Localization of the C terminus of the assembly domain of hepatitis B virus capsid protein: Implications for morphogenesis and organization of encapsidated RNA

The capsid protein of hepatitis B virus, consisting of an “assembly” domain (residues 1–149) and an RNA-binding “protamine” domain (residues 150–183), assembles from dimers into icosahedral capsids of two different sizes. The C terminus of the assembly domain (residues 140–149) functions as a morphogenetic switch, longer C termini favoring a higher proportion of the larger capsids, it also connects the protamine domain to the capsid shell. We now have defined the location of this peptide in capsids assembled in vitro by engineering a mutant assembly domain with a single cysteine at its C terminus (residue 150), labeling it with a gold cluster and visualizing the cluster by cryo-electron microscopy. The labeled protein is unimpaired in its ability to form capsids. Our density map reveals a single undecagold cluster under each fivefold and quasi-sixfold vertex, connected to sites at either end of the undersides of the dimers. Considering the geometry of the vertices, the C termini must be more crowded at the fivefolds. Thus, a bulky C terminus would be expected to favor formation of the larger (T = 4) capsids, which have a greater proportion of quasi-sixfolds. Capsids assembled by expressing the full-length protein in Escherichia coli package bacterial RNAs in amounts equivalent to the viral pregenome. Our density map of these capsids reveals a distinct inner shell of density—the RNA. The RNA is connected to the protein shell via the C-terminal linkers and also makes contact around the dimer axes.

Implications of macromolecular crowding for signal transduction and metabolite channeling

The effect of different total enzyme concentrations on the flux through the bacterial phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) in vitro was determined by measuring PTS-mediated carbohydrate phosphorylation at different dilutions of cell-free extract of Escherichia coli. The dependence of the flux on the protein concentration was more than linear but less than quadratic. The combined flux–response coefficient of the four enzymes constituting the glucose PTS decreased slightly from values of ≈1.8 with increasing protein concentrations in the assay. Addition of the macromolecular crowding agents polyethylene glycol (PEG) 6000 and PEG 35000 led to a sharper decrease in the combined flux–response coefficient, in one case to values of ≈1. PEG 6000 stimulated the PTS flux at lower protein concentrations and inhibited the flux at higher protein concentrations, with the transition depending on the PEG 6000 concentration. This suggests that macromolecular crowding decreases the dissociation rate constants of enzyme complexes. High concentrations of the microsolute glycerol did not affect the combined flux–response coefficient. The data could be explained with a kinetic model of macromolecular crowding in a two-enzyme group-transfer pathway. Our results suggest that, because of the crowded environment in the cell, the different PTS enzymes form complexes that live long on the time-scale of their turnover. The implications for the metabolic behavior and control properties of the PTS, and for the effect of macromolecular crowding on nonequilibrium processes, are discussed.

Regulation of the polarization of T cells toward antigen-presenting cells by Ras-related GTPase CDC42.

The mechanisms by which cells rapidly polarize in the direction of external signals are not understood. Helper T cells, when contacted by an antigen-presenting cell, polarize their cytoskeletons toward the antigen-presenting cell within minutes. Here we show that, in T cells, the mammalian Ras-related GTPase CDC42 (the homologue of yeast CDC42, a protein involved in budding polarity) can regulate the polarization of both actin and microtubules toward antigen-presenting cells but is not involved in other T-cell signaling processes such as those which culminate in interleukin 2 production. Although T-cell polarization appears dispensable for signaling leading to interleukin 2 production, polarization may direct lymphokine secretion towards the correct antigen-presenting cell in a crowded cellular environment. Inhibitor experiments suggest that phosphatidylinositol 3-kinase is required for cytoskeletal polarization but that calcineurin activity, known to be important for other aspects of signaling, is not. Apparent conservation of CDC42 function between yeast and T cells suggests that this GTPase is a general regulator of cytoskeletal polarity in many cell types.

Relaciones y prácticas que cambian la realidad alimenticia: una aproximación desde la antropología ecológica y feminista

From the 1990s, through the first decade of the XXI century, the food industry has intensified its production in technologically and genetically sophisticated ways. It has introduced transgenic and genetically modified foods, taking into account an economical push to obtain higher quantities in less time. Today, the foods that we consume seem more like products created in a laboratory than ones that come from working directly with the earth and with animals. These changes in the food industry are just a part of a long and complicated story in which economical interests figure heavily. The single-crop farming era begins in the 1970s in The United States and Europe. In some regions in Spain having a strong agricultural tradition, small private and family-owned farms that provided food to surrounding populations started disappearing, being uprooted in favor of the creation of large, multi-national companies. The market would expand with the growth of production facilities housing large quantities of animals living numbered and crowded. They mainly house cows, chickens, and pigs from which we obtain different products like milk, eggs and meat. The way these “industrial animals” live today does not even come close to what we think of as a balanced ecosystem, seeing as they are surrounded by machines and by the general use of sophisticated techniques to achieve the best return possible...

FOIA, Inc.

Government transparency is imagined as a public good necessary to a robust democracy. Consistent with that vision, Congress enacted the Freedom of Information Act (FOIA) to allow oversight and accountability of governmental activities, imagining the prime intended users to be journalists. But this democracy-enhancing ideal is at odds with FOIA’s reality: at some agencies, commercial—not public—interests dominate the landscape of FOIA requesters. This Article provides the first in-depth academic study of the commercial use of FOIA, drawing on original datasets from six federal agencies. It documents how corporations, in pursuit of private profit, have overrun FOIA’s supremely inexpensive processes and, in so doing, potentially crowded out journalists and other government watchdogs from doing what the law was intended to facilitate: thirdparty oversight of governmental actors. It also reveals a cottage industry of companies whose entire business model is to request federal records under FOIA and resell them at a profit, which distorts the transparency system even further. Counterintuitively, limiting commercial requesting will not solve this problem. Instead, this Article proposes a targeted and aggressive policy of requiring government agencies to affirmatively disclose sets of records that are the subject of routine FOIA requests—a surprisingly large number of the documents sought by commercial requesters. By meeting information needs in a more efficient manner that is available equally to all, affirmative disclosure will enable federal agencies to reclaim public records from the private market and free up resources to better serve FOIA requests that advance its democratic purpose.

Fiscal policy, banks and the financial crisis. National Bank of Belgium Working Paper No. 234, October 2012

This paper studies the effectiveness of Euro Area (EA) fiscal policy, during the recent financial crisis, using an estimated New Keynesian model with a bank. A key dimension of policy in the crisis was massive government support for banks—that dimension has so far received little attention in the macroeconomics literature. We use the estimated model to analyze the effects of bank asset losses, of government support for banks, and other fiscal stimulus measures, in the EA. Our results suggest that support for banks had a stabilizing effect on EA output, consumption and investment. Increased government purchases helped to stabilize output, but crowded out consumption. Higher transfers to households had a positive impact on private consumption, but a negligible effect on output and investment. Banking shocks and increased government spending explain half of the rise in the public debt/GDP ratio since the onset of the crisis.

Hints to pilgrims /

Essays originally appeared in The Century magazine, The Yale review, The Atlantic monthly and The Literary review.

Immigrants eating in relays at Ellis Island, 9-25-20

ELLIS ISLAND CLOSED TO IMMIGRANTS BECAUSE OF THE CONGESTION. Crowded to almost three times it capacity, the immigration station at Ellis Island, New York harbor, has be closed temporarily until the congrestion is releived....." [sic

Emergence of Special Art Exhibits in Shopping Malls in Shanghai

Thesis (Master's)--University of Washington, 2016-06

The role of insulin-like growth factor II and its receptor in mouse preimplantation development

Insulin-like growth factor II (IGF-II) and its receptor, the IGF-II/mannose-6-phosphate (IGF-II/M6P) receptor, are first expressed from the zygotic genome at the two-cell stage of mouse development. However, their role is not clearly defined. Insulin-like growth factor II is believed to mediate growth through the heterologous type 1 IGF and insulin receptors, whereas the IGF-II/M6P receptor is believed to act as a negative regulator of somatic growth by limiting the availability of excess levels of IGF-II. These studies demonstrate that IGF-II does have a role in growth regulation in the early embryo through the IGF-II/M6P receptor. Insulin-like growth factor II stimulated cleavage rate in two-cell embryos in vitro. Moreover, this receptor is required for the glycaemic response of two-cell embryos to IGF-II and for normal progression of early embryos to the blastocyst stage. Improved development of embryos in crowded culture supports the concept of an endogenous embryonic paracrine activity that enhances cell proliferation. These responses indicate that the IGF-II/M6P receptor is functional and likely to participate in such a regulatory circuit. The functional role of IGF-II and its receptor is discussed with reference to regulation of early development.

A potential role for isothermal calorimetry in studies of the effects of thermodynamic non-ideality in enzyme-catalyzed reactions

Attention is drawn to the feasibility of using isothermal calorimetry for the characterization of enzyme reactions under conditions bearing greater relevance to the crowded biological environment, where kinetic parameters are likely to differ significantly from those obtained by classical enzyme kinetic studies in dilute solution. An outline of the application of isothermal calorimetry to the determination of enzyme kinetic parameters is followed by considerations of the nature and consequences of crowding effects in enzyme catalysis. Some of those effects of thermodynamic non-ideality are then illustrated by means of experimental results from calorimetric studies of the effect of molecular crowding on the kinetics of catalysis by rabbit muscle pyruvate kinase. This review concludes with a discussion of the potential of isothermal calorimetry for the experimental determination of kinetic parameters for enzymes either in biological environments or at least in media that should provide reasonable approximations of the crowded conditions encountered in vivo. Copyright (C) 2004 John Wiley Sons, Ltd.

The HIPASS catalogue - III. Optical counterparts and isolated dark galaxies

We present the largest catalogue to date of optical counterparts for H I radio-selected galaxies, HOPCAT. Of the 4315 H I radio-detected sources from the H I Parkes All Sky Survey (HIPASS) catalogue, we find optical counterparts for 3618 (84 per cent) galaxies. Of these, 1798 (42 per cent) have confirmed optical velocities and 848 (20 per cent) are single matches without confirmed velocities. Some galaxy matches are members of galaxy groups. From these multiple galaxy matches, 714 (16 per cent) have confirmed optical velocities and a further 258 (6 per cent) galaxies are without confirmed velocities. For 481 (11 per cent), multiple galaxies are present but no single optical counterpart can be chosen and 216 (5 per cent) have no obvious optical galaxy present. Most of these 'blank fields' are in crowded fields along the Galactic plane or have high extinctions. Isolated 'dark galaxy' candidates are investigated using an extinction cut of A(Bj) < 1 mag and the blank-fields category. Of the 3692 galaxies with an A(Bj) extinction < 1 mag, only 13 are also blank fields. Of these, 12 are eliminated either with follow-up Parkes observations or are in crowded fields. The remaining one has a low surface brightness optical counterpart. Hence, no isolated optically dark galaxies have been found within the limits of the HIPASS survey.

Drifting blooms of the endemic filamentous brown alga Hincksia sordida at Noosa on the subtropical east Australian coast

Since 2002, the usually uncommon endemic filamentous brown alga Hincksia sordida (Harvey) Silva (Ectocarpales, Phaeophyta) has formed nuisance blooms annually during spring/early summer at Main Beach, Noosa on the subtropical east Australian coast. The Hincksia bloom coincides with the normally intensive recreational use of the popular bathing beach by the local population and tourists. The alga forms dense accumulations in the surf zone at Main Beach, giving the seawater a distinct brown coloration and deterring swimmers from entering the water. Decomposing algae stranded by receding tides emit a nauseating sulphurous stench which hangs over the beach. The stranded algal biomass is removed from the beach by bulldozers. During blooms, the usually crowded Main Beach is deserted, bathers preferring to use the many unaffected beaches on the Sunshine Coast to the south of Main Beach. The bloom worsens with north-easterly winds and is cleared from Noosa by south easterly winds, observations which have prompted the untenable proposal by local authorities that the bloom is forming offshore of Fraser Island in the South Pacific Ocean. The Noosa River estuarine system/Laguna Bay is the more probable source of the bloom and the nutrient inputs into this system must be substantial to generate the high bloom biomass. Current mitigation procedures of removing the blooming alga off the beach with bulldozers treat the symptom, not the cause and are proving ineffective. Environmental management must be based on science and the Noosa bloom would benefit greatly from the accurate ecological data on which to base management options. (c) 2006 Elsevier Ltd. All rights reserved.

Foreign and domestic investment:regional development or crowding out?

This paper tests, at the regional and industry level, the extent to which domestic investment is stimulated or crowded out by inward foreign direct investment. The paper develops a model of domestic investment, based on standard models drawn from macroeconomics and industrial economics. The paper then goes on to show that, at a general level, the 'development' or agglomeration hypothesis is confirmed that domestic investment is indeed stimulated by inward investment. However, there is also evidence that, in certain regions, inward investment has crowded out domestic investment. The implications of this from the perspective of regional policy are briefly discussed.

Lobe growth in foliose lichen thalli

The margins of foliose lichen thalli comprise individual lobes which grow radially and divide. This results in a complex marginal structure in which lobes differing in morphology, state of division, and growth pattern are crowded together. Various aspects of the biology of these lobes are reviewed including their carbohydrate supply, morphology, pattern of division and branching, the effect of lobe overcrowding and interactions between neighbouring lobes. As the thalus grows, lobes become increasingly crowded together and this restricts further lateral growth. Restriction of lobe width may be reponsible for the changes in radial growth rate (RGR) with size observed in foliose species. Various aspects of lobe biology may be responsible for lobe growth variation including the physical independence of lobes from their neighbours, the genetic origin of the lobes, and the pattern of lobe branching. Overall symmetry of a thallus is maintained by a fluctuating pattern of growth of individual lobes in successive months together with competition for space at the margin