Differential divalent cation requirements uncouple the assembly and catalytic reactions of human immunodeficiency virus type 1 integrase

Previous in vitro analyses have shown that the human immunodeficiency virus type 1 (HIV-1) integrase uses either manganese or magnesium to assemble as a stable complex on the donor substrate and to catalyze strand transfer. We now demonstrate that subsequent to assembly, catalysis of both 3' end processing and strand transfer requires a divalent cation cofactor and that the divalent cation requirements for assembly and catalysis can be functionally distinguished based on the ability to utilize calcium and cobalt, respectively. The different divalent cation requirements manifest by these processes are exploited to uncouple assembly and catalysis, thus staging the reaction. Staged 3' end processing and strand transfer assays are then used in conjunction with exonuclease III protection analysis to investigate the effects of integrase inhibitors on each step in the reaction. Analysis of a series of related inhibitors demonstrates that these types of compounds affect assembly and not either catalytic process, therefore reconciling the apparent disparate results obtained for such inhibitors in assays using isolated preintegration complexes. These studies provide evidence for a distinct role of the divalent cation cofactor in assembly and catalysis and have implications for both the identification and characterization of integrase inhibitors.     

Copper-zirconium tungstate composites exhibiting low and negative thermal expansion influenced by reinforcement phase transformations

A fully-dense Cu-75 vol pct ZrW₂O₈ metal matrix composite was fabricated by hot isostatic pressing of Cu-coated ZrW₂O₈ particles. A small amount of the high-pressure γ-ZrW₂O₈ phase was created during the cooldown and depressurization following densification; near complete transformation to γ-ZrW₂O₈ was achieved by subsequent cold isostatic pressing. The thermal expansion behavior of the composite between 25°C and 325°C was altered by the cold isostatic pressing treatment, and also depended on the length of time that had passed between thermal cycles. The measured thermal expansion coefficients within specific temperature ranges varied from −6·10⁻⁶ K⁻¹ to far above the thermal expansion coefficient of the copper matrix. The complex temperature-dependent expansion/contraction behavior could be justified by considering the evolution of phase transformations taking place in the ZrW₂O₈ phase, which were observed by in-situ synchrotron X-ray diffraction measurements.     

Case Study of an Aerosol Explosion and a Method to Determine Explosion Temperatures

A failure analysis case study is presented for a two-piece aerosol containing tetrafluoroethane, commonly referred to as Refrigerant 134a. A gentleman was preparing to recharge the air conditioning system of an automobile when the bottom exploded off the aerosol container, propelling the body of the aerosol container like a rocket, which hit the man in the eye and blinded him in that eye. The aerosol was never connected to the air conditioner, therefore backpressure from the air conditioner (AC) compressor was ruled out as a cause for the explosion. The objective of the study was to determine why the aerosol exploded. Several recently developed test methods were used, including two types of heat-to-burst tests and a puncture chamber to measure the pressure-versus-temperature behavior of aerosols. More common test methods were also used, such as water bath pressure tests, hydro pressure burst tests, pneumatic pressure burst tests, hardness measurements, weight measurements, metallography, scanning electron microscopy (SEM), energy-dispersive spectroscopy (EDS), and an accident scenario recreation. A semi-empirical correlation between the hardness and weights of the container bottoms was used to determine the explosion temperature and/or pressure. This semi-empirical correlation agrees in principle with an analysis of the explosion pressures using finite-element analysis (FEA). The root cause for the explosion was determined to be a lack of strength of the bottom of the two-piece aerosol coupled with heating the aerosol to temperatures significantly above room temperature.     

100 mK bolometers for the submillimetre common-user bolometer array (scuba) I. Design and construction

We describe the design and construction of bolometric detectors for SCUBA - the Submillimetre Common-User Bolometer Array for the James Clerk Maxwell Telescope in Hawaii. The instrument contains 131 individual detectors, in two arrays, optimized for the submillimetre atmospheric transmission windows. The detectors are cooled by dilution refrigeration to a temperature of 100 mK, so that the receiver performance will be limited by photon noise from the sky and telescope background in all wavebands. A future paper will describe the performance of the detectors with reference to typical data obtained during the laboratory commissioning period.     

Detection of ultrasonic tones and simulated dolphin echolocation clicks by a teleost fish, the American shad (Alosa sapidissima)

The authors previously reported that American shad (Alosa sapidissima) can detect sounds from 100 Hz to 180 kHz, with two regions of best sensitivity, one from 200 to 800 Hz and the other from 25 to 150 kHz [Mann et al., Nature 389, 341 (1997)]. These results demonstrated ultrasonic hearing by shad, but thresholds at lower frequencies were potentially masked by background noise in the experimental room. In this study, the thresholds of the American shad in a quieter and smaller tank, as well as thresholds for detecting stimulated echolocation sounds of bottlenosed dolphins was determined. Shad had lower thresholds for detection (from 0.2 to 0.8 kHz) in the quieter and smaller tank compared with the previous experiment, with low-frequency background noise but similar thresholds at ultrasonic frequencies. Shad were also able to detect echolocation clicks with a threshold of 171 dB re: 1 microPa peak to peak. If spherical spreading and an absorption coefficient of 0.02 dB/m of dolphin echolocation clicks are assumed, shad should be able to detect echolocating Tursiops truncatus at ranges up to 187 m. The authors propose that ultrasonic hearing evolved in shad in response to selection pressures from echolocating odontocete cetaceans.     

Liquid phase sintering of hydroxyapatite by phosphate and silicate glass additions: structure and properties of the composites

Phosphate- and silicate-based glasses were added to hydroxyapatite in order to improve its mechanical properties and to fabricate composites with different degrees of bioactivity. A strong chemical bonding was obtained between hydroxyapatite and the phosphate-based glasses leading to samples approaching theoretical density, according to density measurements and scanning electron microscopy. Bioglass^® additions led to the formation of a complex calcium phosphate silicate which hampered the reinforcement process. The fracture toughness of the hydroxyapatite-glass composites was shown to be within the 1.1–1.2 MPam^1/2 range, which is double that determined for sintered hydroxyapatite. A 2 m thick apatite layer was observed on the surface of the hydroxyapatite-glass composites after 48 h immersion in a simulated human blood plasma, whereas only a few apatite crystals were detected on sintered hydroxyapatite after 7 days immersion. From the results obtained we anticipate that the composites might show a higher rate of bone bonding, leading to enhanced bioactivity.     

Reactive Structural Materials: Preparation and Characterization

Reactive structural materials, which can serve both as structural elements as well as a source of chemical energy released upon initiation have emerged as an important class of metal‐based composites for use in various energetic systems. Such materials rely on a variety of exothermic reactions, from oxidation to formation of metal‐metalloid and intermetallic phases. The rates of these reactions are as important as the energy that may be released, in order for them to occur at the time scales compatible with the requirements of applications. Therefore, chemical composition, scale at which reactive components are mixed, and the structure and morphology of materials are important and can be controlled by the method of preparation and compaction of the composite materials. Methods of preparation of the composite structures are briefly reviewed as well as methods of characterization of their mechanical and energetic properties. In addition to common thermo‐analytical and static mechanical property measurements, dynamic tests of mechanical properties as well as ignition and combustion experiments are necessary to understand the fragmentation, initiation, and heat release expected for these materials when they are stimulated by an impact, shock, or rapid heating. Reaction mechanisms are studied presently for the thin layers and small samples of reactive materials initiated in carefully designed experiments. In other experiments, impact and explosive initiation are characterized for larger material compacts in the conditions imitating practical scenarios. Examples of results describing thermal, impact, and explosive initiation of some of the reactive materials are presented.

     

A model of cerebral cortex formation during fetal development using reaction-diffusion-convection equations with Turing space parameters

The cerebral cortex is a gray lamina formed by bodies of neurons covering the cerebral hemispheres, varying in thickness from 1.25 mm in the occipital lobe to 4 mm in the anterior lobe. The brain's surface is about 30 times greater that of the skull because of its many folds; such folds form the gyri, sulci and fissures and mark out areas having specific functions, divided into five lobes. Convolution formation may vary between individuals and is an important feature of brain formation; such patterns can be mathematically represented as Turing patterns. This article describes how a phenomenological model was developed by describing the formation pattern for the gyri occurring in the cerebral cortex by reaction diffusion equations with Turing space parameters. Numerical examples for simplified geometries of a brain were solved to study pattern formation. The finite element method was used for the numerical solution, in conjunction with the Newton–Raphson method. The numerical examples showed that the model can represent cerebral cortex fold formation and reproduce pathologies related to gyri formation, such as polymicrogyria and lissencephaly.     

A Distributed Algorithm for Computing the Node Search Number in Trees

We present a distributed algorithm to compute the node search number in trees. This algorithm extends the centralized algorithm proposed by Ellis et al. (Inf. Comput. 113(1):50–79, 1994). It can be executed in an asynchronous environment, requires an overall computation time of O(nlog?n), and n messages of log?₃ n+4 bits each. The main contribution of this work lies in the data structure proposed to design our algorithm, called hierarchical decomposition. This simple and flexible data structure is used for four operations: updating the node search number after addition or deletion of any tree-edges in a distributed fashion; computing it in a tree whose edges are added sequentially and in any order; computing other graph invariants such as the process number and the edge search number, by changing only initialization rules; extending our algorithms for trees and forests of unknown size (using messages of up to 2log?₃ n+5 bits).