KeaA, a Dictyostelium kelch-domain protein that regulates the response to stress and development

Background: The protein kinase YakA is responsible for the growth arrest and induction of developmental processes that occur upon starvation of Dictyostelium cells. yakA-cells are aggregation deficient, have a faster cell cycle and are hypersensitive to oxidative and nitrosoative stress. With the aim of isolating members of the YakA pathway, suppressors of the death induced by nitrosoative stress in the yakA-cells were identified. One of the suppressor mutations occurred in keaA, a gene identical to DG1106 and similar to Keap1 from mice and the Kelch protein from Drosophila, among others that contain Kelch domains. Results: A mutation in keaA suppresses the hypersensitivity to oxidative and nitrosoative stresses but not the faster growth phenotype of yakA-cells. The growth profile of keaA deficient cells indicates that this gene is necessary for growth. keaA deficient cells are more resistant to nitrosoative and oxidative stress and keaA is necessary for the production and detection of cAMP. A morphological analysis of keaA deficient cells during multicellular development indicated that, although the mutant is not absolutely deficient in aggregation, cells do not efficiently participate in the process. Gene expression analysis using cDNA microarrays of wild-type and keaA deficient cells indicated a role for KeaA in the regulation of the cell cycle and pre-starvation responses. Conclusions: KeaA is required for cAMP signaling following stress. Our studies indicate a role for kelch proteins in the signaling that regulates the cell cycle and development in response to changes in the environmental conditions.

A Component of the Xanthomonadaceae Type IV Secretion System Combines a VirB7 Motif with a N0 Domain Found in Outer Membrane Transport Proteins

Type IV secretion systems (T4SS) are used by Gram-negative bacteria to translocate protein and DNA substrates across the cell envelope and into target cells. Translocation across the outer membrane is achieved via a ringed tetradecameric outer membrane complex made up of a small VirB7 lipoprotein (normally 30 to 45 residues in the mature form) and the C-terminal domains of the VirB9 and VirB10 subunits. Several species from the genera of Xanthomonas phytopathogens possess an uncharacterized type IV secretion system with some distinguishing features, one of which is an unusually large VirB7 subunit (118 residues in the mature form). Here, we report the NMR and 1.0 angstrom X-ray structures of the VirB7 subunit from Xanthomonas citri subsp. citri (VirB7(XAC2622)) and its interaction with VirB9. NMR solution studies show that residues 27-41 of the disordered flexible N-terminal region of VirB7(XAC2622) interact specifically with the VirB9 C-terminal domain, resulting in a significant reduction in the conformational freedom of both regions. VirB7(XAC2622) has a unique C-terminal domain whose topology is strikingly similar to that of N0 domains found in proteins from different systems involved in transport across the bacterial outer membrane. We show that VirB7(XAC2622) oligomerizes through interactions involving conserved residues in the N0 domain and residues 42-49 within the flexible N-terminal region and that these homotropic interactions can persist in the presence of heterotropic interactions with VirB9. Finally, we propose that VirB(7XAC2622) oligomerization is compatible with the core complex structure in a manner such that the N0 domains form an extra layer on the perimeter of the tetradecameric ring.

Intercellular transport of epidermis-expressed MADS domain transcription factors and their effect on plant morphology and floral transition

P>During the lifetime of an angiosperm plant various important processes such as floral transition, specification of floral organ identity and floral determinacy, are controlled by members of the MADS domain transcription factor family. To investigate the possible non-cell-autonomous function of MADS domain proteins, we expressed GFP-tagged clones of AGAMOUS (AG), APETALA3 (AP3), PISTILLATA (PI) and SEPALLATA3 (SEP3) under the control of the MERISTEMLAYER1 promoter in Arabidopsis thaliana plants. Morphological analyses revealed that epidermal overexpression was sufficient for homeotic changes in floral organs, but that it did not result in early flowering or terminal flower phenotypes that are associated with constitutive overexpression of these proteins. Localisations of the tagged proteins in these plants were analysed with confocal laser scanning microscopy in leaf tissue, inflorescence meristems and floral meristems. We demonstrated that only AG is able to move via secondary plasmodesmata from the epidermal cell layer to the subepidermal cell layer in the floral meristem and to a lesser extent in the inflorescence meristem. To study the homeotic effects in more detail, the capacity of trafficking AG to complement the ag mutant phenotype was compared with the capacity of the non-inwards-moving AP3 protein to complement the ap3 mutant phenotype. While epidermal expression of AG gave full complementation, AP3 appeared not to be able to drive all homeotic functions from the epidermis, perhaps reflecting the difference in mobility of these proteins.

Plasma Kallikrein and Angiotensin I-converting enzyme N- and C-terminal domain activities are modulated by the insertion/deletion polymorphism

Angiotensin I-converting enzyme (ACE) is recognized as one of the main effector molecules involved in blood pressure regulation. In the last few years some polymorphisms of ACE such as the insertion/deletion (I/D) polymorphism have been described, but their physiologic relevance is poorly understood. In addition, few studies investigated if the specific activity of ACE domain is related to the I/D polymorphism and if it can affect other systems. The aim of this study was to establish a biochemical and functional characterization of the I/D polymorphism and correlate this with the corresponding ACE activity. For this purpose, 119 male brazilian army recruits were genotyped and their ACE plasma activities evaluated from the C- and N-terminal catalytic domains using fluorescence resonance energy transfer (FRET) peptides, specific for the C-domain (Abz-LFK(Dnp)OH), N-domain (Abz-SDK(Dnp)P-OH) and both C- and N-domains (Abz-FRK(Dnp)P-OH). Plasma kallikrein activity was measured using Z-Phe-Arg-AMC as substrate and inhibited by selective plasma kallikrein inhibitor (PKSI). Some physiological parameters previously described related to the I/D polymorphism such as handgrip strength, blood pressure, heart rate and BMI were also evaluated. The genotype distribution was II n = 27, ID n = 64 and DD n = 28. Total plasma ACE activity of both domains in II individuals was significantly lower in comparison to ID and DD. This pattern was also observed for C- and N-domain activities. Difference between ID and DD subjects was observed only with the N-domain specific substrate. Blood pressure, heart rate, handgrip strength and BMI were similar among the genotypes. This polymorphism also affected the plasma kallikrein activity and DD group presents high activity level. Thus, our data demonstrate that the I/D ACE polymorphism affects differently both ACE domains without effects on handgrip strength. Moreover, this polymorphism influences the kallikrein-kinin system of normotensive individuals. (C) 2009 Elsevier Ltd. All rights reserved.

Homogenization in a thin domain with an oscillatory boundary

In this paper we analyze the behavior of the Laplace operator with Neumann boundary conditions in a thin domain of the type R(epsilon) = {(x(1), x(2)) is an element of R(2) vertical bar x(1) is an element of (0, 1), 0 < x(2) < epsilon G(x(1), x(1)/epsilon)} where the function G(x, y) is periodic in y of period L. Observe that the upper boundary of the thin domain presents a highly oscillatory behavior and, moreover, the height of the thin domain, the amplitude and period of the oscillations are all of the same order, given by the small parameter epsilon. (C) 2011 Elsevier Masson SAS. All rights reserved.

Behavior of Socket Base Connections Emphasizing Pedestal Walls

To evaluate the main design models for socket base connections of precast concrete structures, an experimental investigation was carried out on specimens of this connection with smooth and rough interfaces in contact with cast-in-place concrete. The specimens consisted of pedestal walls and were submitted to loads with large eccentricities. Based on the experimental results, two rational design models are proposed for this connection. One of these models accounts for the friction and is applied to socket bases with smooth interfaces. The main behavior model was verified for sockets with this type of interface and the design of the longitudinal walls as corbels is also suggested in this case. Because the behavior of the rough interface specimens was very close to a monolithic connection, the other proposed model is an adaptation of the bending theory to calculate the vertical reinforcement of socket bases with rough interfaces.

Analysis of the behavior of transverse walls of socket base connections

This paper presents a theoretical and experimental analysis of socket base connections of precast concrete structures with regard to the behavior of transverse walls. The experimental program included seven specimens, for which the type of interface in contact with cast-in-place concrete, the load eccentricities and the embedded lengths were all varied, A design model was proposed to calculate the reinforcements of the transverse walls. Based on the obtained results, some conclusions can be drawn: (a) The top of the transverse wall on the compression side of the smooth connections and the top of the two transverse walls of the rough connections are submitted to a bending-tension and this tension prevails on the bending; (b) The design model proposed for the calculation of the reinforcement of the transverse wall on the compression side provides the best prediction of the experimental results for all specimens when compared to the current design models; (c) For rough interfaces, the top of the transverse wall on the tension side is more requested than the top of transverse wall on the compression side; (d) The results of the proposed design model for the reinforcement of the transverse wall on the tension side of rough connections were in close agreement with the experimental results. (C) 2008 Elsevier Ltd. All rights reserved.

Proposal of a test specimen to evaluate the shear strength of vertical interfaces of running bond masonry walls

There is no normalized test to assess the shear strength of vertical interfaces of interconnected masonry walls. The approach used to evaluate this strength is normally indirect and often unreliable. The aim of this study is to propose a new test specimen to eliminate this deficiency. The main features of the proposed specimen are failure caused by shear stress on the vertical interface and a small number of units (blocks). The paper presents a numerical analysis based on the finite element method, with the purpose of showing the theoretical performance of the designed specimen, in terms of its geometry, boundary conditions, and loading scheme, and describes an experimental program using the specimen built with full- and third-scale clay blocks. The main conclusions are that the proposed specimen is easy to build and is appropriate to evaluate the sheaf strength of vertical interfaces of masonry walls.

A numerical model for the description of the nonlinear behaviour of multi-leaf masonry walls

A nonlinear finite element model was developed to simulate the nonlinear response of three-leaf masonry specimens, which were subjected to laboratory tests with the aim of investigating the mechanical behaviour of multiple-leaf stone masonry walls up to failure. The specimens consisted of two external leaves made of stone bricks and mortar joints, and an internal leaf in mortar and stone aggregate. Different loading conditions, typologies of the collar joints, and stone types were taken into account. The constitutive law implemented in the model is characterized by a damage tensor, which allows the damage-induced anisotropy accompanying the cracking process to be described. To follow the post-peak behaviour of the specimens with sufficient accuracy it was necessary to make the damage model non-local, to avoid mesh-dependency effects related to the strain-softening behaviour of the material. Comparisons between the predicted and measured failure loads are quite satisfactory in most of the studied cases. (c) 2007 Elsevier Ltd. All rights reserved.

Parametrical study of masonry walls subjected to in-plane loading through numerical modeling

This paper deals with the numerical assessment of the influence of parameters such as pre-compression level, aspect ratio, vertical and horizontal reinforcement ratios and boundary conditions on the lateral strength of masonry walls under in-plane loading. The numerical study is performed through the software DIANA (R) based on the Finite Element Method. The validation of the numerical model is carried out from a database of available experimental results on masonry walls tested under cyclic lateral loading. Numerical results revealed that boundary conditions play a central role on the lateral behavior of masonry walls under in-plane loading and determine the influence of level of pre-compression as well as the reinforcement ratio on the wall strength. The lateral capacity of walls decreases with the increase of aspect ratio and with the decrease of pre-compression. Vertical steel bars appear to have almost no influence in the shear strength of masonry walls and horizontal reinforcement only increases the lateral strength of masonry walls if the shear response of the walls is determinant for failure, which is directly related to the boundary conditions. (C) 2011 Elsevier Ltd. All rights reserved.

Study of the influence of simultaneous variation of magnetic material microstructural features on domain wall dynamics

The magnetic Barkhausen noise (MBN) is a phenomenon sensitive to several kinds of magnetic material microstructure changes, as well as to variations in material plastic deformation and stress. This fact stimulates the development of MBN-based non-destructive testing (NDT) techniques for analyzing magnetic materials, being the proposition of such a method, the main objective of the present study. The behavior of the MBN signal envelope, under simultaneous variations of carbon content and plastic deformation, is explained by the domain wall dynamics. Additionally, a non-destructive parameter for the characterization of each of these factors is proposed and validated through the experimental results. (C) 2010 Elsevier B.V. All rights reserved.

Effect of the domain spanwise periodic length on the flow around a circular cylinder

We assess the effect of the choice of spanwise periodic length on simulations of the flow around a fixed circular cylinder. The Reynolds number is set to 400 because, at this value, both lift coefficient and shedding frequency show significant drop due to three-dimensional flow structures. From the analysis of the three-dimensionalities of the wake and of the integral quantities such as Strouhal number, RMS of lift coefficient and energy contained in the three-dimensional portion of the flow we obtain an estimate of the minimum spanwise length to satisfactorily represent the flow. Furthermore, we observe a distinct wake behavior when the spanwise length is approximately the mode B instability wavelength. (C) 2011 Elsevier Ltd. All rights reserved.

Stable MPC for tracking with maximal domain of attraction

In this work, a stable MPC that maximizes the domain of attraction of the closed-loop system is proposed. The proposed approach is suitable to real applications in the sense that it accounts for the case of output tracking, it is offset free if the output target is reachable and minimizes the offset if some of the constraints are active at steady state. The new approach is based on the definition of a Minkowski functional related to the input and terminal constraints of the stable infinite horizon MPC. It is also shown that the domain of attraction is defined by the system model and the constraints, and it does not depend on the controller tuning parameters. The proposed controller is illustrated with small order examples of the control literature. (C) 2011 Elsevier Ltd. All rights reserved.

Enlarging the domain of attraction of stable MPC controllers, maintaining the output performance

This work presents an alternative way to formulate the stable Model Predictive Control (MPC) optimization problem that allows the enlargement of the domain of attraction, while preserving the controller performance. Based on the dual MPC that uses the null local controller, it proposed the inclusion of an appropriate set of slacked terminal constraints into the control problem. As a result, the domain of attraction is unlimited for the stable modes of the system, and the largest possible for the non-stable modes. Although this controller does not achieve local optimality, simulations show that the input and output performances may be comparable to the ones obtained with the dual MPC that uses the LQR as a local controller. (C) 2009 Elsevier Ltd. All rights reserved.