Structural rearrangements near the chromophore influence the maturation speed and brightness of DsRed variants

The red fluorescent protein DsRed has been extensively engineered for use as an in vivo research tool. In fast maturing DsRed variants, the chromophore maturation half-time is approximately 40 min, compared to approximately 12 h for wild-type DsRed. Further, DsRed has been converted from a tetramer into a monomer, a task that entailed mutating approximately 20% of the amino acids. These engineered variants of DsRed have proven extremely valuable for biomedical research, but the structural basis for the improved characteristics has not been thoroughly investigated. Here we present a 1.7 A crystal structure of the fast maturing tetrameric variant DsRed.T4. We also present a biochemical characterization and 1.6 A crystal structure of the monomeric variant DsRed.M1, also known as DsRed-Monomer. Analysis of the crystal structures suggests that rearrangements of Ser69 and Glu215 contribute to fast maturation, and that positioning of the Lys70 side chain modulates fluorescence quantum yield. Despite the 45 mutations in DsRed.M1 relative to wild-type DsRed, there is a root-mean-square deviation of only 0.3 A between the two structures. We propose that novel intramolecular interactions in DsRed.M1 partially compensate for the loss of intermolecular interactions found in the tetramer.     

Vascular access‐related infection in nocturnal home hemodialysis

Frequent hemodialysis is associated with increased vascular access adverse events. We hypothesized that bacteremia would be more frequent in patients with central venous catheter (CVC) than arteriovenous fistula or arteriovenous graft (AVF/AVG) in nocturnal home hemodialysis (NHHD). We reviewed blood culture reports and concurrent clinical data for a cohort of one hundred eighty‐seven NHHD patients between January 1, 2006 and June 30, 2012. The primary outcome was time to first bacteremia, technique failure, or death after commencing NHHD. Types of bacteremia and clinical consequences were analyzed. Analyses were adjusted for a priori defined confounders. One hundred eighty‐seven patients were included with a total follow up of six hundred five patient years. Initial vascular access was AVF in seventy‐eight (42%) patients, AVG in eleven (6%) patients, and CVC in ninety‐eight (52%) patients. A total of 79.3% of patients with a CVC reached the composite endpoint of bacteremia, technique failure, or death in the study period; 44.5% of patients with an AVF or AVG reached this composite endpoint. Adjusted time to first bacteremia, technique failure, or death was significantly shorter in patients with initial CVC access (hazard ratio 2.42, 95% confidence interval 1.50–3.90, p < 0.001). Risk factors for bacteremia were comorbid status quantified by the Charlson Comorbidity Index (p < 0.001) and diabetes (p < 0.001). Coagulase negative staphylococcus was the commonest organism cultured accounting for 51.4% bacteremias. The second commonest organism was staphylococcus aureus (20.3% bacteremias). Patients undergoing NHHD with a CVC have a shorter duration to first infection, technique failure, or death than those with permanent vascular access.     

Experimental and finite element analysis of residual stress and distortion in GTA welding of modified 9Cr-1Mo steel

Journal of Mechanical Science and Technology - In this paper, investigation of residual stress and distortion induced in 3 mm thick Modified 9Cr-1Mo steel plates during GTA welding is carried out....     

Fatigue of 7075-T651 aluminum alloy under constant and variable amplitude loadings

The fatigue crack growth (FCG) behavior of 7075-T651 aluminum alloy was studied under constant and variable amplitude loadings in vacuum, air and 1% NaCl solution. In the study of constant amplitude loading fatigue, the stress ratios were 0.1 and 0.85 and the loading frequency was 10 Hz. In the study of variable amplitude loading fatigue, the load spectrums were tension type and tension–compression type, and the average loading frequency was about 5 Hz. The results of FCG tests, under constant and variable amplitude loadings, validated the unified two parameter driving force model, accounting for the residual stress and stress ratio effects on fatigue crack growth.     

Delayed yielding of oil/water emulsions in presence of stabilizing biopolymer

This study investigates the stabilization of oil/water emulsions as a function of addition of a biopolymer (scleroglucan) which acts as an emulsion stabilizer. Rheological characterization in the form of controlled stress creep measurements has been carried out and it reveals the colloidal gel exhibiting a delayed yielding in a certain applied stress window. The delay time and stresses that an emulsion can withstand depend strongly on the concentration of added scleroglucan. Increasing polymer concentration, however, is limited to a maximum value, above which a limited effect on the delay time is observed. Investigating of the emulsion under study was visualized by means of cryo transmission electron microscopy which shows adsorption of scleroglucan onto the surface of the oil particles and a gel-like structure that connects the oil phases. The results mentioned in this study support that scleroglucan-surfactant interactions play a key role in the stabilization of the oil/water emulsion.     

Challenges associated in stability of food grade nanoemulsions

Food grade nanoemulsions are being increasingly used in the food sector for their physico-chemical properties towards efficient encapsulation, entrapment of bioactive compounds, solubilization, targeted delivery, and bioavailability. Nanoemulsions are considered as one of the important vehicles for the sustained release of food bioactive compounds due to their smaller size (nm), increased surface area, and unique morphological characteristics. Nanoemulsification is an ideal technique for fabricating the bioactive compounds in a nano form. Formation and stabilization of nanoemulsion depends on the physi-cochemical characteristics of its constituents including oil phase, aqueous phase, and emulsifiers. This review is mainly focused on the instability mechanisms of nanoemulsion such as flocculation, Ostwald ripening, creaming, phase separation, coalescence, and sedimentation. Further, the major factors associated with these instability mechanisms like ionic strength, temperature, solubilization, particle size distribution, particle charge, pH strength, acid stability, and heat treatment are also discussed. Finally, safety issues of food grade nanoemulsions are highlighted.     

Manipulating oral behaviour of food emulsions using different emulsifiers

This study aims to investigate the assumption that the presence of α-amylase in the human saliva will interact instantly with starch and will lead to very different oral behaviour and enhanced flavour release. Hence, orange oil flavoured emulsion was prepared with whey protein isolates (WPI) and modified starch (MS). The stability and flavour release of emulsions were examined through in vitro and in vivo studies. MS emulsion mixed with artificial saliva containing α-amylase resulted in more pronounced changes in mean particle size from 0.185 to 2.35 μm and a significant increase in viscosity. Morphology and turbidity revealed strong flocculation, coalescence and creaming. However, WPI emulsion exhibited very little changes in stability and behaviour. Similar results were observed during oral digestion (in vivo) for both emulsion systems. Moreover, a higher intensity of flavour release (37%) was observed in MS emulsion than WPI. This work demonstrated that a starch-stabilised emulsion has a very different oral behaviour to that of a protein-stabilised emulsion.     

The effect of lubricant additives on friction and wear studies of polyphenylene sulphide

Polyphenylene sulphide is a polymer with good thermal stability and high crystallinity. This paper summarizes the results of friction and wear studies of polyphenylene sulphide and its composites made with conventional solid lubricants to ascertain the suitability of the material as a matrix for solid lubricant additives. The polymer itself has a high coefficient of friction. Wear rate increases with load and speed. Addition of solid lubricant additives helps in improving the friction and wear of the polymer. Composites with MoS₂-Sb₂O₃ and PTFE gave better results than composites made by the addition of graphite and MoS₂ graphite. Wear rate of these composites increased with load and speed; but load and speed had little effect on friction.