Recombinant human erythrocyte cytochrome b5

The gene encoding the human erythrocyte form of cytochrome b5 (97 residues in length) has been prepared by mutagenesis of an expression vector encoding lipase-solubilized bovine liver microsomal cytochrome b5 (93 residues in length) (Funk et al., 1990). Efficient expression of this gene in Escherichia coli has provided the first opportunity to obtain this protein in quantities sufficient for physical and functional characterization. Comparison of the erythrocytic cytochrome with the trypsin-solubilized bovine liver cytochrome b5 by potentiometric titration indicates that the principal electrostatic difference between the two proteins results from two additional His residues present in the human erythrocytic protein. The midpoint reduction potential of this protein determined by direct electrochemistry is -9 +/- 2 mV vs SHE at pH 7.0 (mu = 0.10 M, 25.0 degrees C), and this value varies with pH in a fashion that is consistent with the presence of a single ionizable group that changes pKa from 6.0 +/- 0.1 in the ferricytochrome to 6.3 +/- 0.1 in the ferrocytochrome with delta H degrees = -3.2 +/- 0.1 kcal/mol and delta S degrees = -11.5 +/- 0.3 eu (pH 7.0, mu = 0.10). The 1D 1H NMR spectrum of the erythrocytic ferricytochrome indicates that 90% of the protein binds heme in the "major" orientation and 10% of the protein binds heme in the "minor" orientation (pH 7.0, 25 degrees C) with delta H degrees = -2.9 +/- 0.3 kcal/mol and delta S degrees = -5.4 +/- 0.9 eu for this equilibrium.     

GraftFast Surface Engineering to Improve MOF Nanoparticles Furtiveness

Controlling the outer surface of nanometric metal–organic frameworks (nanoMOFs) and further understanding the in vivo effect of the coated material are crucial for the convenient biomedical applications of MOFs. However, in most studies, the surface modification protocol is often associated with significant toxicity and/or lack of selectivity. As an alternative, how the highly selective and general grafting GraftFast method leads, through a green and simple process, to the successful attachment of multifunctional biopolymers (polyethylene glycol (PEG) and hyaluronic acid) on the external surface of nanoMOFs is reported. In particular, effectively PEGylated iron trimesate MIL‐100(Fe) nanoparticles (NPs) exhibit suitable grafting stability and superior chemical and colloidal stability in different biofluids, while conserving full porosity and allowing the adsorption of bioactive molecules (cosmetic and antitumor agents). Furthermore, the nature of the MOF–PEG interaction is deeply investigated using high‐resolution soft X‐ray spectroscopy. Finally, a cell penetration study using the radio‐labeled antitumor agent gemcitabine monophosphate (³H‐GMP)‐loaded MIL‐100(Fe)@PEG NPs shows reduced macrophage phagocytosis, confirming a significant in vitro PEG furtiveness.     

Biogenic amines in Spanish beers: differences among breweries

Ten biogenic amines in Spanish beers were studied using HPLC. Agmatine, tyramine and putrescine were the prevailing amines, while histamine,-phenylethylamine, tryptamine, cadaverine, spermine and spermidine were detected at relatively low levels (in general <2 mg/l). On the basis of the wide range of levels observed for tyramine (from 1.90 to 31.55 mg/l), the consumption of beer requires restriction in patients receiving monoamine oxidase inhibitor drugs. Biogenic amine levels in beers of the same type from the same company and from different companies were monitored every month over 1 year. Agmatine and putrescine levels showed minimal fluctuations in beers from the same company as well as from different breweries. Raw materials and brewing conditions would not affect the levels of those amines. However, tyramine levels were subject to wide fluctuations in beers produced by particular breweries irrespective of whether they were produced by the same company. Relatively high levels of tyramine were specific to some breweries.     

A generic framework for median graph computation based on a recursive embedding approach

The median graph has been shown to be a good choice to obtain a representative of a set of graphs. However, its computation is a complex problem. Recently, graph embedding into vector spaces has been proposed to obtain approximations of the median graph. The problem with such an approach is how to go from a point in the vector space back to a graph in the graph space. The main contribution of this paper is the generalization of this previous method, proposing a generic recursive procedure that permits to recover the graph corresponding to a point in the vector space, introducing only the amount of approximation inherent to the use of graph matching algorithms. In order to evaluate the proposed method, we compare it with the set median and with the other state-of-the-art embedding-based methods for the median graph computation. The experiments are carried out using four different databases (one semi-artificial and three containing real-world data). Results show that with the proposed approach we can obtain better medians, in terms of the sum of distances to the training graphs, than with the previous existing methods.     

Methodology for capturing and formalizing DFM Knowledge

Design for manufacturing (DFM) practices lead to more competitive products from the point of view of cost, development time and quality. However, the success of considering manufacturing issues during design process would be higher if manufacturing information was more readily available and designers needed less experience to select information relevant to DFM.     

Surface plasmon resonance imaging (SPRi) as an alternative technique for rapid and quantitative screening of small molecules, useful in drug discovery

Surface plasmon resonance imaging (SPRi) is a label free technology for biomolecular interaction, which gives access to binding kinetic parameters from real time acquisition. It offers the possibility to test in a single run a large number of interactions, allowing rapid identification of the most suitable compounds toward a given biological entity. Until now, this technique has proven to be relevant for interaction between relatively large molecules (protein, antibodies, DNA) but has not been challenged yet for the screening of small molecules that can be of interested in the field of drug discovery. As a proof a principle, we have used SPRi to screen for interaction of several small molecules, referred to as G4-ligands on G-quadruplex DNA. This technology allowed to easy discrimination of the binding properties of four G4-ligands on quadruplex DNA models.     

The effects of the number of links and navigation support on cognitive load and learning with hypertext: The mediating role of reading order

Problems in learning with hypertext systems have been claimed to be caused by high levels of disorientation and cognitive load. This was recognized by DeStefano and LeFevre [DeStefano, D., & LeFevre, J. -A., (2007). Cognitive load in hypertext reading: A review. Computers in Human Behavior, 23(3), 1616–1641.] who predicted an increase of cognitive load and impairment of learning for hypertexts with a higher number of links per page. From a practical perspective, several navigation support techniques, such as providing link suggestions, have been proposed for guiding learners and reducing cognitive overload. In an experiment, we tested DeStefano and LeFevre’s predictions as well as the usefulness of link suggestions. Participants used different versions of a hypertext, either with 3-links or 8-links per page, presenting link suggestions or not. We tested their cognitive load and learning outcomes. Results showed that there was a benefit of using link suggestions for learning, but no effect of number of links on learning was found. Moreover, the effects of our manipulations on cognitive load were mediated by the reading order that participants selected. Implications for research and the design of navigation support systems are discussed.