A subunit interaction in chloroplast ATP synthase determined by genetic complementation between chloroplast and bacterial ATP synthase genes

F1F0-ATP synthases utilize protein conformational changes induced by a transmembrane proton gradient to synthesize ATP. The allosteric cooperativity of these multisubunit enzymes presumably requires numerous protein-protein interactions within the enzyme complex. To correlate known in vitro changes in subunit structure with in vivo allosteric interactions, we introduced the beta subunit of spinach chloroplast coupling factor 1 ATP into a bacterial F1 ATP synthase. A cloned atpB gene, encoding the complete chloroplast beta subunit, complemented a chromosomal deletion of the cognate uncD gene in Escherichia coli and was incorporated into a functional hybrid F1 ATP synthase. The cysteine residue at position 63 in chloroplast beta is known to be located at the interface between alpha and beta subunits and to be conformationally coupled, in vitro, to the nucleotide binding site > 40 A away. Enlarging the side chain of chloroplast coupling factor 1 beta residue 63 from Cys to Trp blocked ATP synthesis in vivo without significantly impairing ATPase activity or ADP binding in vitro. The in vivo coupling of nucleotide binding at catalytic sites to transmembrane proton movement may thus involve an interaction, via conformational changes, between the amino-terminal domains of the alpha and beta subunits.     

Geometry modeling in cellular network planning

To design a cellular radio network (GSM, UMTS, WiFi …) it is indispensable today to use a radio planning software. In current commercialized tools, radio network planning is based on a numerical evaluation of a set of radio metrics, directly linked to quality of service notion, such as interference, throughput, coverage, capacity … The network configurations produced by such tools are generally faced with the human expert judgment who attaches great importance to the organizational aspects such as cells size and the topology. In other words, all those concepts related to the visual cells shape and geometry. In this article we emphasize the simplicity and the strength by which the geometrical concepts helps to bring out good network properties which are not taken into account by conventional radio criteria. On one hand, well-formed cells lead to reduce the number of handover calls, and to form transition zones between cells allowing proper completion of the handover procedure. On the other hand, cell geometry optimization produce performing network schemes in which the frequencies reuse (frequencies planning in GSM, WiFi) is simplified. We describe in this paper an original and practical modeling of the cellular geometry criteria granting the integration of this concept into the automatic planning process of mobile radio networks. The relevance of this criterion is assessed on three different levels. On a local level, we first establish the correlation between the geometric criteria and the improvement of point-by-point radio quality indicators. We secondly show geometry impact on radio frequency planning and the improving of the handover zones. Finally we analyze the impact of geometric criterion on improving indoor positioning systems.     

Abschätzung der Schwingfestigkeit nitrierter bauteilähnlicher Proben mit Hilfe normierter Wöhlerstreubänder

Estimation of fatigue strength of nitrided samples by means of standardized Wöhler diagram Based on results of rotation bending tests of the annealed and nitrided steel 31CrMoV9 statistic verified S-N-curves were estimated. The location of the failure initiation point, surface or subsurface, has an importened influence on the parameters of the S-N-curve. The origin of failure mainly depends of the specimen geometry, the structure of the nitrided case and the load conditions. For smooth and notched specimens with subsurface failure the fatigue strength can be calculated by Kogaev and Serensen and the standardized Wöhler diagram can be used for fatigue limit calculation independently of the structure of nitrided core.     

An Atomistic Understanding of Allosteric Inhibition of Glutamate Racemase: a Dampening of Native Activation Dynamics

Glutamate racemases (GR) are members of the family of bacterial enzymes known as cofactor-independent racemases and epimerases and catalyze the stereoinversion of glutamate. D-amino acids are universally important for the proper construction of viable bacterial cell walls, and thus have been repeatedly validated as attractive targets for novel antimicrobial drug design. Significant aspects of the mechanism of this challenging stereoinversion remain unknown. The current study employs a combination of MD and QM/MM computational approaches to show that the GR from H. pylori must proceed via a pre-activation step, which is dependent on the enzyme's flexibility. This mechanism is starkly different from previously proposed mechanisms. These findings have immediate pharmaceutical relevance, as the H. pylori GR enzyme is a very attractive allosteric drug target. The results presented in this study offer a distinctly novel understanding of how AstraZeneca's lead series of inhibitors cripple the H. pylori GR's native motions, via prevention of this critical chemical pre-activation step. Our experimental studies, using SPR, fluorescence and NMR WaterLOGSY, show that H. pylori GR is not inhibited by the uncompetitive mechanism originally put forward by Lundqvist et al.. The current study supports a deep connection between native enzyme motions and chemical reactivity, which has strong relevance to the field of allosteric drug discovery.     

Mammalian Exocrine Secretions. XII: Constituents of Interdigital Secretions of Bontebok, Damaliscus dorcas dorcas, and Blesbok, D. d. phillipsi

In addition to the nine compounds identified in the interdigital secretion of the bontebok, Damaliscus dorcas dorcas, in a previous study, 76 compounds belonging to different compound types, were identified in the interdigital secretions of the bontebok and the blesbok, D. d. phillipsi. These compounds include alkanes, alcohols, aldehydes, ketones, fatty acids, terpenoids, -lactones, an isopropyl ester, long-chain hydroxyesters, 2-substituted pyridines, phenols, steroids, and dimethylsulfone. No qualitative differences were found between secretions from the two sexes or from animals from different habitats. Although no attempt was made to correlate territorial behavior or other behavioral phenomena with the qualitative composition of interdigital secretions from individual animals, available information seems to indicate that quantitative differences probably do not have a major semiochemical function. Only two species of bacteria, Bacillus brevis and Planococcus citreus, were found in the interdigital pouches of male and female members of the two subspecies, regardless of the habitat of the animals. B. brevis synthesized, among other unidentified constituents, (Z)-3-penten-2-ol, 2-hexanone, 2-octanone, 2-nonanone, tetradecanoic acid, pentadecanoic acid, heptadecanoic acid, octadecanoic acid, (Z)-9-hexadecenoic acid, and isopropyl hexadecanoate in vitro, while P. citreus produced, among others, the -lactones dodecan-4-olide and (Z)-6-dodecen-4-olide, which is one of the major constituents of the interdigital secretions of both subspecies. Some components of the interdigital secretions are not present in the interdigital glandular tissue, and the possibility is discused that these compounds could be produced by microbiological activity in the interdigital pouch.     

Einfluß von Sauerstoffzusätzen beim Gasnitrieren auf den strukturellen Aufbau von Nitrierschichten

Influence of oxygen additions during gas nitriding on the structure of the nitrided layers The influence of controlled oxygen additions during gas nitriding of steels on the structure and built-up of the nitrided layers as well as on the nitriding reactions was investigated with metallurgical methods and Mössbauer spectroscopy. The results show that oxygen additions below the oxidising threshold slightly improve the nitriding behaviour of unalloyed steels. Furthermore, it is shown that for alloyed steel, which are tending to passivation starting form chromium concentrations of 3%, the nitriding can be considerably improved by oxygen additions. The passivation layers present for these steels are destroyed by external oxidation during the oxinitriding process. This external oxidation and the internal nitriding occurred parallel in the early stages of the oxinitriding.     

The urinary ratio of 5-hydroxytryptophol to 5-hydroxyindole-3-acetic acid in surgical patients with chronic alcohol misuse

Interactions among growth factors are important in a variety of physiological and pathological processes. The regulation of IGF-I mRNA expression by bFGF was investigated in cultured rat Müller cells and the mechanism of regulation studied. Müller cells from 1- to 3-day-old Sprague-Dawley rats were isolated and cultured with Eagle MEM+10% FCS. Cultured cells were identified by immunocytochemistry using antibodies against vimentin, carbonic anhydrase C, and glutamine synthetase. Cells of passage 1-4 were treated with bFGF, the PKC inhibitor H-7, calphostin C, the PKC activator PMA or the PKA inhibitor H-89, as well as the adenylate cyclase activator forskolin, or adenylate cyclase inhibitor SQ22536. IGF-I and bFGF expression levels were assessed by Northern blot analysis. The addition of bFGF to culture medium down-regulated IGF-I expression in a dose- and time-dependent manner. Decrease of IGF-I expression started at a bFGF concentration of 1 ng ml-1. IGF-I mRNA level declined to 44% of baseline level at 10 ng ml-1 of bFGF, and reached a trough of 40% at 50 ng ml-1. At 10 ng ml-1 of bFGF, down-regulation of IGF-I expression was observed as early as 4 hr (60%) after treatment, and reached a trough of 42% by 8 hr. The temporal and concentration dependence of IGF-I expression by addition of the PKC activator PMA, to culture medium was similar to that due to the addition of bFGF. The down-regulation of IGF-I expression by bFGF (10 ng ml-1) and PMA (0.1 microM) was blocked by the PKC inhibitors H-7 (30 microM) and calphostin C (1 microM). Forskolin (5 microM), an adenylate cyclase activator, had activator, had no effect on IGF-I expression. SQ22536 (100 microM), an adenylate cyclase inhibitor, and H-89, a PKA inhibitor, had no inhibitory effect on bFGF-induced down-regulation of IGF-I expression. These results indicate that bFGF down-regulates IGF-I expression in cultured rat M uller cells through PKC activation.     

Uncertainty in biomass supply estimates: Lessons from a Yakama Nation case study

The increasing demand for bioenergy in the United States necessitates detailed case studies of cost and supply to assess its feasibility. We have developed supply curves based on six feedstocks in five counties surrounding the Yakama Nation in central Washington using spatially explicit estimates of supply and transportation cost. The supply curves were used to examine a base case supply for a bioenergy plant, to analyze the effects of land ownership, and examine the impacts of uncertainty in parameters used to determine cost and supply. The results show that existing industries produce the cheapest supply of feedstock as a byproduct of their operations, while supplies harvested specifically for bioenergy are considerably more expensive. Fragmented land ownerships lead to the necessity of cooperation between owners and highlight the importance of a strong anchor supply close to the plant. Lastly, uncertainty in supply and cost parameters leads to larger ranges in available biomass, leading to reluctant investment in large plants.     

Texture of metal-matrix composites by ultrasonic velocity measurements

An ultrasonic method is developed for the nondestructive characterization of texture in metal-matrix composites. In this approach, it is assumed that the presence of reinforcement particles changes the elastic properties of the composite but only the texture of the matrix. The method utilizes the measurements of the six independent ultrasonic velocitiesV _ij and the formulation given by Bunge. The examined composites are the silicon carbide (SiC)-particle-reinforced aluminum 8091, 7064, and 6061 metal-matrix composites. The fourth-order expansion coefficients of the orientation distribution function are determined as a function of the SiC content in these composites. The results show that the expansion coefficients change with the presence of SiC where the coefficients C ₄ ¹¹ and C ₄ ¹³ increase as the volume fraction of SiC is increased and the coefficient C ₄ ¹² is zero in all composites examined. The analysis of these results indicates that ultrasonics can provide a promising technique for the texture characterization of metal-matrix composites.