Transcriptional repression by ApiAP2 factors is central to chronic toxoplasmosis

Tachyzoite to bradyzoite development in Toxoplasma is marked by major changes in gene expression resulting in a parasite that expresses a new repertoire of surface antigens hidden inside a modified parasitophorous vacuole called the tissue cyst. The factors that control this important life cycle transition are not well understood. Here we describe an important transcriptional repressor mechanism controlling bradyzoite differentiation that operates in the tachyzoite stage. The ApiAP2 factor, AP2IV-4, is a nuclear factor dynamically expressed in late S phase through mitosis/cytokinesis of the tachyzoite cell cycle. Remarkably, deletion of the AP2IV-4 locus resulted in the expression of a subset of bradyzoite-specific proteins in replicating tachyzoites that included tissue cyst wall components BPK1, MCP4, CST1 and the surface antigen SRS9. In the murine animal model, the mis-timing of bradyzoite antigens in tachyzoites lacking AP2IV-4 caused a potent inflammatory monocyte immune response that effectively eliminated this parasite and prevented tissue cyst formation in mouse brain tissue. Altogether, these results indicate that suppression of bradyzoite antigens by AP2IV-4 during acute infection is required for Toxoplasma to successfully establish a chronic infection in the immune-competent host

Geophysical anomalies and quartz deformation of the Warburton West structure, central Australia

This paper reports geophysical anomalies and intra-crystalline quartz lamellae in drill cores from the Warburton West Basin overlapping the border of South Australia and the Northern Territory. The pre-Upper Carboniferous ~450×300km-large Warburton Basin, north-eastern South Australia, is marked by distinct eastern and western magnetic, gravity and low-velocity seismic tomography anomalies. Quartz grains from arenite core samples contain intra-crystalline lamellae in carbonate-quartz veins and in clastic grains, similar to those reported earlier from arenites, volcanic rocks and granites from the Warburton East Basin. Universal Stage measurements of quartz lamellae in both sub-basins define Miller-Bravais indices of {10-12} and {10-13}. In-situ quartz lamellae occur only in pre-Late Carboniferous rocks whereas lamellae-bearing clastic quartz grains occur in both pre-Late Carboniferous and post-Late Carboniferous rocks - the latter likely redeposited from the pre-Late Carboniferous basement. Quartz lamellae in clastic quartz grains are mostly curved and bent either due to tectonic deformation or to re-deformation of impact-generated planar features during crustal rebound or/and post-impact tectonic deformation. Seismic tomography low-velocity anomalies in both Warburton West Basin and Warburton East Basin suggest fracturing of the crust to depths of more than 20km. Geophysical modelling of the Cooper Basin, which overlies the eastern Warburton East Basin, suggests existence of a body of high-density (~2.9-3.0gr/cm) and high magnetic susceptibility (SI~0.012-0.037) at a depth of ~6-10km at the centre of the anomalies. In the Warburton West Basin a large magnetic body of SI=0.030 is modelled below ~10km, with a large positive gravity anomaly offset to the north of the magnetic anomaly. In both the Warburton East and Warburton West the deep crustal fracturing suggested by the low velocity seismic tomography complicates interpretations of the gravity data. Universal Stage measurements of quartz lamellae suggest presence of both planar deformation features of shock metamorphic derivation and deformed planar lamella. The latter may be attributed either to re-deformation of impact-generated lamella, impact rebound deformation or/and post impact tectonic deformation. The magnetic anomalies in the Warburton East and West sub-basins are interpreted in terms of (1) presence of deep seated central mafic bodies; (2) deep crustal fracturing and (3) removal of Devonian and Carboniferous strata associated with rebound of a central uplift consequent on large asteroid impact. Further tests of the Warburton structures require deep crustal seismic transects

Toxoplasma gondii AP2IX-4 Regulates Gene Expression during Bradyzoite Development

Toxoplasma gondii is a protozoan parasite of great importance to human and animal health. In the host, this obligate intracellular parasite persists as a tissue cyst that is imperceptible to the immune response and unaffected by current therapies. The tissue cysts facilitate transmission through predation and give rise to chronic cycles of toxoplasmosis in immunocompromised patients. Transcriptional changes accompany conversion of the rapidly replicating tachyzoites into the encysted bradyzoites, and yet the mechanisms underlying these alterations in gene expression are not well defined. Here we show that AP2IX-4 is a nuclear protein exclusively expressed in tachyzoites and bradyzoites undergoing division. Knockout of AP2IX-4 had no discernible effect on tachyzoite replication but resulted in a reduced frequency of tissue cyst formation following alkaline stress induction-a defect that is reversible by complementation. AP2IX-4 has a complex role in regulating bradyzoite gene expression, as the levels of many bradyzoite mRNAs dramatically increased beyond those seen under conditions of normal stress induction in AP2IX-4 knockout parasites exposed to alkaline media. The loss of AP2IX-4 also resulted in a modest virulence defect and reduced cyst burden in chronically infected mice, which was reversed by complementation. These findings illustrate that the transcriptional mechanisms responsible for tissue cyst development operate across the intermediate life cycle from the dividing tachyzoite to the dormant bradyzoite. IMPORTANCEToxoplasma gondii is a single-celled parasite that persists in its host as a transmissible tissue cyst. How the parasite converts from its replicative form to the bradyzoites housed in tissue cysts is not well understood, but the process clearly involves changes in gene expression. Here we report that parasites lacking a cell cycle-regulated transcription factor called AP2IX-4 display reduced frequencies of tissue cyst formation in culture and in a mouse model of infection. Parasites missing AP2IX-4 lose the ability to regulate bradyzoite genes during tissue cyst development. Expressed in developing bradyzoites still undergoing division, AP2IX-4 may serve as a useful marker in the study of transitional forms of the parasite

Coordinated progression through two subtranscriptomes underlies the tachyzoite cycle of Toxoplasma gondii

BACKGROUND: Apicomplexan parasites replicate by varied and unusual processes where the typically eukaryotic expansion of cellular components and chromosome cycle are coordinated with the biosynthesis of parasite-specific structures essential for transmission. METHODOLOGY/PRINCIPAL FINDINGS: Here we describe the global cell cycle transcriptome of the tachyzoite stage of Toxoplasma gondii. In dividing tachyzoites, more than a third of the mRNAs exhibit significant cyclical profiles whose timing correlates with biosynthetic events that unfold during daughter parasite formation. These 2,833 mRNAs have a bimodal organization with peak expression occurring in one of two transcriptional waves that are bounded by the transition into S phase and cell cycle exit following cytokinesis. The G1-subtranscriptome is enriched for genes required for basal biosynthetic and metabolic functions, similar to most eukaryotes, while the S/M-subtranscriptome is characterized by the uniquely apicomplexan requirements of parasite maturation, development of specialized organelles, and egress of infectious daughter cells. Two dozen AP2 transcription factors form a series through the tachyzoite cycle with successive sharp peaks of protein expression in the same timeframes as their mRNA patterns, indicating that the mechanisms responsible for the timing of protein delivery might be mediated by AP2 domains with different promoter recognition specificities. CONCLUSION/SIGNIFICANCE: Underlying each of the major events in apicomplexan cell cycles, and many more subordinate actions, are dynamic changes in parasite gene expression. The mechanisms responsible for cyclical gene expression timing are likely crucial to the efficiency of parasite replication and may provide new avenues for interfering with parasite growth

Hyperboloidal slices for the wave equation of Kerr-Schild metrics and numerical applications

We present new results from two open source codes, using finite differencing and pseudo-spectral methods for the wave equations in (3+1) dimensions. We use a hyperboloidal transformation which allows direct access to null infinity and simplifies the control over characteristic speeds on Kerr-Schild backgrounds. We show that this method is ideal for attaching hyperboloidal slices or for adapting the numerical resolution in certain spacetime regions. As an example application, we study late-time Kerr tails of sub-dominant modes and obtain new insight into the splitting of decay rates. The involved conformal wave equation is freed of formally singular terms whose numerical evaluation might be problematically close to future null infinity.Comment: 15 pages, 12 figure

Characteristic extraction in numerical relativity: binary black hole merger waveforms at null infinity

The accurate modeling of gravitational radiation is a key issue for gravitational wave astronomy. As simulation codes reach higher accuracy, systematic errors inherent in current numerical relativity wave-extraction methods become evident, and may lead to a wrong astrophysical interpretation of the data. In this paper, we give a detailed description of the Cauchy-characteristic extraction technique applied to binary black hole inspiral and merger evolutions to obtain gravitational waveforms that are defined unambiguously, that is, at future null infinity. By this method we remove finite-radius approximations and the need to extrapolate data from the near zone. Further, we demonstrate that the method is free of gauge effects and thus is affected only by numerical error. Various consistency checks reveal that energy and angular momentum are conserved to high precision and agree very well with extrapolated data. In addition, we revisit the computation of the gravitational recoil and find that finite radius extrapolation very well approximates the result at \scri. However, the (non-convergent) systematic differences to extrapolated data are of the same order of magnitude as the (convergent) discretisation error of the Cauchy evolution hence highlighting the need for correct wave-extraction.Comment: 41 pages, 8 figures, 2 tables, added references, fixed typos. Version matches published version

Foundations of character: methodological aspects of a study of character development in three- to six-year-old children with a focus on sharing behaviours

This article focuses on methodological issues arising in a study of character development, using illustrations of ‘sharing behaviours.’ Based primarily in six early years settings in southeast England the research records naturalistic observations of peer interactions for 55 children aged three to six years. Applying grounded theory to the processes of observing, analysing and interpreting evidence required a cautious and collectively reflective approach. The methodology sought to moderate the influence of the researchers' prior knowledge of ‘grand theories’ of moral development and assumptions about relevance to the observation records. The study's originality lay in the exploration of moral development without reference to any particular grand theory as an explanatory framework; and in the reluctance to be drawn to potentially simplistic rationalisations of the children's intentions on the basis of their observed behaviours. Exploring young children's subjective experiences, this research provides insights into the intricacy of this process, steering away from ‘neat’ findings and attempting to reflect the sophistication of the children's skilful and sometimes surprising negotiations of moral dilemmas. Implications for practice relate to the complexities involved in attempts to unravel the developing moral characters of young children and the practice through which this may be nurtured

Gas emissions, minerals, and tars associated with three coal fires, Powder River Basin, USA.

Ground-based surveys of three coal fires and airborne surveys of two of the fires were conducted near Sheridan, Wyoming. The fires occur in natural outcrops and in abandoned mines, all containing Paleocene-age subbituminous coals. Diffuse (carbon dioxide (CO(2)) only) and vent (CO(2), carbon monoxide (CO), methane, hydrogen sulfide (H(2)S), and elemental mercury) emission estimates were made for each of the fires. Additionally, gas samples were collected for volatile organic compound (VOC) analysis and showed a large range in variation between vents. The fires produce locally dangerous levels of CO, CO(2), H(2)S, and benzene, among other gases. At one fire in an abandoned coal mine, trends in gas and tar composition followed a change in topography. Total CO(2) fluxes for the fires from airborne, ground-based, and rate of fire advancement estimates ranged from 0.9 to 780mg/s/m(2) and are comparable to other coal fires worldwide. Samples of tar and coal-fire minerals collected from the mouth of vents provided insight into the behavior and formation of the coal fires