Identification and analysis of vaccinia virus acylproteins

Vaccinia virus (VV) encodes at least six proteins that are modified by the addition of a 14-carbon saturated fatty acid through an amide linkage and at least eight proteins that are modified post-translationally by the addition of a 16-carbon saturated fatty acid through linkage to cysteine residues. These post-translational modifications are referred to as myristylation and palmitylation, respectively. The purpose of this work was to further characterize the known myristylproteins and to define a consensus motif for the palmitylation of a protein so that we could identify and begin the characterization of new palmitylproteins. Through this work we have identified a loosely conserved consensus motif that directs the palmitylation of a protein. Using the VV palmitylprotein p37, we characterized this motif and then used it in the identification three new VV palmitylproteins. We have also determined the membrane orientation of the VV myristylproteins, L1R, within the intracellular mature virus (IMV) particle. Hydrophobicity plot analysis identified two possible membrane orientations based on two putative transmembrane domains. Through transient expression data, L1R was determined to span the IMV membrane twice, with both the amino and carboxy termini being on the lumen side. Three lac recombinant viruses which are inducible for the A16L, E7R, and G9R open reading frames were created and analyzed using a newly developed vector system that fuses the green fluorescent protein to the neomycin resistance gene. Propagation of these viruses in the absence of the inducer IPTG determined that these genes are essential to VV replication. We have identified and characterized the primary structural determinants specifying the modification of a protein by palmitate, and have identified three new VV palmitylproteins. In addition, the membrane orientation of the VV myristylprotein L1R was deduced, which can enable the construction of better recombinant vaccines through efficient antigen presentation. Lastly, we have developed a technique and vector system to easily create and isolate VV recombinants for characterization. This system enabled us to further characterize the A16L, E7R, and G9R myristylproteins

Misdiagnosis, Mistreatment, and Harm - When Medical Care Ignores Social Forces.

The Case Studies in Social Medicine demonstrate that when physicians use only biologic or individual behavioral interventions to treat diseases that stem from or are exacerbated by social factors, we risk harming the patients we seek to serve

Mean field theory of hard sphere glasses and jamming

Hard spheres are ubiquitous in condensed matter: they have been used as models for liquids, crystals, colloidal systems, granular systems, and powders. Packings of hard spheres are of even wider interest, as they are related to important problems in information theory, such as digitalization of signals, error correcting codes, and optimization problems. In three dimensions the densest packing of identical hard spheres has been proven to be the FCC lattice, and it is conjectured that the closest packing is ordered (a regular lattice, e.g, a crystal) in low enough dimension. Still, amorphous packings have attracted a lot of interest, because for polydisperse colloids and granular materials the crystalline state is not obtained in experiments for kinetic reasons. We review here a theory of amorphous packings, and more generally glassy states, of hard spheres that is based on the replica method: this theory gives predictions on the structure and thermodynamics of these states. In dimensions between two and six these predictions can be successfully compared with numerical simulations. We will also discuss the limit of large dimension where an exact solution is possible. Some of the results we present here have been already published, but others are original: in particular we improved the discussion of the large dimension limit and we obtained new results on the correlation function and the contact force distribution in three dimensions. We also try here to clarify the main assumptions that are beyond our theory and in particular the relation between our static computation and the dynamical procedures used to construct amorphous packings.Comment: 59 pages, 25 figures. Final version published on Rev.Mod.Phy

Pyroelectric detectors

The multi-agency, long-term Global Change programs, and specifically NASA's Earth Observing system, will require some new and advanced photon detector technology which must be specifically tailored for long-term stability, broad spectral range, cooling constraints, and other parameters. Whereas MCT and GaAs alloy based photovoltaic detectors and detector arrays reach most impressive results to wavelengths as long as 12 microns when cooled to below 70 K, other materials, such as ferroelectrics and pyroelectrics, appear to offer special opportunities beyond 12 microns and above 70 K. These materials have found very broad use in a wide variety of room temperature applications. Little is known about these classes of materials at sub-room temperatures and no photon detector results have been reported. From the limited information available, researchers conclude that the room temperature values of D asterisk greater than or equal to 10(exp 9) cm Hz(exp 1/2)/W may be improved by one to two orders of magnitude upon cooling to temperatures around 70 K. Improvements of up to one order of magnitude appear feasible for temperatures achievable by passive cooling. The flat detector response over a wavelength range reaching from the visible to beyond 50 microns, which is an intrinsic advantage of bolometric devices, makes for easy calibration. The fact that these materials have been developed for reduced temperature applications makes ferro- and pyroelectric materials most attractive candidates for serious exploration

Acute Infection and Subsequent Subclinical Reactivation of Herpes Simplex Virus 2 after Vaginal Inoculation of Rhesus Macaques.

Herpes simplex virus 2 (HSV-2) is a common sexually transmitted infection with a highly variable clinical course. Many infections quickly become subclinical, with episodes of spontaneous virus reactivation. To study host-HSV-2 interactions, an animal model of subclinical HSV-2 infection is needed. In an effort to develop a relevant model, rhesus macaques (RM) were inoculated intravaginally with two or three HSV-2 strains (186, 333, and/or G) at a total dose of 1 × 107 PFU of HSV-2 per animal. Infectious HSV-2 and HSV-2 DNA were consistently shed in vaginal swabs for the first 7 to 14 days after each inoculation. Proteins associated with wound healing, innate immunity, and inflammation were significantly increased in cervical secretions immediately after HSV-2 inoculation. There was histologic evidence of acute herpesvirus pathology, including acantholysis in the squamous epithelium and ballooning degeneration of and intranuclear inclusion bodies in epithelial cells, with HSV antigen in mucosal epithelial cells and keratinocytes. Further, an intense inflammatory infiltrate was found in the cervix and vulva. Evidence of latent infection and reactivation was demonstrated by the detection of spontaneous HSV-2 shedding post-acute inoculation (102 to 103 DNA copies/swab) in 80% of RM. Further, HSV-2 DNA was detected in ganglia in most necropsied animals. HSV-2-specifc T-cell responses were detected in all animals, although antibodies to HSV-2 were detected in only 30% of the animals. Thus, HSV-2 infection of RM recapitulates many of the key features of subclinical HSV-2 infection in women but seems to be more limited, as virus shedding was undetectable more than 40 days after the last virus inoculation.IMPORTANCE Herpes simplex virus 2 (HSV-2) infects nearly 500 million persons globally, with an estimated 21 million incident cases each year, making it one of the most common sexually transmitted infections (STIs). HSV-2 is associated with increased human immunodeficiency virus type 1 (HIV-1) acquisition, and this risk does not decline with the use of antiherpes drugs. As initial acquisition of both HIV and HSV-2 infections is subclinical, study of the initial molecular interactions of the two agents requires an animal model. We found that HSV-2 can infect RM after vaginal inoculation, establish latency in the nervous system, and spontaneously reactivate; these features mimic some of the key features of HSV-2 infection in women. RM may provide an animal model to develop strategies to prevent HSV-2 acquisition and reactivation

TIGIT Marks Exhausted T Cells, Correlates with Disease Progression, and Serves as a Target for Immune Restoration in HIV and SIV Infection.

HIV infection induces phenotypic and functional changes to CD8+ T cells defined by the coordinated upregulation of a series of negative checkpoint receptors that eventually result in T cell exhaustion and failure to control viral replication. We report that effector CD8+ T cells during HIV infection in blood and SIV infection in lymphoid tissue exhibit higher levels of the negative checkpoint receptor TIGIT. Increased frequencies of TIGIT+ and TIGIT+ PD-1+ CD8+ T cells correlated with parameters of HIV and SIV disease progression. TIGIT remained elevated despite viral suppression in those with either pharmacological antiretroviral control or immunologically in elite controllers. HIV and SIV-specific CD8+ T cells were dysfunctional and expressed high levels of TIGIT and PD-1. Ex-vivo single or combinational antibody blockade of TIGIT and/or PD-L1 restored viral-specific CD8+ T cell effector responses. The frequency of TIGIT+ CD4+ T cells correlated with the CD4+ T cell total HIV DNA. These findings identify TIGIT as a novel marker of dysfunctional HIV-specific T cells and suggest TIGIT along with other checkpoint receptors may be novel curative HIV targets to reverse T cell exhaustion

The Outbursts and Orbit of the Accreting Pulsar GS 1843-02 = 2S 1845-024

We present observations of a series of 10 outbursts of pulsed hard X-ray flux from the transient 10.6 mHz accreting pulsar GS 1843-02, using the Burst and Transient Source Experiment on the Compton Gamma Ray Observatory. These outbursts occurred regularly every 242 days, coincident with the ephemeris of the periodic transient GRO J1849-03 (Zhang et al. 1996), which has recently been identified with the SAS 3 source 2S 1845-024 (Soffitta et al. 1998). Our pulsed detection provides the first clear identification of GS 1843-02 with 2S 1845-024. We present a pulse timing analysis which shows that the 2S 1845-024 outbursts occur near the periastron passage of the neutron star's highly eccentric (e = 0.88+-0.01) 242.18+-0.01 day period binary orbit about a high mass (M > 7 solar masses) companion. The orbit and transient outburst pattern strongly suggest the pulsar is in a binary system with a Be star. Our observations show a long-term spin-up trend, with most of the spin-up occurring during the outbursts. From the measured spin-up rates and inferred luminosities we conclude that an accretion disk is present during the outbursts.Comment: Accepted for publication in Astrophysical Journa

Towards a formalism for mapping the spacetimes of massive compact objects: Bumpy black holes and their orbits

Observations have established that extremely compact, massive objects are common in the universe. It is generally accepted that these objects are black holes. As observations improve, it becomes possible to test this hypothesis in ever greater detail. In particular, it is or will be possible to measure the properties of orbits deep in the strong field of a black hole candidate (using x-ray timing or with gravitational-waves) and to test whether they have the characteristics of black hole orbits in general relativity. Such measurements can be used to map the spacetime of a massive compact object, testing whether the object's multipoles satisfy the strict constraints of the black hole hypothesis. Such a test requires that we compare against objects with the ``wrong'' multipole structure. In this paper, we present tools for constructing bumpy black holes: objects that are almost black holes, but that have some multipoles with the wrong value. The spacetimes which we present are good deep into the strong field of the object -- we do not use a large r expansion, except to make contact with weak field intuition. Also, our spacetimes reduce to the black hole spacetimes of general relativity when the ``bumpiness'' is set to zero. We propose bumpy black holes as the foundation for a null experiment: if black hole candidates are the black holes of general relativity, their bumpiness should be zero. By comparing orbits in a bumpy spacetime with those of an astrophysical source, observations should be able to test this hypothesis, stringently testing whether they are the black holes of general relativity. (Abridged)Comment: 16 pages + 2 appendices + 3 figures. Submitted to PR

Reservoir hosts for Gyrodactylus salaris may play a more significant role in epidemics than previously thought

Background: Gyrodactylus salaris Malmberg, 1957 has had a devastating impact on wild Norwegian stocks of Atlantic salmon Salmo salar L., and it is the only Office International des Epizooties (OIE) listed parasitic pathogen of fish. The UK is presently recognised as G. salaris-free, and management plans for its containment and control are currently based on Scandinavian studies. The current study investigates the susceptibility of British salmonids to G. salaris, and determines whether, given the host isolation since the last glaciation and potential genetic differences, the populations under test would exhibit different levels of susceptibility, as illustrated by the parasite infection trajectory over time, from their Scandinavian counterparts. Methods: Populations of S. salar, brown trout Salmo trutta L., and grayling Thymallus thymallus (L.), raised from wild stock in UK government hatcheries, were flown to Norway and experimentally challenged with a known pathogenic strain of G. salaris. Each fish was lightly anaesthetised and marked with a unique tattoo for individual parasite counting. A single Norwegian population of S. salar from the River Lærdalselva was used as a control. Parasite numbers were assessed every seven days until day 48 and then every 14 days. Results: Gyrodactylus salaris regularly leads to high mortalities on infected juveniles S. salar. The number of G. salaris on British S. salar rose exponentially until the experiment was terminated at 33 days due to fish welfare concerns. The numbers of parasites on S. trutta and T. thymallus increased sharply, reaching a peak of infection on days 12 and 19 post-infection respectively, before declining to a constant low level of infection until the termination of the experiment at 110 days. Conclusions: The ability of S. trutta and T. thymallus to carry an infection for long periods increases the window of exposure for these two hosts and the potential transfer of G. salaris to other susceptible hosts. This study demonstrates that G. salaris can persist on S. trutta for longer periods than previously thought, and that the role that S. trutta could play in disseminating G. salaris needs to be considered carefully and factored into management plans and epidemics across Europe