Alteration of the Route to Menaquinone towards Isochorismate-Derived Metabolites

Chorismate and isochorismate constitute branch-point intermediates in the biosynthesis of many aromatic metabolites in microorganisms and plants. To obtain unnatural compounds, we modified the route to menaquinone in Escherichia coli. We propose a model for the binding of isochorismate to the active site of MenD ((1R,2S, 5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxycyclohex-3-ene-1-carboxylate (SEPHCHC) synthase) that explains the outcome of the native reaction with α-ketoglutarate. We have rationally designed variants of MenD for the conversion of several isochorismate analogues. The double-variant Asn117Arg–Leu478Thr preferentially converts (5S,6S)-5,6-dihydroxycyclohexa-1,3-diene-1-carboxylate (2,3-trans-CHD), the hydrolysis product of isochorismate, with a >70-fold higher ratio than that for the wild type. The single-variant Arg107Ile uses (5S,6S)-6-amino-5-hydroxycyclohexa-1,3-diene-1-carboxylate (2,3-trans-CHA) as substrate with >6-fold conversion compared to wild-type MenD. The novel compounds have been made accessible in vivo (up to 5.3 g L⁻¹). Unexpectedly, as the identified residues such as Arg107 are highly conserved (>94 %), some of the designed variations can be found in wild-type SEPHCHC synthases from other bacteria (Arg107Lys, 0.3 %). This raises the question for the possible natural occurrence of as yet unexplored branches of the shikimate pathway.     

Simulation of the submerged energy nozzle-mold water model system using laser-optical and computational fluid dynamics methods

The present work describes quantitative digital particle image velocimetry measurements of a full-scale water model of a thin slab mold. Different casting speeds and two submerged entry nozzles with one and two outlet ports have been investigated. The flow pattern of the single-port nozzle shows a counterclockwise-rotating double vortex that is nearly steady-state but leads to high stationary surface waves. The flow jets out of the two-port nozzle oscillate and produce a transient flow pattern with low wave amplitudes. The amplitudes for the one-port nozzle show a linear variation with the volumetric flow rate. The experimental results lead to a good interpretation of the flow phenomena and are used to validate steady-state numerical simulations with the commercial program, CFX, on the basis of the Reynolds equations. To describe anisotropic turbulence effects, the Reynolds stress model (RSM) is used for the flat single-port nozzle and the standard k-ɛ model for the mold flow. The calculated mean velocities and wave amplitudes, predicted from pressure distribution at the water surface, are generally in the consensus of the experimental data. An erratum to this article is available at .     

The Israeli-Palestinian peace process and its vicissitudes: Insights from attitude theory.

The vicissitudes of the Israeli-Palestinian peace process since 1967 are analyzed using attitudes and related concepts where relevant. The 1967 war returned the two peoples' zero-sum conflict around national identity to its origin as a conflict within the land both peoples claim. Gradually, new attitudes evolved regarding the necessity and possibility of negotiations toward a two-state solution based on mutual recognition, which became the building stones of the 1993 Oslo agreement. Lacking a commitment to a final outcome, the Oslo-based peace process was hampered by reserve options, which increased avoidance at the expense of approach tendencies as the parties moved toward a final agreement. The resulting breakdown of the process in 2000 produced clashing narratives, reflecting different anchors for judgment and classical mirror images. Public support for violence increased, even as public opinion continued to favor a negotiated two-state solution. Reviving the peace process requires mutual reassurance about the availability of a partner for negotiating a principled peace based on a historic compromise that meets the basic needs and validates the identities of both peoples. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Treatment of depressive symptoms during short-term rehabilitation: An attempted replication of the DOUR project.

Objective: To replicate and extend P. A. Lichtenberg and colleagues' (1996) cross-disciplinary intervention to improve physical and mental health among older adults. Participants: 14 depressed older adults (6 treatment, 8 control). Setting: The short-term rehabilitation unit of an urban nursing home. Intervention: Occupational therapists were trained to treat depression using pleasant events and cognitive-behavioral therapies. Outcome Measures: Geriatric Depression Scale, the Short Form-12, and the Multi-Level Assessment Instrument: Activities of Daily Living. Results: No significant group differences were found in physical or mental health. However, more control group members (75%) than treatment group members (33%) were depressed at study completion. Conclusions: The treatment of depressive symptoms can be integrated with a nonmental health treatment modality. (PsycINFO Database Record (c) 2011 APA, all rights reserved)     

Development of calcium signalling mechanisms during maturation of human oocytes

Sperm-induced Ca2+ signals mediate the events of oocyte activation at fertilization. In this study, the development of mechanisms involved in the generation of Ca2+ signals in human oocytes was investigated. The thiol reagent, thimerosal, which induces oscillations of intracellular Ca2+ ([Ca2+]i) similar to those seen during fertilization, was used to mobilize Ca2+ in in-vivo matured, immature and in-vitro matured human oocytes. There was an increase in the sensitivity to thimerosal during maturation of human oocytes, with oocytes from small antral follicles being relatively insensitive, compared with those from luteinized follicles, which displayed a large spike followed by sustained oscillations in [Ca2+]i. These oscillations were inhibited by caffeine which suggests that they were mediated by the inositol trisphosphate receptor Ca2+ release system. When immature oocytes were cultured in vitro they acquired the capacity to undergo a single large spike in [Ca2+]i, however, subsequent sustained oscillations were not observed, indicating that these oocytes failed to develop fully competent Ca2+ signalling mechanisms during culture in vitro. This finding may be a key factor in the poor developmental competence of in-vitro matured human oocytes.