Structure of metal site in Cd-substituted His117Gly mutant of azurin with and without addition of imidazole derivatives

The present work uses 111mCd-perturbed angular correlations of gamma-rays (PAC) to investigate the structure of the metal site of the His117Gly mutant of Pseudomonas aeruginosa azurin in aqueous solution and the effect on the structure upon addition of the following exogenous ligands: imidazole, 4-methyl imidazole, 1-methyl imidazole, 2-methyl imidazole and histidine. The nuclear quadrupole interaction of cadmium bound to the mutant without addition of exogenous ligands shows a strong pH dependence with three different nuclear quadrupole interactions consistent with two pKa values at about 7.2 and 8.6 at 2 degrees C. Addition of the imidazole derivatives resulted in a significant change in the PAC spectrum showing that they coordinate. This is in accordance with observations by EPR for the same mutant with copper at the metal site [den Blaauwen, T. & Canters, G. W. (1993) J. Am. Chem. Soc. 115, 1121-1129]. However, whereas EPR and ultraviolet/visual absorption show that the characteristics of the wild-type copper protein are regained by addition of the imidazole derivatives with the exception of the possible bidentates (histidine and histamine), the comparison of the PAC results to model calculations shows that the cadmium ion must be fourfold coordinated in most cases, probably binding an additional water or hydroxide ligand. A fourfold coordination is in contrast to cadmium-substituted wild-type azurin where PAC data inferred a threefold coordination by a Cys and two His residues [Danielsen, E. Bauer, R., Hemmingsen, L., Andersen. M., Bjerrum, M. J., Butz, T., Tr?ger, W., Canters, G. W., Hoitink, C. W. G., Karlsson, G., Hansson, O. & Messerschmidt, A. (1995) J. Biol. Chem. 270, 573-580]     

Learning local linear Jacobians for flexible and adaptive robot arm control

Successful planning and control of robots strongly depends on the quality of kinematic models, which define mappings between configuration space (e.g. joint angles) and task space (e.g. Cartesian coordinates of the end effector). Often these models are predefined, in which case, for example, unforeseen bodily changes may result in unpredictable behavior. We are interested in a learning approach that can adapt to such changes—be they due to motor or sensory failures, or also due to the flexible extension of the robot body by, for example, the usage of tools. We focus on learning locally linear forward velocity kinematics models by means of the neuro-evolution approach XCSF. The algorithm learns self-supervised, executing movements autonomously by means of goal-babbling. It preserves actuator redundancies, which can be exploited during movement execution to fulfill current task constraints. For detailed evaluation purposes, we study the performance of XCSF when learning to control an anthropomorphic seven degrees of freedom arm in simulation. We show that XCSF can learn large forward velocity kinematic mappings autonomously and rather independently of the task space representation provided. The resulting mapping is highly suitable to resolve redundancies on the fly during inverse, goal-directed control.     

Mutant p53 Mediates Sensitivity to Cancer Treatment Agents in Oesophageal Adenocarcinoma Associated with MicroRNA and SLC7A11 Expression

TP53 gene mutations occur in 70% of oesophageal adenocarcinomas (OACs). Given the central role of p53 in controlling cellular response to therapy we investigated the role of mutant (mut-) p53 and SLC7A11 in a CRISPR-mediated JH-EsoAd1 TP53 knockout model. Response to 2 Gy irradiation, cisplatin, 5-FU, 4-hydroxytamoxifen, and endoxifen was assessed, followed by a TaqMan OpenArray qPCR screening for differences in miRNA expression. Knockout of mut-p53 resulted in increased chemo- and radioresistance (2 Gy survival fraction: 38% vs. 56%, p < 0.0001) and in altered miRNA expression levels. Target mRNA pathways analyses indicated several potential mechanisms of treatment resistance. SLC7A11 knockdown restored radiosensitivity (2 Gy SF: 46% vs. 73%; p = 0.0239), possibly via enhanced sensitivity to oxidative stress. Pathway analysis of the mRNA targets of differentially expressed miRNAs indicated potential involvement in several pathways associated with apoptosis, ribosomes, and p53 signaling pathways. The data suggest that mut-p53 in JH-EsoAd1, despite being classified as non-functional, has some function related to radio- and chemoresistance. The results also highlight the important role of SLC7A11 in cancer metabolism and redox balance and the influence of p53 on these processes. Inhibition of the SLC7A11-glutathione axis may represent a promising approach to overcome resistance associated with mut-p53.     

Constructing the dependency structure of a multiagent probabilisticnetwork

A probabilistic network consists of a dependency structure and corresponding probability tables. The dependency structure is a graphical representation of the conditional independencies that are known to hold in the problem domain. We propose an automated process for constructing the combined dependency structure of a multiagent probabilistic network. Each domain expert supplies any known conditional independency information and not necessarily an explicit dependency structure. Our method determines a succinct representation of all the supplied independency information called a minimal cover. This process involves detecting all inconsistent information and removing all redundant information. A unique dependency structure of the multiagent probabilistic network can be constructed directly from this minimal cover. The main result is that the constructed dependency structure is a perfect-map of the minimal cover. That is, every probabilistic conditional independency logically implied by the minimal cover can be inferred from the dependency structure and every probabilistic conditional independency inferred from the dependency structure is logically implied by the minimal cover     

Micro-fluidic target chamber machined by proton beam writing for the in situ analysis of gas absorption in synthetic crystals

Many synthetic crystals used for chemical and industrial applications have special internal structures, e.g. nano-pores, which allow separating different gases and fluids. Ion beam analytical methods can be used to study the gas diffusion and absorption in these materials in situ and to visualize their inner surfaces which affect these processes. For this purpose, a small target chamber was constructed in PMMA (polymethyl methacrylate) using proton beam writing (PBW). This micro-fluidic structure enables the establishment of a defined atmosphere around a crystal and allows the simultaneous ion beam analysis. In order to confine the gas from the high vacuum in the measurement chamber Si₃N₄-windows of 200 nm thickness were thermally bonded to the structured PMMA block yielding a closed target chamber with the possibility to accomplish PIXE and RBS measurements. In addition, two capillaries were connected to the chamber for gas inlet and evacuation. First tests showed that the construction is leak-proof and allows to establish a defined atmosphere. After that, the Argon gas diffusion into Zn(tbip)-crystals was studied. These measurements have shown unexpectedly high nickel concentrations in the host crystal which reduces the Argon density in these areas after absorption, because the Ni atoms decrease the pore size by replacing Zn-atoms in the Zn(tbip)-lattice. It could be demonstrated that gas diffusion and absorption in organic crystals can be studied in situ with high lateral resolution using ion beam analysis in a dedicated target chamber machined by PBW.