An Engineered Mannoside Presenting Platform: Escherichia coli Adhesion under Static and Dynamic Conditions

The study of the adhesion mechanisms of pathogens to host tissues has gained increased interest as bacterial adhesion is involved in the early stages of surface colonization and infection. Here we describe a platform to study the specific binding of the bacterium Escherichia coli (E. coli) K‐12 strain to molecularly well‐defined surfaces mimicking cellular interfaces. This approach uses a poly(ethylene glycol) brush interface, which displays synthetic determinants of the high mannose N‐linked glycans in a range of densities (3.8 × 10⁴–1.6 × 10⁵ mannosides µm^?2) for the investigation of multivalent interactions with bacteria. The bacterial attachment is mediated by specific interactions between the adhesive protein FimH located on the tip of the bacterial type 1 pili and the mannosylated surfaces. With synthetically engineered mannoses, it is found that the number of strongly adhering bacteria is co‐regulated by many structural physical parameters. Beyond the dependency on carbohydrate density, higher numbers of E. coli attach to the branched trimannose Man(α1–3)(Man(α1–6))Man compared to the monomannose, while larger oligomannoses exposing Man(α1–2) Man at their non reducing end show low binding capacity. The linker used between the mannose moiety and PEG is also affecting the binding efficacy of E. coli. The (hydrophobic) propyl linker results in higher bacteria numbers in comparison to the (hydrophilic) tri(EG), likely a consequence of additional stabilization of the binding complex by hydrophobic interactions. Furthermore, differences are observed in bacteria attachment between stagnant and flow conditions that depend on the type of mannose ligand. Finally, a photolithographic resist lift‐off combined with site‐selective assembly of the glycopolymers is used to produce micropatterns with bacteria colonies confined to defined areas and at controlled colony numbers.     

Comparative studies of a real-time PCR method and three enzyme-linked immunosorbent assays for the detection of central nervous system tissues in meat products

The removal of certain central nervous system (CNS) tissues (part of the bovine spongiform encephalopathy risk material) from the food chain is one of the highest priority tasks associated with avoiding contamination of the human food chain with the agent of bovine spongiform encephalopathy. A recently developed real-time PCR assay and three commercially available enzyme-linked immunosorbent assays (ELISAs) for the detection of CNS tissues in minced meat and three types of heat-treated sausages were evaluated. Bovine brain was used for spiking of internal reference material, and its detectability was examined during storage times of 12 months (for frozen minced meat and liver sausage) and 24 months (for sausages treated with medium and high heat). The real-time PCR method and both ELISA kits detected 0.1% CNS tissue in frozen minced meat and 0.1 or 1% CNS tissue in heat-treated meat products. The detectability of the amplified mRNA target region with the PCR assay was similar to the detectability of antigen by the ELISAs. Because the real-time PCR method also can be used to distinguish cattle, ovine, and caprine CNS tissues from porcine CNS tissues, it seems to be suitable as a routine diagnostic test for the sensitive and specific detection of CNS tissues in meat and meat products.     

Sorption of pyrene to dissolved humic substances and related model polymers. 2. Solid-phase microextraction (SPME) and fluorescence quenching technique (FHT) as analytical methods

Sorption coefficients for pyrene on dissolved humic substances and on poly(acrylic acid) esters as well-defined model polymers were determined using solid-phase microextraction (SPME) and the fluorescence quenching technique (FQT). The results of both analytical methods were compared and theoretically evaluated, which led to the conclusion that the sorption coefficients measured by SPME and FQT are inevitably different: SPME measures activity-based and FQT concentration-based sorption coefficients. The environmental relevance of the two types of sorption coefficients is discussed. FQT is inappropriate to measure sorption coefficients for pyrene with the synthetic sorbents. Inspection of the vibrational structure of the fluorescence spectra of those solutions indicates a highly hydrophobic microenvironment of pyrene. This can be explained by an intra- or intermolecular agglomeration of hydrophobic moieties forming a favorable host for hydrophobic solutes.     

Studies on acrylamide levels in roasting, storage and brewing of coffee

The content of acrylamide in coffee reaches a peak early in the roasting process, reflecting occurrence of both formation and destruction of acrylamide during roasting. Levels of acrylamide in the fully roasted product are a small fraction of the peak reached earlier. Glucose and moisture in green coffee do not show a significant correlation with acrylamide in roasted coffee. Pre-roasting levels of asparagine show a correlation only in Arabica coffee. The main factors affecting the level of acrylamide in roasted coffee appear to be the Arabica/Robusta ratio, with Robusta giving higher levels; time and degree of roast, with both shorter and lighter roasting at the edges of the normal roasting range giving higher levels; storage condition and time, with clear reduction at ambient storage. This storage reduction of acrylamide followed second order reaction kinetics with an activation energy of 73 KJ/mole. The acrylamide in roasted coffee is largely extracted into the brew and stable within usual time of consumption. As these four main factors also substantially affect the sensorial characteristics of the brew, and as modifications of the process have to comply with the consumer-accepted boundaries of taste profiles, only small effects on the acrylamide level are expected to be achievable.     

Influence of store dimensions and auxiliary volume configuration on the performance of medium-sized solar combisystems

To increase the fractional energy savings achieved with solar thermal combisystems the store volume may be increased. Installation of large stores in single-family houses is, however, often limited by space constraints. In this article the influence of the store dimensions, as well as internal and external auxiliary volume configurations, are investigated for large solar water stores by annual dynamic TRNSYS simulations. The results show that store sizes up to 4 m³ may be used in solar heating systems with 30 m² collector area. It is further shown that well-insulated stores are rather insensitive to the geometry. Stores deviating from the conventional dimensions still yield high fractional energy savings. Furthermore, the simulations show that the performance of an internal auxiliary volume configuration in most cases exceeds that of a solution with an external auxiliary unit. The practical limitations of very thin auxiliary volumes must, however, be further investigated.     

Biomass production,water use and photosynthesis of Salix clones grown in a wastewater purification system

An objective of the study was to identify leaf morphophysiological characteristics indicative of high aboveground woody biomass production of Salix clones growing in the vegetation filter of a wastewater purification system. It was hypothesized, that in these extra irrigated stands, the characteristics of photosynthesis are stronger determinants of the production than the characteristics of leaf water relations. The biomass production of the clones was correlated with numerous characteristics of leaf water relations and photosynthesis. Significant correlation was found between the biomass production and the following characteristics of top foliage leaves: daily decrease in water potential, stomatal conductance, stomatal sensitivity to increase in light intensity, and hydraulic conductance (positive); daily minimum and maximum water potentials, instantaneous water use efficiency, and stomatal sensitivity to decrease in leaf water potential (negative); maximum rate of photosynthetic electron transport, chlorophyll content, and the partitioning coefficient for leaf nitrogen into bioenergetics associated with electron transport (positive). In the most productive clones, the vertical gradient (through foliage) of values of almost all the characteristics was the smallest. The biomass correlated more strongly with the characteristics of photosynthesis light stage than with the characteristics of water relations. We conclude that the characteristics of photosynthesis light stage – high maximum rate of photosynthetic electron transport, high chlorophyll content, and also flat vertical gradient (through foliage) in values of the characteristics could be the strongest indicators of high biomass production of Salix clones growing in vegetation filter.     

Heat and mass transfer during the coffee drying process

For a better comprehension of heat and mass transfer during the coffee drying process and optimization of the industrial application transport coefficients and coffee properties were determined. Heat transfer coefficients were measured for different air velocities and were found to follow the known dimensionless equations for the flow surrounding a sphere. Thermal conductivities and effective diffusion coefficients were measured as a function of moisture content as well as volume and densities of the coffee beans. The mentioned properties depend directly on the humidity of the coffee beans rather than on the drying conditions. Sorption behaviour was investigated and temperature dependent parameters for the Guggenheimer–Anderson–deBoer-isotherm (GAB) were determined according to the Arrhenius relationship.     

Enzymatic formation of styrene during wheat beer fermentation is dependent on pitching rate and cinnamic acid content

Styrene is formed by the thermal decarboxylation of cinnamic acid during wort boiling or by enzymatic decarboxylation during fermentation. The enzymatic reaction processes simultaneously to the decarboxylation of ferulic‐ and p‐cumaric acid to clove‐like 4‐vinylguaiacol and phenolic 4‐vinylphenol by the same PAD1 and FDC1 decarboxylase enzymes. However, the formation of styrene occurs much faster within the first hours of fermentation. In addition, the conversion of cinnamic acid starts immediately after pitching without an adaption of yeast on the new medium. Only after 120 min does the level of transposition decrease. Moreover, high cinnamic acid content in pitching wort, in combination with an open fermentation management, causes faster and higher styrene formation during this period. In contrast to the formation of 4‐vinylguaiacol, a correlation between pitching rate and styrene formation during open fermentation could be shown. The resulting time interval between styrene and 4‐vinylguaiacol formation provides scope for minimization strategies for styrene, while maintaining the typical wheat beer flavours. Copyright © 2012 The Institute of Brewing & Distilling