Cellulase and hemicellulase enzymes as single molecular nanobiocomposites

We have worked out two different stabilization procedures for glycolytic enzymes with significant biotechnology relevance: for the commercial Novozyme cellulase complex (Celluclast) and for recombinant Thermobifida fusca hemicellulases (β-D-xylosidase and β-D-mannosidase). In the applied methods, individual cellulase and hemicellulase enzyme molecules were conjugated with a polymer nano-layer providing nanobiocomposites (single enzyme nanoparticles, SENs) that exihibit an excellent stability under extreme conditions. The first method that we have been successfully used earlier on chymotrypsin [3], creates a polymer layer around the enzyme molecules with trimethoxysilyl (TMS) functionalities in three steps [4]. The second stabilization method is a novel one pot reaction resulting in an acrylamide-bisacrylamide (AA) copolymer layer around the enzyme molecules in two steps. The heat stability of the developed single enzyme nanoparticles with TMS and AA nanolayers were tested at two different temperatures (+4 °C; 80 °C) following the residual activities of the modified enzymes in regular time interval. Upon incubation of the cellulase complex nanoparticles with TMS and the non-modified cellulase at 80 °C, the nanoparticle has kept its 40% of its starting cellulase activity after 6 hours, whereas the non-modified enzyme lost its activity completely in half an hour. The relative activity of SENs with AA is about 50% of the initial activity. After 12 hours under 80 °C the activity of NCK is reduced to 14%, while the activity of NCKA is 24% of the activity of the native enzyme at the start of the incubation. The nanoparticles of the Thermobifida fusca hemicellulases (β-D-xylosidase and β-D-mannosidase) obtained from both TMS and AA nanolayers have exhibited concomitant stability increase. TMS nanoparticles of β-xylosidase has lost 60% of its activity after 120 days during an incubation procedure at +4 °C, whereas the non-modified β-xylosidase lost all its activity in 40 days during the same conditions. The 40% of the starting enzyme activity of β-D mannosidase with AA has remained after 6 hours, during an incubation process at 80 °C, the activity of the non-modified enzyme has lost in one hour

Preference-Based Monte Carlo Tree Search

Monte Carlo tree search (MCTS) is a popular choice for solving sequential anytime problems. However, it depends on a numeric feedback signal, which can be difficult to define. Real-time MCTS is a variant which may only rarely encounter states with an explicit, extrinsic reward. To deal with such cases, the experimenter has to supply an additional numeric feedback signal in the form of a heuristic, which intrinsically guides the agent. Recent work has shown evidence that in different areas the underlying structure is ordinal and not numerical. Hence erroneous and biased heuristics are inevitable, especially in such domains. In this paper, we propose a MCTS variant which only depends on qualitative feedback, and therefore opens up new applications for MCTS. We also find indications that translating absolute into ordinal feedback may be beneficial. Using a puzzle domain, we show that our preference-based MCTS variant, wich only receives qualitative feedback, is able to reach a performance level comparable to a regular MCTS baseline, which obtains quantitative feedback.Comment: To be publishe

Serum osteoprotegerin level, carotid-femoral pulse wave velocity and cardiovascular survival in haemodialysis patients

BACKGROUND: Osteoprotegerin (OPG) is a marker and regulator of arterial calcification, and it is related to cardiovascular survival in haemodialysis patients. The link between OPG and aortic stiffening--a consequence of arterial calcification--has not been previously evaluated in this population, and it is not known whether OPG-related mortality risk is mediated by arterial stiffening. METHODS: At baseline, OPG and aortic pulse wave velocity (PWV) were measured in 98 chronic haemodialysis patients who were followed for a median of 24 months. The relationship between OPG and PWV was assessed by multivariate linear regression. The role of PWV in mediating OPG related cardiovascular mortality was evaluated by including both OPG and PWV in the same survival model. RESULTS: At baseline mean (standard deviation) PWV was 11.2 (3.3) m/s and median OPG (interquartile range) was 11.1 (7.5-15.9) pmol/L. There was a strong, positive, linear relationship between PWV and lnOPG (P = 0.009, model R(2) = 0.540) independent of covariates. During follow-up 23 patients died of cardiovascular causes. In separate univariate survival models both PWV and lnOPG were related to cardiovascular mortality [hazard ratios 1.31 (1.14-1.50) and 8.96 (3.07-26.16), respectively]. When both PWV and lnOPG were entered into the same model, only lnOPG remained significantly associated with cardiovascular mortality [hazard ratio 1.11 (0.93-1.33) and 7.18 (1.89-27.25), respectively). CONCLUSION: In haemodialysis patients OPG is strongly related to PWV and OPG related cardiovascular mortality risk is, in part, mediated by increased PWV

Vegetation-based landscape regions of Hungary.

he first version of the map of the Hungarian vegetation-based landscape regions were prepared at the scale of 1 : 200,000 (1 km or higher resolution). The primary goal of the map was to provide an exact background for the presentation and evaluation of the data of theMÉTA database. Secondly, we intended to give an up-to-date and detailed vegetation-based division of Hungary with a comprehensive nomenclature of the regions. Regions were primarily defined on the basis of their present zonal vegetation, or their dominant extrazonal or edaphic vegetation. Where this was not possible, abiotic factors that influence the potential vegetation, the flora were taken into consideration, thus, political and economical factors were ignored. All region borders were defined by local expert botanists, mainly based on their field knowledge. The map differs in many features from the currently used, country- wide, flora- or geography-based divisions in many features. We consider our map to be temporary (i.e. a work map), and we plan to refine and improve it after 5 years of testing

Overactivation of Notch1 Signaling Induces Ectopic Hair Cells in the Mouse Inner Ear in an Age-Dependent Manner

Background: During mouse inner ear development, Notch1 signaling first specifies sensory progenitors, and subsequently controls progenitors to further differentiate into either hair cells (HCs) or supporting cells (SCs). Overactivation of NICD (Notch1 intracellular domain) at early embryonic stages leads to ectopic HC formation. However, it remains unclear whether such an effect can be elicited at later embryonic or postnatal stages, which has important implications in mouse HC regeneration by reactivation of Notch1 signaling. Methodology/Principal Findings: We performed comprehensive in vivo inducible overactivation of NICD at various developmental stages. In CAG CreER+; Rosa26-NICD loxp/+ mice, tamoxifen treatment at embryonic day 10.5 (E10.5) generated ectopic HCs in the non-sensory regions in both utricle and cochlea, whereas ectopic HCs only appeared in the utricle when tamoxifen was given at E13. When tamoxifen was injected at postnatal day 0 (P0) and P1, no ectopic HCs were observed in either utricle or cochlea. Interestingly, Notch1 signaling induced new HCs in a non-cell-autonomous manner, because the new HCs did not express NICD. Adjacent to the new HCs were cells expressing the SC marker Sox10 (either NICD+ or NICDnegative). Conclusions/Significance: Our data demonstrate that the developmental stage determines responsiveness of embryonic otic precursors and neonatal non-sensory epithelial cells to NICD overactivation, and that Notch 1 signaling in the wild type, postnatal inner ear is not sufficient for generating new HCs. Thus, our genetic mouse model is suitable to test additiona

Bmp4 Is Essential for the Formation of the Vestibular Apparatus that Detects Angular Head Movements

Angular head movements in vertebrates are detected by the three semicircular canals of the inner ear and their associated sensory tissues, the cristae. Bone morphogenetic protein 4 (Bmp4), a member of the Transforming growth factor family (TGF-β), is conservatively expressed in the developing cristae in several species, including zebrafish, frog, chicken, and mouse. Using mouse models in which Bmp4 is conditionally deleted within the inner ear, as well as chicken models in which Bmp signaling is knocked down specifically in the cristae, we show that Bmp4 is essential for the formation of all three cristae and their associated canals. Our results indicate that Bmp4 does not mediate the formation of sensory hair and supporting cells within the cristae by directly regulating genes required for prosensory development in the inner ear such as Serrate1 (Jagged1 in mouse), Fgf10, and Sox2. Instead, Bmp4 most likely mediates crista formation by regulating Lmo4 and Msx1 in the sensory region and Gata3, p75Ngfr, and Lmo4 in the non-sensory region of the crista, the septum cruciatum. In the canals, Bmp2 and Dlx5 are regulated by Bmp4, either directly or indirectly. Mechanisms involved in the formation of sensory organs of the vertebrate inner ear are thought to be analogous to those regulating sensory bristle formation in Drosophila. Our results suggest that, in comparison to sensory bristles, crista formation within the inner ear requires an additional step of sensory and non-sensory fate specification

A mathematical model of quorum sensing regulated EPS production in biofilm communities

<p>Abstract</p> <p>Background</p> <p>Biofilms are microbial communities encased in a layer of extracellular polymeric substances (EPS). The EPS matrix provides several functional purposes for the biofilm, such as protecting bacteria from environmental stresses, and providing mechanical stability. Quorum sensing is a cell-cell communication mechanism used by several bacterial taxa to coordinate gene expression and behaviour in groups, based on population densities.</p> <p>Model</p> <p>We mathematically model quorum sensing and EPS production in a growing biofilm under various environmental conditions, to study how a developing biofilm impacts quorum sensing, and conversely, how a biofilm is affected by quorum sensing-regulated EPS production. We investigate circumstances when using quorum-sensing regulated EPS production is a beneficial strategy for biofilm cells.</p> <p>Results</p> <p>We find that biofilms that use quorum sensing to induce increased EPS production do not obtain the high cell populations of low-EPS producers, but can rapidly increase their volume to parallel high-EPS producers. Quorum sensing-induced EPS production allows a biofilm to switch behaviours, from a colonization mode (with an optimized growth rate), to a protection mode.</p> <p>Conclusions</p> <p>A biofilm will benefit from using quorum sensing-induced EPS production if bacteria cells have the objective of acquiring a thick, protective layer of EPS, or if they wish to clog their environment with biomass as a means of securing nutrient supply and outcompeting other colonies in the channel, of their own or a different species.</p