Reactivity of intein thioesters: appending a functional group to a protein

The success of genome sequencing has heightened the demand for new means to manipulate proteins. An especially desirable goal is the ability to modify a target protein at a specific site with a functional group of orthogonal reactivity. Here, we achieve that goal by exploiting the intrinsic electrophilicity of the thioester intermediate formed during intein-mediated protein splicing. Detailed kinetic analyses of the reaction of nitrogen nucleophiles with a chromogenic small-molecule thioester revealed that the alpha-hydrazino acetyl group was the optimal nucleophile for attacking a thioester at neutral pH to form a stable linkage. A bifunctional reagent bearing an alpha-hydrazino acetamido and azido group was synthesized in high overall yield. This reagent was used to attack the thioester linkage between a target protein and intein, and thereby append an azido group to the target protein in a single step. The azido protein retained full biological activity. Furthermore, its azido group was available for chemical modification by Huisgen 1,3-dipolar azide-alkyne cycloaddition. Thus, the mechanism of intein-mediated protein splicing provides the means to install a useful functional group at a specific site-the C terminus-of virtually any protein.     

Dynamics of Consolidation and Crack Growth in Nanocluster-Assembled Amorphous Silicon Nitride

Consolidation and fracture dynamics in nanophase amorphous Si3N4 are investigated using 106-atom molecular-dynamics simulations. At a pressure of 15 GPa and 2000 K, the nanophase system is almost fully consolidated within a fraction of a nanosecond. The consolidation process is well-described by the classical theory of sintering. Under an applied strain the consolidated system develops several cracks which propagate parallel to each other, causing failure at multiple sites. The critical strain at which the nanophase system fractures is much larger than that for crystalline Si3N4.     

Hierarchical parallelization and optimization of high-order stencil computations on multicore clusters

We present a scalable parallelization scheme for high-order stencil computations that also optimizes memory behavior on multicore clusters. Our multilevel approach combines: (i)?inter-node parallelization via spatial decomposition; (ii)?inter-core parallelization via multithreading and explicit non-uniform memory access (NUMA) control; (iii)?data locality optimizations through auto-tuned tiling for efficient use of hierarchical memory; and (iv)?register blocking and data parallelism via single-instruction multiple-data techniques to utilize registers and exploit data locality. The scheme is applied to a sixth-order stencil based finite-difference time-domain code. Weak-scaling parallel efficiency is over 98?% on 32,768 BlueGene/P processors. Multithreading with explicit NUMA control attains 9.9-fold speedup on a dual 12-core AMD Opteron system. Data locality optimizations achieve 7.7-fold reduction of the last level cache miss rate of Intel Nehalem, whereas register blocking increases data parallelism and thereby achieves 5.9 Gflops performance on a single core. Register blocking?+ multithreading optimizations achieve 5.8-fold speedup on a single quadcore Nehalem.     

The electrical characteristics of human skin in vivo

PURPOSE: The objectives of this study were to investigate the impedance properties of human skin in vivo and to examine the effect of iontophoresis upon them. METHODS: Having established the intra- and inter-individual variation in basal values of skin impedance, the effect of varying iontophoretic current density, ionic strength and counter-ion on the rate of recovery of skin impedance after iontophoresis was investigated. RESULTS: Passage of an iontophoretic current caused a significant reduction in the magnitude of the skin impedance. Increasing the current density caused an even greater reduction in the value of the skin impedance and slowed the rate of recovery. Reduction of the ionic strength resulted in an increase in the rate of recovery following iontophoresis. A significant increase in the rate of recovery was observed when CaCl2 replaced NaCl as the electrolyte. Although visual inspection revealed the presence of greater erythema when CaCl2 was used, there was an absence of the mild sensation experienced by volunteers when using NaCl. The last part of the study established a correlation between transepidermal water loss and impedance analysis as complementary methods for probing skin barrier function in vivo. The data were fitted to an equivalent circuit consisting of a resistor in parallel with a constant-phase element and a mechanistic model proposed to explain the electrical properties of the skin. CONCLUSIONS: The first comprehensive investigation of the effect of iontophoresis on the electrical properties of human skin in vivo has been described. It would appear from the results, and from their interpretation, that impedance spectroscopy may be an effective method to quantify the impact of iontophoresis on the skin, and to determine the extent to which proposed drug delivery regimens will perturb skin barrier function.     

A preliminary study of the corrosiveness of methyl fuel/gasoline blends on materials used in automotive fuel systems

A preliminary study was made of the corrosiveness of 15% by volume methyl fuel/gasoline blends on materials conventionally used for gasoline service in automotive fuel systems. The presence of water, salt and air dramatically increased the corrosiveness of the fuel. Of the metals studied, aluminum was relatively unaffected, while copper, brass, steel (types 1181 and 1008), zinc and template all showed signs of corrosion. Non-metallic samples, particularly expanded polyurethane and Buna-N. underwent greater attack in the presence of methyl fuel. Conductivity measurements showed that the presence of water and salt in the fuel can cause galvanic corrosion to occur.     

Biomolecule-entrapped SiO2 nanoparticles for ultrafast green synthesis of silver nanoparticle–decorated hybrid nanostructures as effective catalysts

Noble-metal hybrid nanostructures have gained tremendous attention due to their potential roles in biomedical and catalytic applications. In this study, for the synthesis of silver nanoparticles (Ag NPs)–silica (SiO₂) NPs, a novel green chemistry approach was employed, in which green tea biomolecule–encapsulated SiO₂ nanostructures were used for the reduction of silver ions to produce hybrid nanostructures within 300?s. The high-resolution transmission electron microscopy (HrTEM) revealed the formation of uniform ultrafine spherical Ag NPs that were evenly distributed in the nanostructures. The formed nanohybrid structures showed efficient catalytic activity for the formation of derivatives of dihydroquinoline, and retained 91% of their reusability capacity, even after 5 repeated cycles. Hence, this work provides a novel synthesis method not only for the synthesis of biomolecule-entrapped SiO₂ nanostructures, but also for the rapid formation of catalytically active hybrid nanostructures.     

Enhancement in hydrogen production by co-cultures of Bacillus and Enterobacter

Defined co-cultures of hydrogen (H₂) producers belonging to Citrobacter, Enterobacter, Klebsiella and Bacillus were used for enhancing the efficiency of biological H₂ production. Out of 11 co-cultures consisting of 2–4 strains, two co-cultures composed of Bacillus cereus EGU43, Enterobacter cloacae HPC123, and Klebsiella sp. HPC793 resulted in H₂ yield up to 3.0 mol mol⁻¹ of glucose. Up-scaling of the reactor by 16-fold resulted in a corresponding increase in H₂ production with an actual evolution of 7.44 L of H₂. It constituted 58.2% of the total biogas. Continuous culture evolution of H₂ by co-cultures (B. cereus EGU43 and E. cloacae HPC123) immobilized on ligno-cellulosic materials resulted in 6.4-fold improvement in H₂ yield compared to free floating bacteria. This synergistic influence of B. cereus and E. cloacae can offer a better strategy for H₂ production than undefined or mixed cultures.