Coupling the phosphotransferase system and the methyl-accepting chemotaxis protein-dependent chemotaxis signaling pathways of Escherichia coli.

Chemotactic responses in Escherichia coli are typically mediated by transmembrane receptors that monitor chemoeffector levels with periplasmic binding domains and communicate with the flagellar motors through two cytoplasmic proteins, CheA and CheY. CheA autophosphorylates and then donates its phosphate to CheY, which in turn controls flagellar rotation. E. coli also exhibits chemotactic responses to substrates that are transported by the phosphoenolpyruvate (PEP)-dependent carbohydrate phosphotransferase system (PTS). Unlike conventional chemoreception, PTS substrates are sensed during their uptake and concomitant phosphorylation by the cell. The phosphoryl groups are transferred from PEP to the carbohydrates through two common intermediates, enzyme I (EI) and phosphohistidine carrier protein (HPr), and then to sugar-specific enzymes II. We found that in mutant strains HPr-like proteins could substitute for HPr in transport but did not mediate chemotactic signaling. In in vitro assays, these proteins exhibited reduced phosphotransfer rates from EI, indicating that the phosphorylation state of EI might link the PTS phospho-relay to the flagellar signaling pathway. Tests with purified proteins revealed that unphosphorylated EI inhibited CheA autophosphorylation, whereas phosphorylated EI did not. These findings suggest the following model for signal transduction in PTS-dependent chemotaxis. During uptake of a PTS carbohydrate, EI is dephosphorylated more rapidly by HPr than it is phosphorylated at the expense of PEP. Consequently, unphosphorylated EI builds up and inhibits CheA autophosphorylation. This slows the flow of phosphates to CheY, eliciting an up-gradient swimming response by the cell.

Muscarinic acetylcholine receptor down-regulation limits the extent of inhibition of cell cycle progression in Chinese hamster ovary cells.

Cellular desensitization is believed to be important for growth control but direct evidence is lacking. In the current study we compared effects of wild-type and down-regulation-resistant mutant m3 muscarinic receptors on Chinese hamster ovary (CHO-K1) cell desensitization, proliferation, and transformation. We found that down-regulation of m3 muscarinic acetylcholine receptors was the principal mechanism of desensitization of receptor-activated inositol phosphate phospholipid hydrolysis in these cells. Activation of wild-type and mutant receptors inhibited anchorage-independent growth as assayed by colony formation in agar. However, the potency for inhibition of anchorage-independent growth was greater for cells expressing the mutant receptor. Activation of either receptor also initially inhibited anchorage-dependent cell proliferation in randomly growing populations. Rates of DNA synthesis and cell division were profoundly reduced by carbachol in cells expressing either receptor at early time points. Analysis of cell cycle parameters indicated that cell cycle progression was inhibited at transitions from G1 to S and G2/M to G1 phases. However, mutant receptor effects on anchorage-dependent growth were sustained, whereas wild-type receptor effects were transient. Thus, receptor down-regulation restored cell cycle progression. In contrast, activation of either receptor blocked entry into the cell cycle from quiescence, and this response was not reduced by receptor down-regulation. Therefore, activation of m3 muscarinic acetylcholine receptors inhibited CHO cell anchorage-dependent and -independent growth. In anchored cells carbachol inhibited the cell cycle at three distinct points. Inhibitions at two of these points were eliminated by wild-type receptor down-regulation while the other was not. These results directly demonstrate that desensitization mechanisms can act as principal determinants of cellular growth responses.

Expression and signaling specificity of the IFNAR chain of the type I interferon receptor complex.

The IFNAR chain of the type I interferon (IFN) receptor (IFNIR) undergoes rapid ligand-dependent tyrosine phosphorylation and acts as a species-specific transducer for type I IFN action. Using the vaccinia/T7 expression system to amplify IFNAR expression, we found that human HeLa-S3 cells transiently express high levels of cell surface IFNAR chains (approximately 250,000 chains per cell). Metabolic labeling and immunoblot analysis of transfected HeLa cells show that the IFNAR chain is initially detected as 65-kDa and 98-kDa precursors, and then as the 130-kDa mature protein. Due to variation in N-glycosylation, the apparent molecular mass of the mature IFNAR chain varies from 105 to 135 kDa in different cells. IFNIR structure was characterized in various human cell lines by analyzing 125I-labeled IFN cross-linked complexes recognized by various antibodies against IFNIR subunits and JAK protein-tyrosine kinases. Precipitation of cross-linked material from Daudi cells with anti-IFNAR antibodies showed that IFNAR was present in a 240-kDa complex. Precipitation of cross-linked material from U937 cells with anti-TYK2 sera revealed a 240-kDa complex, which apparently did not contain IFNAR and was not present in IFN-resistant HEC1B cells. The tyrosine phosphorylation and down-regulation of the IFNAR chain were induced by type I IFN in several human cell lines of diverse origins but not in HEC1B cells. However, of type I IFNs, IFN-beta uniquely induced the tyrosine phosphorylation of a 105-kDa protein associated with the IFNAR chain in two lymphoblastoid cell lines (Daudi and U266), demonstrating the specificity of transmembrane signaling for IFN-beta and IFN-alpha through the IFNAR chain.

The vesicular monoamine transporter 2 is present in small synaptic vesicles and preferentially localizes to large dense core vesicles in rat solitary tract nuclei.

In central neurons, monamine neurotransmitters are taken up and stored within two distinct classes of regulated secretory vesicles: small synaptic vesicles and large dense core vesicles (DCVs). Biochemical and pharmacological evidence has shown that this uptake is mediated by specific vesicular monamine transporters (VMATs). Recent molecular cloning techniques have identified the vesicular monoamine transporter (VMAT2) that is expressed in brain. This transporter determines the sites of intracellular storage of monoamines and has been implicated in both the modulation of normal monoaminergic neurotransmission and the pathogenesis of related neuropsychiatric disease. We used an antiserum against VMAT2 to examine its ultrastructural distribution in rat solitary tract nuclei, a region that contains a dense and heterogeneous population of monoaminergic neurons. We find that both immunoperoxidase and immunogold labeling for VMAT2 localize to DCVs and small synaptic vesicles in axon terminals, the trans-Golgi network of neuronal perikarya, tubulovesicles of smooth endoplasmic reticulum, and potential sites of vesicular membrane recycling. In axon terminals, immunogold labeling for VMAT2 was preferentially associated with DCVs at sites distant from typical synaptic junctions. The results provide direct evidence that a single VMAT is expressed in two morphologically distinct types of regulated secretory vesicles in central monoaminergic neurons.

Triassic redbeds in the Malaguide Complex (Betic Cordillera — Spain): petrography, geochemistry and geodynamic implications

Sandstone petrography and mudstone mineralogy and geochemistry of Triassic mudstones and sandstones from continental redbeds of the Malaguide Complex (Betic Cordillera, southern Spain) provide useful information on provenance, palaeoclimate and geodynamics during the early stages of the Pangea break-up, and on their diagenetic evolution. The sandstones are quartzarenites to sub-litharenites, with minor lithic fragments and rare feldspars. The mudstone samples show a PAAS like elemental distribution. The samples likely record recycling processes from their metasedimentary basement rocks that significantly affected the weathering indices, and monitors cumulative effects, including a first cycle of weathering at the source rocks. Sandstone composition and chemical–mineralogical features of mudstones record a provenance derived from continental block and recycled orogen that were weathered under warm and episodically wet climate. Source areas were located towards the east of the present-day Malaguide outcrops, and were formed by fairly silicic rock types, made up mainly of Palaezoic metasedimentary rocks, similar to those of the Paleozoic underlying series, with subordinate contributions from magmatic–metamorphic sources, and a rare supply from mafic metavolcanic rocks. Clay-mineral distribution of mudstones is dominated by illite and illite/smectite mixed-layer that result from differences in provenance, weathering, and burial/temperature history. Illite crystallinity values, illitization of kaolinite, occurrence of typical authigenic minerals and apatite fission-track studies, coupled with a subsidence analysis of the whole Malaguide succession suggest burial depths of at least 4–6 km with temperatures of 140–160 °C, typical of the burial diagenetic stage, and confirm the Middle Miocene exhumation of the Betic Internal Domain tectonic stack topped by the Malaguide Complex.

Architectural Technology and the BIM Acronym 3: Getting to Grips with BIM

Among Small and Medium Sized Enterprises (SMEs) in particular, the UK Government’s ambitions regarding BIM uptake and diffusion across the construction sector may be tempered by a realpolitik shaped in part by interactions between the industry, Higher Education (HE) and professional practice. That premise also has a global perspective. Building on the previous 2 papers, Architectural technology and the BIM Acronym 1 and 2, this third iteration is a synthesis of research and investigations carried out over a number of years directly related to the practical implementation of BIM and its impact upon BE SMEs. First challenges, risks and potential benefits for SMEs and micros in facing up to the necessity to engage with digital tools in a competitive and volatile marketplace are discussed including tailoring BIM to suit business models, and filtering out achievable BIM outcomes from generic and bespoke aspects of practice. Second the focus is on setting up and managing teams engaging with BIM scenarios, including the role of clients; addresses a range of paradigms including lonely BIM and collaborative working. The significance of taking a whole life view with BIM is investigated including embedding soft landings principles into project planning and realisation. Thirdly procedures for setting up and managing common data environments are identified and the value of achieving smooth information flow is addressed. The overall objective of this paper is to provide SMEs with a practical strategy to develop a toolkit to BIM implementation.

Three types of transnational players: differing women's football mobility projects in core and developing countries

Mobile players in men's football are highly skilled professionals who move to a country other than the one where they grew up and started their careers. They are commonly described as migrants or expatriate players. Due to a much less advanced stage of professionalism and production of the game in women's football mobility projects are different. At describing the cases of Brazil, Equatorial Guinea, Mexico, Colombia and Portugal, the aim of this paper is to conceptualise an umbrella category for mobile players that can include current realities in the women's game, namely the transnational player who has gained and displays transnational football experience in different countries and socio-culturally contexts. Furthermore, analyses allow introducing two new subcategories besides the “expatriate”, namely diaspora players and new citizens.

Report on democracy assistance from the European Union to the Middle East and North Africa. EUSpring Report, 18 May 2016

Since the Arab uprisings of 2011, European Union (EU) assistance has nominally targeted more resources to supporting democracy movements in the Middle East and North Africa (MENA) region. The EU has better equipped itself institutionally, financially and conceptually, by strengthening its bottom up grassroots approach to democracy support; resources earmarked for supporting civil society have been increased, the budget for the European Instrument for Democracy and Human Rights (EIDHR) has been beefed up, and the strengthened EU Delegations have be come more empowered to reach out to groups at the local grass roots level behind democracy activities; The European Endowment for Democracy (EED) was created with the mandate to support individuals and organizations in neighbouring countries that work for democracy. Whether this translates into a more effective strategy for democracy support, however, remains to be seen. In this report, Rosa Balfour, Francesca Fabbri and Richard Youngs present a detailed overview of the support given to civil society in the MENA region by the EU, with a special focus on the various financial instruments used.

3D models created from ROV videos and their corresponding subtransect dimensions

Underwater video transects have become a common tool for quantitative analysis of the seafloor. However a major difficulty remains in the accurate determination of the area surveyed as underwater navigation can be unreliable and image scaling does not always compensate for distortions due to perspective and topography. Depending on the camera set-up and available instruments, different methods of surface measurement are applied, which make it difficult to compare data obtained by different vehicles. 3-D modelling of the seafloor based on 2-D video data and a reference scale can be used to compute subtransect dimensions. Focussing on the length of the subtransect, the data obtained from 3-D models created with the software PhotoModeler Scanner are compared with those determined from underwater acoustic positioning (ultra short baseline, USBL) and bottom tracking (Doppler velocity log, DVL). 3-D model building and scaling was successfully conducted on all three tested set-ups and the distortion of the reference scales due to substrate roughness was identified as the main source of imprecision. Acoustic positioning was generally inaccurate and bottom tracking unreliable on rough terrain. Subtransect lengths assessed with PhotoModeler were on average 20% longer than those derived from acoustic positioning due to the higher spatial resolution and the inclusion of slope. On a high relief wall bottom tracking and 3-D modelling yielded similar results. At present, 3-D modelling is the most powerful, albeit the most time-consuming, method for accurate determination of video subtransect dimensions.

Cholesterol and C30 of sediment profile MD90-963

In a deep-sea sediment core recovered from a site lying well above the local lysocline, several organic geochemical proxies, and two different calcite dissolution indicators, are compared in order to evaluate the relationship between calcite dissolution and paleoproductivity over the past three glacial-interglacial cycles. The degree of foraminiferal break-up, and the CaCO3 particle size distribution, both point to significant periods of dissolution every 22 kyr during glacial stages and substages. These dissolution events are concomitant with periods of enhanced primary productivity, as indicated by the abundance of several biomarkers (alkenones, cholesterol, brassicasterol, keto-ol), used here to indicate changes in paleoproductivity. Dissolution fluctuations are highly coherent and in phase with the estimated paleoproductivity variations providing strong evidence that the observed dissolution is due to organic matter remineralization within the sediments rather, than to changes in CO32? concentration in the overlying water column.

Cyclopedia of American horticulture, comprising suggestions for cultivation of horticultural plants, descriptions of the species of fruits, vegetables, flowers and ornamental plants sold in the United States and Canada, together with geographical and biographical sketches /

Illustrated with two thousand eight hundred original engravings.

The improvement of the moral qualities ; an ethical treatise of the eleventh century by Solomon ibn Gabirol, printed from an unique Arabic manuscript, together with a translation, and an essay on the place of Gabirol in the history of the development of Jewish ethics,

From a manuscript in the Bodleian library, Oxford.

How Tyson came home : a story of England and America /

Advertisements on p. [2]-[5] at end.

Dollars and cents /

Frontispiece and plates facing p. 96 and 280.

Following the conquistadores ...

Vol. 1-2, written under the author's pseud., H.J. Mozans.