Double Mutant Cycles as a Tool to Address Folding,Binding, and Allostery

Quantitative measurement of intramolecular and intermolecular interactions in protein structure is an elusive task, not easy to address experimentally. The phenomenon denoted ‘energetic coupling’ describes short- and long-range interactions between two residues in a protein system. A powerful method to identify and quantitatively characterize long-range interactions and allosteric networks in proteins or protein–ligand complexes is called double-mutant cycles analysis. In this review we describe the thermodynamic principles and basic equations that underlie the double mutant cycle methodology, its fields of application and latest employments, and caveats and pitfalls that the experimentalists must consider. In particular, we show how double mutant cycles can be a powerful tool to investigate allosteric mechanisms in protein binding reactions as well as elusive states in protein folding pathways.     

Pectins from Aloe Vera: Extraction and production of gels for regenerative medicine

Extraction, purification, and gel preparation of Aloe Vera pectin and the evaluation of the biocompatibility of the pectin gels were studied, considering as end use as implantable materials for regenerative medicine. A. Vera was chosen as source of pectin, as this pectin was described to possess high molecular weight and a low degree of esterification. As the properties of pectins are strictly dependent upon the extraction methods in combination with the natural source, the extraction method was modified in order to optimize the yield of the final product, its purity, the duration of the process and the selection of non‐toxic chemical reagents. Changing the experimental conditions resulted in four different extraction processes and products with different physical and chemical characteristics. The optimal extraction resulted to be the process: with enzimatic deactivation by microwave and the use of sodium citrate as chelating agent the molecular weight of the pectin extracted was estimated to be 118 kDa and the 2.93% esterification degree. Cytocompatibility of pectin gels, prepared by ionotropic gelation, showing an improved cell adhesion if compared to commercial pectin. The results suggest that the extracted A. Vera pectins possess interesting properties to be exploited for the production of mechanically stable gels by ionotropic gelation and high rhamnose content matrices for application in regenerative medicine. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 39760.     

Metal Oxide Nanowires for Gas Sensor Applications

We report on the synthesis of SnO₂ and CuO nanowires and their application as gas sensing components. The fabrication of SnO₂ and CuO single nanowire devices by optical and electron beam lithography is described, and sensing performance to the toxic gas carbon monoxide is demonstrated. We briefly present the development of CMOS fabricated micro-hotplates as platforms for gas sensors and show our approach for nanowire implementation. Finally, we demonstrate a fully CMOS integrated CuO multi nanowire device and present our roadmap for a fully integrated multi-parameter smart sensor device which could be implemented as safety feature in smart phones.     

Linear matrix inequality conditions for robustness and control design

In this paper, well‐known stability conditions for linear, time‐invariant systems subject to structured uncertainty are extended to encompass matrix valued and affine integral constraints on the system inputs and outputs. Necessary and sufficient analysis conditions are derived in terms of linear matrix inequalities. For a large class of these problems, the synthesis question can also be expressed as a linear matrix inequality. Copyright © 2001 John Wiley & Sons, Ltd.     

Changes in free amino acids and biogenic amines of Egyptian salted-fermented fish (Feseekh) during ripening and storage

The aim of this study was to investigate changes in the formation of amino acids and biogenic amines in Egyptian salted-fermented fish (Feseekh) during ripening (20 days) and storage (40–60 days). The total concentration of free amino acids increased from 8 (dry weight; DW) to 72 g/kg (DW) after 60 days of storage. The predominant free amino acids were leucine, glutamic acid, lysine, alanine, valine, aspartic acid, isoleucine and citrulline. Their concentrations accounted for 68% of the total concentration of amino acids after 60 days. The total contents of biogenic amines ranged from 84 to 1633 mg/kg (DW) during the investigated period. Cadaverine was the major amine detected in Feseekh at all sampling stages and its concentration varied between 21 and 997 mg/kg (DW). The histamine content (211 mg/kg DW) only exceeded the maximum tolerance level (200 mg/kg) after 60 days. It could be concluded that Feseekh can be consumed without any health risks between 20 and 40 days but it can be hazardous after 60 days due to the biogenic amine content.     

Biochemical changes in cut vs. intact lamb's lettuce (Valerianella olitoria) leaves during storage

Consumers are oriented towards fresh-cut vegetables that provide phytonutrients useful for preventing stress-related diseases. The aim of this work was to evaluate the cut operations on the quality changes of lamb's lettuce ( Valerianella olitoria L.) cv. Trofy during storage at 4 °C for 8 days. Results showed that chlorophyll and carotenoids reduction was observed after 8 days of storage. In both treatments, total carotenoids after 8 days decreased from 20 to 16 mg g⁻¹ FW. Free and total phenols increased with storage in both treatments. Total phenols were 23% higher in control (32 μmol g⁻¹ FW) compared to cut leaves (25 μmol g⁻¹ FW) after 8 days of storage. Anthocyanins increased after 8 days and reached 30 mg 100 g⁻¹ FW without significant difference between treatments. Ascorbic acid (AsA) and dehydroascorbic (DHA) acid increased in cut leaves compared to control. After 1 day AsA concentration was 3 300 nmol g⁻¹ FW in cut leaves, while in control leaves was 1 500 nmol g⁻¹ FW. Analogously AsA + DHA was higher in cut leaves, 4 100 nmol g⁻¹ FW, while in control leaves the mean was 3 000 nmol g⁻¹ FW. After 5 days of storage the values of AsA returned to initial values, while AsA + DHA were lower.     

Endemic goiter in school children of Salta Province, Argentina

The Province of Salta, located in the northwest of Argentina, showed severe endemic goiter-cretinism. The purpose of our work, therefore, was to evaluate the results of the iodized salt prophylaxis measure implemented since 1963. In 1980-1981 the prevalence of goiter was determined in 16,935 school children whose ages ranged from four to 15 years. The urinary iodine/creatinine index was also determined in a subsample of 401 children. Findings revealed that the goiter prevalence in the Province was 16.1%. In 10 Departments of the Province, those with greater socioeconomic development and denser population, the prevalences found were below 10%, limit established as endemic. In the remaining 13 Departments, especially in those less developed--where mountainous regions make their access difficult and hence, their populations consume non-iodized salt obtained from natural salt basins--values exceeded this limit. The average ioduria was 104.0 micrograms I/g Cr, with 4.5% presenting values below 25 micrograms. After 20 years of prophylaxis, endemic goiter has therefore been partially eradicated. It is suggested, however, that consumption of iodized salt and, even the supply of iodized oil in those areas where prevalence is high, be recommended.