Secretion and assembly of functional mini-cellulosomes from synthetic chromosomal operons in Clostridium acetobutylicum ATCC 824.

Background: Consolidated bioprocessing (CBP) is reliant on the simultaneous enzyme production, saccharification of biomass, and fermentation of released sugars into valuable products such as butanol. Clostridial species that produce butanol are, however, unable to grow on crystalline cellulose. In contrast, those saccharolytic species that produce predominantly ethanol, such as Clostridium thermocellum and Clostridium cellulolyticum, degrade crystalline cellulose with high efficiency due to their possession of a multienzyme complex termed the cellulosome. This has led to studies directed at endowing butanol-producing species with the genetic potential to produce a cellulosome, albeit by localising the necessary transgenes to unstable autonomous plasmids. Here we have explored the potential of our previously described Allele-Coupled Exchange (ACE) technology for creating strains of the butanol producing species Clostridium acetobutylicum in which the genes encoding the various cellulosome components are stably integrated into the genome. Results: We used BioBrick2 (BB2) standardised parts to assemble a range of synthetic genes encoding C. thermocellum cellulosomal scaffoldin proteins (CipA variants) and glycoside hydrolases (GHs, Cel8A, Cel9B, Cel48S and Cel9K) as well as synthetic cellulosomal operons that direct the synthesis of Cel8A, Cel9B and a truncated form of CipA. All synthetic genes and operons were integrated into the C. acetobutylicum genome using the recently developed ACE technology. Heterologous protein expression levels and mini-cellulosome self-assembly were assayed by western blot and native PAGE analysis. Conclusions: We demonstrate the successful expression, secretion and self-assembly of cellulosomal subunits by the recombinant C. acetobutylicum strains, providing a platform for the construction of novel cellulosomes. © 2013 Kovács et al.; licensee BioMed Central Ltd

The dynamic relationships between the active and catabolic vitamin D metabolites, their ratios, and associations with PTH

Vitamin D status, assessed by serum concentration of 25(OH)D, is the prime candidate marker for many disease-association studies, but the interplay between the subsequent 1,25-dihydroxyvitamin D (1,25(OH)2D) and 24,25-dihydroxyvitamin D (24,25(OH)2D) metabolites is unclear. In this study, we conducted an analysis from a large cohort of healthy, physically fit, young army recruits (n = 940). We found a significant, inverse relationship between serum 25(OH)D and 1,25(OH)2D:24,25(OH)2D vitamin D metabolite ratio (VMR) (r(2)Exp = 0.582, p /=35 to be the threshold value for vitamin D insufficiency, and >/=51 to be predictive of vitamin D deficiency. Our three-dimensional model provides mechanistic insight into the vitamin D-PTH endocrine system, and further substantiates the role of 24,25(OH)2D in human physiology. The model sets a new paradigm for vitamin D treatment strategy, and may help the establishment of vitamin D-adjusted PTH reference intervals. The study was approved by the UK Ministry of Defence research ethics committee (MODREC 165/Gen/10 and 692/MoDREC/15). ClinicalTrials.gov Identifier NCT02416895

The performance of plant species in removing nutrients from stormwater in biofiltration systems in Cape Town

In 2009, the City of Cape Town (CoCT) adopted a stormwater policy which mandates that new and existing developments should reduce the concentration of phosphorus and suspended solids in stormwater runoff by 45% and 80%, respectively, but offered no explicit guidance about how these water quality targets might be achieved. This study aims to contribute to the limited knowledge that exists about the performance of local plant species to treat stormwater. A large nursery-based study was conducted to investigate the performance of 9 locally-occurring plant species to remove orthophosphate (PO4-3), ammonia (NH3) and nitrate (NO3-) found in urban stormwater. Synthetic stormwater was applied to each species as well as a control consisting only of soil (Malmesbury shale). The discharge was collected from a drainage pipe at the base of each of the 150 containers. The results show that all species (excluding Ficinia) reduced the average concentrations of PO4-3 by 81% and NH3 by 90%. By contrast, NO3- was reduced by an average of 69% (excluding by Elegia and Phragmites) with 8 of the 9 species removing significantly more than the control. The species that performed well for all three nutrients include Agapanthus and turf grasses, Stenotaphrum and Pennisetum. The results of the study highlight three important factors in the design of biofilters: that a substantial proportion of nutrients can be captured or absorbed by plants; that the soil medium is an important factor in the removal of PO4-3 and NH3; and that plant choice is essential in the removal of NO3-. Future research should test plant species in both the laboratory and field settings, and should include additional contaminants such as household detergents, heavy metals and bacteria

Beyond Gross-Pitaevskii Mean Field Theory

A large number of effects related to the phenomenon of Bose-Einstein Condensation (BEC) can be understood in terms of lowest order mean field theory, whereby the entire system is assumed to be condensed, with thermal and quantum fluctuations completely ignored. Such a treatment leads to the Gross-Pitaevskii Equation (GPE) used extensively throughout this book. Although this theory works remarkably well for a broad range of experimental parameters, a more complete treatment is required for understanding various experiments, including experiments with solitons and vortices. Such treatments should include the dynamical coupling of the condensate to the thermal cloud, the effect of dimensionality, the role of quantum fluctuations, and should also describe the critical regime, including the process of condensate formation. The aim of this Chapter is to give a brief but insightful overview of various recent theories, which extend beyond the GPE. To keep the discussion brief, only the main notions and conclusions will be presented. This Chapter generalizes the presentation of Chapter 1, by explicitly maintaining fluctuations around the condensate order parameter. While the theoretical arguments outlined here are generic, the emphasis is on approaches suitable for describing single weakly-interacting atomic Bose gases in harmonic traps. Interesting effects arising when condensates are trapped in double-well potentials and optical lattices, as well as the cases of spinor condensates, and atomic-molecular coupling, along with the modified or alternative theories needed to describe them, will not be covered here.Comment: Review Article (19 Pages) - To appear in 'Emergent Nonlinear Phenomena in Bose-Einstein Condensates: Theory and Experiment', Edited by P.G. Kevrekidis, D.J. Frantzeskakis and R. Carretero-Gonzalez (Springer Verlag

Arsenic, Organic Foods, and Brown Rice Syrup

Background: Rice can be a major source of inorganic arsenic (Asi) for many sub-populations. Rice products are also used as ingredients in prepared foods, some of which may not be obviously rice based. Organic brown rice syrup (OBRS) is used as a sweetener in organic food products as an alternative to high-fructose corn syrup. We hypothesized that OBRS introduces As into these products

The Hall Technique 10 years on: its effect and influence

Dental caries in early childhood can have a very significant effect not only on the oral health of young children but on their quality of life and that of their families. Added to this are the long-term infective risks to the host of untreated caries in primary teeth but also the risk of damage to successor permanent teeth. Traditional restoration of damaged primary teeth has been shown to have only moderate outcomes depending on the techniques and materials used and the ability of children to cooperate because of age or other factors. The Hall technique has outperformed other techniques in randomised clinical trials. In this Opinion Article I will take a look back at the Innes et al. paper, first published in this Journal in 2006, that introduced the Hall Technique and the subsequent scientific literature which provided us with high quality scientific evidence of the efficacy of the technique. I will evaluate how this new technique has affected the management of early childhood caries and hence its influence on wider oral and general health

Probabilistic analysis of degradation of façade claddings using Markov chain models

In this study, the time-dependent stochastic degradation of three types of claddings is analysed. For this purpose, 203 fac¸ades with stone claddings(directly adhered to the substrate), 195 with adhered ceramic claddings and 220 with painted surfaces were analysed. All the fac¸ades are located in Lisbon, Portugal. Their degradation condition was assessed through an extensive field work. Based on the data gathered, Markov chains are used to predict the degradation of claddings and to understand, in some detail, how the characteristics of the claddings contribute to the overall degradation. The results show that the distance from the sea and exposure to damp are significant to the degradation of all types of cladding. The type and size of stone plates also influence the degradation of stone claddings. The exposure to wind-rain action has a high impact on the degradation of ceramic claddings. The models proposed provide useful information on the probability of failure of the claddings; these results are fundamental in the context of insurance policies and in the definition of building maintenance plans

Capsular profiling of the Cronobacter genus and the association of specific Cronobacter sakazakii and C. malonaticus capsule types with neonatal meningitis and necrotizing enterocolitis

Background: Cronobacter sakazakii and C. malonaticus can cause serious diseases especially in infants where they are associated with rare but fatal neonatal infections such as meningitis and necrotising enterocolitis. Methods: This study used 104 whole genome sequenced strains, covering all seven species in the genus, to analyse capsule associated clusters of genes involved in the biosynthesis of the O-antigen, colanic acid, bacterial cellulose, enterobacterial common antigen (ECA), and a previously uncharacterised K-antigen. Results: Phylogeny of the gnd and galF genes flanking the O-antigen region enabled the defining of 38 subgroups which are potential serotypes. Two variants of the colanic acid synthesis gene cluster (CA1 and CA2) were found which differed with the absence of galE in CA2. Cellulose (bcs genes) were present in all species, but were absent in C. sakazakii sequence type (ST) 13 and clonal complex (CC) 100 strains. The ECA locus was found in all strains. The K-antigen capsular polysaccharide Region 1 (kpsEDCS) and Region 3 (kpsMT) genes were found in all Cronobacter strains. The highly variable Region 2 genes were assigned to 2 homology groups (K1 and K2). C. sakazakii and C. malonaticus isolates with capsular type [K2:CA2:Cell+] were associated with neonatal meningitis and necrotizing enterocolitis. Other capsular types were less associated with clinical infections. Conclusion: This study proposes a new capsular typing scheme which identifies a possible important virulence trait associated with severe neonatal infections. The various capsular polysaccharide structures warrant further investigation as they could be relevant to macrophage survival, desiccation resistance, environmental survival, and biofilm formation in the hospital environment, including neonatal enteral feeding tubes

How many motoric body representations can we grasp?

At present there is a debate on the number of body representations in the brain. The most commonly used dichotomy is based on the body image, thought to underlie perception and proven to be susceptible to bodily illusions, versus the body schema, hypothesized to guide actions and so far proven to be robust against bodily illusions. In this rubber hand illusion study we investigated the susceptibility of the body schema by manipulating the amount of stimulation on the rubber hand and the participant’s hand, adjusting the postural configuration of the hand, and investigating a grasping rather than a pointing response. Observed results showed for the first time altered grasping responses as a consequence of the grip aperture of the rubber hand. This illusion-sensitive motor response challenges one of the foundations on which the dichotomy is based, and addresses the importance of illusion induction versus type of response when investigating body representations

Good perceived sleep quality protects against the raised risk of respiratory infection during sleep restriction in young adults.

Study Objectives: Prospectively examine the association between sleep restriction, perceived sleep quality (PSQ) and upper respiratory tract infection (URTI). Methods: In 1318 military recruits (68% males) self-reported sleep was assessed at the beginning and end of a 12-week training course. Sleep restriction was defined as an individualized reduction in sleep duration of ≥2 hours/night compared with civilian life. URTIs were retrieved from medical records. Results: On commencing training, approximately half of recruits were sleep restricted (52%; 2.1 ± 1.6 h); despite the sleep debt, 58% of recruits with sleep restriction reported good PSQ. Regression adjusted for covariates showed that recruits commencing training with sleep restriction were more likely to suffer URTI during the course (OR = 2.93, 95% CI 1.29–6.69, p = .011). Moderation analysis showed this finding was driven by poor PSQ (B = −1.12, SE 0.50, p = .023), as no significant association between sleep restriction and URTI was observed in recruits reporting good PSQ, despite a similar magnitude of sleep restriction during training. Associations remained in the population completing training, accounting for loss to follow-up. Recruits reporting poor PSQ when healthy at the start and end of training were more susceptible to URTI (OR = 3.16, 95% CI 1.31–7.61, p = .010, vs good PSQ). Conclusion: Good perceived sleep quality was associated with protection against the raised risk of respiratory infection during sleep restriction. Studies should determine whether improvements in sleep quality arising from behavioral sleep interventions translate to reduced respiratory infection during sleep restriction