Development and application of new nucleic acid-based technologies for microbial community analyses in foods

Several challenges still persist in the analysis of microorganisms in foods, particularly in studies of complex communities. Nucleic acid-based methods are promising tools in addressing new questions concerning microbial communities. We have developed several new methods in the field of nucleic acid-based microbial community analyses. These methods cover both sample preparation and detection approaches. The sample preparation method involves simplified DNA purification using paramagnetic beads. As an extension of this method, the same paramagnetic beads are used for both cell separation and DNA purification. This enables full automation. The separate detection of viable and dead bacteria is a major issue in nucleic acid-based diagnostics. We have applied a living/dead dye that binds covalently to DNA and inhibits the PCR from dead cells. In addition, a DNA array-based detection assay has been developed. The assay combines the specificity obtained by enzymatic labeling of DNA probes with the possibility of detecting several targets simultaneously by DNA array hybridization. In combination with 16S rDNA amplification, this is a promising tool for community analyses. Also, we have developed a novel approach for multiplex quantitative PCR. The multiplex PCR has been combined with our DNA array-based detection method. Finally, we are now in the process of adapting a system for monitoring microbial growth and death in real-time through the tagging of bacteria with green fluorescent protein (GFP) combined with fluorescence detection using a high-resolution confocal laser scanner.     

Design criteria for liners of concrete vessels of future HTRs

The paper summarizes the work on design criteria for liners of Prestressed Concrete Reactor Vessels (PCRVs), which has been carried out by several German institutions from 1984 to 1988 within a future High Temperature Reactor (HTR) research project.The liner is discussed as part of the concept guaranteeing the integrity of the safe enclosure of the cooling medium. Its main function is leak-tightness during the whole lifetime of the power station. On the basis of the minor safety function of the composite liner compared to that of the prestressed concrete structure with its pressure bearing function and its integrity requirements the combined action and its effects between the liner and the concrete structure has been worked out. As it is shown the composite structure ‘steel liner + anchorage + concrete structure’ guarantees a high safety of the liner.The results deal with general and special requirements to the liner-anchor-system with regard to material, design details, analytical methods, fabrication and testing.     

Solid-state Reactions and Phase Relations in the Ti-Si-O System at 1373 K

Reactions between Ti and SiO₂ were studied at 1373 K (1100°C) under vacuum conditions using planar diffusion couples. A method to correct for the presence of surface oxide was developed which led to improved oxygen measurements with the electron probe microanalyzer. An isothermal section through the Ti-Si-O phase diagram at 1373 K was determined using measured diffusion paths and phase compositions in equilibrated alloys. The experimentally determined isothermal section was compared to isothermal sections calculated using thermodynamic data. In addition the sequence of reaction layers formed in the diffusion couples is discussed in terms of thermodynamic activity diagrams.     

Cloning and sequencing of the URA3 gene of Kluyveromyces marxianus CBS 6556

The URA3 gene, coding for orotidine-5′-phosphate decarboxylase, from Kluyveromyces marxianus CBS 6556, was isolated from a genomic DNA library. The K. marxianus URA3 gene encodes a protein of 267 amino acids with a calculated molecular weight of 29·3 kDa. Comparison of the K. marxianus protein with the corresponding enzymes of Saccharomyces cerevisiae and Kluyveromyces lactis showed amino acid sequence identifies of 81% and 88%, respectively. Using contour-clamped homogeneous electric field gel electrophoresis, the genomic copy was found to be located on chromosome VI. We have used the cloned gene for the construction of a K. marxianus leu2 mutant. This mutant contains no heterologous sequences, which is essential to make it acceptable for application in the food industry.     

The effect of processing conditions on glucosinolates in cruciferous vegetables

Summary Glucosinolates are found naturally in cruciferous plants, such asbrassica vegetables, which are important in the human diet. Glucosinolates are readily converted into a range of physiologically active compounds by hydrolysis under the influence of an endogenous thioglucosidase, which becomes active when plant cells are disrupted. Cruciferous vegetables are eaten raw, they are also consumed in the processed form. The influence of processing conditions on glucosinolate degradation and on the type and levels of the reaction products formed is reviewed.

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Der Einfluß von Prozeßbedingungen auf Glucosinolate in Cruciferen-Gemüse

Zusammenfassung Glucosinolate kommen naturgemäß in Kreuzblütlern vor, wie z. B. inBrassica-Gemüse, welche einen wesentlichen Bestandteil der Nahrung bilden. Glucosinolate werden leicht umgesetzt in eine Reihe physiologisch aktiver Verbindungen unter Einfluß einer endogenen Thioglucosidase, welche aktiv wird, wenn das pflanzliche Gewebe zerstört wird. Die Cruciferen-Gemüse werden roh, aber auch in verarbeiteter Form, gegessen. Der Einfluß der Verarbeitung auf den Abbau der Glucosinolate, sowie auf Art und Menge der gebildeten Reaktionssprodukte, wird besprochen.

     

Induction of Gushing with Recombinant Class II Hydrophobin FcHyd5p from Fusarium culmorum and the Impact of Hop Compounds on its Gushing Potential

Malt‐induced gushing is a problem that has been known for many years. Mechanisms and inducing agents are still not fully understood and identified. Hydrophobins produced by various filamentous fungi are currently under discussion as biological gushing‐inducing compounds. In the current study the class II hydrophobin FcHyd5p, from the cereal pathogen Fusarium culmorum, was employed in beer and other carbonated beverages for gushing experiments and the influence of hop compounds on gushing potential was examined. It was demonstrated that this protein strongly induces gushing in various carbonated beverages, including beer. It was further demonstrated that the resulting gushing volume is susceptible to certain hop compounds and can be decreased significantly by the addition of these substances.     

Ligand-Tuning of the Stability of Pd(II) Conjugates with Cyanocobalamin

Besides the well-known functions performed by vitamin B₁₂ (CblCN) in biochemical processes of the human body, an increasing interest has been raised by the possibility of its use as a transmembrane drug carrier, capable, among others, of enhancing the accumulation of inorganic cytostatics in cancer cells. The present study was aimed at determining the possibility of the formation of CblCN conjugates with Pd(II) complexes. A key aspect was their stability, which we attempted to tune by appropriate choice of ligands. Syntheses, spectroscopic analysis of postreaction systems and kinetic investigations of conjugate formation reactions, have been complemented by DFT modelling. The obtained results showed that ligand charge, geometry and electron affinity may have a significant impact on carrier binding and release leading to the activation of the Pd(II) complex. This provides a rationale to expect that with appropriate composition of the coordination sphere, it will be possible to extend the spectrum of less toxic inorganic chemotherapeutics.     

Multilayer assembly of ionic starches on old corrugated container recycled cellulosic fibers

In this study, old corrugated container recycled fibers were treated with polyelectrolyte multilayers consisting of biopolymer cationic starch with two degrees of substitution (DS) each in combination with one anionic starch. Pulp zeta potential, paper strength and the thin layer ellipsometry technique were applied to examine the influence of cationic starch DS on the formation of polyelectrolyte multilayers. The results indicated a significant interaction between the DS of cationic starch and the number of ionic starch layers formed. When low‐DS cationic starch was used, the pulp zeta potential and the paper strength increased significantly in assembling the first cationic layer. However, in depositing high‐DS cationic starch a greater zeta potential and a stronger influence on the paper strength were observed with a larger number of starch layers. This was confirmed by thin layer ellipsometry when a greater thickness of multilayers was achieved by employing high‐DS cationic starch to form a higher number of layers. © 2017 Society of Chemical Industry