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OBJECTIVE: To evaluate the clinical outcome of root canal treatment by obturation technique, root canal filling quality, and tooth- and patient-related variables. METHODS: This clinical study investigated the endodontic outcome of 114 teeth treated between the years 2009 and 2012. Three different obturation methods were used: (1) adhesive obturation using the continuous wave of condensation technique with Resilon® (CWR), (2) matching-taper single-cone technique with gutta-percha and AH Plus® (SCGP), and (3) matching-taper single-cone technique with gutta-percha and GuttaFlow® (SCGF). Pre- and postoperative periapical radiographs were performed to detect the presence of endodontic lesions (PAI classification) and to assess the quality of both the obturation and the restoration. Tooth- and patient-related data were collected. RESULTS: The overall endodontic success rate was 75.4% after a mean observation period of 6.3 years. There were no significant correlations between the type or overall quality of obturation and the treatment outcome. Teeth with preoperative lesions had the highest odds ratio (factor of 4.98) for endodontic failure. Tooth- and patient-related variables had no significant effect on endodontic outcome. CONCLUSIONS: The preoperative periapical status of teeth requiring endodontic treatment was a substantial prognostic factor for endodontic outcome, whereas the type of obturation material or technique did not affect it.

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Acetic acid bacteria (AAB) play an important role during cocoa fermentation, as their main product, acetate, is a major driver for the development of the desired cocoa flavors. Here, we investigated the specialized metabolism of these bacteria under cocoa pulp fermentation-simulating conditions. A carefully designed combination of parallel 13C isotope labeling experiments allowed the elucidation of intracellular fluxes in the complex environment of cocoa pulp, when lactate and ethanol were included as primary substrates among undefined ingredients. We demonstrate that AAB exhibit a functionally separated metabolism during coconsumption of two-carbon and three-carbon substrates. Acetate is almost exclusively derived from ethanol, while lactate serves for the formation of acetoin and biomass building blocks. Although this is suboptimal for cellular energetics, this allows maximized growth and conversion rates. The functional separation results from a lack of phosphoenolpyruvate carboxykinase and malic enzymes, typically present in bacteria to interconnect metabolism. In fact, gluconeogenesis is driven by pyruvate phosphate dikinase. Consequently, a balanced ratio of lactate and ethanol is important for the optimum performance of AAB. As lactate and ethanol are individually supplied by lactic acid bacteria and yeasts during the initial phase of cocoa fermentation, respectively, this underlines the importance of a well-balanced microbial consortium for a successful fermentation process. Indeed, AAB performed the best and produced the largest amounts of acetate in mixed culture experiments when lactic acid bacteria and yeasts were both present.

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Anaerobic digestion of organic waste generated by households, businesses, agriculture, and industry is an important approach as method of waste treatment - especially with regard to its potential as an alternative energy source and its cost-effectiveness. Separate collection of biowaste from households or vegetal waste from public green spaces is already established in some EU-27 countries. The material recovery in composting plants is common for biowaste and vegetal waste. Brewery waste fractions generated by beer production are often used for animal feeding after a suitable preparation. Waste streams from paper industry generated by pulp and paper production such as black liquor or paper sludge are often highly contaminated with toxic substances. Recovery of chemicals and the use in thermal processes like incineration, pyrolysis, and gasification are typical utilization paths. The current utilization of organic waste from households and institutions (without agricultural waste) was investigated for EU-27 countries with Germany as an in-depth example. Besides of biowaste little is known about the suitability of waste streams from brewery and paper industry for anaerobic digestion. Therefore, an evaluation of the most important biogas process parameters for different substrates was carried out, in order to calculate the biogas utilization potential of these waste quantities. Furthermore, a calculation of biogas energy potentials was carried out for defined waste fractions which are most suitable for anaerobic digestion. Up to 1% of the primary energy demand can be covered by the calculated total biogas energy potential. By using a "best-practice-scenario" for separately collected biowaste, the coverage of primary energy demand may be increased above 2% for several countries. By using sector-specific waste streams, for example the German paper industry could cover up to 4.7% and the German brewery industry up to 71.2% of its total energy demand.

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In the present work, simulated cocoa fermentation was investigated at the level of metabolic pathway fluxes (fluxome) of lactic acid bacteria (LAB), which are typically found in the microbial consortium known to convert nutrients from the cocoa pulp into organic acids. A comprehensive (13)C labeling approach allowed to quantify carbon fluxes during simulated cocoa fermentation by (i) parallel (13)C studies with [(13)C6]glucose, [1,2-(13)C2]glucose, and [(13)C6]fructose, respectively, (ii) gas chromatography-mass spectrometry (GC/MS) analysis of secreted acetate and lactate, (iii) stoichiometric profiling, and (iv) isotopomer modeling for flux calculation. The study of several strains of L. fermentum and L. plantarum revealed major differences in their fluxes. The L. fermentum strains channeled only a small amount (4 to 6%) of fructose into central metabolism, i.e., the phosphoketolase pathway, whereas only L. fermentum NCC 575 used fructose to form mannitol. In contrast, L. plantarum strains exhibited a high glycolytic flux. All strains differed in acetate flux, which originated from fractions of citrate (25 to 80%) and corresponding amounts of glucose and fructose. Subsequent, metafluxome studies with consortia of different L. fermentum and L. plantarum strains indicated a dominant (96%) contribution of L. fermentum NCC 575 to the overall flux in the microbial community, a scenario that was not observed for the other strains. This highlights the idea that individual LAB strains vary in their metabolic contribution to the overall fermentation process and opens up new routes toward streamlined starter cultures. L. fermentum NCC 575 might be one candidate due to its superior performance in flux activity.

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A dynamic metabolic model is presented for Pediococcus pentosaceus producing lactic acid from lignocellulose-derived mixed sugars including glucose, mannose, galactose, arabinose, and xylose. Depending on the pairs of mixed sugars, P. pentosaceus exhibits diverse (i.e., sequential, simultaneous or mixed) consumption patterns. This regulatory behavior of P. pentosaceus is portrayed using the hybrid cybernetic model (HCM) framework which views elementary modes of the network as metabolic options dynamically modulated. Comprehensive data are collected for model identification and validation through fermentation experiments involving single substrates and various combinations of mixed sugars. Most sugars are metabolized rather sequentially while co-consumption of galactose and arabinose is observed. It is demonstrated that the developed HCM successfully predicts mixed sugar data based on the parameters identified mostly from single substrate data only. Further, we discuss the potential of HCMs as a tool for predicting intracellular flux distribution with comparison with flux balance analysis.

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An unstructured model for an integrated fermentation/membrane extraction process for the production of the aroma compounds 2-phenylethanol and 2-phenylethylacetate by Kluyveromyces marxianus CBS 600 was developed. The extent to which this model, based only on data from the conventional fermentation and separation processes, provided an estimation of the integrated process was evaluated. The effect of product inhibition on specific growth rate and on biomass yield by both aroma compounds was approximated by multivariate regression. Simulations of the respective submodels for fermentation and the separation process matched well with experimental results. With respect to the in situ product removal (ISPR) process, the effect of reduced product inhibition due to product removal on specific growth rate and biomass yield was predicted adequately by the model simulations. Overall product yields were increased considerably in this process (4.0 g/L 2-PE+2-PEA vs. 1.4 g/L in conventional fermentation) and were even higher than predicted by the model. To describe the effect of product concentration on product formation itself, the model was extended using results from the conventional and the ISPR process, thus agreement between model and experimental data improved notably. Therefore, this model can be a useful tool for the development and optimization of an efficient integrated bioprocess.

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