An atomic model of the outer layer of the bluetongue virus core derived from X-ray crystallography and electron cryomicroscopy

BACKGROUND: Bluetongue virus (BTV), which belongs to the Reoviridae family and orbivirus genus, is a non-enveloped, icosahedral, double-stranded RNA virus. Several protein layers enclose its genome; upon cell entry the outer layer is stripped away leaving a core, the surface of which is composed of VP7. The structure of the trimeric VP7 molecule has previously been determined using X-ray crystallography. The articulated VP7 subunit consists of two domains, one which is largely alpha-helical and the other, smaller domain, is a beta barrel with jelly-roll topology. The relative orientations of these two domains vary in different crystal forms. The structure of VP7 and the organizations of 780 subunits of this molecule in the core of virus is central to the assembly and function of BTV. RESULTS: A 23 A resolution map of the core, determined using electron cryomicroscopy (cryoEM) data, reveals that the 260 trimers of VP7 are organized on a rather precise T = 13 laevo icosahedral lattice, in accordance with the theory of quasi-equivalence. The VP7 layer occupies a shell that is between 260 A and 345 A from the centre of the core. Below this radius (230-260 A) lies the T = 1 layer of 120 molecules of VP3. By fitting the X-ray structure of an individual VP7 trimer onto the cryoEM BTV core structure, we have generated an atomic model of the VP7 layer of BTV. This demonstrates that one of the molecular structures seen in crystals of the isolated VP7 corresponds to the in vivo conformation of the molecule in the core. CONCLUSIONS: The beta-barrel domains of VP7 are external to the core and interact with protein in the outer layer of the mature virion. The lower, alpha-helical domains of VP7 interact with VP3 molecules which form the inner layer of the BTV core. Adjacent VP7 trimer-trimer interactions in the T = 13 layer are mediated principally through well-defined regions in the broader lower domains, to form a structure that conforms well with that expected from the theory of quasi-equivalence with no significant conformational changes within the individual trimers. The VP3 layer determines the particle size and forms a rather smooth surface upon which the two-dimensional lattice of VP7 trimers is laid down.     

An automated, parallel processing approach to biomolecular sample preparation

Sample preparation for DNA and RNA assays is a prime candidate for laboratory automation. A novel, parallel processing device that performs the three separate liquid-handling functions necessary for such sample preparation-dispensing, pipetting, and pressurizing-is presented. The device comprises an array of fine nozzles connected by fluidic channels to automatically and precisely distribute flow between one source and an array of points. The design principles, as well as the experimental and computational methods used to develop the device, are described. Test results, including accuracy, uniformity, volume range, and timing, are presented. The functionality of the device is demonstrated by performing a solid-phase extraction of DNA with two types of microcolumns.     

Absolute stability of an implicit method based on third-order off-step discretization for the initial-value problem on a graded mesh

Engineering with Computers - We studied a new implicit method of order three based on two off-step points for the numerical solution of second-order nonlinear initial-value problems...     

Effects of temperature and blank holding force on biaxial forming behavior of aluminum sheet alloys

Biaxial forming behavior is investigated for three aluminum sheet alloys (Al 5182 containing 1% Mn (5182+Mn), Al 5754, and 6111-T4) using a heated die and punch in the warm forming temperature range of 200–350 °C. It is found that, while all three alloys exhibit significant improvement in their formability compared with that at room temperature, the non-heat-treatable alloys 5182 + Mn and 5754 give higher part depths than that of heat-treatable 6111-T4. The formability generally increases with decreasing BHP (BHP), but increasing the forming temperature and/or BHP minimizes the wrinkling tendency and improves the forming performance. The stretchability of the sheet alloys increase with increasing temperature and increasing BHP. For the alloys and forming conditions involved in the current study, the formability, measured in terms of part depth, comes mainly from the drawing of metal into the die cavity, although stretching effects do influence the overall forming behavior. The optimum formability is achieved by setting the die temperature 50 °C higher than the punch temperature to enhance the drawing component. Setting the die temperature higher than the punch temperature also improves the strain distribution in a part in such a manner that postpones necking and fracture by altering the location of greatest thinning.     

FORCED CONVECTIVE MASS TRANSFER IN ANNULI

Mass transfer in annuli has been critically examined for various flow situations. The overall mass transfer rate depends on the hydrodynamic regions prevailing in the annular channel as well as on its dimensions. Theoretically consistent correlations are proposed and recommended for both developed and developing boundary layers under laminar and turbulent flow conditions.     

Activation of rat liver cholesterol ester hydrolase by cAMP-dependent protein kinase and protein kinase C

Short term regulation of hepatic cholesterol ester hydrolase by reversible phosphorylation is described. Two different kinase systems seem to be involved in this regulation. The addition of ATP, cyclic AMP and Mg²⁺ to rat liver 104,000× g supernatant (S104) produced a 100–140% increase in cholesterol ester hydrolase activity. This stimulation was abolished when protein kinase inhibitor was added prior to the addition of ATP, cyclic AMP and Mg²⁺. Cholesterol ester hydrolase activity was also stimulated when calcium ions, phosphatidylserine, and diolein were added to S104 along with ATP and Mg²⁺. Diolein in this reaction could be substituted by phorbol 12-myristate 13-acetate. Preincubation of S104 with alkaline phosphatase resulted in a deactivation of cholesterol ester hydrolase. The addition of increasing concentrations of Mg²⁺ to S104 produced increasing inhibition of cholesterol ester hydrolase activity, and this effect was blocked by NaF. It is suggested that rat liver cholesterol ester hydrolase is activated by cyclic AMP dependent protein kinase and protein kinase C. Deactivation is accomplished by dephosphorylation catalyzed by a phosphoprotein phosphatase, dependent on Mg²⁺. This work was presented at the Twenty-Third Southeastern Regional Lipid Conference, held October 26–28, in Cashiers, North Carolina.     

Natural rubber–ethylene‐propylene‐diene rubber covulcanization: Effect of reinforcing fillers

Filled covulcanizates of elastomer blend comprising natural rubber (NR) and ethylene‐propylene‐diene rubber (EPDM) of commercial importance were successfully prepared by using a multifunctional rubber additive; namely, bis(diisopropyl)thiophosphoryl disulfide (DIPDIS). A Two‐stage vulcanization technique further improved the physicochemical properties of the blend vulcanizates by restricting, through the formation of polar rubber bound intermediates, the migration of curative and filler from lower to highly unsaturated rubber. Scanning electron microscopy studies indicate homogeneity and coherency in the morphology of the two‐stage vulcanizates. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 84: 1001–1010, 2002; DOI 10.1002/app.10361     

Conducting blends of poly(o‐toluidine) and ethylene–propylene–diene terpolymer

Poly(o‐toluidine) (POT) is an electroactive polymer with poor mechanical and thermal characteristics. We examined the scope for improving such properties by making blends of POT with ethylene–propylene–diene rubber (EPDM). We prepared POT–EPDM blends containing different weight fractions of POT by intimately mixing known volumes of separate solutions of the two polymers (POT in THF and EPDM in toluene). Films of EPDM and POT–EPDM blends in solution were obtained by spreading, solvent evaporation, and film casting techniques. POT, EPDM, and their blends were characterized in solution by ultraviolet‐visible spectroscopy, and the respective dried samples were analyzed by Fourier transform infrared spectroscopy and thermogravimetry. The polymer samples were further analyzed morphologically by scanning electron microscopy, and their tensile strengths were also evaluated. Spectroscopic and thermal studies of the blends indicated some sort of interaction between the two constituent polymers. The direct current electrical conductivity of the blends in increasing order of POT loading (12.5–100%) was in the range 9.9 × 10^?5 to 11.6 × 10^?2 S cm^?1. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 88: 2550–2555, 2003     

Predictive models for multibiometric systems

Recognizing a subject given a set of biometrics is a fundamental pattern recognition problem. This paper builds novel statistical models for multibiometric systems using geometric and multinomial distributions. These models are generic as they are only based on the similarity scores produced by a recognition system. They predict the bounds on the range of indices within which a test subject is likely to be present in a sorted set of similarity scores. These bounds are then used in the multibiometric recognition system to predict a smaller subset of subjects from the database as probable candidates for a given test subject. Experimental results show that the proposed models enhance the recognition rate beyond the underlying matching algorithms for multiple face views, fingerprints, palm prints, irises and their combinations.     

An Experimental Study to Evaluate the Effect of Ambient Temperature during Manual Lifting and Design of Optimal Task Parameters

In the present study, the effect of lifting task parameters on the heart rate and oxygen uptake of workers during manual lifting tasks in different ambient conditions was evaluated. The experiments conducted in two different temperature conditions showed a significantly higher oxygen uptake and heart rate in colder conditions as compared to warmer conditions. Three other factors, namely, load, lifting frequency, and vertical distance were found to significant affect the responses. Various combinations of significant factors were used to calculate oxygen uptake and heart rate. These were then compared with the safe limits as per the maximum aerobic capacity of workers. Based on these comparisons, the safe combinations were identified that can be used to design lifting tasks in varied ambient conditions. The study further concluded that lifting tasks performed in winter should have different relaxation or fatigue allowances built into the cycle time of the task to compensate for higher exertion. © 2011 Wiley Periodicals, Inc.