The effect of ripening and storage conditions on the distribution of tyramine,putrescine and cadaverine in Edam-cheese

The aim of the work was to describe the development of selected biogenic amines (histamine, tyramine, putrescine and cadaverine) in 4 layers of Dutch-type cheese (Edam-cheese) depending on 3 ripening/storage regimes during a 98-day period. Biogenic amines were analysed by means of ion-exchange chromatography. A further goal was to identify microbial sources of biogenic amines in the material analysed. Phenotype characterization and repetitive sequence-based PCR fingerprinting were used to identify the isolated bacteria. The highest content of tyramine, putrescine and cadaverine was determined in cheeses stored in a ripening cellar at a temperature of 10 °C during the whole observation period. Lower biogenic amines content was determined in samples which were moved into a cold storage device (5 °C) after 38 days of storage in a ripening cellar (10 °C). The lowest concentrations of biogenic amines were detected in cheeses which were moved into a cold storage device (5 °C) after 23 days of storage in a ripening cellar (10 °C). During the 98-day period, histamine was not detected in any of the regimes. Within the cheeses analysed, non-starter lactic acid bacteria Lactobacillus curvatus, Lactobacillus casei/paracasei and Lactobacillus plantarum were detected as the main producers of the biogenic amines tested. In starter bacteria Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris the decarboxylase activity tested was not detected.     

Gold-enhanced biomolecular surface imaging of cells and tissue by SIMS and MALDI mass spectrometry

Surface metallization by plasma coating enhances desorption/ionization of membrane components such as lipids and sterols in imaging time-of-flight secondary ion mass spectrometry (TOF-SIMS) of tissues and cells. High-resolution images of cholesterol and other membrane components were obtained for neuroblastoma cells and revealed subcellular details (resolving power 1.5 mum). Alternatively, in matrix-enhanced SIMS, 2,5-dihydroxybenzoic acid electrosprayed on neuroblastoma cells allowed intact molecular ion imaging of phosphatidylcholine and sphingomyelin at the cellular level. Gold deposition on top of matrix-coated rat brain tissue sections strongly enhanced image quality and signal intensity in stigmatic matrix-assisted laser desorption/ionization imaging mass spectrometry. High-quality total ion count images were acquired, and the neuropeptide vasopressin was localized in the rat brain tissue section at the hypothalamic area around the third ventricle. Although the mechanism of signal enhancement by gold deposition is under debate, the results we have obtained for cells and tissue sections illustrate the potential of this sample preparation technique for biomolecular surface imaging by mass spectrometry.     

Key to analyte migration and retention in electrochromatography

This work identifies electrical field-induced concentration polarization (CP) as a key physical mechanism influencing the retention behavior of charged analytes in electrochromatography with fixed beds of porous adsorbent particles. Due to an insufficient screening of intraparticle surface charge, under most general conditions the porous (permeable) particles become charge-selective. CP is caused by coupled mass and charge transport normal to the charge-selective external surface of the permeable particles, which leads to concentration gradients of ionic species in the adjoining interparticle electrolyte solution. Cation-exchange (cation-selective) particles were employed to investigate the influence of applied voltage on the retention factor of counterionic, i.e., positively charged, analytes. It is demonstrated by macroscopic retention data and microscopic studies resolving the CP phenomenon on a particle scale that the dependence of CP on electrical field and mobile-phase ionic strengths is directly reflected in concomitant changes of analyte retention. The CP zones that develop at the interface between interparticle and intraparticle pore space are recognized by charged, but not electroneutral analytes while entering or leaving the particles. The intensity of these convective-diffusion boundary layers (CP zones) depends on the applied field strength and charge selectivity of a particle. Thus, it is the charge-selective transport between the interparticle and intraparticle pore space in packed beds that prevails under typical experimental conditions in electrochromatography and that forms the physical basis for a general electrical field dependence of the retention factor of charged analytes.     

Broad-specificity immunoassays for sulfonamide detection: immunochemical strategy for generic antibodies and competitors

Development of antibodies with broad specificity recognition for sulfonamide drugs was found to be surprisingly difficult when conventional immunochemical strategies were applied to hapten design. To improve the cross-reactivity pattern of antibodies for the family of sulfonamide drugs, a novel strategy based on the single-ring (fragment-derived) hapten moieties with different spacer substituent lengths was employed for the preparation of immunogens, coating conjugates, and enzyme competitors. The rabbit antibodies raised against a common (one-ring) p-aminobenzenesulfonamide hapten moiety (attached to a carrier protein through the N-1 position) in combination with a homologous hapten-peroxidase tracer allowed the detection of 15 sulfonamide species at the maximum residue limit level using direct ELISA. The two-ring 6-(4-aminobenzensulfonylamino)hexanoic hapten mimics, previously reported in the literature as a weak generic antigen, generated surprisingly superior immune responses in rabbits. The antibodies raised against this two-ring hapten were capable of detecting at least 19 and 17 sulfonamides in a direct ELISA system at the regulatory level with sensitivities corresponding to 20 and 50% binding inhibition, respectively. A negligible cross-reaction with N4 metabolites makes it possible to measure responses of parent sulfonamides in the presence of their metabolized forms. In skimmed milk, the highest limit of detection (LOD) for sulfacetamide defined as 20% inhibition was 65.2 microg x L(-1) (IC20 value), whereas the additional 18 sulfonamides tested exhibited LODs in the range of 0.2-36.8 microg x L(-1). This sensitivity allows simple multisulfonamide tests to be established for use in the laboratory or on site.     

A Comprehensive Survey of Multiagent Reinforcement Learning

Multiagent systems are rapidly finding applications in a variety of domains, including robotics, distributed control, telecommunications, and economics. The complexity of many tasks arising in these domains makes them difficult to solve with preprogrammed agent behaviors. The agents must, instead, discover a solution on their own, using learning. A significant part of the research on multiagent learning concerns reinforcement learning techniques. This paper provides a comprehensive survey of multiagent reinforcement learning (MARL). A central issue in the field is the formal statement of the multiagent learning goal. Different viewpoints on this issue have led to the proposal of many different goals, among which two focal points can be distinguished: stability of the agents' learning dynamics, and adaptation to the changing behavior of the other agents. The MARL algorithms described in the literature aim---either explicitly or implicitly---at one of these two goals or at a combination of both, in a fully cooperative, fully competitive, or more general setting. A representative selection of these algorithms is discussed in detail in this paper, together with the specific issues that arise in each category. Additionally, the benefits and challenges of MARL are described along with some of the problem domains where the MARL techniques have been applied. Finally, an outlook for the field is provided.     

On the Enzymatic Formation of Metal Base Pairs with Thiolated and pKa-Perturbed Nucleotides

The formation of artificial metal base pairs is an alluring and versatile method for the functionalization of nucleic acids. Access to DNA functionalized with metal base pairs is granted mainly by solid-phase synthesis. An alternative, yet underexplored method, envisions the installation of metal base pairs through the polymerization of modified nucleoside triphosphates. Herein, we have explored the possibility of using thiolated and pK_a-perturbed nucleotides for the enzymatic construction of artificial metal base pairs. The thiolated nucleotides S2C, S6G, and S4T as well as the fluorinated analogue 5FU are readily incorporated opposite a templating S4T nucleotide through the guidance of metal cations. Multiple incorporation of the modified nucleotides along with polymerase bypass of the unnatural base pairs are also possible under certain conditions. The thiolated nucleotides S4T, S4T, S2C, and S6G were also shown to be compatible with the synthesis of modified, high molecular weight single-stranded (ss)DNA products through TdT-mediated tailing reactions. Thus, sulfur-substitution and pK_a perturbation represent alternative strategies for the design of modified nucleotides compatible with the enzymatic construction of metal base pairs.     

Switching between negative and positive electrorheological effect of g-C3N4 by copper ions doping

Investigation of pristine g-C₃N₄ and copper ions doped g-C₃N₄ as dispersed phases in silicone-oil based electrorheological (ER) fluids is reported for the first time. Pristine g-C₃N₄ was prepared via standard thermal polycondensation of urea. Introduction of Cu ions into g-C₃N₄ polymeric network was performed by treatment of pristine powder by a solution containing tetraammoniumdiaquacopper(II) complex ions followed by annealing. The structure and properties of products were revealed with the aid of XRD, FTIR, UV–Vis and XPS, and complemented by SEM investigation of morphology, pycnometry, BET analysis, and conductivity measurements. The response of prepared ER fluids to an external electric field was examined by rheometry and the effects visualized with optical microscopy. While ER fluids based on g-C₃N₄ exhibited negative ER effect (decrease in viscosity when the field is switched-on), ER fluids based on a g-C₃N₄/Cu-doped analogue exhibited positive ER effect. Dielectric spectroscopy using Havriliak-Negami model revealed that introduction of the dopant almost doubled the dielectric relaxation strength while the relaxation time was halved. The ability of g-C₃N₄/Cu as a candidate for further studies necessary to increase the ER effect was demonstrated. Thereto, described modification of g-C₃N₄ by copper ions shall be applicable also for other metals forming ammonia complexes.     

Digitally reconstructing Van Gogh's Field with Irises near Arles part 3: Determining the original colors

In earlier articles, we determined the spatial distributions and concentrations of all pigments used by Van Gogh in his painting Field with Irises near Arles. The colors of some pigments are expected to have changed over time, especially those of chrome yellow, cochineal, and eosin lake. For all pigments in this painting, we made physical paint reconstructions by following historical sources on raw materials and production processes, and we determined their optical properties. We combined this with pigment concentration maps to reconstruct the original colors of the painting digitally. When substituting the reconstruction paints into the calculations, we found that technical‐scientific data was not sufficient to resolve several issues. In those cases, discussions within the broad interdisciplinary team allowed us to make informed decisions. These issues refer to the representation of the sky area, and the original contributions of the red lake pigments to local colors. The digitally reconstructed colors of the painting show that due to discoloring of red lake pigments, the irises in the field have changed from a warm purple to purplish blue, and many pink spots in the field have turned to white. The range of yellows in the field has decreased and partly turned to dark brown. The digital reconstruction gives a better understanding of the color scheme used by Van Gogh when compared to remarks the artist made in letters when describing this painting. Also, the original color composition is seen to be aligned with color theories on which Van Gogh based his work.