Identification and analysis of genes from Streptomyces pristinaespiralis encoding enzymes involved in the biosynthesis of the 4-dimethylamino-L-phenylalanine precursor of pristinamycin I

Dextran has been used as a carrier molecule for the synthesis of monofunctional peptide-dextran conjugates. The immunodetection of such carrier immobilized peptides on ELISA plates was compared to that of peptides adsorbed directly to immunoplates. The main features observed with peptide-dextran conjugates were as follows: only small amounts of peptide (1-2 mg) were necessary for coupling via alpha- or epsilon-amino groups to NaIO4-activated dextran (4 mg); the coupling yield was up to 68%; an amino acid analysis of the conjugate enabled the amount of carrier immobilized peptide to be calculated; an estimated 15-17 peptides were bound per dextran molecule (MW 73,500); using a carbohydrate as carrier reduces the possibility of non-specific interactions because no hydrophobic or ionic sites and no protein-like epitopes exist on the carrier apart from the peptide ligand. It can be assumed that some peptide ligands provide the forces for an interaction with the plate surface whereas other remain free for the interaction with the antibody. Thus, the detection with monoclonal anti-peptide antibodies allowed peptide-dextran conjugates to be used at coating concentrations of 1-3 nM peptide, corresponding to 0.6-2.6 ng peptide-dextran per well. In contrast, concentrations of 150-500 nM were required for coating with peptides. The applicability of monofunctional peptide-dextran conjugates was demonstrated by investigating the titer and specificity of a polyclonal anti-peptide serum developed against human gastrointestinal glutathione peroxidase. The introduction of biotin as a second ligand of the dextran conjugate permitted its capture on streptavidin coated plates. This synthesis of bifunctional peptide-biotin-dextran conjugates opens up additional possibilities for applications.     

The Esterase PfeE,the Achilles’ Heel in the Battle for Iron between Pseudomonas aeruginosa and Escherichia coli

Bacteria access iron, a key nutrient, by producing siderophores or using siderophores produced by other microorganisms. The pathogen Pseudomonas aeruginosa produces two siderophores but is also able to pirate enterobactin (ENT), the siderophore produced by Escherichia coli. ENT-Fe complexes are imported across the outer membrane of P. aeruginosa by the two outer membrane transporters PfeA and PirA. Iron is released from ENT in the P. aeruginosa periplasm by hydrolysis of ENT by the esterase PfeE. We show here that pfeE gene deletion renders P. aeruginosa unable to grow in the presence of ENT because it is unable to access iron via this siderophore. Two-species co-cultures under iron-restricted conditions show that P. aeruginosa strongly represses the growth of E. coli as long it is able to produce its own siderophores. Both strains are present in similar proportions in the culture as long as the siderophore-deficient P. aeruginosa strain is able to use ENT produced by E. coli to access iron. If pfeE is deleted, E. coli has the upper hand in the culture and P. aeruginosa growth is repressed. Overall, these data show that PfeE is the Achilles’ heel of P. aeruginosa in communities with bacteria producing ENT.     

The Effect of Organic Diluent on the Extraction of Eu(III) by HEH[EHP]

ABSTRACT

Because there are fewer tools available to probe the interactions therein, the effect of the fundamental chemistry of the organic diluent on solvent extraction equilibria has been under-characterized relative to the aqueous. As a result, diluents for solvent extraction are often selected for an application not for their utility as a medium for reaction, but for other (often equally) important reasons (like low flammability). To begin to improve this imbalance in the science, twenty different diluents have been used in a study of the extraction of radiotracer ^152/154Eu³⁺ from dilute nitric acid solutions using the extractant 2-ethylhexylphosphonic acid mono-2-ethylhexyl ester (HEH[EHP]). To increase the utility of the study and to honor the memory of Professor Jan Rydberg, this investigation was conducted by a cadre of comparatively inexperienced separation scientists (who are as a result no longer inexperienced separation scientists) as a radioanalytical chemistry and solvent extraction educational exercise. Slope analysis was used to determine the apparent stoichiometry of the extracted metal complex. The results discussed in the following indicate that, while the pH dependence exhibits the expected three H⁺ exchanged per metal ion extracted, the extractant dependence suggests that the number of protonated extractant molecules in the extracted complex changes with the organic diluent. The experimentally observed “extractant dependency” ranges from 2.5 to 3.0 dimer equivalent molecules per extracted metal ion. Ironically, in the diluents exhibiting the highest apparent M:(HA)₃ stoichiometry, HEH[EHP] extracts Eu³⁺ less efficiently. Europium luminescence spectroscopy was used to probe for changes in the first coordination sphere of the complex in different diluents. A model and conceptual framework for understanding these observations is described.     

Choquet optimal set in biobjective combinatorial optimization

We study in this paper the generation of the Choquet optimal solutions of biobjective combinatorial optimization problems. Choquet optimal solutions are solutions that optimize a Choquet integral. The Choquet integral is used as an aggregation function, presenting different parameters, and allowing to take into account the interactions between the objectives. We develop a new property that characterizes the Choquet optimal solutions. From this property, a general method to easily generate these solutions in the case of two objectives is defined. We apply the method to two classical biobjective optimization combinatorial optimization problems: the biobjective knapsack problem and the biobjective minimum spanning tree problem. We show that Choquet optimal solutions that are not weighted sum optimal solutions represent only a small proportion of the Choquet optimal solutions and are located in a specific area of the objective space, but are much harder to compute than weighted sum optimal solutions.     

Content Management Systems: Enriched learning opportunities for all?

This article examines the popular claim of Content Management Systems (CMSs) that providing a rich toolset and leaving the use under learner control is beneficial to learning. By means of a literature review, the current contribution examines whether all students are capable of using CMS tools so that their learning is enhanced. In contrast to what is assumed, the study conceptualizes tool use as a complex self-regulation strategy that cannot be taken for granted. Specifically, the article reviews empirical studies in relation to three topics: (a) personal agency in tool use, (b) performance effects of tool use and (c) influencing tool use variables. Findings reveal that not every student profited from the CMS learning opportunities; in multiple studies students differed in their tool use, and these differences had significant performance effects. Hence, these findings suggest that the pedagogical claim CMSs make is problematic. Besides this accumulated corpus of knowledge, the review revealed serious limitations in the retrieved studies which could hamper our findings. As a consequence, the review establishes a need for further research into students’ CMS tool use from an instructional design perspective. In addition to the theoretical framework, several directions for future research are given.     

The multiobjective multidimensional knapsack problem: a survey and a new approach

The knapsack problem (KP) and its multidimensional version (MKP) are basic problems in combinatorial optimization. In this paper, we consider their multiobjective extension (MOKP and MOMKP), for which the aim is to obtain or approximate the set of efficient solutions. In the first step, we classify and briefly describe the existing works that are essentially based on the use of metaheuristics. In the second step, we propose the adaptation of the two‐phase Pareto local search (2PPLS) to the resolution of the MOMKP. With this aim, we use a very large scale neighborhood in the second phase of the method, that is the PLS. We compare our results with state‐of‐the‐art results and show that the results we obtained were never reached before by heuristics for biobjective instances. Finally, we consider the extension to three‐objective instances.     

Patterns in melanocytic lesions: impact of the geometry on growth and transport inside the epidermis

In glabrous skin, nevi and melanomas exhibit pigmented stripes during clinical dermoscopic examination. They find their origin in the basal layer geometry which periodically exhibits ridges, alternatively large (limiting ridges) and thin (intermediate ridges). However, nevus and melanoma lesions differ by the localization of the pigmented stripes along furrows or ridges of the epidermis surface. Here, we propose a biomechanical model of avascular tumour growth which takes into account this specific geometry in the epidermis where both kinds of lesions first appear. Simulations show a periodic distribution of tumour cells inside the lesion, with a global contour stretched out along the ridges. In order to be as close as possible to clinical observations, we also consider the melanin transport by the keratinocytes. Our simulations show that reasonable assumptions on melanocytic cell repartition in the ridges favour the limiting ridges of the basal compared with the intermediate ones in agreement with nevus observations but not really with melanomas. It raises the question of cell aggregation and repartition of melanocytic cells in acral melanomas and requires further biological studies of these cells in situ.     

Selective adsorption of multivalent ions into TiC-derived nanoporous carbon

A TiC-derived carbon (TiC-CDC) was prepared, and the adsorption of large hexacyanocobaltate and tetrabutylammonium ions of approximately same size was examined on this carbon. While selectively absorbing these large ions, it rejects smaller chloride and ammonium ions in mixed electrolyte solutions. The result demonstrates the important role of electrostatic repulsive forces, space-efficient charge packing and hydrophobic ion interactions with the pore walls of TiC-CDC, similar to what is known for a variety of biological membranes.     

Comparison of the wound-activated transformation of caulerpenyne by invasive and noninvasive Caulerpa species of the Mediterranean

The invasive green alga, Caulerpa taxifolia, that has spread rapidly after its introduction into the Mediterranean and the North American Pacific, reacts to wounding by transforming its major metabolite caulerpenyne (1). This wound-activated reaction involves the transformation of the bis-enol acetate moiety of 1, releasing reactive 1,4-dialdehydes. The ability to perform this transformation is found also in both the noninvasive Mediterranean C. prolifera and the invasive C. racemosa. Trapping experiments, as well as transformation of the model substrate geranyl acetate, suggest that all three investigated Caulerpa spp. rely on esterases that act upon wounding of the algae by subsequently removing the three acetate residues of caulerpenyne. The resulting reactive 1,4-dialdehyde oxytoxin 2 (9) can be identified by liquid chromatography–mass spectrometry and is unstable in the wounded tissue. Caulerpenyne transformation occurs rapidly, and severe tissue damage caused degradation of more than 50% of the stored caulerpenyne within 1 min in all three algae. Prevention of the enzymatic reaction before extraction, by shock freezing the tissue with liquid nitrogen, was used for the determination of the caulerpenyne content in intact algae. It gives about twofold higher values compared to an established methanol extraction protocol. The speed and mechanism of the wound-activated transformation, as well as the caulerpenyne content in intact tissue of invasive and noninvasive Caulerpa spp., are comparable. Thus, this enzymatic , transformation, despite being fast and efficient, is likely not the key for the success of the investigated invasive species.     

Students’ tool-use within a web enhanced course: Explanatory mechanisms of students’ tool-use pattern

The popularity of content management systems (CMSs) in today’s higher education is driven by the assumption that providing a rich toolset and leaving the use of this toolset under learner control will stimulate self-regulated and deeper learning. Current evidence on students’ tool-use within CMS supported courses however tackles this assumption and indicates that CMSs may empower students’ learning only under particular learner-related conditions. The current study addresses this concern and investigates how students’ tool-use within a CMS supported course can be explained in terms of (a) students’ conceptions on the tool functionalities, (b) self-efficacy beliefs for self-regulated learning and (c) goal orientation. Data were collected within a first year undergraduate course ‘Learning and Instruction’. Students’ (n = 182) tool-use within the course was logged throughout the course episode and the influencing variables were measured through questionnaires. K-means cluster analyses revealed four clusters that reflected differences in students’ tool-choice and tool-use throughout the course. Multinominal regression analyses revealed that these tool-use differences could be explained in terms of students’ goal orientation. The study provides thus perspectives in order to capture students’ academic motivation through unobtrusive, behavioral, measures. Furthermore, questions are raised regarding the parallel between students’ tool-use pattern and study strategy use.