211 research outputs found
Satellite lifetime routine user's manual
A FORTRAN coded computer program which determines secular variations in mean orbital elements of earth satellites and the lifetime of the orbit is described. The dynamical model treats a point mass satellite subject to solar and lunar disturbing gravitational fields, second, third and fourth harmonics of the earth's oblate potential, earth's atmospheric drag, and solar radiation pressure. Each of these disturbing functions may be selectively simulated. Data preparation instructions, a sample problem, and definitions of output quantities are included
Ab-initio calculations of the optical properties of the Si(113)3x2ADI surface
We investigated the stable silicon (113) surface with a 3x2ADI reconstruction
by ab-initio methods. The ground state properties have been obtained using the
density-functional theory. We present the dispersion of the electronic band
structure, where the surface bands have been distinguished from the projected
bulk bands by calculating their localization in the slab. The optical spectra,
here the reflectance anisotropy (RAS), have been obtained within the
independent particle random phase approximation. We identified surface features
in the spectra tracing them back to the responsible electronic states and,
studied their localization in the slab. A comparison with available
experimental data for the band structure and the RAS shows a good agreement.Comment: 10 pages, 10 figure
Micromechanical finite element analyses of fire retardant woven fabric composites at elevated temperatures using unit cells at multiple length scales
This paper presents a micromechanical Finite Element (FE) model developed to predict the effective mechanical properties of glass fibre-reinforced (woven fabric) polymer composites with/without fire retardant particulate additives at elevated temperatures. The elevated mechanical properties of glass fibre-reinforced epoxy composites with/without fire retardants were predicted using three unit cells of varying length scales in micromechanical FE analysis. Theoretically predictions of flexural behaviour of these fibre-reinforced polymer composites at elevated temperatures were satisfactorily validated against experimentally measured data. The numerical model developed herein was then used for the prediction of other mechanical properties of fibre-reinforced polymer composites that would have been difficult to collect at elevated temperatures. Micromechanical FE models such as the one contained in this paper are useful to architectural engineers as they can be used to guide the design and qualification of new engineering composites that satisfy stringent Building codes in fire prone engineering applications
Histogram specification of 24-bit color images in the color difference (C-Y) color space
Histogram equalization and specification have been widely used to enhance information in a gray scale image, with histogram specification having the advantage of allowing the output histogram to be specified as compared to histogram equalization, which attempts to produce an output histogram that is uniform Unfortunately, expanding histogram techniques to color images is not very straightforward Since humans are sensitive to chromatic changes, care must be taken to ensure that incorrect colors are not produced. Additionally expanding the one-dimensional histogram used in gray level histogram techniques to a joint histogram (usually of three variables representing the primary colors of red, green, and blue) can yield specified histograms which have no physical meaning, hence making if difficult to determine the set of histograms required for a desired enhancement in this paper, we describe a method of extending gray level histogram specification to color images by performing histogram specification an the luminance, saturation, and hue components in the color difference C-Y color space. These methods take into account the relationship between the luminance and saturation components while yielding specified histograms that produce natural-looking results
Towards the Construction of Expressed Proteomes Using a Leishmania tarentolae Based Cell-Free Expression System
The adaptation of organisms to a parasitic life style is often accompanied by the emergence of novel biochemical pathways absent in free-living organisms. As a result, the genomes of specialized parasitic organisms often code for a large number (>50%) of proteins with no detectable homology or predictable function. Although understanding the biochemical properties of these proteins and their roles in parasite biogenesis is the next challenge of molecular parasitology, analysis tools developed for free-living organisms are often inadequate for this purpose. Here we attempt to solve some of these problems by developing a methodology for the rapid production of expressed proteomes in cell-free systems based on parasitic organisms. To do so we take advantage of Species Independent Translational Sequences (SITS), which can efficiently mediate translation initiation in any organism. Using these sequences we developed a single-tube in vitro translation system based on the parasitic protozoan Leishmania tarentolae. We demonstrate that the system can be primed directly with SITS containing templates constructed by overlap extension PCR. To test the systems we simultaneously amplified 31 of L. tarentolae's putative translation initiation factors and phosphatases directly from the genomic DNA and subjected them to expression, purification and activity analysis. All of the amplified products produced soluble recombinant proteins, and putative phosphatases could be purified to at least 50% purity in one step. We further compared the ability of L. tarentolae and E. coli based cell-free systems to express a set of mammalian, L. tarentolae and Plasmodium falciparum Rab GTPases in functional form. We demonstrate that the L. tarentolae cell-free system consistently produced higher quality proteins than E. coli-based system. The differences were particularly pronounced in the case of open reading frames derived from P. falciparum. The implications of these developments are discussed
Structural Insight into How Bacteria Prevent Interference between Multiple Divergent Type IV Secretion Systems
Prokaryotes use type IV secretion systems (T4SSs) to translocate substrates (e.g., nucleoprotein, DNA, and protein) and/or elaborate surface structures (i.e., pili or adhesins). Bacterial genomes may encode multiple T4SSs, e.g., there are three functionally divergent T4SSs in some Bartonella species (vir, vbh, and trw). In a unique case, most rickettsial species encode a T4SS (rvh) enriched with gene duplication. Within single genomes, the evolutionary and functional implications of cross-system interchangeability of analogous T4SS protein components remains poorly understood. To lend insight into cross-system interchangeability, we analyzed the VirB8 family of T4SS channel proteins. Crystal structures of three VirB8 and two TrwG Bartonella proteins revealed highly conserved C-terminal periplasmic domain folds and dimerization interfaces, despite tremendous sequence divergence. This implies remarkable structural constraints for VirB8 components in the assembly of a functional T4SS. VirB8/TrwG heterodimers, determined via bacterial two-hybrid assays and molecular modeling, indicate that differential expression of trw and vir systems is the likely barrier to VirB8-TrwG interchangeability. We also determined the crystal structure of Rickettsia typhi RvhB8-II and modeled its coexpressed divergent paralog RvhB8-I. Remarkably, while RvhB8-I dimerizes and is structurally similar to other VirB8 proteins, the RvhB8-II dimer interface deviates substantially from other VirB8 structures, potentially preventing RvhB8-I/RvhB8-II heterodimerization. For the rvh T4SS, the evolution of divergent VirB8 paralogs implies a functional diversification that is unknown in other T4SSs. Collectively, our data identify two different constraints (spatiotemporal for Bartonella trw and vir T4SSs and structural for rvh T4SSs) that mediate the functionality of multiple divergent T4SSs within a single bacterium.
IMPORTANCE:
Assembly of multiprotein complexes at the right time and at the right cellular location is a fundamentally important task for any organism. In this respect, bacteria that express multiple analogous type IV secretion systems (T4SSs), each composed of around 12 different components, face an overwhelming complexity. Our work here presents the first structural investigation on factors regulating the maintenance of multiple T4SSs within a single bacterium. The structural data imply that the T4SS-expressing bacteria rely on two strategies to prevent cross-system interchangeability: (i) tight temporal regulation of expression or (ii) rapid diversification of the T4SS components. T4SSs are ideal drug targets provided that no analogous counterparts are known from eukaryotes. Drugs targeting the barriers to cross-system interchangeability (i.e., regulators) could dysregulate the structural and functional independence of discrete systems, potentially creating interference that prevents their efficient coordination throughout bacterial infection.</p
Using student data: Student-staff collaborative development of compassionate pedagogic interventions based on learning analytics and mentoring
UK Universities are increasingly being ‘encouraged’ to focus on student engagement, retention and performance, with learning analytics becoming commonplace. Based on inter-related student-staff partnerships, this study adopted a human and compassionate approach to the use of student data and subsequent interventions. Analysis of focus group and interview data from 86 student participants explored key themes: peer-mentoring increasing engagement with the communal-habitus; increased confidence and engagement; and the demystification and humanisation of the university environment. Findings highlight the importance of emphasising human and compassionate support for students within rapidly developing learning analytics approaches, with subject-specific peer-mentoring found here to be beneficial
Telomeric expression sites are highly conserved in trypanosoma brucei
Subtelomeric regions are often under-represented in genome sequences of eukaryotes. One of the best known examples of the use of telomere proximity for adaptive purposes are the bloodstream expression sites (BESs) of the African trypanosome Trypanosoma brucei. To enhance our understanding of BES structure and function in host adaptation and immune evasion, the BES repertoire from the Lister 427 strain of T. brucei were independently tagged and sequenced. BESs are polymorphic in size and structure but reveal a surprisingly conserved architecture in the context of extensive recombination. Very small BESs do exist and many functioning BESs do not contain the full complement of expression site associated genes (ESAGs). The consequences of duplicated or missing ESAGs, including ESAG9, a newly named ESAG12, and additional variant surface glycoprotein genes (VSGs) were evaluated by functional assays after BESs were tagged with a drug-resistance gene. Phylogenetic analysis of constituent ESAG families suggests that BESs are sequence mosaics and that extensive recombination has shaped the evolution of the BES repertoire. This work opens important perspectives in understanding the molecular mechanisms of antigenic variation, a widely used strategy for immune evasion in pathogens, and telomere biology
Biochemical and Structural Characterization of Selective Allosteric Inhibitors of the Plasmodium falciparum Drug Target, Prolyl-tRNA-synthetase
Plasmodium falciparum (<i>Pf</i>) prolyl-tRNA
synthetase (ProRS) is one of the few chemical-genetically validated
drug targets for malaria, yet highly selective inhibitors have not
been described. In this paper, approximately 40,000 compounds were
screened to identify compounds that selectively inhibit <i>Pf</i>ProRS enzyme activity versus Homo sapiens (<i>Hs</i>) ProRS. X-ray crystallography structures were
solved for apo, as well as substrate- and inhibitor-bound forms of <i>Pf</i>ProRS. We identified two new inhibitors of <i>Pf</i>ProRS that bind outside the active site. These two allosteric inhibitors
showed >100 times specificity for <i>Pf</i>ProRS compared
to <i>Hs</i>ProRS, demonstrating this class of compounds
could overcome the toxicity related to <i>Hs</i>ProRS inhibition
by halofuginone and its analogues. Initial medicinal chemistry was
performed on one of the two compounds, guided by the cocrystallography
of the compound with <i>Pf</i>ProRS, and the results can
instruct future medicinal chemistry work to optimize these promising
new leads for drug development against malaria
Biological and Structural Characterization of a Host-Adapting Amino Acid in Influenza Virus
Two amino acids (lysine at position 627 or asparagine at position 701) in the polymerase subunit PB2 protein are considered critical for the adaptation of avian influenza A viruses to mammals. However, the recently emerged pandemic H1N1 viruses lack these amino acids. Here, we report that a basic amino acid at position 591 of PB2 can compensate for the lack of lysine at position 627 and confers efficient viral replication to pandemic H1N1 viruses in mammals. Moreover, a basic amino acid at position 591 of PB2 substantially increased the lethality of an avian H5N1 virus in mice. We also present the X-ray crystallographic structure of the C-terminus of a pandemic H1N1 virus PB2 protein. Arginine at position 591 fills the cleft found in H5N1 PB2 proteins in this area, resulting in differences in surface shape and charge for H1N1 PB2 proteins. These differences may affect the protein's interaction with viral and/or cellular factors, and hence its ability to support virus replication in mammals
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