an integrated luminosity of 454 $\mathrm{fb^{-1}}$. We investigate the $J/\psi \pi^{+}\pi^{-}$ mass distribution in the region from 3.5 to 5.5 $\mathrm{GeV/c^{2}}$. Below 3.7 $\mathrm{GeV/c^{2}}$ the $\psi(2S)$ signal dominates, and above 4 $\mathrm{GeV/c^{2}}$ there is a significant peak due to the Y(4260). A fit to the data in the range 3.74 -- 5.50 $\mathrm{GeV/c^{2}}$ yields a mass value $4244 \pm 5$ (stat) $\pm 4$ (syst)$\mathrm{MeV/c^{2}}$ and a width value $114 ^{+16}_{-15}$ (stat)$\pm 7$(syst)$\mathrm{MeV}$ for this state. We do not confirm the report from the Belle collaboration of a broad structure at 4.01 $\mathrm{GeV/c^{2}}$. In addition, we investigate the $\pi^{+}\pi^{-}$ system which results from Y(4260) decay

Predicting Spatial Patterns of Plant Recruitment Using Animal-Displacement Kernels

For plants dispersed by frugivores, spatial patterns of recruitment are primarily influenced by the spatial arrangement and characteristics of parent plants, the digestive characteristics, feeding behaviour and movement patterns of animal dispersers, and the structure of the habitat matrix. We used an individual-based, spatially-explicit framework to characterize seed dispersal and seedling fate in an endangered, insular plant-disperser system: the endemic shrub Daphne rodriguezii and its exclusive disperser, the endemic lizard Podarcis lilfordi. Plant recruitment kernels were chiefly determined by the disperser's patterns of space utilization (i.e. the lizard's displacement kernels), the position of the various plant individuals in relation to them, and habitat structure (vegetation cover vs. bare soil). In contrast to our expectations, seed gut-passage rate and its effects on germination, and lizard speed-of-movement, habitat choice and activity rhythm were of minor importance. Predicted plant recruitment kernels were strongly anisotropic and fine-grained, preventing their description using one-dimensional, frequency-distance curves. We found a general trade-off between recruitment probability and dispersal distance; however, optimal recruitment sites were not necessarily associated to sites of maximal adult-plant density. Conservation efforts aimed at enhancing the regeneration of endangered plant-disperser systems may gain in efficacy by manipulating the spatial distribution of dispersers (e.g. through the creation of refuges and feeding sites) to create areas favourable to plant recruitment

Reduced Reactivation from Dormancy but Maintained Lineage Choice of Human Mesenchymal Stem Cells with Donor Age

Mesenchymal stem cells (MSC) are promising for cell-based regeneration therapies but up to date it is still controversial whether their function is maintained throughout ageing. Aim of this study was to address whether frequency, activation in vitro, replicative function, and in vitro lineage choice of MSC is maintained throughout ageing to answer the question whether MSC-based regeneration strategies should be restricted to younger individuals. MSC from bone marrow aspirates of 28 donors (5–80 years) were characterized regarding colony-forming unit-fibroblast (CFU-F) numbers, single cell cloning efficiency (SSCE), osteogenic, adipogenic and chondrogenic differentiation capacity in vitro. Alkaline phosphatase (ALP) activity, mineralization, Oil Red O content, proteoglycan- and collagen type II deposition were quantified. While CFU-F frequency was maintained, SSCE and early proliferation rate decreased significantly with advanced donor age. MSC with higher proliferation rate before start of induction showed stronger osteogenic, adipogenic and chondrogenic differentiation. MSC with high osteogenic capacity underwent better chondrogenesis and showed a trend to better adipogenesis. Lineage choice was, however, unaltered with age. Conclusion: Ageing influenced activation from dormancy and replicative function of MSC in a way that it may be more demanding to mobilize MSC to fast cell growth at advanced age. Since fast proliferation came along with high multilineage capacity, the proliferation status of expanded MSC rather than donor age may provide an argument to restrict MSC-based therapies to certain individuals

Seamounts

Definition: Seamounts are literally mountains rising from the seafloor. More specifically, they are “any geographically isolated topographic feature on the seafloor taller than 100 m, including ones whose summit regions may temporarily emerge above sea level, but not including features that are located on continental shelves or that are part of other major landmasses” (Staudigel et al., 2010). The term “guyot” can be used for seamounts having a truncated cone shape with a flat summit produced by erosion at sea level (Hess, 1946), development of carbonate reefs (e.g., Flood, 1999), or partial collapse due to caldera formation (e.g., Batiza et al., 1984). Seamounts <1,000 m tall are sometimes referred to as “knolls” (e.g., Hirano et al., 2008). “Petit spots” are a newly discovered subset of sea knolls confined to the bulge of subducting oceanic plates of oceanic plates seaward of deep-sea trenches (Hirano et al., 2006)

Taxonomic distribution and origins of the extended LHC (light-harvesting complex) antenna protein superfamily

<p>Abstract</p> <p>Background</p> <p>The extended light-harvesting complex (LHC) protein superfamily is a centerpiece of eukaryotic photosynthesis, comprising the LHC family and several families involved in photoprotection, like the LHC-like and the photosystem II subunit S (PSBS). The evolution of this complex superfamily has long remained elusive, partially due to previously missing families.</p> <p>Results</p> <p>In this study we present a meticulous search for LHC-like sequences in public genome and expressed sequence tag databases covering twelve representative photosynthetic eukaryotes from the three primary lineages of plants (Plantae): glaucophytes, red algae and green plants (Viridiplantae). By introducing a coherent classification of the different protein families based on both, hidden Markov model analyses and structural predictions, numerous new LHC-like sequences were identified and several new families were described, including the red lineage chlorophyll <it>a/b</it>-binding-like protein (RedCAP) family from red algae and diatoms. The test of alternative topologies of sequences of the highly conserved chlorophyll-binding core structure of LHC and PSBS proteins significantly supports the independent origins of LHC and PSBS families via two unrelated internal gene duplication events. This result was confirmed by the application of cluster likelihood mapping.</p> <p>Conclusions</p> <p>The independent evolution of LHC and PSBS families is supported by strong phylogenetic evidence. In addition, a possible origin of LHC and PSBS families from different homologous members of the stress-enhanced protein subfamily, a diverse and anciently paralogous group of two-helix proteins, seems likely. The new hypothesis for the evolution of the extended LHC protein superfamily proposed here is in agreement with the character evolution analysis that incorporates the distribution of families and subfamilies across taxonomic lineages. Intriguingly, stress-enhanced proteins, which are universally found in the genomes of green plants, red algae, glaucophytes and in diatoms with complex plastids, could represent an important and previously missing link in the evolution of the extended LHC protein superfamily.</p