The nuclease activities of both the Smr domain and an additional LDLK motif are required for an efficient anti-recombination function of Helicobacter pyloriMutS2

Helicobacter pylori, a human pathogen, is a naturally and constitutively competent bacteria, displaying a high rate of intergenomic recombination. While recombination events are essential for evolution and adaptation of H.pylori to dynamic gastric niches and new hosts, such events should be regulated tightly to maintain genomic integrity. Here, we analyze the role of the nuclease activity of MutS2, a protein that limits recombination during transformation in H.pylori. In previously studied MutS2 proteins, the C-terminal Smr domain was mapped as the region responsible for its nuclease activity. We report here that deletion of Smr domain does not completely abolish the nuclease activity of HpMutS2. Using bioinformatics analysis and mutagenesis, we identified an additional and novel nuclease motif (LDLK) at the N-terminus of HpMutS2 unique to Helicobacter and related epsilon-proteobacterial species. A single point mutation (D30A) in the LDLK motif and the deletion of Smr domain resulted in approximate to 5-10-fold loss of DNA cleavage ability of HpMutS2. Interestingly, the mutant forms of HpMutS2 wherein the LDLK motif was mutated or the Smr domain was deleted were unable to complement the hyper-recombination phenotype of a mutS2(-) strain, suggesting that both nuclease sites are indispensable for an efficient anti-recombinase activity of HpMutS2.

Biochemical Characterization of Nonamer Binding Domain of RAG1 Reveals its Thymine Preference with Respect to Length and Position

RAG complex consisting of RAG1 and RAG2 is a site-specific endonuclease responsible for the generation of antigen receptor diversity. It cleaves recombination signal sequence (RSS), comprising of conserved heptamer and nonamer. Nonamer binding domain (NBD) of RAG1 plays a central role in the recognition of RSS. To investigate the DNA binding properties of the domain, NBD of murine RAG1 was cloned, expressed and purified. Electrophoretic mobility shift assays showed that NBD binds with high affinity to nonamer in the context of 12/23 RSS or heteroduplex DNA. NBD binding was specific to thymines when single stranded DNA containing poly A, C, G or T were used. Biolayer interferometry studies showed that poly T binding to NBD was robust and comparable to that of 12RSS. More than 23 nt was essential for NBD binding at homothymidine stretches. On a double-stranded DNA, NBD could bind to A:T stretches, but not G:C or random sequences. Although NBD is indispensable for sequence specific activity of RAGs, external supplementation of purified nonamer binding domain to NBD deleted cRAG1/cRAG2 did not restore its activity, suggesting that the overall domain architecture of RAG1 is important. Therefore, we define the sequence requirements of NBD binding to DNA.

Energy principle of ferroelectric ceramics and single domain mechanical model

Many physical experiments have shown that the domain switching in a ferroelectric material is a complicated evolution process of the domain wall with the variation of stress and electric field. According to this mechanism, the volume fraction of the domain switching is introduced in the constitutive law of ferroelectric ceramic and used to study the nonlinear constitutive behavior of ferroelectric body in this paper. The principle of stationary total energy is put forward in which the basic unknown quantities are the displacement u (i) , electric displacement D (i) and volume fraction rho (I) of the domain switching for the variant I. Mechanical field equation and a new domain switching criterion are obtained from the principle of stationary total energy. The domain switching criterion proposed in this paper is an expansion and development of the energy criterion. On the basis of the domain switching criterion, a set of linear algebraic equations for the volume fraction rho (I) of domain switching is obtained, in which the coefficients of the linear algebraic equations only contain the unknown strain and electric fields. Then a single domain mechanical model is proposed in this paper. The poled ferroelectric specimen is considered as a transversely isotropic single domain. By using the partial experimental results, the hardening relation between the driving force of domain switching and the volume fraction of domain switching can be calibrated. Then the electromechanical response can be calculated on the basis of the calibrated hardening relation. The results involve the electric butterfly shaped curves of axial strain versus axial electric field, the hysteresis loops of electric displacement versus electric filed and the evolution process of the domain switching in the ferroelectric specimens under uniaxial coupled stress and electric field loading. The present theoretic prediction agrees reasonably with the experimental results given by Lynch.

Design, Construction and Testing of a Hydraulic Power Take-Off for Wave Energy Converters

This paper presents the construction, mathematical modeling and testing of a scaled universal hydraulic Power Take-Off (PTO) device for Wave Energy Converters (WECs). A specific prototype and test bench were designed and built to carry out the tests. The results obtained from these tests were used to adjust an in-house mathematical model. The PTO was initially designed to be coupled to a scaled wave energy capture device with a low speed and high torque oscillating motion and high power fluctuations. Any Energy Capture Device (ECD) that fulfils these requirements can be coupled to this PTO, provided that its scale is adequately defined depending on the rated power of the full scale prototype. The initial calibration included estimation of the pressure drops in the different components, the pressurization time of the oil inside the hydraulic cylinders and the volumetric efficiency of the complete circuit. Since the overall efficiency measured during the tests ranged from 0.69 to 0.8 and the dynamic performance of the PTO was satisfactory, the results are really promising and it is believed that this solution might prove effective in real devices.

Measures of tuna abundance from purse-seine operations in the Eastern Pacific Ocean, adjusted for fleet-wide evolution of increased fishing power, 1960-1971

ENGLISH: The fishing power of the tuna purse-seine fleet of the eastern Pacific Ocean has increased since the early 1960's. Because the entire fleet seems to have adopted equipment and techniques to increase its efficiency in capturing tunas, traditional methods of adjusting catch rates to a reference vessel type of fixed efficiency to index tuna abundance from fishing success are inapplicable. Instead, a methodology for such adjustment based on a mathematical representation of purse seining activities is developed. Observed changes in efficiency in subprocesses of purse seining are then used to adjust catch rates when computing abundance histories for yellowfin and skipjack in large regions of the eastern Pacific Ocean. SPANISH: La eficacia de pesca de la flota de cerco atunera en el Océano Pacífico oriental ha aumentado desde el comienzo del decenio de 1960. Como toda la flota parece haber adoptado equipo y métodos para incrementar su eficaciaen capturar atunes, no se pueden aplicar los métodos tradicionales de ajustar los índices de captura a un tipo normalizado de barco (es decir de eficacia fija) para indicar la abundancia del atún según los resultados de pesca. En su lugar se ha desarrollado un método para realizar tal ajuste basado en una representación matemática de las actividades de las embarcaciones de cerco. Cuando se calcula la abundancia histórica del atún aleta amarilla y barrilete en grandes regiones del Océano Pacífico oriental, se usan entonces los cambios observados en la eficacia de los subprocesos cerqueros para ajustar los índices de captura. (PDF contains 120 pages.)

Proteomic analysis of the Cdc48/Ubx network identifies a role for Ubx2 in the regulation of lipid biosynthesis

Cdc48/p97 is an essential, highly abundant hexameric member of the AAA (ATPase associated with various cellular activities) family. It has been linked to a variety of processes throughout the cell but it is best known for its role in the ubiquitin proteasome pathway. In this system it is believed that Cdc48 behaves as a segregase, transducing the chemical energy of ATP hydrolysis into mechanical force to separate ubiquitin-conjugated proteins from their tightly-bound partners.

Current models posit that Cdc48 is linked to its substrates through a variety of adaptor proteins, including a family of seven proteins (13 in humans) that contain a Cdc48-binding UBX domain. As such, due to the complexity of the network of adaptor proteins for which it serves as the hub, Cdc48/p97 has the potential to exert a profound influence on the ubiquitin proteasome pathway. However, the number of known substrates of Cdc48/p97 remains relatively small, and smaller still is the number of substrates that have been linked to a specific UBX domain protein. As such, the goal of this dissertation research has been to discover new substrates and better understand the functions of the Cdc48 network. With this objective in mind, we established a proteomic screen to assemble a catalog of candidate substrate/targets of the Ubx adaptor system.

Here we describe the implementation and optimization of a cutting-edge quantitative mass spectrometry method to measure relative changes in the Saccharomyces cerevisiae proteome. Utilizing this technology, and in order to better understand the breadth of function of Cdc48 and its adaptors, we then performed a global screen to identify accumulating ubiquitin conjugates in cdc48-3 and ubxΔ mutants. In this screen different ubx mutants exhibited reproducible patterns of conjugate accumulation that differed greatly from each other, pointing to various unexpected functional specializations of the individual Ubx proteins.

As validation of our mass spectrometry findings, we then examined in detail the endoplasmic-reticulum bound transcription factor Spt23, which we identified as a putative Ubx2 substrate. In these studies ubx2Δ cells were deficient in processing of Spt23 to its active p90 form, and in localizing p90 to the nucleus. Additionally, consistent with reduced processing of Spt23, ubx2Δ cells demonstrated a defect in expression of their target gene OLE1, a fatty acid desaturase. Overall, this work demonstrates the power of proteomics as a tool to identify new targets of various pathways and reveals Ubx2 as a key regulator lipid membrane biosynthesis.

Upconversion luminescence and optical power limiting effect based on two- and three-photon absorption processes of ZnO crystal

Near-infrared to UV and visible upconversion luminescence was observed in single-crystalline ZnO under an 800 nm infrared femtosecond laser irradiation. The optical properties of the crystal reveal that the UV and VIS emission band are due to the exciton transition (D0X) bound to neutral donors and the deep luminescent centers in ZnO, respectively. The relationship between the upconversion luminescence intensity and the pump power of the femtosecond laser reveals that the UV emission belongs to three-photon sequential band-to-band excitation and the VIS emission belongs to two-photon simultaneous defect-absorption induced luminescence. A saturation phenomenon and polarization-dependent effect are also observed in the upconversion process of ZnO. A very good optical power limiting performance at 800 nm has been demonstrated. The two- and three-photon absorption coefficients of ZnO crystal were measured to be 0.2018 cm GW(-1) and 7.102 x 10(-3) cm(3) GW(-2), respectively. The two- and three-photon cross sections were calculated to be 1.189 x 10(-51) cm(4) s and 1.040 x 10(-80) cm(6) s(2), respectively. (c) 2007 Elsevier B.V. All rights reserved.

Dynamic phase-mapping of domain nucleation on MgO : LiNbO3 crystal by digital holographic interferometry

The quantitative phase-mapping of the domain nucleation in MgO:LiNbO3 crystals is presented by using the digital holographic interferometry. An unexpected peak phase at the beginning of the domain nucleation is observed and it is lowered as the spreading of the domain nucleus. The existence of the nucleus changes the moving speed of the domain wall by pinning it for 3s. Such in-situ quantitative analysis of the domain nucleation process is a key to optimizing domain structure fabrication.

Phase mapping of domain kinetics in lithium niobate by digital holographic interferometry

The phase mapping of domain kinetics under the uniform steady-state electric field is achieved and investigated in the LiNbO3 crystals by digital holographic interferometry. We obtained the sequences of reconstructed three-dimensional and two-dimensional wave-field phase distributions during the electric poling in the congruent and near stoichiometric LiNbO3 crystals. The phase mapping of individual domain nucleation and growth in the two crystals are obtained. It is found that both longitudinal and lateral domain growths are not linear during the electric poling. The phase mapping of domain wall motions in the two crystals is also obtained. Both the phase relaxation and the pinning-depinning mechanism are observed during the domain wall motion. The residual phase distribution is observed after the high-speed domain wall motion. The corresponding analyses and discussions are proposed to explain the phenomena.

Nd : YAG ceramic laser obtained high slope-efficiency of 62% in high power applications

By using quite uniformly nine-stacks side-around arranged compact pumping system, a high power Nd:YAG ceramic quasi-CW laser with high slope efficiency of 62% has been demonstrated. With 450 W quasi-CW stacked laser diode bars pumping at 808 nm, performance of the Nd: YAG ceramic laser with different output coupling mirrors has been investigated. Optimum output power of 236 W at 1064 nm was obtained and corresponding optical-to-optical conversion efficiency was as high as 52.5%. The laser system operated quite stably and no saturation phenomena have been observed, which means higher output laser power could be obtained if injecting higher pumping power. The still-evolving Nd: YAG ceramics are potential super excellent media for high power practical laser applications. (c) 2005 Optical Society of America.

High power single transverse mode operation of a tapered large-mode-area fiber laser

To get high output power with good beam quality, a tapered section is introduced to large-mode-area (LMA) Yb-doped fiber laser. Output characteristics of the fiber laser without tapered section and with tapered section are compared experimentally. When the launched pump power is 119.1 W, 77.9 W with M-2 3.08 and 56.4 W with M-2 1.14 can be obtained, respectively. The corresponding slope efficiencies are 71.8% and 54.1%, respectively. Although output power of the tapered fiber laser has 30.6% penalty, brightness of it is as much as 5.28 times of the fiber laser without tapered section. Moreover, spectra of them are measured. It is found that tapered section makes lasing wavelength of the fiber laser shorter. (c) 2007 Elsevier B.V. All rights reserved.

7.3 W of single-frequency output power at 2.09 μm from an Ho:YAG monolithic nonplanar ring laser

Single-frequency output power of 7.3 W at 2.09 mu m from a monolithic Ho:YAG nonplanar ring oscillator (NPRO) is demonstrated. Resonantly pumped by a Tm-doped fiber laser at 1.91 mu m, the Ho:YAG NPRO produces 71% of slope efficiency with respect to absorbed pump power and nearly diffraction-limited output with a beam quality parameter of M-2 approximate to 1.1. (c) 2008 Optical Society of America

Application of domain decomposition and transmission line modelling techniques to 2D, time-domain, finite element problems

In this paper a recently published finite element method, which combines domain decomposition with a novel technique for solving nonlinear magnetostatic finite element problems is described. It is then shown how the method can be extended to, and optimised for, the solution of time-domain problems. © 1999 IEEE.

Characterize dynamic conformational space of human CCR5 extracellular domain by molecular modeling and molecular dynamics simulation

The chemokine receptor CCR5 is the receptor for several chemokines and major coreceptor for R5 human immunodeficiency virus type-1 strains entry into cell. Three-dimensional models of CCR5 were built by using homology modeling approach and 1 ns molecular dynamics (MD) simulation, because studies of site-directed mutagenesis and chimeric receptors have indicated that the N-terminus (Nt) and extracellular loops (ECLs) of CCR5 are important for ligands binding and viral fusion and entry, special attention was focused on disulfide bond function, conformational flexibility, hydrogen bonding, electrostatic interactions, and solvent-accessible surface area of Nt and ECLs of this protein part. We found that the extracellular segments of CCR5 formed a well-packet globular domain with complex interactions occurred between them in a majority of time of MID simulation, but Nt region could protrude from this domain sometimes. The disulfide bond Cys20-Cys269 is essential in controlling specific orientation of Nt region and maintaining conformational integrity of extracellular domain. RMS comparison analysis between conformers revealed the ECL1 of CCR5 stays relative rigid, whereas the ECL2 and Nt are rather flexible. Solvent-accessible surface area calculations indicated that the charged residues within Nt and ECL2 are often exposed to solvent. Integrating these results with available experimental data, a two-step gp120-CCR5 binding mechanism was proposed. The dynamic interaction of CCR5 extracellular domain with gp120 was emphasized. (C) 2004 Elsevier B.V. All rights reserved.

The evolution of information while building cross-domain models of a design: A video experiment

Models capturing the connectivity between different domains of a design, e.g. between components and functions, can provide a tool for tracing and analysing aspects of that design. In this paper, video experiments are used to explore the role of cross-domain modelling in building up information about a design. The experiments highlight that cross-domain modelling can be a useful tool to create and structure design information. Findings suggest that consideration of multiple domains encourages discussion during modelling, helps identify design aspects that might otherwise be overlooked, and can help promote consideration of alternative design options. Copyright © 2002-2012 The Design Society. All rights reserved.