992 resultados para Two-domain architecture


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En route from Birmingham to Syria in 2013, British-Jihadi neophytes aged 22, Yusuf Sarwar and Mohammed Ahmed purchased two books via Amazon to prepare for their mission in Syria after joining ISIS: The Koran for Dummies and Islam for Dummies. Journalists were swift to disparage their reading. The book’s author, Princeton University campus imam, Sohaib Nazeer Sultan remarked “Even though they may have ordered it, I don't think they read it.” In 1933, aged 27, Adolf Eichmann moved to Berlin to join the Sicherheitsdienst SD whereupon he read Immanuel Kant’s book the Kritik der praktischen Vernunft (The Critique of Practical Reason) for the first time. After his trial in Jerusalem, Hannah Arendt of course dismissed Eichmann’s reading of the German philosopher as thoroughly vacuous. Ever since, writers have sought to undermine the veracity of Eichmann’s account. The global Jihadis are illiterate, a journalist recently commented: they’re not well read in the Qur’an, and if they have read it, they have thoroughly misunderstood it. He cited as evidence Abdul Raqib Amin’s YouTube rhetorical: Forget everyone. Read the Koran, read the instruction of life. Find out what is jihad. Eichmann on the other hand was not illiterate in his youth. Before Berlin, he had already read Kant’s Groundwork of the Metaphysics of Morals ; he would also re-read the Critique of Practical Reason, and from his testimony and terminology we can infer he was familiar with Kantian concepts that extend beyond both books...

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Enzymes belonging to the M1 family play important cellular roles and the key amino acids (aa) in the catalytic domain are conserved. However, C-terminal domain aa are highly variable and demonstrate distinct differences in organization. To address a functional role for the C-terminal domain, progressive deletions were generated in Tricorn interacting factor F2 from Thermoplasma acidophilum (F2) and Peptidase N from Escherichia coli (PepN). Catalytic activity was partially reduced in PepN lacking 4 C-terminal residues (PepNΔC4) whereas it was greatly reduced in F2 lacking 10 C-terminal residues (F2ΔC10) or PepN lacking eleven C-terminal residues (PepNΔC11). Notably, expression of PepNΔC4, but not PepNΔC11, in E. coliΔpepN increased its ability to resist nutritional and high temperature stress, demonstrating physiological significance. Purified C-terminal deleted proteins demonstrated greater sensitivity to trypsin and bound stronger to 8-amino 1-napthalene sulphonic acid (ANS), revealing greater numbers of surface exposed hydrophobic aa. Also, F2 or PepN containing large aa deletions in the C-termini, but not smaller deletions, were present in high amounts in the insoluble fraction of cell extracts probably due to reduced protein solubility. Modeling studies, using the crystal structure of E. coli PepN, demonstrated increase in hydrophobic surface area and change in accessibility of several aa from buried to exposed upon deletion of C-terminal aa. Together, these studies revealed that non-conserved distal C-terminal aa repress the surface exposure of apolar aa, enhance protein solubility, and catalytic activity in two soluble and distinct members of the M1 family.

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The spot or strip application of poisoned protein bait is a lure-and-kill technique used for the management of fruit flies. Knowledge of where flies occur in the crop environment is an important part of maximizing the efficacy of this tool. Bactrocera tryoni is a polyphagous pest of horticulture for which very little is known about its distribution within crops. With particular reference to edge effects, we monitored the abundance of B. tryoni in two crops of different architecture; strawberry and apple. In strawberries, we found more flies on the crop edge early in the fruiting season, which lessened gradually and eventually disappeared as the season progressed. In apple orchards, no such edge effect was observed and flies were found equally throughout the orchard. We postulated these differences may be due to differences in crop height (high vs. short) and/or crop canopy architecture (opened and branched in apple, dense and closed in strawberry). In a field cage trial, we tested these predictions using artificial plants of different height and canopy condition. Height and canopy structure type had no significant effects on fly oviposition and protein feeding, but the 'apple' type canopy significantly influenced resting. We thus postulate that there was an edge effect in strawberry because the crop was not providing resting sites and flies were doing so in vegetation around the field margins. The finding that B. tryoni shows different resting site preferences based on plant architecture offers the potential for strategic manipulation of the fly through specific border or inter-row plantings. © 2013 Blackwell Verlag GmbH.

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With the intent of probing the feasibility of employing annulation as a tactic to engender axial rich conformations in nucleoside analogues, two adenine-derived, ``conformationally restricted'' nucleocylitols, 9 and 10, have been conceptualized as representatives of a hitherto unexplored class of nucleic acid base-cyclitol hybrids. A general synthetic strategy, with an inherent scope for diversification, allowed rapid functionalization of indane and tetralin to furnish 9 and 10 respectively in fair yield. Single-crystal X-ray diffraction analysis revealed that the two nucleocyclitols under study, though homologous, present completely dissimilar modes of molecular packing, marked, in particular, by the nature of involvement of the adenynyl NH2 group in the supramolecular assembly. In addition, the crystal structures of 9 and 10 also exhibit two different conformations of the functionalized cyclohexane ring. Thus, while the six-membered carbocycle in cyclopenta-annulated 9 exists in the expected chair (C) conformation that in cyclohexaannulated 10, which crystallizes as a dihydrate, shows an unusual twist-boat (TB) conformation. From a close analysis of the (HNMR)-H-1 spectroscopic data recorded for 9 and 10 in CD3OD, it was possible to put forth a putative explanation for the uncanny conformational preferences of crystalline 9 and 10.

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Replication and transcription of the RNA genome of alphaviruses relies on a set of virus-encoded nonstructural proteins. They are synthesized as a long polyprotein precursor, P1234, which is cleaved at three processing sites to yield nonstructural proteins nsP1, nsP2, nsP3 and nsP4. All the four proteins function as constitutive components of the membrane-associated viral replicase. Proteolytic processing of P1234 polyprotein is precisely orchestrated and coordinates the replicase assembly and maturation. The specificity of the replicase is also controlled by proteolytic cleavages. The early replicase is composed of P123 polyprotein intermediate and nsP4. It copies the positive sense RNA genome to complementary minus-strand. Production of new plus-strands requires complete processing of the replicase. The papain-like protease residing in nsP2 is responsible for all three cleavages in P1234. This study addressed the mechanisms of proteolytic processing of the replicase polyprotein in two alphaviruses Semliki Forest virus (SFV) and Sindbis virus (SIN) representing different branches of the genus. The survey highlighted the functional relation of the alphavirus nsP2 protease to the papain-like enzymes. A new structural motif the Cys-His catalytic dyad accompanied with an aromatic residue following the catalytic His was described for nsP2 and a subset of other thiol proteases. Such an architecture of the catalytic center was named the glycine specificity motif since it was implicated in recognition of a specific Gly residue in the substrate. In particular, the presence of the motif in nsP2 makes the appearance of this amino acid at the second position upstream of the scissile bond a necessary condition for the cleavage. On top of that, there were four distinct mechanisms identified, which provide affinity for the protease and specifically direct the enzyme to different sites in the P1234 polyprotein. Three factors RNA, the central domain of nsP3 and the N-terminus of nsP2 were demonstrated to be external modulators of the nsP2 protease. Here I suggest that the basal nsP2 protease specificity is inherited from the ancestral papain-like enzyme and employs the recognition of the upstream amino acid signature in the immediate vicinity of the scissile bond. This mechanism is responsible for the efficient processing of the SFV nsP3/nsP4 junction. I propose that the same mechanism is involved in the cleavage of the nsP1/nsP2 junction of both viruses as well. However, in this case it rather serves to position the substrate, whereas the efficiency of the processing is ensured by the capability of nsP2 to cut its own N-terminus in cis. Both types of cleavages are demonstrated here to be inhibited by RNA, which is interpreted as impairing the basal papain-like recognition of the substrate. In contrast, processing of the SIN nsP3/nsP4 junction was found to be activated by RNA and additionally potentiated by the presence of the central region of nsP3 in the protease. The processing of the nsP2/nsP3 junction in both viruses occurred via another mechanism, requiring the exactly processed N-terminus of nsP2 in the protease and insensitive to RNA addition. Therefore, the three processing events in the replicase polyprotein maturation are performed via three distinct mechanisms in each of two studied alphaviruses. Distinct sets of conditions required for each cleavage ensure sequential maturation of P1234 polyprotein: nsP4 is released first, then the nsP1/nsP2 site is cut in cis, and liberation of the nsP2 N-terminus activates the cleavage of the nsP2/nsP3 junction at last. The first processing event occurs differently in SFV and SIN, whereas the subsequent cleavages are found to be similar in the two viruses and therefore, their mechanisms are suggested to be conserved in the genus. The RNA modulation of the alphavirus nonstructural protease activity, discovered here, implies bidirectional functional interplay between the alphavirus RNA metabolism and protease regulation. The nsP2 protease emerges as a signal transmitting moiety, which senses the replication stage and responds with proteolytic cleavages. A detailed hypothetical model of the alphavirus replicase core was inferred from the data obtained in the study. Similar principles in replicase organization and protease functioning are expected to be employed by other RNA viruses.

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The PRP17 gene product is required for the second step of pre-mRNA splicing reactions. The C-terminal half of this protein bears four repeat units with homology to the beta transducin repeat. Missense mutations in three temperature-sensitive prp17 mutants map to a region in the N-terminal half of the protein. We have generated, in vitro, 11 missense alleles at the beta transducin repeat units and find that only one affects function in vivo. A phenotypically silent missense allele at the fourth repeat unit enhances the slow-growing phenotype conferred by an allele at the third repeat, suggesting an interaction between these domains. Although many missense mutations in highly conserved amino acids lack phenotypic effects, deletion analysis suggests an essential role for these units. Only mutations in the N-terminal nonconserved domain of PRP17 are synthetically lethal in combination with mutations in PRP16 and PRP18, two other gene products required for the second splicing reaction. A mutually allele-specific interaction between Prp17 and snr7, with mutations in U5 snRNA, was observed. We therefore suggest that the functional region of Prp17p that interacts with Prp18p, Prp16p, and U5 snRNA is the N terminal region of the protein.

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Double-stranded RNA (dsRNA) viruses encode only a single protein species that contains RNA-dependent RNA polymerase (RdRP) motifs. This protein is a central component in the life cycle of a dsRNA virus, carrying out both RNA transcription and replication. The architecture of viral RdRPs resembles that of a 'cupped right hand' with fingers, palm and thumb domains. Those applying de novo initiation have additional structural features, including a flexible C-terminal domain that constitutes the priming platform. Moreover, viral RdRPs must be able to interact with the incoming 3'-terminus of the template and position it so that a productive binary complex is formed. Bacteriophage phi6 of the Cystoviridae family is to date one of the best studied dsRNA viruses. The purified recombinant phi6 RdRP is highly active in vitro and possesses both RNA replication and transcription activities. The extensive biochemical observations and the atomic level crystal structure of the phi6 RdRP provides an excellent platform for in-depth studies of RNA replication in vitro. In this thesis, targeted structure-based mutagenesis, enzymatic assays and molecular mapping of phi6 RdRP and its RNA were used to elucidate the formation of productive RNA-polymerase binary complexes. The positively charged rim of the template tunnel was shown to have a significant role in the engagement of highly structured ssRNA molecules, whereas specific interactions further down in the template tunnel promote ssRNA entry to the catalytic site. This work demonstrated that by aiding the formation of a stable binary complex with optimized RNA templates, the overall polymerization activity of the phi6 RdRP can be greatly enhanced. Furthermore, proteolyzed phi6 RdRPs that possess a nick in the polypeptide chain at the hinge region, which is part of the extended loop, were better suited for catalysis at higher temperatures whilst favouring back-primed initiation. The clipped C-terminus remains associated with the main body of the polymerase and the hinge region, although structurally disordered, is involved in the control of C-terminal domain displacement. The accumulated knowhow on bacteriophage phi6 was utilized in the development of two technologies for the production of dsRNA: (i) an in vitro system that combines the T7 RNA polymerase and the phi6 RdRP to generate dsRNA molecules of practically unlimited length, and (ii) an in vivo RNA replication system based on restricted infection with phi6 polymerase complexes in bacterial cells to produce virtually unlimited amounts of dsRNA. The pools of small interfering RNAs derived from dsRNA produced by these systems were validated and shown to efficiently decrease the expression of both exogenous and endogenous targets.

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For the first time, we find the complex solitons for a quasi-one-dimensional Bose-Einstein condensate with two-and three-body interactions. These localized solutions are characterized by a power law behaviour. Both dark and right solitons can be excited in the experimentally allowed parameter domain, when two-and three-body interactions are,respectively, repulsive and attractive. The dark solitons travel with a constant speed, which is quite different from the Lieb mode, where profiles with different speeds, bounded above by sound velocity, can exist for specified interaction strengths. We also study the properties of these solitons in the presence of harmonic confinement with time-dependent nonlinearity and loss. The modulational instability and the Vakhitov-Kolokolov criterion of stability are also studied.

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Single-stranded DNA-binding proteins (SSB) play an important role in most aspects of DNA metabolism including DNA replication, repair, and recombination. We report here the identification and characterization of SSB proteins of Mycobacterium smegmatis and Mycobacterium tuberculosis. Sequence comparison of M. smegmatis SSB revealed that it is homologous to M. tuberculosis SSB, except for a small spacer connecting the larger amino-terminal domain with the extreme carboxyl-terminal tail. The purified SSB proteins of mycobacteria bound single-stranded DNA with high affinity, and the association and dissociation constants were similar to that of the prototype SSB. The proteolytic signatures of free and bound forms of SSB proteins disclosed that DNA binding was associated with structural changes at the carboxyl-terminal domain. Significantly, SSB proteins from mycobacteria displayed high affinity for cognate RecA, whereas Escherichia coli SSB did not under comparable experimental conditions. Accordingly, SSB and RecA were coimmunoprecipitated from cell lysates, further supporting an interaction between these proteins in vivo. The carboxyl-terminal domain of M. smegmatis SSB, which is not essential for interaction with ssDNA, is the site of binding of its cognate RecA. These studies provide the first evidence for stable association of eubacterial SSB proteins with their cognate RecA, suggesting that these two proteins might function together during DNA repair and/or recombination.

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Superhydrophobic and superhydrophilic surfaces have been extensively investigated due to their importance for industrial applications. It has been reported, however, that superhydrophobic surfaces are very sensitive to heat, ultraviolet (UV) light, and electric potential, which interfere with their long-term durability. In this study, we introduce a novel approach to achieve robust superhydrophobic thin films by designing architecture-defined complex nanostructures. A family of ZnO hollow microspheres with controlled constituent architectures in the morphologies of 1D nanowire networks, 2D nanosheet stacks, and 3D mesoporous nanoball blocks, respectively, was synthesized via a two-step self-assembly approach, where the oligomers or the constituent nanostructures with specially designed structures are first formed from surfactant templates, and then further assembled into complex morphologies by the addition of a second co-surfactant. The thin films composed of two-step synthesized ZnO hollow microspheres with different architectures presented superhydrophobicities with contact angles of 150°-155°, superior to the contact angle of 103° for one-step synthesized ZnO hollow microspheres with smooth and solid surfaces. Moreover, the robust superhydrophobicity was further improved by perfluorinated silane surface modification. The perfluorinated silane treated ZnO hollow microsphere thin films maintained excellent hydrophobicity even after 75 h of UV irradiation. The realization of environmentally durable superhydrophobic surfaces provides a promising solution for their long-term service under UV or strong solar light irradiations.

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It is a challenge to increase the visible-light photoresponses of wide-gap metal oxides. In this study, we proposed a new strategy to enhance the visible-light photoresponses of wide-gap semiconductors by deliberately designing a multi-scale nanostructure with controlled architecture. Hollow ZnO microspheres with constituent units in the shape of one-dimensional (1D) nanowire networks, 2D nanosheet stacks, and 3D mesoporous nanoball blocks are synthesized via an approach of two-step assembly, where the oligomers or the constituent nanostructures with specially designed structures are first formed, and then further assembled into complex morphologies. Through deliberate designing of constituent architectures allowing multiple visible-light scattering, reflections, and dispersion inside the multiscale nanostructures, enhanced wide range visible-light photoresponses of the ZnO hollow microspheres were successfully achieved. Compared to the one-step synthesized ZnO hollow microspheres, where no nanostructured constituents were produced, the ZnO hollow microspheres with 2D nanosheet stacks presented a 50 times higher photocurrent in the visible-light range (λ > 420 nm). The nanostructure induced visible-light photoresponse enhancement gives a direction to the development of novel photosensitive materials.

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Cities of Light is the first global overview of modern urban illumination, a development that allows human wakefulness to colonize the night, doubling the hours available for purposeful and industrious activities. Urban lighting is undergoing a revolution due to recent developments in lighting technology, and increased focus on sustainability and human-scaled environments. Cities of Light is expansive in coverage, spanning two centuries and touching on developments on six continents, without diluting its central focus on architectural and urban lighting. Covering history, geography, theory, and speculation in urban lighting, readers will have numerous points of entry into the book, finding it easy to navigate for a quick reference and or a coherent narrative if read straight through. With chapters written by respected scholars and highly-regarded contemporary practitioners, this book will delight students and practitioners of architectural and urban history, area and cultural studies, and lighting design professionals and the institutional and municipal authorities they serve. At a moment when the entire world is being reshaped by new lighting technologies and new design attitudes, the longer history of urban lighting remains fragmentary. Cities of Light aims to provide a global framework for historical studies of urban lighting and to offer a new perspective on the fast-moving developments of lighting today.

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Copper(I) complexes with {Cu(μ2-S)N}4 and {Cu(μ3-S)N}12 core portions of butterfly-shaped or double wheel architectures have been isolated in the reaction of Cu(I) with the Schiff base ligand C6H4(CHNC6H4S)2, aiso-abtâ, under different conditions. View the MathML source containing the tetranuclear electroneutral complex View the MathML source is formed by the reaction of CuI in acetonitrilic solution and recrystallization from DMF, whereas View the MathML source containing dodecanuclear View the MathML source wheels is accessible starting from CuBF4. Complexes 2 and 4 represent the first examples of cyclic complexes with the same overall stoichiometry but different ring sizes. The ligand induces two different coordination environments around copper(I) by switching between μ2- and μ3-sulfur bridging modes.

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This study examines the properties of Generalised Regression (GREG) estimators for domain class frequencies and proportions. The family of GREG estimators forms the class of design-based model-assisted estimators. All GREG estimators utilise auxiliary information via modelling. The classic GREG estimator with a linear fixed effects assisting model (GREG-lin) is one example. But when estimating class frequencies, the study variable is binary or polytomous. Therefore logistic-type assisting models (e.g. logistic or probit model) should be preferred over the linear one. However, other GREG estimators than GREG-lin are rarely used, and knowledge about their properties is limited. This study examines the properties of L-GREG estimators, which are GREG estimators with fixed-effects logistic-type models. Three research questions are addressed. First, I study whether and when L-GREG estimators are more accurate than GREG-lin. Theoretical results and Monte Carlo experiments which cover both equal and unequal probability sampling designs and a wide variety of model formulations show that in standard situations, the difference between L-GREG and GREG-lin is small. But in the case of a strong assisting model, two interesting situations arise: if the domain sample size is reasonably large, L-GREG is more accurate than GREG-lin, and if the domain sample size is very small, estimation of assisting model parameters may be inaccurate, resulting in bias for L-GREG. Second, I study variance estimation for the L-GREG estimators. The standard variance estimator (S) for all GREG estimators resembles the Sen-Yates-Grundy variance estimator, but it is a double sum of prediction errors, not of the observed values of the study variable. Monte Carlo experiments show that S underestimates the variance of L-GREG especially if the domain sample size is minor, or if the assisting model is strong. Third, since the standard variance estimator S often fails for the L-GREG estimators, I propose a new augmented variance estimator (A). The difference between S and the new estimator A is that the latter takes into account the difference between the sample fit model and the census fit model. In Monte Carlo experiments, the new estimator A outperformed the standard estimator S in terms of bias, root mean square error and coverage rate. Thus the new estimator provides a good alternative to the standard estimator.

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Let Ohm be a bounded domain in IRN, N greater than or equal to 2, lambda > 0, q is an element of (0, N - 1) and alpha is an element of (1, N/N-1 In this article we show the existence of at least two positive solutions for the following quasilinear elliptic problem with an exponential type nonlinearity: